Evaluation of Indigenous Yeasts from Local Drinks (Korefe, Areki and Tej) and Dough as Potential Fermenters for Bioethanol Production

Evaluation of Indigenous Yeasts from Local Drinks (Korefe, Areki and Tej) and Dough as Potential Fermenters for Bioethanol Production | 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 Evaluation of Indigenous Yeasts from Local Drinks ( Korefe, Areki and Tej ) and Dough as Potential Fermenters for Bioethanol Production Goiteom Senay Niguse, Melaku Mekonen Kasegn, Birhanu Kahsay Meresa, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9532904/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The depletion of fossil fuels forced researchers to find alternative, cheap, locally variable, and renewable sources of energy. Molasses is among the promised sources of fuel-grade ethanol. However, yeast strains that can produce large amounts of ethanol while resisting high sugar concentrations are difficult. Therefore, this study aims to investigate the potential of yeasts from Tigray’s local drinks and their potential for bioethanol production. Accordingly, yeasts were isolated from Korefe, Areki, teff dough, and Tej . Isolation, morphological characterization, and biochemical tests were carried out to identify potential yeasts. Then the yeasts were applied in molasses fermentation to examine their potential for bioethanol production. Bioethanol was estimated using the estimation of ethanol content using potassium dichromate solution and a standard curve-generated linear equation (absorbance = -0.09700 + 0.3691x). Results showed that the yeasts have the capability of osmotolerance, and the yeasts isolated from Areki (A-pen-3) produce a high amount of ethanol (8.5%) from molasses. Hence, production of ethanol from molasses using current yeasts has moderate potential and contributes to the effort of creating a sustainable and green environment to make the earth suitable for living. This study provides a good insight into using wild yeasts, and conducting further comprehensive research is appreciated. bioethanol fermentation local drinks molasses yeasts 1. Introduction Ethiopia has a wide variety of traditional indigenous fermented foods and beverages that are prepared and consumed. Alcoholic fermentation, induced by indigenous microbes, is associated with particular fermentable substances and gives a distinctive style and quality to that wine or beer ( 1 ). Yeast can grow at temperatures of 30 and 37°C, and some of them grow at 45°C. Yeasts that show positive assimilation of glucose, sucrose, maltose, galactose, fructose, and raffinose and negative for lactose, xylose, lysine, nitrate, and urea are considered Saccharomyces cerevisiae . Palm wine is the main source of Saccharomyces cerevisiae for industrial applications. The yeast cells from palm have high viability, high leavening (82-94.7%) capacity, CO₂ production, temperature tolerance, ethanol tolerance, and osmotic tolerance ( 2 ). Locally available substances are the main sources of industrial ethanol fermentation because of reduced production costs ( 3 ). Some yeast strains contain necessary properties that are used as fermentation starters for various food or beverage industries ( 4 ). Yeasts that are obtained from food crops have extracellular enzymatic activity and a high level of fermentation ability. Yeasts from local sources like food crops could increase ethanol production by fermenting carbohydrates efficiently, and this reduces the cost for importing yeast from abroad ( 5 ). Using yeasts isolated from readily available sources enables the production of fermentation products with a special taste and aroma. Unlike Saccharomyces cerevisiae , which could produce pleasant, fruity aromas when fermenting barley malt extract, yeasts like Brettanomyces from wine barrels are wine spoilage or give unfavorable beer tastes ( 6 ). In order to ferment wine efficiently and make superior-quality wine, using yeast strains isolated from the winery itself is best. This is because the yeast strains are more familiar with the wine’s climatic conditions and adapted to their own climatic conditions ( 7 ). Saccharomyces cerevisiae strains from indigenous sources like Murcha, grapes, and sugarcane provide special flavor to bread and are good strains for the bakery industry. However, yeasts that can be obtained from a plant called “Dar” are not important for baking because they lack the leavening properties that are required for yeast to be bakery yeast ( 8 ). Yeast isolates from different fruit peels, like pineapple and orange, were found to be temperature, pH, ethanol, and osmotolerant. Furthermore, it was reported that yeasts obtained from fruit peels were resistant to chloramphenicol. So, these yeast strains are important to ferment carbohydrates and enable them to produce bioethanol, which serves people as an alternative source of energy/fuel ( 9 ). Yeasts are an important component of insect gut microflora that are used to degrade toxic materials and produce vitamins, hormones, and different digestive enzymes ( 10 ). Arabidopsis is the habitat of various yeast strains that are beneficial, such as the ability of the yeasts to activate auxin responses in plants ( 11 ). Critical selection of appropriate yeast strains with needed characteristics is important to develop the required flavor and other regional characteristics of wine. For example, a yeast strain that is able to produce a large amount of hydrogen sulfide is not needed for wine production because it results in wine with an unfavorable flavor. Saccharomyces cerevisiae strains obtained from berries and spontaneously fermented musts are able to grow up to 16% alcohol and 40–50% of glucose concentration ( 12 ). According to Iwona Gientka et al. , kefir (fermented milk) is a source of Candida inconspicua, Debaryomyces hansenii, Kluyveromyces marxianus, Kazachstania unispora , and Zygotorulaspora Florentina; about 20% of lipid substances can be accumulated in yeast cells by using carbon-rich deproteinated potato wastewater, a starch processing industry waste, as a medium for the yeast ( 13 ). Candida adriatica and Candida taoyuanica , isolated from palm wine, a sugar-rich substrate from a non-oil-impacted environment using Bacto Bushnell-Haas broth, were found to be hydrocarbon - utilizing yeasts. This assists bioremediation by serving as biodegradable ( 14 ). Well-adapted yeast strains with low polysaccharolytic activity and desirable biochemical properties derived from brine olives are able to produce high-quality green table olives with consistent organoleptic characteristics. Furthermore, they have the potential to degrade the polysaccharides of olives and be used as a starter culture ( 15 ). The predominant microbes found in cracked green table olives, Manzanilla variety (a fermented food of Portugal), were yeasts with a large population of the S. cerevisiae species. Native strains that were selected from the food were proposed to act as starter cultures with the purpose of preventing spoilage during the storage of the food. The identification of the defined native strains helps to get superior-quality food products via fermentation, and this supports food-producing companies to be profitable ( 16 ). Phenolic compounds generated from industrial wastewater, like petrochemical plants, are environmental pollutants. Yeasts from soil and wastewater are the most important phenol biodegraders. It was reported that the yeast from soil samples had higher rates of phenol-degrading capacity than that of yeast isolated from water. Strong yeasts in phenol-degrading yeasts were identified using amplification of the 18S rRNA gene region, and these isolated yeasts were found to be Candida tropicalis , Pichia guilliermondii , Meyerozyma guilliermondii , and C. tropicalis ( 17 ). Yeasts obtained from water bodies have several good features: higher osmosis tolerance, higher special chemical productivity, and production of industrial enzymes compared to yeasts from terrestrial sources. Marine yeasts are very important in the production of bioethanol and pharmaceutical products ( 18 ). The use of mixed starters of selected strains of non- Saccharomyces yeasts with strains of Saccharomyces cerevisiae enhances wine fermentations and is crucial for better organoleptic characteristics of wine. Yeasts other than Saccharomyces cerevisiae yeasts are used to improve the primary and secondary aroma of wines and produce important enzymes ( 19 ). People have been using yeasts to make fermented beer, wine, sake, and bread via the fermentation process for many years. Furthermore, yeasts are important for biotechnological applications such as the production of biofuels and for bioremediation purposes. Modification of yeasts by mutagenesis, protoplast fusion, breeding, genome shuffling, and global transcription machinery engineering provides new improved strains with higher quality and fermentation efficiency than the wild type ( 20 ). Yeast strains that are psychro-tolerant and able to assimilate carbon sources efficiently were isolated from King George Island, the sub-Antarctic region. These yeasts are of potential use in industrial applications requiring high enzyme activities at low temperatures ( 21 ). To our knowledge, there are scanty reports in the area of wild yeast generated from indigenous sources. Therefore, the objective of the current work was to isolate and characterize yeast from local drinks ( Tej, Areki , and Korefe) and Teff dough and to determine their potential for bioethanol production. 1.1. Significance of the study The current study already opened the door to the idea that the local drinks and dough are important sources of yeast, which can serve to produce bioethanol. Since the local drink Korefe is found only at Korem , a study of the type of yeast in this type of local drink is a sole for researchers to study the yeast in a multidirectional way. Superior quality yeast strains can be found from the Korefe . 