Exploring the profile of amino acids and ethnography study in the biodiversity of tempeh as fermented foods of Indonesia

Exploring the profile of amino acids and ethnography study in the biodiversity of tempeh as fermented foods of Indonesia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Exploring the profile of amino acids and ethnography study in the biodiversity of tempeh as fermented foods of Indonesia Umi Purwandari, Darimiyya Hidayati, Askur Rahman, Enung Siti Nurhidayah, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5724562/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 06 Jun, 2025 Read the published version in Journal of Ethnic Foods → Version 1 posted 5 You are reading this latest preprint version Abstract Tempeh has been world widely known as a functional food for several health benefits such as ACE inhibitory, hypotensive, and antioxidative. Although soybeans contained opioids, a peptide that affects mental state, there was no report on the potential of tempeh as a source of peptides with affective or cognitive function. This study examined the protein quality of tempeh as indicated by the amino acids profile and its possible role in improving depression, as indicated by the concentration of tryptophan and its ratio to large neutral amino acids (LNAA). Fresh tempeh types of different raw materials (soybean, okara, velvet bean, lablab bean, fresh coconut press cake) were collected from production areas. Samples were run for high-performance liquid chromatography. Results showed that protein and amino acid concentrations in tempeh ranged respectively from 13.97 and 8% ( gembus tempeh) and 45.5 and 18% (Malang tempeh). The amino acid score in all tempeh met the WHO standard for nine amino acids, indicating good quality of amino acids. Tempeh made from peanut press cake showed all amino acids scored higher than 100, while gembus tempeh showed three amino acids with a score of less than 100. Methionine was the limiting amino acid while glutamate was the predominant amino acid of tempeh. Except that in koro , gembus , or dage tempeh , tryptophan in tempeh samples showed the ability to serve as an anti-depressant activity. Amino acid fermented foods Indonesia tempeh Figures Figure 1 Introduction Fermented food or beverages have evolved from preserved into functional foods or drinks. Most fermented foods or drinks were acquired from ethnic treasures. One of the successful worldwide fermented foods was tempeh (or tempe), which originated in Indonesia. The health benefits of tempeh have been extensively reported or reviewed [ 1 ]. Different types of tempeh were created based on their raw materials, which are commercially available in traditional markets in Indonesia. These include peanut press cake, okara, velvet bean, and coconut press cake. The main fungus used was Rhizopus spp. Wrapping material used in tempeh production in Indonesia was mostly plastic bags and banana leaves, and sometimes teak or canna leaves were used. Tempeh fermented in plastic bag showed a higher number of total bacteria but less concentration of soluble protein (0.17 to 0.45 mg.L − 1 ) than those wrapped in banana (0.18 to 0.61 mg.L − 1 ) or teak leaves (0.18 to 0.60%), or slightly lower (66.42%) than those wrapped in banana leaves (67.58%) or teak leaves [ 2 ]. However, tempeh wrapped in plastic bags became stinky faster and had less preference toward flavour than those wrapped in banana leaves [ 2 ]. Fermentation of tempeh has been reported to alter protein and amino acids profile. Prolonged fermentation to 96 hours increased protein content from 34.9 to 53.43% and pH from 5.4 to 6.1 [ 3 ]. Similarly, total amino acids increased to 72 hours of fermentation, from 69.3 to 200.7 g.L − 1 , but then decreased to 190.1 g.L − 1 [ 3 ]. Amino acid composition determined the sensory characteristics of tempeh. Umami taste was correlated to aspartic and glutamic acids [ 3 ]. The sweetness of tempeh was correlated to alanine, glycine, serine, and threonine [ 3 ], bitterness was correlated to arginine, histidine, isoleucine, leucine, methionine, phenylalanine, valine, and tyrosine [ 3 ]. Proteolysis of soybean protein during fermentation of tempeh was likely to result in amino acids and peptides. Nevertheless, prolonged fermentation might degrade amino acids to reduce their concentration, especially those susceptible to microbial degradation [ 4 ]. Therefore, fermentation might increase or reduce amino acid concentration during fermentation. Due to fermentation, nitrogen in soybean seed was reduced (from 6.65 to 2.98 gN), protein was increased (from 8 to 17 g), threonine was reduced (from 1603 to 742 mg), tryptophan was reduced (from 532 to 235 mg), and lysine was reduced (from 2653 to 1120 mg) per 100 g [ 4 ]. One of the most studied amino acids in relation to mood or mental state was tryptophan. Tryptophan was a precursor of serotonin, a hormone -also known as 5-HT (contraction of 5-hydroxytryptamine) related to mood, cognition, reward, learning, and memory, where the synthesis occurs in the brain [ 5 ]. Although serotonin was also produced in the large intestine by the influence of gut microbiota, this plasma serotonin could not pass the blood-brain barrier (BBB) to get into the central nervous system and affect mood [ 6 ]. Therefore, serotonin synthesis in the brain relies mostly on dietary tryptophan and is strongly related to dietary tryptophan [ 6 ]. Similarly, in healthy subjects, high tryptophan food resulted in reduced depression and stress as well as improved mood [ 7 ]. Although there were reports on a positive correlation between dietary tryptophan and brain serotonin [ 5 ], tryptophan uptake in brain cells was influenced by several factors, including the presence of other large neutral amino acids (LNAA) in the plasma. Tryptophan was one of LNAA, with others, namely phenylalanine, tyrosine, threonine, methionine, valine, isoleucine, leucine, and histidine. There was a competition for transporter among LNAA during passing BBB [ 5 ]. Consequently, the ratio of tryptophan to other LNAA was important to determine the concentration of tryptophan to pass over BBB. The proposed ratio of tryptophan to other LNAA was 0.09 or above [ 8 ]. Another factor that influenced tryptophan uptake was dietary carbohydrates. Consumption of carbohydrates after fasting was reported to induce secretion of insulin, which increases tryptophan availability to cross over BBB by removing another amino acid in the plasma. Tryptophan transport to the brain was also affected by albumin since tryptophan is lipophilic and needs albumin to allow it to solubilize in the plasma. This implied the important role of the food matrix in the formation of serotonin from dietary tryptophan. Nutrient-based treatment had been proposed previously in order to support alleviating manic-depressive disorder, especially referring to high tryptophan concentration in the diet [ 9 ]. Different types of protein or their concentration in the diet affected tryptophan/LNAA differently, and the effect was also affected by simple carbohydrates in the diet and their concentration [ 8 ]. Tryptophan was not synthesized in the human body, so it had to be provided in the diet. The daily requirement of tryptophan in adults was 250 to 425 mg.day-1 or 3.5-6 mg.kg-1 body weight a day, while in children, it was 12 mg.kg − 1 body weight a day [ 10 ]. Tryptophan in tempeh might be considered a minor amino acid, being the lowest concentration compared to others, ranging from 0.13% (wb) [ 11 ] to around 2% [ 12 ]. The amino acid score of tryptophan in benguk tempeh was 59.01 [ 13 ], and thus was considered very low and not adequate for good nutrition. This work studied the amino acid profile and ethnographic aspects of several types of tempeh from Java, Indonesia. Subsequently, it analysed them for their possible effect on alleviating depression as expressed by tryptophan content. Methodology Materials Samples were collected from traditional markets in production areas in Malang (fresh Malang tempeh, peanut tempeh, dan gembus tempeh), Manis market in Purwokerto ( dage tempeh), and Bantul ( koro tempeh ), and Yogyakarta province ( benguk tempeh ). About 0.5 kg of samples were bought from the sellers, kept in an iced-containing container, dried, and kept in a sealed plastic bag until used for analysis. Benguk tempeh was bought from an online shop. Information on the process of making the product was collected from the seller or the maker(s). Methods Samples were first digested using the Kjeldhal method to determine protein concentration and then reacted with O-phtalalhdehide (OPA) [ 12 ]. They were then analysed for amino acid content using HPLC (Thermos Scientific ODS-1 Hyersil) with a fluorescent detector. The mobile phase was Na-acetate (pH 6.5, 0.02%), Na-EDTA (0.005%), methanol (9%), and THF (1,5%) [ 12 ]. The nutritional value of protein was determined by calculating the amino acid score. Amino acid scores were obtained by comparing essential amino acids (mg.g − 1 protein) with the WHO amino acid standard [ 10 ]. This score was based on the FAO-WHO's nine amino acid requirements for preschool children. Result and Discussion The History, Ethnography and Philosophy of Tempeh Tempeh, a fermented soy product, has a long and rich history rooted in Southeast Asia, particularly in Indonesia. This nutritious and versatile food has grown from being a regional specialty to gaining international recognition as a staple in plant-based diets. Its history is deeply intertwined with the cultural, agricultural, and culinary practices of the regions where it originated. Tempeh's history can be traced back over 400 years to Java, Indonesia, where it is believed to have first been developed. The exact origins remain unclear, as tempeh was not explicitly mentioned in ancient texts until the 19th century. However, oral traditions and historical records suggest that tempeh-making techniques were passed down through generations long before they were documented. The development of tempeh was likely a result of resourceful innovation. Soybeans, introduced to Indonesia from China, became a common crop in the region. Javanese farmers discovered that fermenting soybeans with Rhizopus mould not only enhanced the nutritional profile of the beans but also made them more digestible and flavourful. This process also allowed for longer storage, making tempeh a practical solution in tropical climates [ 2 ]. In Indonesia, tempeh is more than just a food—it is a cultural symbol and a dietary staple. It plays a vital role in Javanese cuisine, where it is often fried, steamed, or cooked in various dishes. Its affordability, high protein content, and versatility have made it an essential source of nutrition for generations of Indonesians, especially in rural areas where access to meat and fish might be limited. Tempeh is commonly prepared with traditional spices and served alongside rice, the staple carbohydrate of Indonesian meals. Dishes such as tempeh goreng (fried tempeh), tempeh orek (sweet and spicy tempeh), and sambal goreng tempeh (tempeh in spicy chili sauce) are beloved across the archipelago [ 7 ]. As tempeh was an important part of Javanese daily meals, several types of tempeh had been developed, either using different processes or raw materials instead of soybeans. For example, Tempe koro is made from lablab beans, and Tempe koro benguk is made from velvet beans. Tempe koro is widely produced and consumed by people in Yogyakarta province. The reason for making these types of tempeh might be the availability of beans in the area. However, later, people may find that their taste was different from soybean tempeh, and the koro and koro benguk tempeh became a specialty of the area. Creativity seemed to lead to the use of residue from some products, such as peanut oil, tofu, or coconut milk. Tempeh is made from peanut press cake, locally called “ tempe kacang ”, also known as “ tempe bungkil ”, and is only available now mainly in Malang, but also to a lesser extent in Yogyakarta and some areas in Central Java [ 7 ]. Tempeh made from ‘okara’ a residue of tofu, is called “ tempe gembus ” which is widely available. Another tempeh is made from the residue of coconut milk; this tempeh is called “ tempe bongkrek ”, and its consumption in the past led to fatalities due to highly toxic bongkrek acid. This made tempe bongkrek banned since the 80’s. Tempe dage is reported to be made from coconut milk residue, but the exact raw material and the making process is not extensively reported. Another type of tempeh uses different species of fungi, called oncom , which uses Chrysonilia sitophila (orange oncom ) or Rhizopus spp. (black oncom ), and raw material of okara or peanut press cake. Fermentation of tempeh can occur inside wrap leaves or in open spaces. The leaves used as tempeh wrap vary, depending on availability. The most commonly used is a banana leaf, which is considered the finest wrapping material. The less commonly used are canna leaves, sea hibiscus leaves, or teak leaves. Bamboo logs (about 30 cm long) were also used as wrapping material, but their use is very rare now. The perforated plastic bag is now a very common wrapping material [ 7 ]. The spread of tempeh beyond Indonesia began in the 20th century, driven by globalization and increasing interest in vegetarian and vegan diets. Dutch colonization played a significant role in introducing tempeh to Europe. During their time in Indonesia, the Dutch developed a taste for local cuisine, and tempeh was among the foods that found a place in Dutch kitchens. In the 1960s and 1970s, tempeh began gaining popularity in the United States and other Western countries. The growing health food movement and interest in plant-based protein sources fuelled this surge. Researchers and food enthusiasts started studying tempeh, recognizing its high protein content, probiotics, and ability to fit into various cuisines [ 2 , 7 ]. Today, tempeh is produced and consumed worldwide, with manufacturers adapting traditional methods for large-scale production. Its health benefits, including being a rich source of protein, vitamins, and dietary fibre, have made it a favourite among health-conscious consumers and vegetarians. Tempeh’s texture and ability to absorb flavours make it an excellent meat substitute in recipes ranging from stir-fries to burgers. Countries such as the United States, Japan, and the Netherlands now produce tempeh commercially, often tailoring it to suit local tastes. In some regions, other legumes and grains like chickpeas, black beans, and quinoa are used to create variations of traditional tempeh [ 2 , 7 ]. Despite its global spread, tempeh remains deeply tied to its Indonesian roots. In recent years, efforts have been made to preserve traditional tempeh-making techniques and recognise their cultural importance. Festivals, workshops, and culinary events in Indonesia celebrate tempeh as a national treasure and a symbol of sustainable eating. As awareness grows about the environmental impact of meat production, tempeh's role as a sustainable protein source is increasingly acknowledged. Its production requires fewer resources compared to animal agriculture, making it an eco-friendly choice for a growing global population. The history of tempeh is a testament to the ingenuity and resourcefulness of the Javanese people. From its humble beginnings in rural Indonesia to its position as a global health food, tempeh continues to inspire innovation in both culinary and agricultural practices. Its journey reflects not only the evolution of food but also the interconnectedness of culture, sustainability, and nutrition in our modern world [ 2 , 7 ]. Tempeh, a traditional Indonesian food made from fermented soybeans, is much more than a culinary delight. It embodies a philosophy rooted in transformation, patience, sustainability, and interconnectedness [ 2 , 7 ]. At its core, fermentation represents transformation. In the case of tempeh, humble soybeans undergo a profound change through the action of Rhizopus oryzae mould. This microorganism binds the beans into a cohesive cake, creating a product that is more flavourful, nutritious, and digestible than its original form. This transformation is a metaphor for the potential within all living things. It shows how patience and the right conditions can turn the ordinary into the extraordinary. Life, like tempeh, often requires time and care to reveal its full potential [ 2 , 7 ]. Tempeh fermentation is an interconnected process. It relies on a delicate balance of soybeans, mould, air, and warmth, each playing an essential role. This interdependence reflects the broader truth that nothing exists in isolation. Ecosystems thrive through collaboration in nature, and the same principle applies to human relationships. Tempeh