2. Materials and Methods 2.1. Description of the study area This study was conducted in Korem town , Tigray regional state, located about 619 km from Addis Ababa, the capital of Ethiopia. Located on the eastern edge of the Ethiopian highlands in the Southern Zone of the Tigray Region, this town has a latitude and longitude of 12°30′N 39°31′E with an elevation of 2539 meters above sea level. It has an annual average temperature of 17.4°C. Major crops that are produced at Korem are wheat, barley, and maize. 2.2. Sample collection The samples of Korefe, Areki, Tej , and dough, which were fermented for a week, were collected using sterile plastic bags from Korem town, southern Tigray, Ethiopia. The samples put in plastic bags were brought aseptically using an isotherm icebox to the molecular biotechnology laboratory, College of Veterinary Medicine, Mekelle University, Ethiopia, for analysis and placed in a refrigerator until use. 2.3. Sample preparation Four test tubes were prepared in a rack, and 9 ml of 9% (w/v) saline water was added to each test tube. Then 1 ml from each sample ( Korefe, Areki, Tej , and Dough) measured by micropipette was added to the respective test tube and homogenized using a vortex. 2.4. Isolation of yeast Aseptically, 0.5 ml of each dilution was taken via pipette and spread using an inoculating loop on the petri dish containing Potato Dextrose Agar (PDA), to which tetracycline and oxytetracycline (not simultaneously) were added to inhibit the growth of bacteria. Then the petri dishes were incubated at 30°C for 48 hours. Separate yeast colonies from the agar plate were transferred using the streaking technique on Sabouraud Dextrose Agar (SDA) and incubated at 30°C for 48 h. It was then inoculated by streaking on YPD medium ( 13 ). To preserve the yeast, a slant was prepared using test tubes. YPD media was prepared, and 5 ml of YPD was added to each test tube. The test tubes were put slanting (45°C) and incubated at 30°C for 24 hours to check whether they were contaminated or not. Yeast isolates were inoculated into slanted test tubes using an inoculating loop and incubated at 30°C for 48 hours. Then the yeast grown in the slant was preserved in a refrigerator at 4°C for further use ( 25 , 29 ). 2.5. Staining and microscopic observation of yeast isolates A smear of the sample from the colony was prepared on a slide after a drop of distilled water was added and fixed with flame. A few drops of methylene blue were applied to the slide and stayed for two minutes. Then it was decolorized by tap water ( 22 ). The smear was prepared on a slide and fixed with flame. A malachite green concentration of 5% was applied to the slide. The slide was exposed to flame for 3 minutes. The slide was decolorized with tap water and counterstained with safranin for 30 seconds. Then the slide was washed with tap water ( 23 ). Finally, yeast isolates that were stained using both methods were examined under oil immersion in a 100x microscope after the slide was dried in air. 2.6. Determination of physiological characteristics of yeasts through fermentation of sugars Of basal medium was prepared by adding enough powder to the required distilled water. Sugars and the broth were mixed in 1:10 (one part sugar and 9 parts broth). Three ml of mixture was measured with a pipette and added to each test tube. One hundred microliters of penicillin were added to each test tube. A colony of yeast was inoculated into the test tubes except for the controls (a control for each sugar), and tubes in the rack were incubated for 24–96 hours until a color change from red to yellow was observed ( 24 ). 2.7. Antimicrobial activity test Test tubes were prepared, and 1 ml of saline water was added to each test tube. Half a microliter of Staphylococcus aureus was added. The turbidity of the test tubes was compared with McFarland. Then the bacteria spread on the petri dish containing the PDA. After a while, yeast was inoculated at a fixed place on the Petri dish ( 26 ). 2.8. Estimation of Ethanol Content Using Potassium Dichromate Solution In order to determine the amount of ethanol present in the molasses, a potassium dichromate solution was made using the ( 31 ) method, which involved combining 325 milliliters of 96% H₂SO₄ with 33.8 grams of K₂Cr₂O₇ in 1000 milliliters of distilled water. Subsequently, the unknown concentration of ethanol generated from the molasses was estimated using this solution. An initial standard curve was created using known absolute ethanol concentrations in order to estimate the unknown ethanol content. Finally, the known concentrations generated from a standard curve were used to estimate the concentration of unknown materials using Eq. 2. The linear regression equation for the ethanol standard curve was as follows: \(\:\text{A}\text{b}\text{s}\text{o}\text{r}\text{b}\text{a}\text{n}\text{c}\text{e}=-0.09700+0.3691\text{x}\:\:\) Eq. (1) The rearrangement of the equation for the estimated ethanol concentration (x) gives \(\:\text{x}=\:\frac{\text{a}\text{b}\text{s}\text{o}\text{r}\text{b}\text{a}\text{n}\text{c}\text{e}\:+\:0.09700}{0.3691}\) Eq. (2) Considering a dilution factor of 10, we get the percentage of ethanol (y). \(\:\text{y}=\:\text{x}\text{*}\text{D}\text{F}\) Eq. (3) Where x is the estimated ethanol concentration, y is the ethanol concentration percentage (v/v), and DF is the dilution factor. 2.9. Data Analysis All the data were collected and analyzed using the Minitab software (v. 20). Results were displayed in tables. ANOVA was used to determine if the yeasts from different samples have significant differences in their ethanol concentration, which was produced from molasses and presented via tables. 3. Results 3.1. Cultural characteristics of yeast isolates The yeast isolate that was plated on PDA and SDA had different colony structures (Table 1 ). Isolates from Korefe and Areqi were visible, creamy white, and mucoid colonies with raised surfaces. The other isolates were shown to be filamentous, and a yellowish colony structure was observed. The yeast colony isolated from Tej had an irregular form, a convex margin, and an undulate elevation. The yeast colony isolated from dough had a circular form, raised elevation, and an entire margin. The form and margin of yeast isolated from Korefe and Areqi were similar to that of dough, but it had a convex margin. 3.2. Microscopic morphology of the yeast isolates The isolate showed active growth on screening for its ability to grow in yeast extract peptone dextrose medium (YPD), PDA, and SDA. The isolated colonies stained by malachite green and methylene blue were circular, ellipsoid, and rod-shaped structures (Table 1 ). Table 1 Cultural and microscopic features of the yeast colonies N o Yeast isolates Cultural and microscopic features 1 K-oxy-1 creamy white and mucoid colonies 2 K-oxy-2 creamy white and mucoid colonies 3 K-oxy-3 creamy white, mucoid colonies, ellipsoid 4 K-oxy-4 creamy white, mucoid colonies, ellipsoid 5 A-oxy-1 creamy white and mucoid colonies 6 A-oxy-2 creamy white, mucoid colonies, rod-shape 7 A-oxy-3 creamy white, mucoid colonies, ellipsoid 8 K-pen-1 creamy white, mucoid colonies, ellipsoid 9 K-pen-2 creamy white, mucoid colonies, rod-shape 10 K-pen-3 creamy white and mucoid colonies 11 K-pen-4 creamy white, mucoid colonies, circular 12 A-pen-1 creamy white and mucoid colonies 13 A-pen-2 creamy white and mucoid colonies 14 A-pen-3 creamy white and mucoid colonies 15 D-pen-1 raised elevation, margin, circular 16 D-pen-2 raised elevation, margin, circular 17 D-oxy-1 raised elevation, margin, circular 18 D-ox-2 raised elevation, margin, circular 19 T-oxy-1 irregular, a convex margin, undulate elevation 20 T-oxy-2 yellowish colony, filamentous 21 T-oxy-3 yellowish colony, filamentous 22 T-oxy-4 yellowish colony, filamentous 23 T-oxy-5 Irregular, a convex margin, undulate elevation 24 T-pen-6 Irregular, a convex margin, undulate elevation 25 T-pen-7 Irregular, a convex margin, undulate elevation 26 T-pen-8 Irregular, a convex margin, undulate elevation 27 T-pen-9 Irregular, a convex margin, undulate elevation 28 T-pen-10 Irregular, a convex margin, undulate elevation 29 T-pen-11 Irregular, a convex margin, undulate elevation 30 T-pen-12 Irregular, a convex margin, undulate elevation K- korefe, A- areqi, D- dough, T- tela , Oxy- oxy-tetracycline, Pen- penicillin 3.3. Physiological characteristics of yeasts through fermentation of sugars The physiological characteristics of the normal bakery yeast cells were identified by the carbohydrate fermentation tests (Table 2 ). All yeast isolates were able to ferment glucose, galactose, maltose, fructose, and sucrose at different incubation periods, as observed by color change from red to yellow. But none of the yeast isolates were able to ferment sorbitol and xylose. Table 2 Isolates from Korefe, Areqi , Teff dough, and Tej and the result of fermentation N o Yeast Isolates Types of sugars tested Glucose Galactose Maltose Fructose Sucrose Xylose Sorbitol Control 1 K-oxy-1 + + + + + - - - 2 K-oxy-2 + + + + + - - - 3 K-oxy-3 + + + + + - - - 4 K-oxy-4 + + + + + - - - 5 A-oxy-1 + + + + + - - - 6 A-oxy-2 + + + + + - - - 7 A-oxy-3 + + + + + - - - 8 K-pen-1 + + + + + - - - 9 K-pen-2 + + + + + - - - 10 K-pen-3 + + + + + - - - 11 K-pen-4 + + + + + - - - 12 A-pen-1 + + + + + - - - 13 A-pen-2 + + + + + - - - 14 A-pen-3 + + + + + - - - 15 D-pen-1 + + + + - 16 D-pen-2 + + + + + - - - 17 D-oxy-1 + + + + + - - - 18 D-ox-2 + + + + + - - 19 T-oxy-1 + + + + + - - - 20 T-oxy-2 + + + + + - - - 21 T-oxy-3 + + + + + - - - 22 T-oxy-4 + + + + + - - - 23 T-oxy-5 + + + + + - - - 24 T-pen-6 + + + + + - - - 25 T-pen-7 + + + + + - - - 26 T-pen-8 + + + + + - - - 27 T-pen-9 + + + + + - - - 28 T-pen-10 + + + + + - - - 29 T-pen-11 + + + + + - - - 30 T-pen-12 + + + + + - - - K- korefe, A- areqi, D- dough, T- tela , Oxy- oxy-tetracycline, Pen- penicillin 3.4. Antimicrobial activity test Yeast isolate did not show any antimicrobial activity against Staphylococcus aureus; as a result, there was not a clear zone observed. 3.5. Bioethanol production from molasses Yeast isolates did show varied potential for the production of bioethanol. The result for the effect of temperature, pH, and incubation period on ethanol production from molasses using isolated yeast investigated how the parameters significantly affect the fermentation process (Table 3 ). Due to the effect of temperature, pH, and incubation period, higher ethanol content (8.50%) was obtained at pH 5.5 and 30°C and incubated in four days with the isolate of A-pen-3. It is non-significant with ethanol produced at 26°C at pH 3.5. In contrast, lower ethanol content (2%) was recorded at pH 5.5 and 37°C and incubated for three days with the isolate of A-oxy-3. Table 3 Ethanol production from molasses using yeasts isolated from local drinks No. Treatments/ Trials Factors Absorbance X y % (v/v) Temp pH IP 1 K-oxy-1 26 4.5 48 0.134 0.397 3.97 2 K-oxy-2 37 4.5 72 0.140 0.47 4.70 3 K-oxy-3 26 5.5 72 0.246 0.76 7.60 4 K-oxy-4 30 5.5 48 0.150 0.50 5.00 5 A-oxy-1 37 4.5 96 0.201 0.64 6.40 6 A-oxy-2 37 4.5 72 0.143 0.48 4.80 7 A-oxy-3 26 5.5 48 0.104 0.39 3.90 8 K-pen-1 30 5.5 48 0.151 0.51 5.10 9 K-pen-2 26 5.5 96 0.207 0.66 6.60 10 K-pen-3 30 4.5 48 0.255 0.79 7.90 11 K-pen-4 37 4.5 96 0.195 0.62 6.20 12 A-pen-1 37 3.5 96 0.175 0.57 5.70 13 A-pen-2 30 4.5 48 0.267 0.82 8.20 14 A-pen-3 30 5.5 96 0.279 0.85 8.50 15 D-pen-1 37 3.5 96 0.178 0.579 5.79 16 D-pen-2 26 5.5 96 0.152 0.51 5.10 17 D-oxy-1 37 3.5 72 0.123 0.43 4.30 18 D-ox-2 26 3.5 48 0.220 0.693 6.93 19 K-oxy-1 37 5.5 48 0.105 0.381 3.81 20 K-oxy-2 37 5.5 96 0.049 0.23 2.30 21 K-oxy-3 26 4.5 72 0.19 0.61 6.10 22 K-oxy-4 30 3.5 72 0.202 0.644 6.44 23 A-oxy-1 30 3.5 48 0.198 0.633 6.33 24 A-oxy-2 37 4.5 48 0.091 0.343 3.43 25 A-oxy-3 37 5.5 72 0.039 0.20 2.00 26 K-pen-1 30 5.5 96 0.255 0.788 7.88 27 K-pen-2 30 5.5 72 0.230 0.72 7.20 28 K-pen-3 30 4.5 72 0.180 0.585 5.85 29 K-pen-4 26 3.5 72 0.199 0.64 6.40 30 A-pen-1 26 4.5 48 0.128 0.44 4.40 31 A-pen-2 37 3.5 48 0.040 0.21 2.10 32 A-pen-3 26 3.5 72 0.167 0.549 5.49 33 D-pen-1 30 3.5 96 0.2603 0.798 7.98 IP – Incubation period 4. Discussion In this study, 30 yeast isolates were isolated from local fermented Korefe , Teff dough, Tej , and Areqi . Based on the colony morphology, 14 isolates isolated from local fermented Korefe and Areqi were found to be Saccharomyces cerevisiae. The colonies exhibiting characteristics such as creamy to white color, fluffiness, and smooth margins were selected as the Saccharomyces strain ( 8 ). The yeast isolates were identified as Saccharomyces cerevisiae based on the morphology of the colony (circular) and sugar fermentation tests. Reis et al. found S. cerevisiae that grew on YPD medium had a smooth and a rough colony ( 30 ). Microscopic structure of methylene blue stained the yeast cells blue and showed them as circular ( Korefe and Areqi isolates), irregular (dough), and hyphae ( Tej isolates). Methylene blue stains the dead cells blue. Viable yeast cells also can be stained blue, but the active enzymes within them reduce the dye and make it colorless ( 32 ). The microscopic structure of S. cerevisiae has a characteristic ellipsoid or ovoid shape ( 36 ). The malachite green stain penetrated into the endospores by heating. Malachite green removes easily from the vegetative cells using water but not from endospores. The vegetative yeast cells appeared pink (the color of safranin), and the spores appeared green. The green indicates that the yeasts are spore-forming ( 33 ). Yeast isolates from dough and Tej isolates were found to be hyphae based on the microscopic structure, which was thin and rod-shaped. Yeast isolates obtained from Korefe and Areqi (14 isolates) fermented glucose, galactose, fructose, maltose, and sucrose within 24 hours. S. cerevisiae can be identified by its capability to ferment sucrose, maltose, fructose, glucose, galactose, and raffinose but not lactose ( 35 ). Therefore, yeast isolates obtained from Korefe and Areqi are S. cerevisiae . While these 4 isolates isolated from dough took 48–96 hours to ferment the given sugars. In another way, yeast isolates from local Tej (12 isolates) were able to ferment glucose, galactose, fructose, maltose, and sucrose. The yeast isolates were able to ferment sucrose, maltose, fructose, and glucose because yeasts produce enzymes that break the glycosidic bond that attaches two glucose units and convert sugar monomers into alcohol and CO₂. The sugar fermentation was indicated by a color change from red to yellow due to acid production ( 24 ). But none of the yeast isolates were able to ferment sorbitol and xylose. This result is similar to the work reported by ( 3 ). Fermentation tests indicated that none of the isolates produce ethanol using xylose as a carbon source ( 34 ). A control containing OF basal medium without the addition of yeast isolates was used and remained dark black, which was similar to that of yeast-treated xylose and sorbitol. In the antimicrobial test, the S . aureus cannot inhibit the growth of yeast. So, the pathogen S. aureus did not kill yeast. In contrast to this work, Younis et al. found yeast isolates showed antimicrobial activities against S. aureus, E. coli , and P. aeruginosa , showing remarkable antimicrobial activity against E. coli and S. aureus with a large, clear inhibition zone and low antimicrobial activity against P. aeruginosa ( 27 ). In the current study, the highest ethanol was produced when the fermentation was conducted at pH 5.5 at 30°C and incubated for four days by the isolate from Areqi (A-pen-3). However, the least ethanol was produced when the fermentation was conducted at pH 5.5 at 37°C and incubated for three days by the isolate from Areqi (A-oxy-3). Zabed et al. reported that a pH range of 4.0–5.0 is suitable for the ethanol production process ( 41 ). During the fermentation production of bioethanol, it depends on several factors such as temperature, sugar concentration, pH, and fermentation time ( 37 ). Particularly, temperature directly affected the growth rate of microorganisms. The current fermentation of molasses gave a remarkable ethanol yield at 26°C and 30°C incubated for four days. In line with this, Charoenchai et al. reported that the ideal temperature range for fermentation is between 20 and 35°C, while the optimum temperature of S. cerevisiae is near to 30°C ( 38 ). Ethanol production is influenced by the pH of the broth, as it affects yeast growth and fermentation rate. Moreover, the survival and growth of yeasts are influenced by the pH in the range of 2.75–4.25 ( 39 ). In fermentation for ethanol production, the optimum pH range of S. cerevisiae is 4.0–5.0 ( 40 ). Fermentation time affects the growth of microorganisms. Short fermentation time leads to insufficient microbial growth, which negatively affects the fermentation and results in incomplete conversion of sugars into ethanol ( 37 ). On the other hand, longer fermentation time has a toxic effect on microbial growth, especially in batch mode due to the high concentration of ethanol in the fermented broth. Complete fermentation can be achieved at lower temperatures by using a longer fermentation time, which results in the lowest ethanol yield ( 41 ). The lower ethanol content shown in (A-oxy-3) might be due to the inability to tolerate the produced ethanol. Studies reported by Tikka et al. and Benítez et al. stated that not all yeasts have the potential to survive in the produced ethanol ( 42 , 43 ). 5. Conclusion Generally, it can be concluded that the local drinks ( Korefe and Areki ) could be the major source for Saccharomyces cerevisiae , and the main fermenter yeast for these local drinks is Saccharomyces cerevisiae . Most isolates are excellent fermenters, but Saccharomyces cerevisiae is the main and fastest fermenting yeast that can ferment sugars within a short period of time, i.e., 24 hours. Most samples were capable of fermenting most sugars. Samples from Korefe, Areki , and Tej were able to ferment glucose, maltose, sucrose, fructose, and galactose but not xylose and sorbitol. While those samples from dough all ferment glucose, fructose, and sucrose, some of them ferment galactose and maltose, and others cannot ferment. Yeasts generated from indigenous sources are the most common microorganisms in bioethanol production and play a crucial function in fermenting sugars to ethanol from molasses. It is one of the major global issues in the production of alternative energy used in various sectors. Declarations Acknowledgment The authors thank Mekelle University for the fund . Funding This work was financially supported by Mekelle University (RDPDO/MU/CDANR/Generaljunior/020/13). Disclosure statement No potential conflict of interest was reported by the author(s). Author Contributions Sample collection , conceptualization, investigation, writing of the original draft, and visualization were performed by MMK and GSN. ATY achieved validation and project administration. BKM and SZH performed data analysis and interpretation. GSN, MMK, GM, and BW accomplished writing the review & editing and supervision. All authors read and approved the final. Data Availability All the data generated or analyzed during this study are included in this published article. Ethical approval Not applicable References Cappello MS, Bleve G, Grieco F, Dellaglio F, Zacheo G (2004) Characterization of Saccharomyces cerevisiae strains isolated from must of grape grown in experimental vineyard. J Appl Microbiol 97(6):1274–1280 Olowonibi OO (2017) Isolation and Characterization of Palm Wine Strains of Saccharomyces cerevisiae Potentially Useful as Bakery Yeasts. European Journal of Experimental Biology , 2017, 7:11 Nwachukwu IN, Ibekwe VI, Nwabueze RN, Anyanwu BN (2006) Characterization of palm wine yeast isolates for industrial utilization. Afr J Biotechnol 5(19):1725–1728 Tristezza, M., Fantastico, L., Vetrano, C., Bleve, G., Corallo, D., Grieco, F., …Grieco, F. (2014). Molecular and technological characterization of Saccharomyces cerevisiae strains isolated from natural fermentation of Susumaniello grape must in Apulia, Southern Italy. International Journal of Microbiology, 2014 (1), 897 – 428 Ebabhi AM, Adekunle AA, Okunowo WO, Osuntoki AA (2013) Isolation and characterization of yeast strains from local food crops. J Yeast Fungal Res 4(4):38–43 DeLong LN, Noone B, Erickson L (2017) Isolation, Identification, and Characterization of Local Wild Yeasts for Use in Fermentation. FASEB J 31:943–944 Guimarães TM, Moriel DG, Machado IP, Picheth CM, Bonfim T (2006) Isolation and characterization