teaches us the value of working together harmoniously, where each element contributes to a greater whole. It is a celebration of unity between microorganisms, humans, and the environment that results in a nourishing outcome [ 2 , 7 ]. Fermentation cannot be rushed. The process requires time, a stable environment, and a degree of uncertainty as nature takes its course. This patience mirrors the rhythms of life, where meaningful outcomes often require trust in the journey rather than immediate results. Tempeh teaches us to accept life’s natural pace and to appreciate the unseen work happening beneath the surface. In a world increasingly driven by instant gratification, the philosophy of fermentation offers a counterbalance, encouraging mindfulness and faith in slow, deliberate progress [ 7 ]. Tempeh is modest in appearance yet profound in its impact. It is not a luxury food, but its simplicity and versatility sustain millions. This humility reflects a philosophy of valuing substance over spectacle. In a society that often prioritizes extravagance, tempeh reminds us of the beauty in simplicity. Its unassuming nature belies its richness—both as a source of nutrition and as a symbol of cultural and environmental mindfulness [ 2 ]. For those who prepare tempeh traditionally, fermentation can take on a spiritual dimension. It demands mindfulness, respect for nature, and a deep connection to the process. Observing the transformation of soybeans into tempeh is a meditation on life’s cycles, the interplay of chaos and order, and the miracles that emerge from natural processes. In Javanese culture, where tempeh originated, it is more than food. It is a reflection of the people's spiritual and communal values. The act of making tempeh ties individuals to their heritage and to the rhythms of the natural world [ 7 ]. Tempeh nurtures in multiple ways. It sustains the body with its rich protein content and the soul with its deep cultural significance. Its versatility in the kitchen symbolizes adaptability—a willingness to absorb and enhance the flavours it encounters. This culinary philosophy extends to how we approach life: being open to new experiences, embracing change, and finding ways to contribute to the well-being of others. Tempeh is a living example of nourishment in its broadest sense—providing sustenance while connecting us to tradition, nature, and community [ 2 ]. The philosophy of tempeh fermentation offers profound lessons in transformation, patience, interconnectedness, and sustainability. From its humble beginnings as soybeans to its status as a global health food, tempeh’s journey reflects the principles of harmony and mindful living. It invites us to see the world as a collaborative process where growth arises from balance and care. Tempeh is not only a source of nourishment but also a metaphor for life’s potential—a reminder that with time, simplicity, and respect for nature, the most remarkable transformations can unfold. Tempeh is a testament to the quiet wisdom of fermentation and the enduring beauty of simplicity [ 2 , 7 ]. Characteristics of tempeh Malang fresh soybean tempeh Malang soybean tempeh was like other soybean tempeh, fermented using Rhizopus spp . Malang tempeh was considered to be one of the types of soybean tempeh with superior taste in East Java; others were Trenggalek tempeh from Trenggalek regency. This tempeh was collected from the traditional Lawang Market in Malang Regency, East Java Province, Indonesia. Visually, this Malang soybean tempeh was shaped like a white cake with a firm texture (Fig. 1 . a). The white colour of soybean tempeh comes from the mycelium of Rhizopus spp . Evenly distributed mycelium growth was a key indicator of good tempeh quality [ 7 ]. Malang soybean tempeh was widely used as a main meal or snack. Tempeh was often used as a treat at religious ceremonies, weddings, and other social events. Soybean tempeh had significant nutritional, cultural and economic value to the people of Indonesia. Traditionally, tempeh was consumed in various forms, such as sliced and fried, to which spices, such as coriander, salt, and onion, were added. Tempeh was also used in soups and stews and as a meat substitute in vegetarian dishes [ 7 ]. The process of making Malang soybean tempeh was the same as making soybean tempeh in general. The steps of the process included washing, soaking, peeling, boiling, cooling, and inoculating with tempeh starter, packaging, and fermentation at room temperature for 48 hours. When the plastic bag was used as wrapping material, inoculated soybean was put into a plastic bag before fermentation. Otherwise, the inoculated soybean was spread in a flat container made from woven bamboo or wood, covered with cloth, and let stand for 48 hours or until mycelia covered the whole soybean. This fermentation produces tempeh shaped like a solid cake bound by the mycelium of the mould [ 7 ]. Malang overripe soybean tempeh Malang overripe soybean tempeh was prepared by allowing fresh Malang tempeh to stay at room temperature for another 2–3 days. Visually, Malang overripe soybean tempeh was darker (light brownish) in colour compared to fresh tempeh, and there was an increase in pH value as reported previously [ 3 ], as well as strong aroma or flavour (Fig. 1 .b). The taste of Malang soybean overripe tempeh was more savoury than fresh tempeh. The savoury taste of soybean overripe tempeh was due to increased glutamate [ 14 ]. Overripe soybean tempeh was used in a spicy condiment called “ sambal tumpang ”, made by cooking crushed overripe tempeh in thick coconut milk with coriander, chilli, galanga, bay leaves, onion, and shallot. Malang overripe soybean tempeh had a softer texture compared to fresh tempeh. In addition to improving taste, overripe soybean tempeh had another advantage: increasing levels of bioactive compounds, such as antioxidants and isoflavones [ 15 ]. The changes in overripe tempeh were due to increased microbial activity and protein breakdown [ 15 ]. Malang stinky soybean tempeh Stinky soybean tempeh (Fig. 1 . c) was consumed as a flavouring agent by some tempeh users and was perceived to have an umami taste [ 3 ]. Stinky tempeh was not commercially available, and consequently, it was made at home by consumers by letting fresh tempeh stand at room temperature for 3–5 days. Stinky tempeh was made using soybean tempeh but was not used in other types of tempeh, and it was only used in small amounts as a flavouring agent for stir-fry or stew. Stinky tempeh had a strong aroma and a slightly bitter taste [ 3 ]. The aroma was described as a mixture of sulphur, cheese, and earth aroma [ 3 ]. Microorganisms in the advanced fermentation process included Rhizopus oligosporus , lactic acid bacteria, and other microbes [ 3 ]. These microorganisms break down protein components to produce volatile compounds. Prolonged fermentation resulted in a softer texture and increased free amino acid content. Stinky tempeh was also used in the making of “ sambal tumpang ,” but it was mixed with fresh tempeh, seemingly to reduce the strong odour and bitter taste. Peanut tempeh wrapped in banana leaves Peanut press cake tempeh was made from peanut press cake. This peanut tempeh was collected from the Lawang market in Malang Regency, East Java Province, Indonesia. The process of making peanut tempeh was similar to that of soybean tempeh. The fermentation process of peanut press cake tempeh was also carried out using Rhizopus sp . fungus [ 7 ]. Peanut press cake tempeh could be wrapped in banana leaves or a plastic bag (Fig. 1 . d). This type of tempeh might be considered rare, as it is not available in all areas in Java or other areas in Indonesia. In East Java province, it seemed to be available widely only in Malang regency. However, an individual tempeh producer, such as in Yogyakarta, had started using peanut press cake as their raw material. Peanut press cake tempeh showed less mycelium growth on its surface compared to soybean tempeh, resulting in a crumbly texture. Therefore, peanut press cake tempeh had a coarse appearance and black patches due to the seed coat (testa), which usually was not removed during the process. Peanut press cake tempeh was considered to have a tastier flavour than soybean tempeh but with a crumbly texture. Peanut press cake was usually deep-fried, stir-fried, or tempeh cracker as a snack or side dish. It was rarely cooked in other ways, such as with soybean tempeh. Peanut press cake tempeh wrapped in a plastic bag Peanut press cake tempeh wrapped in plastic did not seem different from that wrapped in banana leaves. Plastic wrap offered practicality and better durability, especially for distribution and storage (Fig. 1 .e). The use of plastic packaging did not affect the activity of microorganisms in tempeh, which was shown by the consistency of the total population of microorganisms [ 2 ]. Plastic could protect tempeh from external contamination, such as dust or excess moisture, making it more suitable for modern markets. Peanut press cake tempeh wrapped in plastic showed a more neutral aroma and flavour due to the inert nature of the plastic [ 2 ]. Due to a lack of air circulation, the surface texture tended to be denser and slightly drier. The choice of packaging type was tailored to consumer preferences and product distribution purposes. Benguk Tempeh Benguk tempeh (Fig. 1 . f) was made from velvet bean ( Mucuna pruriens ). It is mostly found in Yogyakarta province and its surrounding area, such as Wonogiri Regency, which belongs to Central Java Province. Benguk tempeh was rarely found in other places, which might be due to the availability of raw materials, technology, and consumers. Benguk tempeh was tastier, had a black or dark appearance, a rather coarser mouthfeel, and a harder texture compared to soybean tempeh. Since the benguk bean was larger (1 to 2 cm in length) than the soybean, benguk tempeh showed a larger lumpy appearance. Benguk tempeh was consumed as a snack or side dish. Benguk tempeh could be deep fried after marinated in salt and garlic, dipped in batter before frying, stir-fried, made into soup with coconut milk (“ lodeh ”), crackers coated in batter (“ kripik ”), and “ bacem ” (cooked about 30 minutes in spices of coriander, galanga, bay leaves, garlic, shallot, palm sugar, and salt, and then deep fried). There was a specialty made from benguk tempeh called “ besengek ” or “ senge k”, where benguk tempeh was cooked in thick coconut milk so that coconut milk curd covered the tempeh. The spices used in besengek included coriander, shallot, garlic, galanga, bay leaves, palm sugar, salt, and might be candle nut. Besengek seemed only available in Yogyakarta province, especially Kulon Progo regency and Wonogiri regency in Central Java. In Yogyakarta city, besengek was likely only sold in main markets such as Kranggan and Beringharjo market, and only available during Sunday. The sellers came from Kulon Progo Regency. This might indicate that the product is becoming rare. Nevertheless, when benguk tempeh was sold in online shops, the sellers were from Kulon Progo or Wonogiri in the form of fresh tempeh, cracker, or frozen bacem . The making process of benguk tempeh was rather lengthy as compared to that of soybean tempeh [36], which seemed to be due to the harder texture of the benguk bean. First, the benguk bean was washed and soaked in water for 24 hours , and then it was cooked in lime solution for 30 minutes until it was easy to peel the seed coat. After removing the seed coat, the bean was washed thoroughly and then soaked for 96 hours, and soaking water was replaced every 12 hours. The bean was then washed until there was no slippery feeling on the surface of the bean. Then, the bean was cooked for about 30 minutes until soft enough to maintain its integrity and not be too soft. The bean was then cooled to reach room temperature, inoculated with tempeh inoculum of Rhizopus spp., and packed in banana leaf every about 50 g, 3 to 5 mm thick, rectangle shape. The packs were then arranged in a single layer and incubated at room temperature for 5 days (120 hours). Koro Tempeh Koro/lablab bean ( Lablab purpureus ) looked similar in size and shape to the benguk bean but had a white seed colour. Koro tempeh (Fig. 1 . g) was similar to benguk tempeh in many ways. It also originated from Kulon Progo and Wonogiri regency. The use and cooking technique was similar to that of benguk tempeh. The product was also getting rare. We bought koro tempeh at the main market in Bantul City, Bantul Regency, in Yogyakarta province. We could not find koro tempeh in Kranggan or Beringharjo market in the city of Yogyakarta. The texture of koro tempeh was softer than that of soybean tempeh. Koro tempeh was also made into besengek . Dage Tempeh Dage tempeh (Fig. 1 . h) originated from the Banyumas regency in Central Java, especially Purwokerto City and its surrounding area. We could not find appropriate information about raw material for dage tempeh either from dage sellers in Pasar Manis (“Manis” market) or the layman that we interviewed. However, Romulo and Surya [ 15 ] mentioned that dage was made from fresh coconut press cake, cooked, cooled, and inoculated by Rhizopus spp., then fermented with inoculum of Rhizopus spp. for two days. A report from a local newspaper mentioned that dage was made from fresh coconut press cake or fresh coconut press cake added with sliced peanuts [ 17 ]. The appearance of dage tempeh (Fig. 1 .h.) looked more like gembus tempeh, a type of tempeh made from okara, where the mycelium was not as fluffy as that of soybean tempeh, and the texture was spongy and soft. However, there were some black spots scattered on the dage tempeh, which might be due to the testa (outer part of the endosperm) of the coconut. Coconut testa had a harder texture and dark brown colour. The process of making dage started with steaming the raw material, cooling, adding tempeh inoculum containing Rhizopus spp., and fermenting for two days [ 15 ]. The appearance and texture of dage tempeh were very similar to that of gembus tempeh, which was porous and softer than soybean tempeh. The price of dage tempeh was about the same as that of soybean tempeh. The way to consume dage tempeh was similar to that of soybean tempeh, which could be deep-fried with or without coating batter made from rice or wheat flour. Dage tempeh was also made into keripik (a cracker-like), where dage tempeh was sliced about 1 mm thick, coated with flour batter, and deep fried. Dage tempeh was also stir fried. The preference towards dage tempeh was probably related to its soft and spongy texture. Amino acids profile of fermented products The protein content of tempeh ranged from 13.97 ( gembus tempeh) to 45.5% (Malang fresh soybean tempeh) (Table 1 ). Tempeh showed good quality of protein as indicated by almost all amino acids scoring over 100 (Table 1 ), except for limiting amino acids. The amino acid score ranges from 36 ( gembus tempeh) to 102% (peanut press cake tempeh wrapped in banana leaves). Most of the samples had methionine and cysteine as limiting amino acids, except benguk tempeh, which also had leucine as a limiting amino acid. In contrast, peanut press cake tempeh had threonine as a limiting amino acid. Fermentation seemed to increase substantially the amino acid score of peanuts, which had an amino acid score of 24 to 28 before fermentation [ 18 ]. Table 1 Essential amino acids and amino acids score of different types of tempeh Amino Acid WHO Std., mg.g − 1 Benguk Tempeh Koro Tempeh Malang Fresh Soybean Tempeh Malang Overripe Soybean Tempeh Malang Stinky Soybean Tempeh Peanut Press Cake Tempeh, in Plastic Wrap Peanut Press Cake Tempeh, in Banana Leaf Wrap Gembus Tempeh Dage Tempeh Conc,mg.g − 1 Amino acid score Conc. mg.g − 1 Amino acid score Conc., mg.g − 1 Amino acid score Conc., mg.g − 1 Amino acid score Conc., mg.g − 1 Amino acid score Conc., mg.g − 1 Amino acid score Conc., mg.g − 1 Amino acid score Conc., mg.g − 1 Amino acid score Conc., mg.g − 1 Amino acid score His 19 30.39 159.94 36.93 194.40 31.12 163.79 27.12 142.74 26.52 139.61 34.06 179.29 35.83 188.58 62.66 329.82 48.18 253.60 Ile 28 12.25 43.75 56.17 200.63 52.68 188.14 56.06 200.23 57.75 206.25 51.30 183.22 47.39 169.25 45.97 164.19 80.73 288.34 Leu 66 178.68 270.73 85.78 129.98 80.26 121.62 84.70 128.33 88.52 134.12 79.86 121.01 71.94 109.01 64.37 97.53 69.27 104.95 Lys 58 58.95 101.65 77.66 133.91 62.81 108.29 58.95 101.64 49.43 85.23 68.91 118.81 65.66 113.21 78.66 135.63 66.97 115.47 Thr 34 39.11 115.03 44.54 131.02 35.86 105.49 34.43 101.27 31.90 93.84 37.57 110.51 34.97 102.87 41.04 120.71 44.33 130.41 Trp 11 29.69 269.91 11.38 103.51 24.89 226.28 24.90 226.40 24.63 223.93 25.66 233.33 25.37 230.63 5.73 52.11 14.60 132.75 Val 35 53.77 153.64 64.93 185.53 52.29 149.40 58.15 