of Saccharomyces cerevisiae strains of winery interest. Revista Brasileira de Ciências Farmacêuticas 42:119–126 Karki, T. B., Timilsina, P. M., Yadav, A., Pandey, G. R., Joshi, Y., Bhujel, S., …Neupane, K. (2017). Selection and characterization of potential baker’s yeast from indigenous resources of Nepal. Biotechnology research international, 2017(1), 1925820. Nasir A, Rahman SS, Hossain MM, Choudhury N (2017) Isolation of Saccharomyces cerevisiae from pineapple and orange and study of metal’s effectiveness on ethanol production. Eur J Microbiol Immunol 7(1):76–91 Matos ITSR, Assunção EN, Carmo EJD, Soares VM, Astolfi-Filho S (2017) Isolation and characterization of yeasts able to assimilate sugarcane bagasse hemicellulosic hydrolysate and produce xylitol associated with veturius transversus (Passalidae, Coleoptera, and Insecta). Int J Microbiol 2017(1):5346741 Wang K, Sipilä TP, Overmyer K (2016) The isolation and characterization of resident yeasts from the phylloplane of Arabidopsis thaliana. Sci Rep 6(1):39403 Šuranská H, Vránová D, Omelková J (2016) Isolation, identification and characterization of regional indigenous Saccharomyces cerevisiae strains. Brazilian J Microbiol 47(1):181–190 Gientka I, Kieliszek M, Jermacz K, Błażejak S (2017) Identification and characterization of oleaginous yeast isolated from kefir and its ability to accumulate intracellular fats in deproteinated potato wastewater with different carbon sources. Biomed Res Int 2017(1):6061042 Okerentugba PO, Ataikiru TL, Ichor T (2016) Isolation and characterization of hydrocarbon utilizing yeast (HUY) isolates from palm wine. Am J Mol Biology 6(2):63–70 Hernández A, Martín A, Aranda E, Pérez-Nevado F, Córdoba MG (2007) Identification and characterization of yeast isolated from the elaboration of seasoned green table olives. Food Microbiol 24(4):346–351 Alves M, Gonçalves T, Quintas C (2012) Microbial quality and yeast population dynamics in cracked green table olives’ fermentations. Food Control 23(2):363–368 Karimi M, Hassanshahian M (2016) Isolation and characterization of phenol degrading yeasts from wastewater in the coking plant of Zarand, Kerman. Brazilian J Microbiol 47(1):18–24 Zaky AS, Tucker GA, Daw ZY, Du C (2014) Marine yeast isolation and industrial application. FEMS Yeast Res 14(6):813–825 Padilla B, Gil JV, Manzanares P (2016) Past and future of non-Saccharomyces yeasts: From spoilage microorganisms to biotechnological tools for improving wine aroma complexity. Front Microbiol 7:411 Steensels J, Snoek T, Meersman E, Nicolino MP, Voordeckers K, Verstrepen KJ (2014) Improving industrial yeast strains: exploiting natural and artificial diversity. FEMS Microbiol Rev 38(5):947–995 Carrasco M, Rozas JM, Barahona S, Alcaíno J, Cifuentes V, Baeza M (2012) Diversity and extracellular enzymatic activities of yeasts isolated from King George Island, the sub-Antarctic region. BMC Microbiol 12(1):251 Baxby D, Blundell N, Hart CA (1984) The development and performance of a simple, sensitive method for the detection of Cryptosporidium oocysts in faces. Epidemiol Infect 93(2):317–323 HiMedia Laboratories Pvt Ltd. A-516, Swastik Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6147 Bertolini MC, Ernandes JR, Laluce C (1991) New yeast strains for alcoholic fermentation at higher sugar concentrations. Biotechnol Lett 13(3):197–202 Roostita R (1993) Occurrence, growth and biochemical properties of yeasts in cheese and milk (Doctoral dissertation, UNSW Sydney) Roostita LB, Fleet GH, Wendry SP, Apon ZM, Gemilang LU (2011) Determination of yeasts antimicrobial activity in milk and meat products. Adv J Food Sci Technol 3(6):442–445 Younis G, Awad A, Dawod RE, Yousef NE (2017) Antimicrobial activity of yeasts against some pathogenic bacteria. Veterinary World 10(8):979 The Shroomery (2005) http://www.shroomery.org/images/23418/green5.jpg , accessed January 14 Martini A (1993) Origin and domestication of the wine yeast Saccharomyces cerevisiae. J Wine Res 4(3):165–176 Reis VR, Bassi APG, Silva JCGD, Ceccato-Antonini SR (2013) Characteristics of Saccharomyces cerevisiae yeasts exhibiting rough colonies and pseudo hyphal morphology with respect to alcoholic fermentation. Brazilian J Microbiol 44:1121–1131 Caputi A, Ueda M, Brown T (1968) Spectrophotometric determination of ethanol in wine. Am J Enol Viticult 19(3):160–165 Bkyeast Dyeing Yeast Cells: Life vs Death (2013) https://bkyeast.wordpress.com/2013/01/26/dyeing-yeast-cells-life-vs-death/ Tankeshwar A (2015) Ziehl-Neelsen technique (AFB Staining): Principle, Procedure and Reporting Hahn-Hägerdal B, Karhumaa K, Fonseca C, Spencer-Martins I, Gorwa-Grauslund MF (2007) Towards industrial pentose-fermenting yeast strains. Appl Microbiol Biotechnol 74(5):937–953 Guimarães TM, Moriel DG, Machado IP, Picheth CM, Bonfim T (2006) Isolation and characterization of Saccharomyces cerevisiae strains of winery interest. Revista Brasileira de Ciências Farmacêuticas 42:119–126 Graeme MW (2005) In: Kevin K (ed) and A.W.Nia, Introduction to Fungal Physiology. Biology and Application, John Wiley and Sons, Ltd, Fungi, pp 1–34 Azhar SHM, Abdulla R, Jambo SA, Marbawi H, Gansau JA, Faik AAM, Rodrigues KF (2017) Yeasts in sustainable bioethanol production: A review. Biochem Biophys Rep 10:52–61 Charoenchai C, Fleet GH, Henschke PA (1998) Effects of temperature, pH, and sugar concentration on the growth rates and cell biomass of wine yeasts. Am J Enol Viticult 49(3):283–288 Fleet GH (1993) Wine microbiology and biotechnology. CRC Lin Y, Zhang W, Li C, Sakakibara K, Tanaka S, Kong H (2012) Factors affecting ethanol fermentation using Saccharomyces cerevisiae BY4742. Biomass Bioenergy 47:395–401 Zabed H, Faruq G, Sahu JN, Azirun MS, Hashim R, Boyce N, A (2014) Bioethanol production from fermentable sugar juice. Sci world J 2014(1):957102 Tikka, C., Osuru, H. P., Atluri, N., Raghavulu, P. C. V., Mannur, I. S., Prasad, U.V., … Bhaskar, M. (2013). Isolation and characterization of ethanol tolerant yeast strains. Bioinformation, 9(8), 421. Benítez T, del Castillo L, Aguilera A, Conde J, Cerdáolmedo E (1983) Selection of wine yeasts for growth and fermentation in the presence of ethanol and sucrose. Appl Environ Microbiol 45(5):1429–1436 Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-9532904","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":629779168,"identity":"5b390023-97b4-404a-8752-1be4d8b5580d","order_by":0,"name":"Goiteom Senay Niguse","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDUlEQVRIiWNgGAWjYFACxgYEO6GiRg5EH3iAX0sjQs+DM8eMwVoSiLWG8WEbcyKYh0+LvHtz+4MPFdvs+aWbn0kksLGlzw87/BBoi52cbgN2LYZnDjY2zjhzO3HmnGNmEgk8Mrkbb6cZALUkG5sdwKFlRmJjM2/b7QSDGwlALRJsuRtnJ4C0HEjchk/L37bb9vY30r9JJBgwpxvOTv+AV4u8BFALY9ttxg0SOUBbEpgT5KVz8NtiwHOwcWYP0C8zbuQUWyQcOGa4QTqn4ECCAW6/yLcDA+xHxW17/hnpG2/+/FcjLz87ffOHDxV2cri0GCCJs0ggRAywKwfb0oBgM39AFxkFo2AUjIJRAAIAbRxrxQnDnRIAAAAASUVORK5CYII=","orcid":"","institution":"Mekelle University","correspondingAuthor":true,"prefix":"","firstName":"Goiteom","middleName":"Senay","lastName":"Niguse","suffix":""},{"id":629779169,"identity":"c64db487-ba0d-4f32-8020-ba55c523a9d7","order_by":1,"name":"Melaku Mekonen Kasegn","email":"","orcid":"","institution":"Mekelle University","correspondingAuthor":false,"prefix":"","firstName":"Melaku","middleName":"Mekonen","lastName":"Kasegn","suffix":""},{"id":629779170,"identity":"eb7ee676-3041-4835-a687-d39084b7e42e","order_by":2,"name":"Birhanu Kahsay Meresa","email":"","orcid":"","institution":"Mekelle University","correspondingAuthor":false,"prefix":"","firstName":"Birhanu","middleName":"Kahsay","lastName":"Meresa","suffix":""},{"id":629779171,"identity":"ed49c825-742f-42ea-8abf-b105f5a39ee4","order_by":3,"name":"Ashenafi Teklay Yaekob","email":"","orcid":"","institution":"Mekelle University","correspondingAuthor":false,"prefix":"","firstName":"Ashenafi","middleName":"Teklay","lastName":"Yaekob","suffix":""},{"id":629779172,"identity":"e9f75fda-8976-44cc-a1e0-ed10186a9699","order_by":4,"name":"Asqual Zeselasie","email":"","orcid":"","institution":"Mekelle University","correspondingAuthor":false,"prefix":"","firstName":"Asqual","middleName":"","lastName":"Zeselasie","suffix":""},{"id":629779173,"identity":"70f2f9ca-9d55-4196-a9fe-77477d0f1112","order_by":5,"name":"Samson Zemikael Haftu","email":"","orcid":"","institution":"Aksum University","correspondingAuthor":false,"prefix":"","firstName":"Samson","middleName":"Zemikael","lastName":"Haftu","suffix":""},{"id":629779174,"identity":"221a5c15-4991-4706-9590-9b2a0a52abbe","order_by":6,"name":"Girmay Mekonen","email":"","orcid":"","institution":"Mekelle University","correspondingAuthor":false,"prefix":"","firstName":"Girmay","middleName":"","lastName":"Mekonen","suffix":""},{"id":629779175,"identity":"f97e8ea7-cef5-4050-ab7f-31a69d992b14","order_by":7,"name":"Birkti Weldegebriel","email":"","orcid":"","institution":"Mekelle University","correspondingAuthor":false,"prefix":"","firstName":"Birkti","middleName":"","lastName":"Weldegebriel","suffix":""}],"badges":[],"createdAt":"2026-04-26 15:02:19","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-9532904/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9532904/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":108007762,"identity":"5cf696d2-7ac6-437c-8fb8-feea33daa97f","added_by":"auto","created_at":"2026-04-28 13:01:47","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":684922,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9532904/v1/84944c6c-c163-4f56-97ba-2d3af5cd80e9.