166.14 61.66 176.18 57.00 162.86 50.46 144.19 68.63 196.09 47.26 135.04 Met + Cys 25 11.97 47.91 11.78 47.14 14.06 56.24 17.38 69.55 16.76 67.05 15.86 63.44 57.00 228.02 9.17 36.68 12.34 49.36 Phe + Tyr 63 92.71 147.17 113.19 179.67 99.49 157.92 109.13 173.22 120.76 191.68 104.72 166.22 99.35 157.70 125.63 199.41 84.47 134.08 The total concentration of non-essential amino acids was greater than that of essential amino acids (Table 2 ). The highest concentration of amino acid was glutamate (a non-essential amino acid) in all tempeh except benguk tempeh, where leucine (an essential amino acid) was the predominant amino acid. The dominant essential amino acid in tempeh was leucine, except for gembus and dage tempeh, where the highest concentration of amino acids was phenylalanine and isoleucine, respectively. The level of essential amino acids in tempeh ranged from 6 ( gembus tempeh) to 18% (fresh Malang tempeh). The total concentration of non-essential amino acids ranged from 53 ( benguk or koro tempeh) to 59% (peanut press cake tempeh wrapped in banana leaves). The dominant non-essential amino acid was glutamate, except in benguk tempeh, where the highest concentration was shown by aspartic acid. Table 2 Amino acid quality of different types of tempeh Amino Acid Quality Parameter Benguk Tempeh Koro Tempeh Malang Fresh Soybean Tempeh Malang Overripe Soybean Tempeh Malang Stinky Soybean Tempeh Peanut press cake tempeh, in plastic wrap Peanut press cake tempeh, in banana leaves wrap Gembus Tempeh Dage Tempeh Protein (%) 33.76 27.61 45.53 40.61 41.60 40.91 41.47 13.97 15.09 EAA (%) 46.61 46.69 41.12 42.59 43.24 42.83 40.19 45.61 43.63 NEAA (%) 53.39 53.31 58.88 57.41 56.76 57.17 59.81 54.39 56.37 EAA/NEAA 0.87 0.87 0.69 0.74 0.76 0.75 0.67 0.84 0.77 Amino acid score 47.91 47.14 56.24 69.55 67.06 63.44 102.87 36.68 49.36 Limiting amino acid Ile, Met, Cys Met, Cys Met, Cys Met, Cys Met, Cys Met, Cys Thr Met, Cys Met, Cys LNAA (including Trp) 6.363 6.785 10.082 10.082 11.016 9.766 9.231 3.438 3.426 Trp/LNAA 0.187 0.048 0.122 0.112 0.103 0.120 0.128 0.023 0.068 Gembus and dage tempeh were made from soybean or coconut residue. Consequently, the nutrition was low (Table 1 ), and so were the parameters of amino acid quality (Table 2 ). Prolonged fermentation seemed to reduce the total concentration of non-essential amino acids (from 58.88% in fresh Malang tempeh to 56.76% in stinky Malang tempeh). It increased that of essential amino acids (from 41.12% in fresh Malang tempeh to 43.24% in stinky tempeh) (Tabel 2). The amino acid profile in tempeh seemed to be affected by the amino acid composition in the raw material. For example, glutamic acid was the predominant amino acid in soybean [ 19 ] or in coconut [ 20 ] and also in the form of tempeh. On the contrary, methionine was low in soybean [ 19 ] or coconut [ 20 ] and also in tempeh. Tempeh could be a candidate for a natural source of glutamate acid. Glutamic acid was suggested as a useful supplement for improving learning and memory performance and neurochemical status of the brain, and in the future, could be effective in the treatment of neurological disorders affecting learning and memory performance, which might be attributed to the improved antioxidant status and cholinergic, monoaminergic and glutamatergic neurotransmission in the brain and hippocampus [ 21 ]. Fermentation in tempeh increased protein content, which might be attributed to fungal biomass. The protein content of unfermented soybean seeds was 35–36% [ 22 ], and it was 40 to 45% in Malang tempeh (Table 2 ). Similarly, peanut, koro and benguk beans had a protein content of 25% [ 18 ], 25% [ 13 ] and 23% [ 23 ], respectively, while in the form of tempeh, their protein concentrations were 40, 27, and 37%, respectively (Table 2 ). Fermentation also altered amino acid composition, as shown by the higher concentration of valine, lysine, and isoleucine in tempeh (Table 1 ) than in raw soybean [ 22 ]. Fermentation of tempeh seemed to involve lactic acid bacteria from the surroundings. The bacteria that play a role in the fermentation of tempeh were the genus Lactobacillus [ 24 ]. Lactic acid bacteria in the tempeh manufacturing process showed the ability to hydrolyse soy protein and subsequently released amino acids such as leucine, phenylalanine, tyrosine, valine and isoleucine [ 24 ]. The amount and composition of tempeh bacteria were influenced by the production process, production environment and raw materials [ 25 ]. Packaging material did not affect the type of bacteria involved in tempeh-making [ 25 ]. Still, it could affect protein levels with higher protein in tempeh wrapped in leaves [ 2 , 25 ] . The ratio between tryptophan and other large neutral amino acids in the plasma determined the ability of tryptophan to cross the blood-brain barrier (BBB) for further synthesis into serotonin. This ratio value started from 0.09 or above [ 26 ]. In all tempeh samples, except koro , gembus , and dage tempeh , the ratio value of tryptophan to LNAA was over 0.102 (Table 2 ), and none was in the suggested range for the ability to cross the blood-brain barrier. Benguk tempeh showed substantially the highest value of this ratio, 0.187 (Table 2 ). Prolonged fermentation seemed to reduce this value, as indicated by Malang fresh tempeh (0.122) and Malang stinky tempeh (0.103) (Table 2 ). There was no report on whether the tryptophan/LNAA ratio in the diet would retain the ratio in the plasma. However, there were reports on the influence of food components such as protein and simple carbohydrates [ 8 ]. Protein hydrolysate was shown to improve tryptophan bioavailability [ 27 ]. Therefore, this technique might be applied to tempeh with a low tryptophan/LNAA ratio. Future, Prospects and Challenges of Development Tempeh Tempeh, a traditional Indonesian soy-based fermented food, has gained global recognition as a versatile, nutritious, and sustainable protein source. With increasing awareness about health and environmental issues, tempeh holds significant promise in addressing the dietary and ecological challenges of the modern world [ 28 ]. Tempeh is rich in protein, dietary fibre, vitamins, and minerals, making it a highly nutritious food option. Unlike unfermented soy products, tempeh’s fermentation process enhances its digestibility and reduces anti-nutritional factors like phytic acid. Moreover, it is a good source of probiotics, which support gut health [ 12 ]. As more people adopt plant-based diets for health reasons, tempeh’s nutritional profile makes it an appealing alternative to meat and other animal products. Scientific advancements in food technology are likely to optimize the nutritional content of tempeh further, potentially fortifying it with additional nutrients to meet specific dietary needs [ 15 ]. The global market for plant-based foods has been expanding rapidly, driven by shifting consumer preferences and increasing investment in sustainable food technologies. Tempeh, with its versatility and adaptability, is well-positioned to capitalize on this trend. According to market research, the global tempeh market is expected to grow at a compound annual growth rate (CAGR) of over 6% in the coming years. Tempeh’s firm texture and ability to absorb flavours make it suitable for a wide range of dishes, from stir-fries to sandwiches and even snacks. This versatility appeals to chefs and home cooks alike. As more people become aware of the environmental and health benefits of plant-based diets, demand for products like tempeh is expected to rise. Companies are developing innovative tempeh-based products, such as pre-seasoned tempeh, tempeh burgers, and tempeh nuggets, to attract a broader audience. Advances in packaging and storage technology are also extending tempeh’s shelf life, making it more accessible to global markets. Encouraging local production of tempeh in regions outside Southeast Asia can reduce costs and promote its adoption in diverse cultures. Startups and small businesses are already leveraging this opportunity [ 28 ]. Despite its potential, tempeh faces challenges that could hinder its growth. One key obstacle is consumer unfamiliarity in certain regions, where it is perceived as an exotic or niche product. Education and marketing campaigns highlighting tempeh’s benefits can address this issue. Additionally, concerns about soy production, including deforestation and pesticide use, may deter environmentally conscious consumers. Promoting the use of organic and non-GMO soybeans, as well as exploring alternative ingredients, can help mitigate these concerns [ 29 ]. Tempeh production is highly dependent on precise conditions such as temperature, humidity, and the quality of the starter culture. Small variations in these factors can result in inconsistent quality and texture, which can affect the final product's consumer appeal. The fermentation process is also sensitive to contamination, which can lead to spoilage or undesirable flavours [ 30 ]. While small-scale artisanal tempeh production can be easily managed, scaling the process to meet increasing demand, particularly in Western markets, introduces complexities. Large-scale production often involves automation, but this can lead to a loss of the traditional artisanal qualities that some consumers prefer. Additionally, the equipment needed for large-scale fermentation is expensive, and maintaining quality control on a larger scale requires substantial investment. For tempeh to be produced on a large scale, reliable sources of high-quality soybeans are essential [ 31 ]. However, soybeans face competition from other crops, and issues like climate change, fluctuating prices, and supply chain disruptions can affect their availability and cost. Furthermore, in some regions, the cost of production and transportation can make tempeh less affordable compared to other protein sources. Despite its long history in Southeast Asia, tempeh faces challenges in terms of consumer acceptance, especially in Western countries. One of the primary barriers to tempeh’s wider acceptance is its taste and texture, which differ significantly from other plant-based proteins such as tofu. Tempeh has a nutty, earthy flavour that may not appeal to all palates, especially those unfamiliar with fermented foods. Its firm and slightly crumbly texture also contrasts with the softer, smoother textures of more familiar plant proteins [ 32 ]. In many Western countries, soy-based products like tofu have been available for decades, but tempeh is still relatively unknown. Cultural unfamiliarity with tempeh often translates into hesitation or lack of interest in trying it. It is commonly perceived as an exotic food, which can create psychological barriers for consumers who prefer familiar and locally sourced options. The challenge of educating consumers about tempeh’s health benefits, versatility, and potential as a protein source is significant. Although plant-based eating has gained popularity, tempeh is often overshadowed by other vegan or vegetarian protein options like lentils, chickpeas, and packaged plant-based meat substitutes. The lack of awareness about the health benefits and sustainability of tempeh makes it difficult to gain widespread consumer traction [ 33 ]. As the demand for plant-based proteins increases globally, tempeh faces significant competition from a variety of alternatives. Products like tofu, seitan, and newer innovations in plant-based meats often receive more attention due to their similarity to meat in taste and texture. These products often dominate the plant-based protein market, leaving tempeh as a niche offering. Tempeh, while generally affordable, can be more expensive than other plant proteins, particularly in regions where it is not widely produced. This price discrepancy can make it harder for tempeh to compete with cheaper, mass-produced alternatives like lentils and beans, especially in budget-conscious markets. Additionally, tempeh may not be as widely available in grocery stores, further limiting its reach. Newer plant-based products, especially those designed to mimic meat, are marketed as convenient, ready-to-eat, or quick-cooking options. In contrast, tempeh typically requires cooking and may be perceived as less convenient. While tempeh can be prepared in various ways, the perceived effort involved in cooking it might deter potential consumers from choosing it over more straightforward alternatives [ 33 ]. While tempeh offers numerous benefits as a nutritious, plant-based protein, it faces a range of challenges that need to be addressed to ensure its widespread adoption. From production hurdles, including quality control and supply chain issues, to consumer perception and competition with other plant-based proteins, many factors limit tempeh’s potential in the global market. However, with increasing awareness of plant-based diets and sustainability concerns, tempeh has the opportunity to gain popularity. Overcoming challenges related to taste, texture, and cultural acceptance, as well as addressing sustainability issues in soybean farming, will be essential to its long-term success and growth in the global food industry. Conclusion This work revealed nutritional and functional health benefits related to the amino acid profile of different types of tempeh available in Indonesian markets. Methionine and cysteine were the limiting amino acids in tempeh samples, except peanut press cake tempeh, which showed threonine as the limiting amino acid. Tryptophan in tempeh had a ratio of tryptophan/LNAA more than 0.1, indicating a probable future role in alleviating depression, except for benguk , gembus, and dage tempeh. Different types of tempeh and prolonged fermentation affected the amino acid profile and protein concentration of tempeh. Abbreviations LNAA: Large neutral amino acids GMO: Genetically modified organism OPA : O-phtalalhdehide 5-HT : 5-hydroxytryptamine CAGR : Compound annual growth rate BBB : blood-brain barrier Declarations Acknowledgement This work was funded by the Ministry of Higher Education, Research, and Technology, the Government of Indonesia, for the KATALIS scheme in 2024, contract no. 054/E5/PG.02.00.PL/2024. Authors contribution All authors contributed equally as the main contributors in this paper. UP, DPM, NN, ECS, and ES formulated and designed the research. DH, AR, and ESN conducted data collection and processing. IP, JJ, ER, SA, CLS, BK, and RHBS contributed to the discussion. RHBS constructed philosophy, history, and future prospect studies. Funding This work was funded by the Ministry of Higher Education, Research, and Technology, the Government of Indonesia, for the KATALIS scheme in 2024, contract no. 054/E5/PG.02.00.PL/2024. Availability of data and materials The author is willing to provide data and materials according to the agreed research needs and ethics. Ethics approval and consent to participate Not applicable Consent for publication Not applicable Competing Interests All authors declared that there is no competing interest in the writing of this manuscript. Authors' information Umi Purwandari 1,8 , Darimiyya Hidayati 1 , Askur Rahman 1 , Enung Siti Nurhidayah 1 , Indah Purwantini 2 , Era Catur Prasetyo 3 , Nurhayati Nurhayati 4 , Dwiyati Puji Mulyani 5 , Jay Jayus 4 , Eka Ruriani 4 , Enny Suswati 4 , Satya Arimurti 4 , Chatarina Lilis Suryani 5 , Bayu Kanetro 5 , R. Haryo Bimo Setiarto 6,7 1 University of Trunojoyo Madura, Jl. Raya Telang, Kamal, Bangkalan, East Java, Indonesia 69162 2 Faculty of Pharmacy, University of Gadjah Mada, Sekip Utara, Yogyakarta, Indonesia 55281 3 Muhammadiyah University of Surabaya, Jl. Sutorejo No. 59 Surabaya, East Java, Indonesia 60113 4 University of Jember, Jl. Kalimantan 37, Kampus Bumi Tegalboto, Kotak Pos 159, Jember, east Java, Indonesia 68121 5 University of Mercu Buana Yogyakarta, Jl. Ring Road Utara, Yogyakarta, Indonesia 55753 6 Research Center for Applied Microbiology, National Research and Innovation Agency (BRIN), Jl. Raya Jakarta-Bogor KM 46, KST Soekarno, Cibinong, Bogor, Indonesia 16911. 7 Research Collaboration Center for Traditional Fermentation, National Research and Innovation Agency (BRIN) 8 Indonesia Tempeh Forum (Forum Tempe Indonesia) East Java Branch. Surabaya References Tamam B, Syah D, Suhartono MT, Kusuma WA, Tachibana S, Lioe HN. Proteomic study of bioactive peptides from tempe. J Biosci Bioeng [Internet]. 2019;128(2):241–8. Available from: https://doi.org/10.1016/j.jbiosc.2019.01.019. Ellent SSC, Dewi L, Tapilouw MC. Quality characteristics of soybean ( Glycine max L .) tempeh packaged with corn husks. 2022;11(1):32–40. Utami R, Wijaya CH, Lioe HN. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5724562","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":432177396,"identity":"af05c3bc-f23c-4391-9b33-2657c6f5a25a","order_by":0,"name":"Umi 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Madura","correspondingAuthor":false,"prefix":"","firstName":"Darimiyya","middleName":"","lastName":"Hidayati","suffix":""},{"id":432177398,"identity":"73f8d2e1-d415-4d44-9271-5a1b805e064a","order_by":2,"name":"Askur Rahman","email":"","orcid":"","institution":"Universitas Trunojoyo Madura","correspondingAuthor":false,"prefix":"","firstName":"Askur","middleName":"","lastName":"Rahman","suffix":""},{"id":432177399,"identity":"247cd419-5a7e-48df-b21c-061544b718f5","order_by":3,"name":"Enung Siti Nurhidayah","email":"","orcid":"","institution":"Universitas Trunojoyo Madura","correspondingAuthor":false,"prefix":"","firstName":"Enung","middleName":"Siti","lastName":"Nurhidayah","suffix":""},{"id":432177400,"identity":"a4e0161f-9b68-4637-8b3d-33b6a5448028","order_by":4,"name":"Indah Purwantini","email":"","orcid":"","institution":"Gadjah Mada University: Universitas Gadjah Mada","correspondingAuthor":false,"prefix":"","firstName":"Indah","middleName":"","lastName":"Purwantini","suffix":""},{"id":432177401,"identity":"35305e79-8385-4a00-b336-bfcc868ab08d","order_by":5,"name":"Era Catur Prasetyo","email":"","orcid":"","institution":"University of Muhammadiyah Surabaya: Universitas Muhammadiyah Surabaya","correspondingAuthor":false,"prefix":"","firstName":"Era","middleName":"Catur","lastName":"Prasetyo","suffix":""},{"id":432177402,"identity":"eea56ae2-17f7-4e9b-a08b-579f70bd920b","order_by":6,"name":"Nurhayati Nurhayati","email":"","orcid":"","institution":"Jember University: Universitas Jember","correspondingAuthor":false,"prefix":"","firstName":"Nurhayati","middleName":"","lastName":"Nurhayati","suffix":""},{"id":432177403,"identity":"011f06c6-a3c2-4471-80d0-9456a3f943a7","order_by":7,"name":"Dwiyati Puji Mulyani","email":"","orcid":"","institution":"Mercu Buana University Yogyakarta: Universitas Mercu Buana Yogyakarta","correspondingAuthor":false,"prefix":"","firstName":"Dwiyati","middleName":"Puji","lastName":"Mulyani","suffix":""},{"id":432177404,"identity":"94ceb6e2-0df9-463b-8d37-8e2b02a49b62","order_by":8,"name":"Jay Jayus","email":"","orcid":"","institution":"Universitas Jember","correspondingAuthor":false,"prefix":"","firstName":"Jay","middleName":"","lastName":"Jayus","suffix":""},{"id":432177405,"identity":"8917af23-f397-4eda-9aa0-4dec8b84a27f","order_by":9,"name":"Eka Ruriani","email":"","orcid":"","institution":"Jember University: Universitas Jember","correspondingAuthor":false,"prefix":"","firstName":"Eka","middleName":"","lastName":"Ruriani","suffix":""},{"id":432177406,"identity":"dd753903-1e0b-454a-9287-0e32d5186251","order_by":10,"name":"Enny Suswati","email":"","orcid":"","institution":"Jember University: Universitas Jember","correspondingAuthor":false,"prefix":"","firstName":"Enny","middleName":"","lastName":"Suswati","suffix":""},{"id":432177407,"identity":"6567028b-b171-4dd6-bfa8-a4b9ce8621b2","order_by":11,"name":"Satya Arimurti","email":"","orcid":"","institution":"Jember University: Universitas Jember","correspondingAuthor":false,"prefix":"","firstName":"Satya","middleName":"","lastName":"Arimurti","suffix":""},{"id":432177408,"identity":"fff3d98b-dc2d-4bf6-9dfd-b9f1ae2c1989","order_by":12,"name":"Chatarina Lilis Suryani","email":"","orcid":"","institution":"Mercu Buana University Yogyakarta: Universitas Mercu Buana Yogyakarta","correspondingAuthor":false,"prefix":"","firstName":"Chatarina","middleName":"Lilis","lastName":"Suryani","suffix":""},{"id":432177409,"identity":"52160924-d063-41c5-966d-d9c06b307995","order_by":13,"name":"Bayu Kanetro","email":"","orcid":"","institution":"Mercu Buana University Yogyakarta: Universitas Mercu Buana Yogyakarta","correspondingAuthor":false,"prefix":"","firstName":"Bayu","middleName":"","lastName":"Kanetro","suffix":""},{"id":432177410,"identity":"863e52ee-f7d9-48bb-919c-ce435c0a6926","order_by":14,"name":"R. Haryo Bimo Setiarto","email":"","orcid":"","institution":"National Agency for Research and Innovation: Agencia Nacional de Investigacion e Innovacion","correspondingAuthor":false,"prefix":"","firstName":"R.","middleName":"Haryo Bimo","lastName":"Setiarto","suffix":""}],"badges":[],"createdAt":"2024-12-28 06:00:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5724562/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5724562/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s42779-025-00285-3","type":"published","date":"2025-06-06T15:57:50+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":79565706,"identity":"720585d3-f6d9-49d4-b20a-3371ff06888c","added_by":"auto","created_at":"2025-03-31 09:28:09","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1415552,"visible":true,"origin":"","legend":"\u003cp\u003eTempeh varieties (a) soybean fresh tempeh, (b) overripe soybean tempeh, (c) stinky soybean tempeh, (d) peanut tempeh wrapped in banana leaves, (e) peanut tempeh wrapped in a plastic bag, (f) \u003cem\u003ebenguk\u003c/em\u003e tempeh, (g) \u003cem\u003ekoro\u003c/em\u003e tempeh, dan (h) \u003cem\u003edage\u003c/em\u003etempeh\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-5724562/v1/8085d5e7cb7e775833f2b9de.png"},{"id":84242614,"identity":"f69cb280-dfe0-41a4-b93f-f800be354468","added_by":"auto","created_at":"2025-06-09 16:10:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3036186,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5724562/v1/0944e7c4-f537-4927-9f7c-b9a1006f99d2.pdf"}],"financialInterests":"","formattedTitle":"Exploring the profile of amino acids and ethnography study in the biodiversity of tempeh as fermented foods of Indonesia","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFermented food or beverages have evolved from preserved into functional foods or drinks. Most fermented foods or drinks were acquired from ethnic treasures. One of the successful worldwide fermented foods was tempeh (or tempe), which originated in Indonesia. The health benefits of tempeh have been extensively reported or reviewed [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Different types of tempeh were created based on their raw materials, which are commercially available in traditional markets in Indonesia. These include peanut press cake, okara, velvet bean, and coconut press cake. The main fungus used was \u003cem\u003eRhizopus\u003c/em\u003e spp. Wrapping material used in tempeh production in Indonesia was mostly plastic bags and banana leaves, and sometimes teak or canna leaves were used. Tempeh fermented in plastic bag showed a higher number of total bacteria but less concentration of soluble protein (0.17 to 0.45 mg.L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) than those wrapped in banana (0.18 to 0.61 mg.L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) or teak leaves (0.18 to 0.60%), or slightly lower (66.42%) than those wrapped in banana leaves (67.58%) or teak leaves [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. However, tempeh wrapped in plastic bags became stinky faster and had less preference toward flavour than those wrapped in banana leaves [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFermentation of tempeh has been reported to alter protein and amino acids profile. Prolonged fermentation to 96 hours increased protein content from 34.9 to 53.43% and pH from 5.4 to 6.1 [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Similarly, total amino acids increased to 72 hours of fermentation, from 69.3 to 200.7 g.L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, but then decreased to 190.1 g.L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Amino acid composition determined the sensory characteristics of tempeh. Umami taste was correlated to aspartic and glutamic acids [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. The sweetness of tempeh was correlated to alanine, glycine, serine, and threonine [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], bitterness was correlated to arginine, histidine, isoleucine, leucine, methionine, phenylalanine, valine, and tyrosine [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eProteolysis of soybean protein during fermentation of tempeh was likely to result in amino acids and peptides. Nevertheless, prolonged fermentation might degrade amino acids to reduce their concentration, especially those susceptible to microbial degradation [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Therefore, fermentation might increase or reduce amino acid concentration during fermentation. Due to fermentation, nitrogen in soybean seed was reduced (from 6.65 to 2.98 gN), protein was increased (from 8 to 17 g), threonine was reduced (from 1603 to 742 mg), tryptophan was reduced (from 532 to 235 mg), and lysine was reduced (from 2653 to 1120 mg) per 100 g [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOne of the most studied amino acids in relation to mood or mental state was tryptophan. Tryptophan was a precursor of serotonin, a hormone -also known as 5-HT (contraction of 5-hydroxytryptamine) related to mood, cognition, reward, learning, and memory, where the synthesis occurs in the brain [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Although serotonin was also produced in the large intestine by the influence of gut microbiota, this plasma serotonin could not pass the blood-brain barrier (BBB) to get into the central nervous system and affect mood [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Therefore, serotonin synthesis in the brain relies mostly on dietary tryptophan and is strongly related to dietary tryptophan [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Similarly, in healthy subjects, high tryptophan food resulted in reduced depression and stress as well as improved mood [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAlthough there were reports on a positive correlation between dietary tryptophan and brain serotonin [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], tryptophan uptake in brain cells was influenced by several factors, including the presence of other large neutral amino acids (LNAA) in the plasma. Tryptophan was one of LNAA, with others, namely phenylalanine, tyrosine, threonine, methionine, valine, isoleucine, leucine, and histidine. There was a competition for transporter among LNAA during passing BBB [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Consequently, the ratio of tryptophan to other LNAA was important to determine the concentration of tryptophan to pass over BBB. The proposed ratio of tryptophan to other LNAA was 0.09 or above [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Another factor that influenced tryptophan uptake was dietary carbohydrates. Consumption of carbohydrates after fasting was reported to induce secretion of insulin, which increases tryptophan availability to cross over BBB by removing another amino acid in the plasma. Tryptophan transport to the brain was also affected by albumin since tryptophan is lipophilic and needs albumin to allow it to solubilize in the plasma. This implied the important role of the food matrix in the formation of serotonin from dietary tryptophan. Nutrient-based treatment had been proposed previously in order to support alleviating manic-depressive disorder, especially referring to high tryptophan concentration in the diet [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Different types of protein or their concentration in the diet affected tryptophan/LNAA differently, and the effect was also affected by simple carbohydrates in the diet and their concentration [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTryptophan was not synthesized in the human body, so it had to be provided in the diet. The daily requirement of tryptophan in adults was 250 to 425 mg.day-1 or 3.5-6 mg.kg-1 body weight a day, while in children, it was 12 mg.kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e body weight a day [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Tryptophan in tempeh might be considered a minor amino acid, being the lowest concentration compared to others, ranging from 0.13% (wb) [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] to around 2% [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The amino acid score of tryptophan in \u003cem\u003ebenguk\u003c/em\u003e tempeh was 59.01 [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], and thus was considered very low and not adequate for good nutrition. This work studied the amino acid profile and ethnographic aspects of several types of tempeh from Java, Indonesia. Subsequently, it analysed them for their possible effect on alleviating depression as expressed by tryptophan content.\u003c/p\u003e"},{"header":"Methodology","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMaterials\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eSamples were collected from traditional markets in production areas in Malang (fresh Malang tempeh, peanut tempeh, dan gembus tempeh), Manis market in Purwokerto (\u003cem\u003edage\u003c/em\u003e tempeh), and Bantul (\u003cem\u003ekoro tempeh\u003c/em\u003e), and Yogyakarta province (\u003cem\u003ebenguk tempeh\u003c/em\u003e). About 0.5 kg of samples were bought from the sellers, kept in an iced-containing container, dried, and kept in a sealed plastic bag until used for analysis. Benguk tempeh was bought from an online shop. Information on the process of making the product was collected from the seller or the maker(s).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMethods\u003c/h3\u003e\n\u003cp\u003eSamples were first digested using the Kjeldhal method to determine protein concentration and then reacted with O-phtalalhdehide (OPA) [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. They were then analysed for amino acid content using HPLC (Thermos Scientific ODS-1 Hyersil) with a fluorescent detector. The mobile phase was Na-acetate (pH 6.5, 0.02%), Na-EDTA (0.005%), methanol (9%), and THF (1,5%) [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe nutritional value of protein was determined by calculating the amino acid score. Amino acid scores were obtained by comparing essential amino acids (mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e protein) with the WHO amino acid standard [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. This score was based on the FAO-WHO's nine amino acid requirements for preschool children.