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eEvaluation of Indigenous Yeasts from Local Drinks (\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eKorefe, Areki and Tej\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e) and Dough as Potential Fermenters for Bioethanol Production\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eEthiopia has a wide variety of traditional indigenous fermented foods and beverages that are prepared and consumed. Alcoholic fermentation, induced by indigenous microbes, is associated with particular fermentable substances and gives a distinctive style and quality to that wine or beer (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). Yeast can grow at temperatures of 30 and 37\u0026deg;C, and some of them grow at 45\u0026deg;C. Yeasts that show positive assimilation of glucose, sucrose, maltose, galactose, fructose, and raffinose and negative for lactose, xylose, lysine, nitrate, and urea are considered \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e. Palm wine is the main source of \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e for industrial applications. The yeast cells from palm have high viability, high leavening (82-94.7%) capacity, CO₂ production, temperature tolerance, ethanol tolerance, and osmotic tolerance (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eLocally available substances are the main sources of industrial ethanol fermentation because of reduced production costs (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). Some yeast strains contain necessary properties that are used as fermentation starters for various food or beverage industries (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e). Yeasts that are obtained from food crops have extracellular enzymatic activity and a high level of fermentation ability. Yeasts from local sources like food crops could increase ethanol production by fermenting carbohydrates efficiently, and this reduces the cost for importing yeast from abroad (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). Using yeasts isolated from readily available sources enables the production of fermentation products with a special taste and aroma. Unlike \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e, which could produce pleasant, fruity aromas when fermenting barley malt extract, yeasts like \u003cem\u003eBrettanomyces\u003c/em\u003e from wine barrels are wine spoilage or give unfavorable beer tastes (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). In order to ferment wine efficiently and make superior-quality wine, using yeast strains isolated from the winery itself is best. This is because the yeast strains are more familiar with the wine\u0026rsquo;s climatic conditions and adapted to their own climatic conditions (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e strains from indigenous sources like Murcha, grapes, and sugarcane provide special flavor to bread and are good strains for the bakery industry. However, yeasts that can be obtained from a plant called \u0026ldquo;Dar\u0026rdquo; are not important for baking because they lack the leavening properties that are required for yeast to be bakery yeast (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). Yeast isolates from different fruit peels, like pineapple and orange, were found to be temperature, pH, ethanol, and osmotolerant. Furthermore, it was reported that yeasts obtained from fruit peels were resistant to chloramphenicol. So, these yeast strains are important to ferment carbohydrates and enable them to produce bioethanol, which serves people as an alternative source of energy/fuel (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). Yeasts are an important component of insect gut microflora that are used to degrade toxic materials and produce vitamins, hormones, and different digestive enzymes (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eArabidopsis is the habitat of various yeast strains that are beneficial, such as the ability of the yeasts to activate auxin responses in plants (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). Critical selection of appropriate yeast strains with needed characteristics is important to develop the required flavor and other regional characteristics of wine. For example, a yeast strain that is able to produce a large amount of hydrogen sulfide is not needed for wine production because it results in wine with an unfavorable flavor. \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e strains obtained from berries and spontaneously fermented musts are able to grow up to 16% alcohol and 40\u0026ndash;50% of glucose concentration (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). According to \u003cem\u003eIwona Gientka et al.\u003c/em\u003e, kefir (fermented milk) is a source of \u003cem\u003eCandida inconspicua, Debaryomyces hansenii, Kluyveromyces marxianus, Kazachstania unispora\u003c/em\u003e, and \u003cem\u003eZygotorulaspora Florentina;\u003c/em\u003e about 20% of lipid substances can be accumulated in yeast cells by using carbon-rich deproteinated potato wastewater, a starch processing industry waste, as a medium for the yeast (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). \u003cem\u003eCandida adriatica\u003c/em\u003e and \u003cem\u003eCandida taoyuanica\u003c/em\u003e, isolated from palm wine, a sugar-rich substrate from a non-oil-impacted environment using Bacto Bushnell-Haas broth, were found to be hydrocarbon\u003cb\u003e-\u003c/b\u003eutilizing yeasts. This assists bioremediation by serving as biodegradable (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e). Well-adapted yeast strains with low polysaccharolytic activity and desirable biochemical properties derived from brine olives are able to produce high-quality green table olives with consistent organoleptic characteristics. Furthermore, they have the potential to degrade the polysaccharides of olives and be used as a starter culture (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe predominant microbes found in cracked green table olives, Manzanilla variety (a fermented food of Portugal), were yeasts with a large population of \u003cem\u003ethe S. cerevisiae\u003c/em\u003e species. Native strains that were selected from the food were proposed to act as starter cultures with the purpose of preventing spoilage during the storage of the food. The identification of the defined native strains helps to get superior-quality food products via fermentation, and this supports food-producing companies to be profitable (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). Phenolic compounds generated from industrial wastewater, like petrochemical plants, are environmental pollutants. Yeasts from soil and wastewater are the most important phenol biodegraders. It was reported that the yeast from soil samples had higher rates of phenol-degrading capacity than that of yeast isolated from water. Strong yeasts in phenol-degrading yeasts were identified using amplification of the 18S rRNA gene region, and these isolated yeasts were found to be \u003cem\u003eCandida tropicalis\u003c/em\u003e, \u003cem\u003ePichia guilliermondii\u003c/em\u003e, \u003cem\u003eMeyerozyma guilliermondii\u003c/em\u003e, and \u003cem\u003eC. tropicalis\u003c/em\u003e (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). Yeasts obtained from water bodies have several good features: higher osmosis tolerance, higher special chemical productivity, and production of industrial enzymes compared to yeasts from terrestrial sources. Marine yeasts are very important in the production of bioethanol and pharmaceutical products (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe use of mixed starters of selected strains of non-\u003cem\u003eSaccharomyces\u003c/em\u003e yeasts with strains of \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e enhances wine fermentations and is crucial for better organoleptic characteristics of wine. Yeasts other than \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e yeasts are used to improve the primary and secondary aroma of wines and produce important enzymes (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e). People have been using yeasts to make fermented beer, wine, sake, and bread via the fermentation process for many years. Furthermore, yeasts are important for biotechnological applications such as the production of biofuels and for bioremediation purposes. Modification of yeasts by mutagenesis, protoplast fusion, breeding, genome shuffling, and global transcription machinery engineering provides new improved strains with higher quality and fermentation efficiency than the wild type (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e). Yeast strains that are psychro-tolerant and able to assimilate carbon sources efficiently were isolated from King George Island, the sub-Antarctic region. These yeasts are of potential use in industrial applications requiring high enzyme activities at low temperatures (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e). To our knowledge, there are scanty reports in the area of wild yeast generated from indigenous sources. Therefore, the objective of the current work was to isolate and characterize yeast from local drinks (\u003cem\u003eTej, Areki\u003c/em\u003e, and \u003cem\u003eKorefe) and Teff\u003c/em\u003e dough and to determine their potential for bioethanol production.\u003c/p\u003e \u003cdiv id=\"Sec2\" class=\"Section2\"\u003e \u003ch2\u003e1.1. Significance of the study\u003c/h2\u003e \u003cp\u003eThe current study already opened the door to the idea that the local drinks and dough are important sources of yeast, which can serve to produce bioethanol. Since the local drink \u003cem\u003eKorefe\u003c/em\u003e is found only at \u003cem\u003eKorem\u003c/em\u003e, a study of the type of yeast in this type of local drink is a sole for researchers to study the yeast in a multidirectional way. Superior quality yeast strains can be found from the \u003cem\u003eKorefe\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Description of the study area\u003c/h2\u003e \u003cp\u003eThis study was conducted in \u003cem\u003eKorem town\u003c/em\u003e, Tigray regional state, located about 619 km from Addis Ababa, the capital of Ethiopia. Located on the eastern edge of the Ethiopian highlands in the Southern Zone of the Tigray Region, this town has a latitude and longitude of 12\u0026deg;30\u0026prime;N 39\u0026deg;31\u0026prime;E with an elevation of 2539 meters above sea level. It has an annual average temperature of 17.4\u0026deg;C. Major crops that are produced at \u003cem\u003eKorem\u003c/em\u003e are wheat, barley, and maize.