\u003c/p\u003e"},{"header":"Result and Discussion","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eThe History, Ethnography and Philosophy of Tempeh\u003c/h2\u003e \u003cp\u003eTempeh, a fermented soy product, has a long and rich history rooted in Southeast Asia, particularly in Indonesia. This nutritious and versatile food has grown from being a regional specialty to gaining international recognition as a staple in plant-based diets. Its history is deeply intertwined with the cultural, agricultural, and culinary practices of the regions where it originated. Tempeh's history can be traced back over 400 years to Java, Indonesia, where it is believed to have first been developed. The exact origins remain unclear, as tempeh was not explicitly mentioned in ancient texts until the 19th century. However, oral traditions and historical records suggest that tempeh-making techniques were passed down through generations long before they were documented. The development of tempeh was likely a result of resourceful innovation. Soybeans, introduced to Indonesia from China, became a common crop in the region. Javanese farmers discovered that fermenting soybeans with \u003cem\u003eRhizopus\u003c/em\u003e mould not only enhanced the nutritional profile of the beans but also made them more digestible and flavourful. This process also allowed for longer storage, making tempeh a practical solution in tropical climates [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn Indonesia, tempeh is more than just a food\u0026mdash;it is a cultural symbol and a dietary staple. It plays a vital role in Javanese cuisine, where it is often fried, steamed, or cooked in various dishes. Its affordability, high protein content, and versatility have made it an essential source of nutrition for generations of Indonesians, especially in rural areas where access to meat and fish might be limited. Tempeh is commonly prepared with traditional spices and served alongside rice, the staple carbohydrate of Indonesian meals. Dishes such as \u003cem\u003etempeh goreng\u003c/em\u003e (fried tempeh), \u003cem\u003etempeh orek\u003c/em\u003e (sweet and spicy tempeh), and \u003cem\u003esambal goreng tempeh\u003c/em\u003e (tempeh in spicy chili sauce) are beloved across the archipelago [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs tempeh was an important part of Javanese daily meals, several types of tempeh had been developed, either using different processes or raw materials instead of soybeans. For example, \u003cem\u003eTempe koro\u003c/em\u003e is made from lablab beans, and Tempe koro benguk is made from velvet beans. Tempe koro is widely produced and consumed by people in Yogyakarta province. The reason for making these types of tempeh might be the availability of beans in the area. However, later, people may find that their taste was different from soybean tempeh, and the koro and koro benguk tempeh became a specialty of the area. Creativity seemed to lead to the use of residue from some products, such as peanut oil, tofu, or coconut milk. Tempeh is made from peanut press cake, locally called \u0026ldquo;\u003cem\u003etempe kacang\u003c/em\u003e\u0026rdquo;, also known as \u0026ldquo;\u003cem\u003etempe bungkil\u003c/em\u003e\u0026rdquo;, and is only available now mainly in Malang, but also to a lesser extent in Yogyakarta and some areas in Central Java [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTempeh made from \u0026lsquo;okara\u0026rsquo; a residue of tofu, is called \u0026ldquo;\u003cem\u003etempe gembus\u003c/em\u003e\u0026rdquo; which is widely available. Another tempeh is made from the residue of coconut milk; this tempeh is called \u0026ldquo;\u003cem\u003etempe bongkrek\u003c/em\u003e\u0026rdquo;, and its consumption in the past led to fatalities due to highly toxic \u003cem\u003ebongkrek\u003c/em\u003e acid. This made \u003cem\u003etempe bongkrek\u003c/em\u003e banned since the 80\u0026rsquo;s. \u003cem\u003eTempe dage\u003c/em\u003e is reported to be made from coconut milk residue, but the exact raw material and the making process is not extensively reported. Another type of tempeh uses different species of fungi, called \u003cem\u003eoncom\u003c/em\u003e, which uses \u003cem\u003eChrysonilia sitophila\u003c/em\u003e (orange \u003cem\u003eoncom\u003c/em\u003e) or \u003cem\u003eRhizopus\u003c/em\u003e spp. (black \u003cem\u003eoncom\u003c/em\u003e), and raw material of okara or peanut press cake. Fermentation of tempeh can occur inside wrap leaves or in open spaces. The leaves used as tempeh wrap vary, depending on availability. The most commonly used is a banana leaf, which is considered the finest wrapping material. The less commonly used are canna leaves, sea hibiscus leaves, or teak leaves. Bamboo logs (about 30 cm long) were also used as wrapping material, but their use is very rare now. The perforated plastic bag is now a very common wrapping material [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe spread of tempeh beyond Indonesia began in the 20th century, driven by globalization and increasing interest in vegetarian and vegan diets. Dutch colonization played a significant role in introducing tempeh to Europe. During their time in Indonesia, the Dutch developed a taste for local cuisine, and tempeh was among the foods that found a place in Dutch kitchens. In the 1960s and 1970s, tempeh began gaining popularity in the United States and other Western countries. The growing health food movement and interest in plant-based protein sources fuelled this surge. Researchers and food enthusiasts started studying tempeh, recognizing its high protein content, probiotics, and ability to fit into various cuisines [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eToday, tempeh is produced and consumed worldwide, with manufacturers adapting traditional methods for large-scale production. Its health benefits, including being a rich source of protein, vitamins, and dietary fibre, have made it a favourite among health-conscious consumers and vegetarians. Tempeh\u0026rsquo;s texture and ability to absorb flavours make it an excellent meat substitute in recipes ranging from stir-fries to burgers. Countries such as the United States, Japan, and the Netherlands now produce tempeh commercially, often tailoring it to suit local tastes. In some regions, other legumes and grains like chickpeas, black beans, and quinoa are used to create variations of traditional tempeh [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDespite its global spread, tempeh remains deeply tied to its Indonesian roots. In recent years, efforts have been made to preserve traditional tempeh-making techniques and recognise their cultural importance. Festivals, workshops, and culinary events in Indonesia celebrate tempeh as a national treasure and a symbol of sustainable eating. As awareness grows about the environmental impact of meat production, tempeh's role as a sustainable protein source is increasingly acknowledged. Its production requires fewer resources compared to animal agriculture, making it an eco-friendly choice for a growing global population. The history of tempeh is a testament to the ingenuity and resourcefulness of the Javanese people. From its humble beginnings in rural Indonesia to its position as a global health food, tempeh continues to inspire innovation in both culinary and agricultural practices. Its journey reflects not only the evolution of food but also the interconnectedness of culture, sustainability, and nutrition in our modern world [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTempeh, a traditional Indonesian food made from fermented soybeans, is much more than a culinary delight. It embodies a philosophy rooted in transformation, patience, sustainability, and interconnectedness [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. At its core, fermentation represents transformation. In the case of tempeh, humble soybeans undergo a profound change through the action of \u003cem\u003eRhizopus oryzae\u003c/em\u003e mould. This microorganism binds the beans into a cohesive cake, creating a product that is more flavourful, nutritious, and digestible than its original form. This transformation is a metaphor for the potential within all living things. It shows how patience and the right conditions can turn the ordinary into the extraordinary. Life, like tempeh, often requires time and care to reveal its full potential [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTempeh fermentation is an interconnected process. It relies on a delicate balance of soybeans, mould, air, and warmth, each playing an essential role. This interdependence reflects the broader truth that nothing exists in isolation. Ecosystems thrive through collaboration in nature, and the same principle applies to human relationships. Tempeh teaches us the value of working together harmoniously, where each element contributes to a greater whole. It is a celebration of unity between microorganisms, humans, and the environment that results in a nourishing outcome [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFermentation cannot be rushed. The process requires time, a stable environment, and a degree of uncertainty as nature takes its course. This patience mirrors the rhythms of life, where meaningful outcomes often require trust in the journey rather than immediate results. Tempeh teaches us to accept life\u0026rsquo;s natural pace and to appreciate the unseen work happening beneath the surface. In a world increasingly driven by instant gratification, the philosophy of fermentation offers a counterbalance, encouraging mindfulness and faith in slow, deliberate progress [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTempeh is modest in appearance yet profound in its impact. It is not a luxury food, but its simplicity and versatility sustain millions. This humility reflects a philosophy of valuing substance over spectacle. In a society that often prioritizes extravagance, tempeh reminds us of the beauty in simplicity. Its unassuming nature belies its richness\u0026mdash;both as a source of nutrition and as a symbol of cultural and environmental mindfulness [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFor those who prepare tempeh traditionally, fermentation can take on a spiritual dimension. It demands mindfulness, respect for nature, and a deep connection to the process. Observing the transformation of soybeans into tempeh is a meditation on life\u0026rsquo;s cycles, the interplay of chaos and order, and the miracles that emerge from natural processes. In Javanese culture, where tempeh originated, it is more than food. It is a reflection of the people's spiritual and communal values. The act of making tempeh ties individuals to their heritage and to the rhythms of the natural world [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTempeh nurtures in multiple ways. It sustains the body with its rich protein content and the soul with its deep cultural significance. Its versatility in the kitchen symbolizes adaptability\u0026mdash;a willingness to absorb and enhance the flavours it encounters. This culinary philosophy extends to how we approach life: being open to new experiences, embracing change, and finding ways to contribute to the well-being of others. Tempeh is a living example of nourishment in its broadest sense\u0026mdash;providing sustenance while connecting us to tradition, nature, and community [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe philosophy of tempeh fermentation offers profound lessons in transformation, patience, interconnectedness, and sustainability. From its humble beginnings as soybeans to its status as a global health food, tempeh\u0026rsquo;s journey reflects the principles of harmony and mindful living. It invites us to see the world as a collaborative process where growth arises from balance and care. Tempeh is not only a source of nourishment but also a metaphor for life\u0026rsquo;s potential\u0026mdash;a reminder that with time, simplicity, and respect for nature, the most remarkable transformations can unfold. Tempeh is a testament to the quiet wisdom of fermentation and the enduring beauty of simplicity [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eCharacteristics of tempeh\u003c/h3\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eMalang fresh soybean tempeh\u003c/h2\u003e \u003cp\u003eMalang soybean tempeh was like other soybean tempeh, fermented using \u003cem\u003eRhizopus spp\u003c/em\u003e. Malang tempeh was considered to be one of the types of soybean tempeh with superior taste in East Java; others were Trenggalek tempeh from Trenggalek regency. This tempeh was collected from the traditional Lawang Market in Malang Regency, East Java Province, Indonesia. Visually, this Malang soybean tempeh was shaped like a white cake with a firm texture (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. a). The white colour of soybean tempeh comes from the mycelium of \u003cem\u003eRhizopus spp\u003c/em\u003e. Evenly distributed mycelium growth was a key indicator of good tempeh quality [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eMalang soybean tempeh was widely used as a main meal or snack. Tempeh was often used as a treat at religious ceremonies, weddings, and other social events. Soybean tempeh had significant nutritional, cultural and economic value to the people of Indonesia. Traditionally, tempeh was consumed in various forms, such as sliced and fried, to which spices, such as coriander, salt, and onion, were added. Tempeh was also used in soups and stews and as a meat substitute in vegetarian dishes [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe process of making Malang soybean tempeh was the same as making soybean tempeh in general. The steps of the process included washing, soaking, peeling, boiling, cooling, and inoculating with tempeh starter, packaging, and fermentation at room temperature for 48 hours. When the plastic bag was used as wrapping material, inoculated soybean was put into a plastic bag before fermentation. Otherwise, the inoculated soybean was spread in a flat container made from woven bamboo or wood, covered with cloth, and let stand for 48 hours or until mycelia covered the whole soybean. This fermentation produces tempeh shaped like a solid cake bound by the mycelium of the mould [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMalang overripe soybean tempeh\u003c/h3\u003e\n\u003cp\u003eMalang overripe soybean tempeh was prepared by allowing fresh Malang tempeh to stay at room temperature for another 2\u0026ndash;3 days. Visually, Malang overripe soybean tempeh was darker (light brownish) in colour compared to fresh tempeh, and there was an increase in pH value as reported previously [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], as well as strong aroma or flavour (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.b). The taste of Malang soybean overripe tempeh was more savoury than fresh tempeh. The savoury taste of soybean overripe tempeh was due to increased glutamate [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Overripe soybean tempeh was used in a spicy condiment called \u0026ldquo;\u003cem\u003esambal tumpang\u003c/em\u003e\u0026rdquo;, made by cooking crushed overripe tempeh in thick coconut milk with coriander, chilli, galanga, bay leaves, onion, and shallot.\u003c/p\u003e \u003cp\u003eMalang overripe soybean tempeh had a softer texture compared to fresh tempeh. In addition to improving taste, overripe soybean tempeh had another advantage: increasing levels of bioactive compounds, such as antioxidants and isoflavones [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. The changes in overripe tempeh were due to increased microbial activity and protein breakdown [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eMalang stinky soybean tempeh\u003c/h3\u003e\n\u003cp\u003eStinky soybean tempeh (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. c) was consumed as a flavouring agent by some tempeh users and was perceived to have an umami taste [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Stinky tempeh was not commercially available, and consequently, it was made at home by consumers by letting fresh tempeh stand at room temperature for 3\u0026ndash;5 days. Stinky tempeh was made using soybean tempeh but was not used in other types of tempeh, and it was only used in small amounts as a flavouring agent for stir-fry or stew. Stinky tempeh had a strong aroma and a slightly bitter taste [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. The aroma was described as a mixture of sulphur, cheese, and earth aroma [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Microorganisms in the advanced fermentation process included \u003cem\u003eRhizopus oligosporus\u003c/em\u003e, lactic acid bacteria, and other microbes [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. These microorganisms break down protein components to produce volatile compounds. Prolonged fermentation resulted in a softer texture and increased free amino acid content. Stinky tempeh was also used in the making of \u0026ldquo;\u003cem\u003esambal tumpang\u003c/em\u003e,\u0026rdquo; but it was mixed with fresh tempeh, seemingly to reduce the strong odour and bitter taste.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003ePeanut tempeh wrapped in banana leaves\u003c/h2\u003e \u003cp\u003ePeanut press cake tempeh was made from peanut press cake. This peanut tempeh was collected from the Lawang market in Malang Regency, East Java Province, Indonesia. The process of making peanut tempeh was similar to that of soybean tempeh. The fermentation process of peanut press cake tempeh was also carried out using \u003cem\u003eRhizopus sp\u003c/em\u003e. fungus [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Peanut press cake tempeh could be wrapped in banana leaves or a plastic bag (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. d). This type of tempeh might be considered rare, as it is not available in all areas in Java or other areas in Indonesia. In East Java province, it seemed to be available widely only in Malang regency. However, an individual tempeh producer, such as in Yogyakarta, had started using peanut press cake as their raw material. Peanut press cake tempeh showed less mycelium growth on its surface compared to soybean tempeh, resulting in a crumbly texture. Therefore, peanut press cake tempeh had a coarse appearance and black patches due to the seed coat (testa), which usually was not removed during the process. Peanut press cake tempeh was considered to have a tastier flavour than soybean tempeh but with a crumbly texture. Peanut press cake was usually deep-fried, stir-fried, or tempeh cracker as a snack or side dish. It was rarely cooked in other ways, such as with soybean tempeh.