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Sample collection\u003c/h2\u003e \u003cp\u003eThe samples of \u003cem\u003eKorefe, Areki, Tej\u003c/em\u003e, and dough, which were fermented for a week, were collected using sterile plastic bags from \u003cem\u003eKorem\u003c/em\u003e town, southern Tigray, Ethiopia. The samples put in plastic bags were brought aseptically using an isotherm icebox to the molecular biotechnology laboratory, College of Veterinary Medicine, Mekelle University, Ethiopia, for analysis and placed in a refrigerator until use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Sample preparation\u003c/h2\u003e \u003cp\u003eFour test tubes were prepared in a rack, and 9 ml of 9% (w/v) saline water was added to each test tube. Then 1 ml from each sample (\u003cem\u003eKorefe, Areki, Tej\u003c/em\u003e, and Dough) measured by micropipette was added to the respective test tube and homogenized using a vortex.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Isolation of yeast\u003c/h2\u003e \u003cp\u003eAseptically, 0.5 ml of each dilution was taken via pipette and spread using an inoculating loop on the petri dish containing Potato Dextrose Agar (PDA), to which tetracycline and oxytetracycline (not simultaneously) were added to inhibit the growth of bacteria. Then the petri dishes were incubated at 30\u0026deg;C for 48 hours. Separate yeast colonies from the agar plate were transferred using the streaking technique on Sabouraud Dextrose Agar (SDA) and incubated at 30\u0026deg;C for 48 h. It was then inoculated by streaking on YPD medium (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). To preserve the yeast, a slant was prepared using test tubes. YPD media was prepared, and 5 ml of YPD was added to each test tube. The test tubes were put slanting (45\u0026deg;C) and incubated at 30\u0026deg;C for 24 hours to check whether they were contaminated or not. Yeast isolates were inoculated into slanted test tubes using an inoculating loop and incubated at 30\u0026deg;C for 48 hours. Then the yeast grown in the slant was preserved in a refrigerator at 4\u0026deg;C for further use (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.5. Staining and microscopic observation of yeast isolates\u003c/h2\u003e \u003cp\u003eA smear of the sample from the colony was prepared on a slide after a drop of distilled water was added and fixed with flame. A few drops of methylene blue were applied to the slide and stayed for two minutes. Then it was decolorized by tap water (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e). The smear was prepared on a slide and fixed with flame. A malachite green concentration of 5% was applied to the slide. The slide was exposed to flame for 3 minutes. The slide was decolorized with tap water and counterstained with safranin for 30 seconds. Then the slide was washed with tap water (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e). Finally, yeast isolates that were stained using both methods were examined under oil immersion in a 100x microscope after the slide was dried in air.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Determination of physiological characteristics of yeasts through fermentation of sugars\u003c/h2\u003e \u003cp\u003eOf basal medium was prepared by adding enough powder to the required distilled water. Sugars and the broth were mixed in \u003cb\u003e1:10\u003c/b\u003e (one part sugar and 9 parts broth). Three ml of mixture was measured with a pipette and added to each test tube. One hundred microliters of penicillin were added to each test tube. A colony of yeast was inoculated into the test tubes except for the controls (a control for each sugar), and tubes in the rack were incubated for 24\u0026ndash;96 hours until a color change from red to yellow was observed (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Antimicrobial activity test\u003c/h2\u003e \u003cp\u003eTest tubes were prepared, and 1 ml of saline water was added to each test tube. Half a microliter of \u003cem\u003eStaphylococcus aureus\u003c/em\u003e was added. The turbidity of the test tubes was compared with McFarland. Then the bacteria spread on the petri dish containing the PDA. After a while, yeast was inoculated at a fixed place on the Petri dish (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e2.8. Estimation of Ethanol Content Using Potassium Dichromate Solution\u003c/h2\u003e \u003cp\u003eIn order to determine the amount of ethanol present in the molasses, a potassium dichromate solution was made using the (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e) method, which involved combining 325 milliliters of 96% H₂SO₄ with 33.8 grams of K₂Cr₂O₇ in 1000 milliliters of distilled water. Subsequently, the unknown concentration of ethanol generated from the molasses was estimated using this solution. An initial standard curve was created using known absolute ethanol concentrations in order to estimate the unknown ethanol content. Finally, the known concentrations generated from a standard curve were used to estimate the concentration of unknown materials using Eq.\u0026nbsp;2.\u003c/p\u003e \u003cp\u003eThe linear regression equation for the ethanol standard curve was as follows:\u003c/p\u003e \u003cp\u003e \u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(\\:\\text{A}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e}=-0.09700+0.3691\\text{x}\\:\\:\\)\u003c/span\u003e \u003c/span\u003e Eq.\u0026nbsp;(1)\u003c/p\u003e \u003cp\u003eThe rearrangement of the equation for the estimated ethanol concentration (x) gives\u003c/p\u003e \u003cp\u003e \u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(\\:\\text{x}=\\:\\frac{\\text{a}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e}\\:+\\:0.09700}{0.3691}\\)\u003c/span\u003e \u003c/span\u003e Eq.\u0026nbsp;(2)\u003c/p\u003e \u003cp\u003eConsidering a dilution factor of 10, we get the percentage of ethanol (y).\u003c/p\u003e \u003cp\u003e \u003cspan class=\"InlineEquation\"\u003e \u003cspan class=\"mathinline\"\u003e\\(\\:\\text{y}=\\:\\text{x}\\text{*}\\text{D}\\text{F}\\)\u003c/span\u003e \u003c/span\u003e Eq.\u0026nbsp;(3)\u003c/p\u003e \u003cp\u003eWhere x is the estimated ethanol concentration, y is the ethanol concentration percentage (v/v), and DF is the dilution factor.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e2.9. Data Analysis\u003c/h2\u003e \u003cp\u003eAll the data were collected and analyzed using the Minitab software (v. 20). Results were displayed in tables. ANOVA was used to determine if the yeasts from different samples have significant differences in their ethanol concentration, which was produced from molasses and presented via tables.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Cultural characteristics of yeast isolates\u003c/h2\u003e \u003cp\u003eThe yeast isolate that was plated on PDA and SDA had different colony structures (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Isolates from \u003cem\u003eKorefe\u003c/em\u003e and \u003cem\u003eAreqi\u003c/em\u003e were visible, creamy white, and mucoid colonies with raised surfaces. The other isolates were shown to be filamentous, and a yellowish colony structure was observed. The yeast colony isolated from \u003cem\u003eTej\u003c/em\u003e had an irregular form, a convex margin, and an undulate elevation. The yeast colony isolated from dough had a circular form, raised elevation, and an entire margin. The form and margin of yeast isolated from \u003cem\u003eKorefe\u003c/em\u003e and \u003cem\u003eAreqi\u003c/em\u003e were similar to that of dough, but it had a convex margin.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Microscopic morphology of the yeast isolates\u003c/h2\u003e \u003cp\u003eThe isolate showed active growth on screening for its ability to grow in yeast extract peptone dextrose medium (YPD), PDA, and SDA. The isolated colonies stained by malachite green and methylene blue were circular, ellipsoid, and rod-shaped structures (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\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\u003eCultural and microscopic features of the yeast colonies\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eo\u003c/span\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eYeast isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCultural and microscopic features\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white and mucoid colonies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white and mucoid colonies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white, mucoid colonies, ellipsoid\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white, mucoid colonies, ellipsoid\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white and mucoid colonies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white, mucoid colonies, rod-shape\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white, mucoid colonies, ellipsoid\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white, mucoid colonies, ellipsoid\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white, mucoid colonies, rod-shape\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white and mucoid colonies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white, mucoid colonies, circular\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white and mucoid colonies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white and mucoid colonies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecreamy white and mucoid colonies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eraised elevation, margin, circular\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eraised elevation, margin, circular\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eraised elevation, margin, circular\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-ox-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eraised elevation, margin, circular\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eirregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eyellowish colony, filamentous\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eyellowish colony, filamentous\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eyellowish colony, filamentous\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIrregular, a convex margin, undulate elevation\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\u003eK- korefe, A- areqi, D- dough, T- tela\u003c/em\u003e, Oxy- oxy-tetracycline, Pen- penicillin\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e3.3. Physiological characteristics of yeasts through fermentation of sugars\u003c/h2\u003e \u003cp\u003eThe physiological characteristics of the normal bakery yeast cells were identified by the carbohydrate fermentation tests (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). All yeast isolates were able to ferment glucose, galactose, maltose, fructose, and sucrose at different incubation periods, as observed by color change from red to yellow. But none of the yeast isolates were able to ferment sorbitol and xylose.