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003ePeanut press cake tempeh wrapped in a plastic bag\u003c/h2\u003e \u003cp\u003ePeanut press cake tempeh wrapped in plastic did not seem different from that wrapped in banana leaves. Plastic wrap offered practicality and better durability, especially for distribution and storage (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.e). The use of plastic packaging did not affect the activity of microorganisms in tempeh, which was shown by the consistency of the total population of microorganisms [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Plastic could protect tempeh from external contamination, such as dust or excess moisture, making it more suitable for modern markets. Peanut press cake tempeh wrapped in plastic showed a more neutral aroma and flavour due to the inert nature of the plastic [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Due to a lack of air circulation, the surface texture tended to be denser and slightly drier. The choice of packaging type was tailored to consumer preferences and product distribution purposes.\u003c/p\u003e \u003cp\u003e \u003cb\u003eBenguk\u003c/b\u003e \u003cb\u003eTempeh\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eBenguk\u003c/em\u003e tempeh (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. f) was made from velvet bean (\u003cem\u003eMucuna pruriens\u003c/em\u003e). It is mostly found in Yogyakarta province and its surrounding area, such as Wonogiri Regency, which belongs to Central Java Province. \u003cem\u003eBenguk\u003c/em\u003e tempeh was rarely found in other places, which might be due to the availability of raw materials, technology, and consumers. \u003cem\u003eBenguk\u003c/em\u003e tempeh was tastier, had a black or dark appearance, a rather coarser mouthfeel, and a harder texture compared to soybean tempeh. Since the benguk bean was larger (1 to 2 cm in length) than the soybean, benguk tempeh showed a larger lumpy appearance. \u003cem\u003eBenguk\u003c/em\u003e tempeh was consumed as a snack or side dish. \u003cem\u003eBenguk\u003c/em\u003e tempeh could be deep fried after marinated in salt and garlic, dipped in batter before frying, stir-fried, made into soup with coconut milk (\u0026ldquo;\u003cem\u003elodeh\u003c/em\u003e\u0026rdquo;), crackers coated in batter (\u0026ldquo;\u003cem\u003ekripik\u003c/em\u003e\u0026rdquo;), and \u0026ldquo;\u003cem\u003ebacem\u003c/em\u003e\u0026rdquo; (cooked about 30 minutes in spices of coriander, galanga, bay leaves, garlic, shallot, palm sugar, and salt, and then deep fried). There was a specialty made from benguk tempeh called \u0026ldquo;\u003cem\u003ebesengek\u003c/em\u003e\u0026rdquo; or \u0026ldquo;\u003cem\u003esenge\u003c/em\u003ek\u0026rdquo;, where \u003cem\u003ebenguk\u003c/em\u003e tempeh was cooked in thick coconut milk so that coconut milk curd covered the tempeh. The spices used in \u003cem\u003ebesengek\u003c/em\u003e included coriander, shallot, garlic, galanga, bay leaves, palm sugar, salt, and might be candle nut. \u003cem\u003eBesengek\u003c/em\u003e seemed only available in Yogyakarta province, especially Kulon Progo regency and Wonogiri regency in Central Java. In Yogyakarta city, \u003cem\u003ebesengek\u003c/em\u003e was likely only sold in main markets such as Kranggan and Beringharjo market, and only available during Sunday. The sellers came from Kulon Progo Regency. This might indicate that the product is becoming rare. Nevertheless, when \u003cem\u003ebenguk\u003c/em\u003e tempeh was sold in online shops, the sellers were from Kulon Progo or Wonogiri in the form of fresh tempeh, cracker, or frozen \u003cem\u003ebacem\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eThe making process of \u003cem\u003ebenguk\u003c/em\u003e tempeh was rather lengthy as compared to that of soybean tempeh [36], which seemed to be due to the harder texture of the \u003cem\u003ebenguk\u003c/em\u003e bean. First, \u003cem\u003ethe benguk bean was washed and soaked in water for 24 hours\u003c/em\u003e, and then it was cooked in lime solution for 30 minutes until it was easy to peel the seed coat. After removing the seed coat, the bean was washed thoroughly and then soaked for 96 hours, and soaking water was replaced every 12 hours. The bean was then washed until there was no slippery feeling on the surface of the bean. Then, the bean was cooked for about 30 minutes until soft enough to maintain its integrity and not be too soft. The bean was then cooled to reach room temperature, inoculated with tempeh inoculum of Rhizopus spp., and packed in banana leaf every about 50 g, 3 to 5 mm thick, rectangle shape. The packs were then arranged in a single layer and incubated at room temperature for 5 days (120 hours).\u003c/p\u003e \u003cp\u003e \u003cb\u003eKoro\u003c/b\u003e \u003cb\u003eTempeh\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eKoro/lablab\u003c/em\u003e bean (\u003cem\u003eLablab purpureus\u003c/em\u003e) looked similar in size and shape to \u003cem\u003ethe benguk bean but had a\u003c/em\u003e white seed colour. \u003cem\u003eKoro\u003c/em\u003e tempeh (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. g) was similar to \u003cem\u003ebenguk\u003c/em\u003e tempeh in many ways. It also originated from Kulon Progo and Wonogiri regency. The use and cooking technique was similar to that of \u003cem\u003ebenguk\u003c/em\u003e tempeh. The product was also getting rare. We bought \u003cem\u003ekoro\u003c/em\u003e tempeh at the main market in Bantul City, Bantul Regency, in Yogyakarta province. We could not find \u003cem\u003ekoro\u003c/em\u003e tempeh in Kranggan or Beringharjo market in the city of Yogyakarta. The texture of koro tempeh was softer than that of soybean tempeh. Koro tempeh was also made into \u003cem\u003ebesengek\u003c/em\u003e.\u003c/p\u003e \u003cp\u003e \u003cb\u003eDage\u003c/b\u003e \u003cb\u003eTempeh\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eDage\u003c/em\u003e tempeh (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. h) originated from the Banyumas regency in Central Java, especially Purwokerto City and its surrounding area. We could not find appropriate information about raw material for \u003cem\u003edage\u003c/em\u003e tempeh either from \u003cem\u003edage\u003c/em\u003e sellers in Pasar Manis (\u0026ldquo;Manis\u0026rdquo; market) or the layman that we interviewed. However, Romulo and Surya [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] mentioned that \u003cem\u003edage\u003c/em\u003e was made from fresh coconut press cake, cooked, cooled, and inoculated by \u003cem\u003eRhizopus\u003c/em\u003e spp., then fermented with inoculum of \u003cem\u003eRhizopus\u003c/em\u003e spp. for two days. A report from a local newspaper mentioned that \u003cem\u003edage\u003c/em\u003e was made from fresh coconut press cake or fresh coconut press cake added with sliced peanuts [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe appearance of \u003cem\u003edage\u003c/em\u003e tempeh (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.h.) looked more like \u003cem\u003egembus\u003c/em\u003e tempeh, a type of tempeh made from okara, where the mycelium was not as fluffy as that of soybean tempeh, and the texture was spongy and soft. However, there were some black spots scattered on the \u003cem\u003edage\u003c/em\u003e tempeh, which might be due to the testa (outer part of the endosperm) of the coconut. Coconut testa had a harder texture and dark brown colour. The process of making \u003cem\u003edage\u003c/em\u003e started with steaming the raw material, cooling, adding tempeh inoculum containing \u003cem\u003eRhizopus\u003c/em\u003e spp., and fermenting for two days [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe appearance and texture of \u003cem\u003edage\u003c/em\u003e tempeh were very similar to that of \u003cem\u003egembus\u003c/em\u003e tempeh, which was porous and softer than soybean tempeh. The price of \u003cem\u003edage\u003c/em\u003e tempeh was about the same as that of soybean tempeh. The way to consume \u003cem\u003edage\u003c/em\u003e tempeh was similar to that of soybean tempeh, which could be deep-fried with or without coating batter made from rice or wheat flour. \u003cem\u003eDage\u003c/em\u003e tempeh was also made into \u003cem\u003ekeripik\u003c/em\u003e (a cracker-like), where \u003cem\u003edage\u003c/em\u003e tempeh was sliced about 1 mm thick, coated with flour batter, and deep fried. \u003cem\u003eDage\u003c/em\u003e tempeh was also stir fried. The preference towards \u003cem\u003edage\u003c/em\u003e tempeh was probably related to its soft and spongy texture.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eAmino acids profile of fermented products\u003c/h2\u003e \u003cp\u003eThe protein content of tempeh ranged from 13.97 (\u003cem\u003egembus\u003c/em\u003e tempeh) to 45.5% (Malang fresh soybean tempeh) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Tempeh showed good quality of protein as indicated by almost all amino acids scoring over 100 (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), except for limiting amino acids. The amino acid score ranges from 36 (\u003cem\u003egembus\u003c/em\u003e tempeh) to 102% (peanut press cake tempeh wrapped in banana leaves). Most of the samples had methionine and cysteine as limiting amino acids, except \u003cem\u003ebenguk\u003c/em\u003e tempeh, which also had leucine as a limiting amino acid. In contrast, peanut press cake tempeh had threonine as a limiting amino acid. Fermentation seemed to increase substantially the amino acid score of peanuts, which had an amino acid score of 24 to 28 before fermentation [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEssential amino acids and amino acids score of different types of tempeh\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"22\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c16\" colnum=\"16\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c17\" colnum=\"17\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c18\" colnum=\"18\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c19\" colnum=\"19\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c20\" colnum=\"20\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c21\" colnum=\"21\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c22\" colnum=\"22\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAmino Acid\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eWHO Std., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u003cem\u003eBenguk\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e\u003cem\u003eKoro\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eMalang Fresh Soybean Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003eMalang Overripe Soybean Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e \u003cp\u003eMalang Stinky Soybean Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c14\" namest=\"c13\"\u003e \u003cp\u003ePeanut Press Cake Tempeh, in Plastic Wrap\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c16\" namest=\"c15\"\u003e \u003cp\u003ePeanut Press Cake Tempeh, in Banana Leaf Wrap\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c19\" namest=\"c17\"\u003e \u003cp\u003e\u003cem\u003eGembus\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c22\" namest=\"c20\"\u003e \u003cp\u003e\u003cem\u003eDage\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eConc,mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eConc.\u003c/p\u003e \u003cp\u003emg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eConc., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eConc., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eConc., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003eConc., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e \u003cp\u003eConc., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c16\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c17\"\u003e \u003cp\u003eConc., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c18\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003eConc., mg.g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c21\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e159.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e36.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e194.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e31.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e163.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e27.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e142.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e26.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e139.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e34.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e179.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e35.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e188.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e62.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e329.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e48.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e253.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIle\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e43.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e56.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e200.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e52.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e188.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e56.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e200.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e57.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e206.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e51.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e183.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e47.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e169.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e45.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e164.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e80.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e288.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLeu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e178.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e270.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e85.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e129.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e80.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e121.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e84.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e128.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e88.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e134.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e79.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e121.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e71.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e109.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e64.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e97.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e69.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e104.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e101.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e77.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e133.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e62.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e108.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e58.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e101.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e49.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e85.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e68.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e118.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e65.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e113.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e78.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e135.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e66.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e115.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eThr\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e39.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e115.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e131.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e35.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e105.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e34.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e101.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e31.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e93.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e37.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e110.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e34.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e102.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e41.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e120.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e44.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e130.