\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\u003eIsolates from \u003cem\u003eKorefe, Areqi\u003c/em\u003e, \u003cem\u003eTeff\u003c/em\u003e dough, and \u003cem\u003eTej\u003c/em\u003e and the result of fermentation\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\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eN\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eo\u003c/span\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eYeast Isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c10\" namest=\"c3\"\u003e \u003cp\u003eTypes of sugars tested\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGlucose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGalactose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMaltose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFructose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSucrose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eXylose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eSorbitol\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e 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align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e 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align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e 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\u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-oxy-5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eT-pen-12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\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\u003eK- korefe, A- areqi, D- dough, T- tela\u003c/em\u003e, Oxy- oxy-tetracycline, Pen- penicillin\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e3.4. Antimicrobial activity test\u003c/h2\u003e \u003cp\u003eYeast isolate did not show any antimicrobial activity against \u003cem\u003eStaphylococcus aureus;\u003c/em\u003e as a result, there was not a clear zone observed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e3.5. Bioethanol production from molasses\u003c/h2\u003e \u003cp\u003eYeast isolates did show varied potential for the production of bioethanol. The result for the effect of temperature, pH, and incubation period on ethanol production from molasses using isolated yeast investigated how the parameters significantly affect the fermentation process (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Due to the effect of temperature, pH, and incubation period, higher ethanol content (8.50%) was obtained at pH 5.5 and 30\u0026deg;C and incubated in four days with the isolate of A-pen-3. It is non-significant with ethanol produced at 26\u0026deg;C at pH 3.5. In contrast, lower ethanol content (2%) was recorded at pH 5.5 and 37\u0026deg;C and incubated for three days with the isolate of A-oxy-3.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEthanol production from molasses using yeasts isolated from local drinks\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNo.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatments/\u003c/p\u003e \u003cp\u003eTrials\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003eFactors\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAbsorbance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eX\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ey % (v/v)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTemp\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eIP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.134\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.397\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3.97\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.140\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e4.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.246\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e7.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.150\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e4.80\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.104\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3.90\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.151\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.207\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.255\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e7.90\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.195\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.175\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.267\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.279\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8.50\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.178\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.579\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.79\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.152\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.123\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e4.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e 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align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.381\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3.81\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.049\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-oxy-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.202\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.644\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.198\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.633\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.091\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.343\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3.43\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-oxy-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.255\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.788\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e7.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.230\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e7.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.585\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK-pen-4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.199\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e4.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.040\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA-pen-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.549\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.49\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eD-pen-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.2603\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.798\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e7.98\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eIP \u0026ndash; Incubation period\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eIn this study, 30 yeast isolates were isolated from local fermented \u003cem\u003eKorefe\u003c/em\u003e, \u003cem\u003eTeff\u003c/em\u003e dough, \u003cem\u003eTej\u003c/em\u003e, and \u003cem\u003eAreqi\u003c/em\u003e. Based on the colony morphology, 14 isolates isolated from local fermented \u003cem\u003eKorefe\u003c/em\u003e and \u003cem\u003eAreqi\u003c/em\u003e were found to be \u003cem\u003eSaccharomyces cerevisiae.\u003c/em\u003e The colonies exhibiting characteristics such as creamy to white color, fluffiness, and smooth margins were selected as the \u003cem\u003eSaccharomyces\u003c/em\u003e strain (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). The yeast isolates were identified as \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e based on the morphology of the colony (circular) and sugar fermentation tests. Reis et al. found \u003cem\u003eS. cerevisiae\u003c/em\u003e that grew on YPD medium had a smooth and a rough colony (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e). Microscopic structure of methylene blue stained the yeast cells blue and showed them as circular (\u003cem\u003eKorefe\u003c/em\u003e and \u003cem\u003eAreqi\u003c/em\u003e isolates), irregular (dough), and hyphae (\u003cem\u003eTej\u003c/em\u003e isolates). Methylene blue stains the dead cells blue. Viable yeast cells also can be stained blue, but the active enzymes within them reduce the dye and make it colorless (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). The microscopic structure of \u003cem\u003eS. cerevisiae\u003c/em\u003e has a characteristic ellipsoid or ovoid shape (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e). The malachite green stain penetrated into the endospores by heating. Malachite green removes easily from the vegetative cells using water but not from endospores. The vegetative yeast cells appeared pink (the color of safranin), and the spores appeared green. The green indicates that the yeasts are spore-forming (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e). Yeast isolates from dough and \u003cem\u003eTej\u003c/em\u003e isolates were found to be hyphae based on the microscopic structure, which was thin and rod-shaped.