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrp\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e29.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e269.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e103.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e24.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e226.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e24.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e226.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e24.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e223.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e25.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e233.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e25.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e230.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e5.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e52.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e14.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e132.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e53.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e153.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e64.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e185.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e52.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e149.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e58.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e166.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e61.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e176.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e57.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e162.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e50.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e144.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e68.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e196.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e47.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e135.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMet\u0026thinsp;+\u0026thinsp;Cys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e47.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e47.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e56.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e17.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e69.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e16.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e67.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e15.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e63.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e57.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e228.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e9.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e36.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e12.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e49.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhe\u0026thinsp;+\u0026thinsp;Tyr\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e92.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e147.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e113.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e179.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e99.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e157.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e109.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e173.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e120.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e191.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e104.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e166.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e99.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e157.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e125.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e199.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c20\" namest=\"c19\"\u003e \u003cp\u003e84.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e134.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c22\" namest=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe total concentration of non-essential amino acids was greater than that of essential amino acids (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The highest concentration of amino acid was glutamate (a non-essential amino acid) in all tempeh except \u003cem\u003ebenguk\u003c/em\u003e tempeh, where leucine (an essential amino acid) was the predominant amino acid. The dominant essential amino acid in tempeh was leucine, except for \u003cem\u003egembus\u003c/em\u003e and \u003cem\u003edage\u003c/em\u003e tempeh, where the highest concentration of amino acids was phenylalanine and isoleucine, respectively. The level of essential amino acids in tempeh ranged from 6 (\u003cem\u003egembus\u003c/em\u003e tempeh) to 18% (fresh Malang tempeh). The total concentration of non-essential amino acids ranged from 53 (\u003cem\u003ebenguk\u003c/em\u003e or \u003cem\u003ekoro\u003c/em\u003e tempeh) to 59% (peanut press cake tempeh wrapped in banana leaves). The dominant non-essential amino acid was glutamate, except in \u003cem\u003ebenguk\u003c/em\u003e tempeh, where the highest concentration was shown by aspartic acid.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAmino acid quality of different types of tempeh\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmino Acid Quality Parameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eBenguk\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eKoro\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMalang Fresh Soybean Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMalang Overripe Soybean Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMalang Stinky Soybean Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePeanut press cake tempeh, in plastic wrap\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003ePeanut press cake tempeh, in banana leaves wrap\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003eGembus\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cem\u003eDage\u003c/em\u003e Tempeh\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtein (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e33.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e27.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e45.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e41.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e40.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e41.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e13.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e15.09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEAA (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e46.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e41.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e42.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e43.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e42.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e40.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e45.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e43.63\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNEAA (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e53.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e53.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e58.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e57.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e56.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e57.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e59.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e54.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e56.37\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEAA/NEAA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmino acid score\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e47.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e56.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e69.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e67.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e63.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e102.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e36.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e49.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLimiting amino acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIle, Met,\u003c/p\u003e \u003cp\u003eCys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMet, Cys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMet, Cys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMet, Cys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMet, Cys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMet, Cys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eThr\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eMet, Cys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eMet, Cys\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLNAA (including Trp)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.363\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.785\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.082\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10.082\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e9.766\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.231\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.438\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e3.426\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrp/LNAA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.048\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.112\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.103\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.068\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cem\u003eGembus\u003c/em\u003e and \u003cem\u003edage\u003c/em\u003e tempeh were made from soybean or coconut residue. Consequently, the nutrition was low (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), and so were the parameters of amino acid quality (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Prolonged fermentation seemed to reduce the total concentration of non-essential amino acids (from 58.88% in fresh Malang tempeh to 56.76% in stinky Malang tempeh). It increased that of essential amino acids (from 41.12% in fresh Malang tempeh to 43.24% in stinky tempeh) (Tabel 2).\u003c/p\u003e \u003cp\u003eThe amino acid profile in tempeh seemed to be affected by the amino acid composition in the raw material. For example, glutamic acid was the predominant amino acid in soybean [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] or in coconut [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] and also in the form of tempeh. On the contrary, methionine was low in soybean [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] or coconut [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] and also in tempeh. Tempeh could be a candidate for a natural source of glutamate acid. Glutamic acid was suggested as a useful supplement for improving learning and memory performance and neurochemical status of the brain, and in the future, could be effective in the treatment of neurological disorders affecting learning and memory performance, which might be attributed to the improved antioxidant status and cholinergic, monoaminergic and glutamatergic neurotransmission in the brain and hippocampus [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFermentation in tempeh increased protein content, which might be attributed to fungal biomass. The protein content of unfermented soybean seeds was 35\u0026ndash;36% [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], and it was 40 to 45% in Malang tempeh (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Similarly, peanut, \u003cem\u003ekoro\u003c/em\u003e and \u003cem\u003ebenguk\u003c/em\u003e beans had a protein content of 25% [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e], 25% [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] and 23% [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], respectively, while in the form of tempeh, their protein concentrations were 40, 27, and 37%, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Fermentation also altered amino acid composition, as shown by the higher concentration of valine, lysine, and isoleucine in tempeh (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) than in raw soybean [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFermentation of tempeh seemed to involve lactic acid bacteria from the surroundings. The bacteria that play a role in the fermentation of tempeh were the genus Lactobacillus [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Lactic acid bacteria in the tempeh manufacturing process showed the ability to hydrolyse soy protein and subsequently released amino acids such as leucine, phenylalanine, tyrosine, valine and isoleucine [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The amount and composition of tempeh bacteria were influenced by the production process, production environment and raw materials [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Packaging material did not affect the type of bacteria involved in tempeh-making [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Still, it could affect protein levels with higher protein in tempeh wrapped in leaves [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] .\u003c/p\u003e \u003cp\u003eThe ratio between tryptophan and other large neutral amino acids in the plasma determined the ability of tryptophan to cross the blood-brain barrier (BBB) for further synthesis into serotonin. This ratio value started from 0.09 or above [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. In all tempeh samples, except \u003cem\u003ekoro\u003c/em\u003e, \u003cem\u003egembus\u003c/em\u003e, and \u003cem\u003edage tempeh\u003c/em\u003e, the ratio value of tryptophan to LNAA was over 0.102 (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), and none was in the suggested range for the ability to cross the blood-brain barrier. Benguk tempeh showed substantially the highest value of this ratio, 0.187 (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Prolonged fermentation seemed to reduce this value, as indicated by Malang fresh tempeh (0.122) and Malang stinky tempeh (0.103) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). There was no report on whether the tryptophan/LNAA ratio in the diet would retain the ratio in the plasma. However, there were reports on the influence of food components such as protein and simple carbohydrates [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Protein hydrolysate was shown to improve tryptophan bioavailability [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Therefore, this technique might be applied to tempeh with a low tryptophan/LNAA ratio.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eFuture, Prospects and Challenges of Development Tempeh\u003c/h2\u003e \u003cp\u003eTempeh, a traditional Indonesian soy-based fermented food, has gained global recognition as a versatile, nutritious, and sustainable protein source. With increasing awareness about health and environmental issues, tempeh holds significant promise in addressing the dietary and ecological challenges of the modern world [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Tempeh is rich in protein, dietary fibre, vitamins, and minerals, making it a highly nutritious food option. Unlike unfermented soy products, tempeh\u0026rsquo;s fermentation process enhances its digestibility and reduces anti-nutritional factors like phytic acid. Moreover, it is a good source of probiotics, which support gut health [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. As more people adopt plant-based diets for health reasons, tempeh\u0026rsquo;s nutritional profile makes it an appealing alternative to meat and other animal products. Scientific advancements in food technology are likely to optimize the nutritional content of tempeh further, potentially fortifying it with additional nutrients to meet specific dietary needs [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe global market for plant-based foods has been expanding rapidly, driven by shifting consumer preferences and increasing investment in sustainable food technologies. Tempeh, with its versatility and adaptability, is well-positioned to capitalize on this trend. According to market research, the global tempeh market is expected to grow at a compound annual growth rate (CAGR) of over 6% in the coming years. Tempeh\u0026rsquo;s firm texture and ability to absorb flavours make it suitable for a wide range of dishes, from stir-fries to sandwiches and even snacks. This versatility appeals to chefs and home cooks alike. As more people become aware of the environmental and health benefits of plant-based diets, demand for products like tempeh is expected to rise. Companies are developing innovative tempeh-based products, such as pre-seasoned tempeh, tempeh burgers, and tempeh nuggets, to attract a broader audience. Advances in packaging and storage technology are also extending tempeh\u0026rsquo;s shelf life, making it more accessible to global markets. Encouraging local production of tempeh in regions outside Southeast Asia can reduce costs and promote its adoption in diverse cultures. Startups and small businesses are already leveraging this opportunity [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDespite its potential, tempeh faces challenges that could hinder its growth. One key obstacle is consumer unfamiliarity in certain regions, where it is perceived as an exotic or niche product. Education and marketing campaigns highlighting tempeh\u0026rsquo;s benefits can address this issue. Additionally, concerns about soy production, including deforestation and pesticide use, may deter environmentally conscious consumers. Promoting the use of organic and non-GMO soybeans, as well as exploring alternative ingredients, can help mitigate these concerns [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTempeh production is highly dependent on precise conditions such as temperature, humidity, and the quality of the starter culture. Small variations in these factors can result in inconsistent quality and texture, which can affect the final product's consumer appeal. The fermentation process is also sensitive to contamination, which can lead to spoilage or undesirable flavours [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. While small-scale artisanal tempeh production can be easily managed, scaling the process to meet increasing demand, particularly in Western markets, introduces complexities. Large-scale production often involves automation, but this can lead to a loss of the traditional artisanal qualities that some consumers prefer. Additionally, the equipment needed for large-scale fermentation is expensive, and maintaining quality control on a larger scale requires substantial investment. For tempeh to be produced on a large scale, reliable sources of high-quality soybeans are essential [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. However, soybeans face competition from other crops, and issues like climate change, fluctuating prices, and supply chain disruptions can affect their availability and cost. Furthermore, in some regions, the cost of production and transportation can make tempeh less affordable compared to other protein sources.\u003c/p\u003e \u003cp\u003eDespite its long history in Southeast Asia, tempeh faces challenges in terms of consumer acceptance, especially in Western countries. One of the primary barriers to tempeh\u0026rsquo;s wider acceptance is its taste and texture, which differ significantly from other plant-based proteins such as tofu. Tempeh has a nutty, earthy flavour that may not appeal to all palates, especially those unfamiliar with fermented foods. Its firm and slightly crumbly texture also contrasts with the softer, smoother textures of more familiar plant proteins [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. In many Western countries, soy-based products like tofu have been available for decades, but tempeh is still relatively unknown. Cultural unfamiliarity with tempeh often translates into hesitation or lack of interest in trying it. It is commonly perceived as an exotic food, which can create psychological barriers for consumers who prefer familiar and locally sourced options. The challenge of educating consumers about tempeh\u0026rsquo;s health benefits, versatility, and potential as a protein source is significant. Although plant-based eating has gained popularity, tempeh is often overshadowed by other vegan or vegetarian protein options like lentils, chickpeas, and packaged plant-based meat substitutes. The lack of awareness about the health benefits and sustainability of tempeh makes it difficult to gain widespread consumer traction [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs the demand for plant-based proteins increases globally, tempeh faces significant competition from a variety of alternatives. Products like tofu, seitan, and newer innovations in plant-based meats often receive more attention due to their similarity to meat in taste and texture. These products often dominate the plant-based protein market, leaving tempeh as a niche offering. Tempeh, while generally affordable, can be more expensive than other plant proteins, particularly in regions where it is not widely produced. This price discrepancy can make it harder for tempeh to compete with cheaper, mass-produced alternatives like lentils and beans, especially in budget-conscious markets. Additionally, tempeh may not be as widely available in grocery stores, further limiting its reach. Newer plant-based products, especially those designed to mimic meat, are marketed as convenient, ready-to-eat, or quick-cooking options. In contrast, tempeh typically requires cooking and may be perceived as less convenient. While tempeh can be prepared in various ways, the perceived effort involved in cooking it might deter potential consumers from choosing it over more straightforward alternatives [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWhile tempeh offers numerous benefits as a nutritious, plant-based protein, it faces a range of challenges that need to be addressed to ensure its widespread adoption. From production hurdles, including quality control and supply chain issues, to consumer perception and competition with other plant-based proteins, many factors limit tempeh\u0026rsquo;s potential in the global market. However, with increasing awareness of plant-based diets and sustainability concerns, tempeh has the opportunity to gain popularity. Overcoming challenges related to taste, texture, and cultural acceptance, as well as addressing sustainability issues in soybean farming, will be essential to its long-term success and growth in the global food industry.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis work revealed nutritional and functional health benefits related to the amino acid profile of different types of tempeh available in Indonesian markets. Methionine and cysteine were the limiting amino acids in tempeh samples, except peanut press cake tempeh, which showed threonine as the limiting amino acid. Tryptophan in tempeh had a ratio of tryptophan/LNAA more than 0.1, indicating a probable future role in alleviating depression, except for \u003cem\u003ebenguk\u003c/em\u003e, gembus, and \u003cem\u003edage\u003c/em\u003e tempeh. Different types of tempeh and prolonged fermentation affected the amino acid profile and protein concentration of tempeh.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eLNAA: Large neutral amino acids\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;GMO: Genetically modified organism\u003c/p\u003e\n\u003cp\u003eOPA\u0026nbsp; \u0026nbsp;\u0026nbsp;: \u0026nbsp; \u0026nbsp;O-phtalalhdehide\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;5-HT : 5-hydroxytryptamine\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCAGR : Compound annual growth rate \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; BBB \u0026nbsp; \u0026nbsp;: blood-brain barrier\u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was funded by the Ministry of Higher Education, Research, and Technology, the Government of Indonesia, for the KATALIS scheme in 2024, contract no. 054/E5/PG.02.00.PL/2024.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed equally as the main contributors in this paper. UP, DPM, NN, ECS, and ES formulated and designed the research. DH, AR, and ESN conducted data collection and processing. IP, JJ, ER, SA, CLS, BK, and RHBS contributed to the discussion. RHBS constructed philosophy, history, and future prospect studies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was funded by the Ministry of Higher Education, Research, and Technology, the Government of Indonesia, for the KATALIS scheme in 2024, contract no. 054/E5/PG.02.00.PL/2024.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author is willing to provide data and materials according to the agreed research needs and ethics.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors declared that there is no competing interest in the writing of this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; information\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUmi Purwandari\u003csup\u003e1,8\u003c/sup\u003e, Darimiyya Hidayati\u003csup\u003e1\u003c/sup\u003e, Askur Rahman\u003csup\u003e1\u003c/sup\u003e, Enung Siti Nurhidayah\u003csup\u003e1\u003c/sup\u003e, Indah Purwantini\u003csup\u003e2\u003c/sup\u003e, Era Catur Prasetyo\u003csup\u003e3\u003c/sup\u003e, Nurhayati Nurhayati\u003csup\u003e4\u003c/sup\u003e, Dwiyati Puji Mulyani\u003csup\u003e5\u003c/sup\u003e, Jay Jayus\u003csup\u003e4\u003c/sup\u003e, Eka Ruriani\u003csup\u003e4\u003c/sup\u003e, Enny Suswati\u003csup\u003e4\u003c/sup\u003e, Satya Arimurti\u003csup\u003e4\u003c/sup\u003e, Chatarina Lilis Suryani\u003csup\u003e5\u003c/sup\u003e, Bayu Kanetro\u003csup\u003e5\u003c/sup\u003e, R. Haryo Bimo Setiarto\u003csup\u003e6,7\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003eUniversity of Trunojoyo Madura, Jl. Raya Telang, Kamal, Bangkalan, East Java, Indonesia 69162\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003eFaculty of Pharmacy, University of Gadjah Mada, Sekip Utara, Yogyakarta, Indonesia 55281\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e3\u003c/sup\u003eMuhammadiyah University of Surabaya, Jl. Sutorejo No. 59 Surabaya, East Java, Indonesia 60113\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e4\u003c/sup\u003eUniversity of Jember, Jl. Kalimantan 37, Kampus Bumi Tegalboto, Kotak Pos 159, Jember, east Java, Indonesia 68121\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e5\u003c/sup\u003eUniversity of Mercu Buana Yogyakarta, Jl. Ring Road Utara, Yogyakarta, Indonesia 55753\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e6\u003c/sup\u003eResearch Center for Applied Microbiology, National Research and Innovation Agency (BRIN), Jl. Raya Jakarta-Bogor KM 46, KST Soekarno, Cibinong, Bogor, Indonesia 16911.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e7\u003c/sup\u003eResearch Collaboration Center for Traditional Fermentation, National Research and Innovation Agency (BRIN)\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e8\u003c/sup\u003eIndonesia Tempeh Forum (Forum Tempe Indonesia) East Java Branch. Surabaya\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eTamam B, Syah D, Suhartono MT, Kusuma WA, Tachibana S, Lioe HN. Proteomic study of bioactive peptides from tempe. J Biosci Bioeng [Internet]. 2019;128(2):241\u0026ndash;8. Available from: https://doi.org/10.1016/j.jbiosc.2019.01.019.\u003c/li\u003e\n\u003cli\u003eEllent SSC, Dewi L, Tapilouw MC. 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Quality of soybean products in terms of essential amino acids composition. Molecules. 2021;26(16):1\u0026ndash;9.\u003c/li\u003e\n\u003cli\u003eHossain S, Ahmed R, Bhowmick S, Mamun A Al, Hashimoto M. Proximate composition and fatty acid analysis of \u003cem\u003eLablab purpureus\u003c/em\u003e (L.) legume seed: implicates to both protein and essential fatty acid supplementation. Springerplus. 2016;5(1):1\u0026ndash;10.\u003c/li\u003e\n\u003cli\u003eSirilun S, Sivamaruthi BS, Kesika P, Peerajan S, Chaiyasut C. Lactic acid bacteria mediated fermented soybean as a potent nutraceutical candidate. Asian Pac J Trop Biomed [Internet]. 2017;7(10):930\u0026ndash;6. Available from: https://doi.org/10.1016/j.apjtb.2017.09.007.\u003c/li\u003e\n\u003cli\u003eMarkus CR, Kl\u0026ouml;pping-Ketelaars WI, Pasman W, Klarenbeek B, Van Den Berg H. Dose-Dependent Effect of \u0026alpha;-Lactalbumin in Combination with Two Different Doses of Glucose on the Plasma Trp/LNAA Ratio. 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Monitoring of fungal biomass changed by \u003cem\u003eRhizopus oryzae\u003c/em\u003e in relation to amino acid and essential fatty acids profile in soybean meal, wheat and rice. CYTA - J Food [Internet]. 2018;16(1):156\u0026ndash;64. Available from: https://doi.org/10.1080/19476337.2017.1359676\u003c/li\u003e\n\u003cli\u003eSirilun S, Sivamaruthi BS, Kesika P, Peerajan S, Chaiyasut C. Lactic acid bacteria mediated fermented soybean as a potent nutraceutical candidate. Asian Pac J Trop Biomed [Internet]. 2017;7(10):930\u0026ndash;6. Available from: https://doi.org/10.1016/j.apjtb.2017.09.007\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-ethnic-foods","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jefo","sideBox":"Learn more about [Journal of Ethnic Foods](http://journalofethnicfoods.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/JEFO/default.aspx","title":"Journal of Ethnic Foods","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Amino acid, fermented foods, Indonesia, tempeh","lastPublishedDoi":"10.21203/rs.3.rs-5724562/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5724562/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eTempeh has been world widely known as a functional food for several health benefits such as ACE inhibitory, hypotensive, and antioxidative. Although soybeans contained opioids, a peptide that affects mental state, there was no report on the potential of tempeh as a source of peptides with affective or cognitive function. This study examined the protein quality of tempeh as indicated by the amino acids profile and its possible role in improving depression, as indicated by the concentration of tryptophan and its ratio to large neutral amino acids (LNAA). Fresh tempeh types of different raw materials (soybean, okara, velvet bean, lablab bean, fresh coconut press cake) were collected from production areas. Samples were run for high-performance liquid chromatography. Results showed that protein and amino acid concentrations in tempeh ranged respectively from 13.97 and 8% (\u003cem\u003egembus\u003c/em\u003e tempeh) and 45.5 and 18% (Malang tempeh). The amino acid score in all tempeh met the WHO standard for nine amino acids, indicating good quality of amino acids. Tempeh made from peanut press cake showed all amino acids scored higher than 100, while \u003cem\u003egembus\u003c/em\u003e tempeh showed three amino acids with a score of less than 100. Methionine was the limiting amino acid while glutamate was the predominant amino acid of tempeh. Except that in \u003cem\u003ekoro\u003c/em\u003e, \u003cem\u003egembus\u003c/em\u003e, or \u003cem\u003edage tempeh\u003c/em\u003e, tryptophan in tempeh samples showed the ability to serve as an anti-depressant activity.\u003c/p\u003e","manuscriptTitle":"Exploring the profile of amino acids and ethnography study in the biodiversity of tempeh as fermented foods of Indonesia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-31 09:12:04","doi":"10.21203/rs.3.rs-5724562/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-03-21T15:19:19+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-03-21T15:17:57+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-03-04T00:50:03+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Ethnic Foods","date":"2025-03-03T05:58:00+00:00","index":"","fulltext":""},{"type":"decision","content":"Major revision","date":"2025-02-06T04:04:02+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-ethnic-foods","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jefo","sideBox":"Learn more about [Journal of Ethnic Foods](http://journalofethnicfoods.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/JEFO/default.aspx","title":"Journal of Ethnic Foods","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"282cf86f-1729-4d42-a8d1-a6f654e1a388","owner":[],"postedDate":"March 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-06-09T16:04:17+00:00","versionOfRecord":{"articleIdentity":"rs-5724562","link":"https://doi.org/10.1186/s42779-025-00285-3","journal":{"identity":"journal-of-ethnic-foods","isVorOnly":false,"title":"Journal of Ethnic Foods"},"publishedOn":"2025-06-06 15:57:50","publishedOnDateReadable":"June 6th, 2025"},"versionCreatedAt":"2025-03-31 09:12:04","video":"","vorDoi":"10.1186/s42779-025-00285-3","vorDoiUrl":"https://doi.org/10.1186/s42779-025-00285-3","workflowStages":[]},"version":"v1","identity":"rs-5724562","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5724562","identity":"rs-5724562","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}