\u003c/p\u003e \u003cp\u003eYeast isolates obtained from \u003cem\u003eKorefe\u003c/em\u003e and \u003cem\u003eAreqi\u003c/em\u003e (14 isolates) fermented glucose, galactose, fructose, maltose, and sucrose within 24 hours. \u003cem\u003eS. cerevisiae\u003c/em\u003e can be identified by its capability to ferment sucrose, maltose, fructose, glucose, galactose, and raffinose but not lactose (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e). Therefore, yeast isolates obtained from \u003cem\u003eKorefe\u003c/em\u003e and \u003cem\u003eAreqi\u003c/em\u003e are \u003cem\u003eS. cerevisiae\u003c/em\u003e. While these 4 isolates isolated from dough took 48\u0026ndash;96 hours to ferment the given sugars. In another way, yeast isolates from local \u003cem\u003eTej\u003c/em\u003e (12 isolates) were able to ferment glucose, galactose, fructose, maltose, and sucrose. The yeast isolates were able to ferment sucrose, maltose, fructose, and glucose because yeasts produce enzymes that break the glycosidic bond that attaches two glucose units and convert sugar monomers into alcohol and CO₂. The sugar fermentation was indicated by a color change from red to yellow due to acid production (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e). But none of the yeast isolates were able to ferment sorbitol and xylose. This result is similar to the work reported by (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFermentation tests indicated that none of the isolates produce ethanol using xylose as a carbon source (\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e). A control containing OF basal medium without the addition of yeast isolates was used and remained dark black, which was similar to that of yeast-treated xylose and sorbitol. In the antimicrobial test, the \u003cem\u003eS\u003c/em\u003e. \u003cem\u003eaureus\u003c/em\u003e cannot inhibit the growth of yeast. So, the pathogen \u003cem\u003eS. aureus\u003c/em\u003e did not kill yeast. In contrast to this work, Younis \u003cem\u003eet al.\u003c/em\u003e found yeast isolates showed antimicrobial activities \u003cem\u003eagainst S. aureus, E. coli\u003c/em\u003e, and \u003cem\u003eP. aeruginosa\u003c/em\u003e, showing remarkable antimicrobial activity against \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e with a large, clear inhibition zone and low antimicrobial activity against \u003cem\u003eP. aeruginosa\u003c/em\u003e (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the current study, the highest ethanol was produced when the fermentation was conducted at pH 5.5 at 30\u0026deg;C and incubated for four days by the isolate from Areqi (A-pen-3). However, the least ethanol was produced when the fermentation was conducted at pH 5.5 at 37\u0026deg;C and incubated for three days by the isolate from Areqi (A-oxy-3). Zabed et al. reported that a pH range of 4.0\u0026ndash;5.0 is suitable for the ethanol production process (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e). During the fermentation production of bioethanol, it depends on several factors such as temperature, sugar concentration, pH, and fermentation time (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e). Particularly, temperature directly affected the growth rate of microorganisms. The current fermentation of molasses gave a remarkable ethanol yield at 26\u0026deg;C and 30\u0026deg;C incubated for four days. In line with this, Charoenchai et al. reported that the ideal temperature range for fermentation is between 20 and 35\u0026deg;C, while the optimum temperature of S. cerevisiae is near to 30\u0026deg;C (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e). Ethanol production is influenced by the pH of the broth, as it affects yeast growth and fermentation rate. Moreover, the survival and growth of yeasts are influenced by the pH in the range of 2.75\u0026ndash;4.25 (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e). In fermentation for ethanol production, the optimum pH range of S. cerevisiae is 4.0\u0026ndash;5.0 (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). Fermentation time affects the growth of microorganisms. Short fermentation time leads to insufficient microbial growth, which negatively affects the fermentation and results in incomplete conversion of sugars into ethanol (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e). On the other hand, longer fermentation time has a toxic effect on microbial growth, especially in batch mode due to the high concentration of ethanol in the fermented broth. Complete fermentation can be achieved at lower temperatures by using a longer fermentation time, which results in the lowest ethanol yield (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e). The lower ethanol content shown in (A-oxy-3) might be due to the inability to tolerate the produced ethanol. Studies reported by Tikka et al. and Ben\u0026iacute;tez et al. stated that not all yeasts have the potential to survive in the produced ethanol (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e).\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eGenerally, it can be concluded that the local drinks (\u003cem\u003eKorefe\u003c/em\u003e and \u003cem\u003eAreki\u003c/em\u003e) could be the major source for \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e, and the main fermenter yeast for these local drinks is \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e. Most isolates are excellent fermenters, but \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e is the main and fastest fermenting yeast that can ferment sugars within a short period of time, i.e., 24 hours. Most samples were capable of fermenting most sugars. Samples from \u003cem\u003eKorefe, Areki\u003c/em\u003e, and \u003cem\u003eTej\u003c/em\u003e were able to ferment glucose, maltose, sucrose, fructose, and galactose but not xylose and sorbitol. While those samples from dough all ferment glucose, fructose, and sucrose, some of them ferment galactose and maltose, and others cannot ferment. Yeasts generated from indigenous sources are the most common microorganisms in bioethanol production and play a crucial function in fermenting sugars to ethanol from molasses. It is one of the major global issues in the production of alternative energy used in various sectors.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgment\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors thank \u003cem\u003eMekelle University for the fund\u003c/em\u003e.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eThis work was financially supported by Mekelle University (RDPDO/MU/CDANR/Generaljunior/020/13).\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclosure statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo potential conflict of interest was reported by the author(s).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSample collection\u003cstrong\u003e,\u0026nbsp;\u003c/strong\u003econceptualization, investigation, writing of the original draft, and visualization were performed by MMK and GSN. ATY achieved validation and project administration. \u0026nbsp;BKM and SZH performed\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003edata analysis and interpretation. GSN, MMK, GM, and BW accomplished writing the review \u0026amp; editing and supervision. \u003cem\u003eAll authors read and approved the final.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the data generated or analyzed during this study are included in this published article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eCappello MS, Bleve G, Grieco F, Dellaglio F, Zacheo G (2004) Characterization of Saccharomyces cerevisiae strains isolated from must of grape grown in experimental vineyard. J Appl Microbiol 97(6):1274\u0026ndash;1280\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlowonibi OO (2017) Isolation and Characterization of Palm Wine Strains of Saccharomyces cerevisiae Potentially Useful as Bakery Yeasts. \u003cem\u003eEuropean Journal of Experimental Biology\u003c/em\u003e, 2017, 7:11\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNwachukwu IN, Ibekwe VI, Nwabueze RN, Anyanwu BN (2006) Characterization of palm wine yeast isolates for industrial utilization. Afr J Biotechnol 5(19):1725\u0026ndash;1728\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTristezza, M., Fantastico, L., Vetrano, C., Bleve, G., Corallo, D., Grieco, F., \u0026hellip;Grieco, F. (2014). Molecular and technological characterization of Saccharomyces cerevisiae strains isolated from natural fermentation of Susumaniello grape must in Apulia, Southern Italy. International Journal of Microbiology,\u003cem\u003e2014\u003c/em\u003e(1), 897\u0026thinsp;\u0026ndash;\u0026thinsp;428\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEbabhi AM, Adekunle AA, Okunowo WO, Osuntoki AA (2013) Isolation and characterization of yeast strains from local food crops. J Yeast Fungal Res 4(4):38\u0026ndash;43\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDeLong LN, Noone B, Erickson L (2017) Isolation, Identification, and Characterization of Local Wild Yeasts for Use in Fermentation. FASEB J 31:943\u0026ndash;944\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuimar\u0026atilde;es TM, Moriel DG, Machado IP, Picheth CM, Bonfim T (2006) Isolation and characterization of Saccharomyces cerevisiae strains of winery interest. Revista Brasileira de Ci\u0026ecirc;ncias Farmac\u0026ecirc;uticas 42:119\u0026ndash;126\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKarki, T. B., Timilsina, P. M., Yadav, A., Pandey, G. R., Joshi, Y., Bhujel, S., \u0026hellip;Neupane, K. (2017). Selection and characterization of potential baker\u0026rsquo;s yeast from indigenous resources of Nepal. Biotechnology research international, 2017(1), 1925820.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNasir A, Rahman SS, Hossain MM, Choudhury N (2017) Isolation of Saccharomyces cerevisiae from pineapple and orange and study of metal\u0026rsquo;s effectiveness on ethanol production. Eur J Microbiol Immunol 7(1):76\u0026ndash;91\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMatos ITSR, Assun\u0026ccedil;\u0026atilde;o EN, Carmo EJD, Soares VM, Astolfi-Filho S (2017) Isolation and characterization of yeasts able to assimilate sugarcane bagasse hemicellulosic hydrolysate and produce xylitol associated with veturius transversus (Passalidae, Coleoptera, and Insecta). Int J Microbiol 2017(1):5346741\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang K, Sipil\u0026auml; TP, Overmyer K (2016) The isolation and characterization of resident yeasts from the phylloplane of Arabidopsis thaliana. Sci Rep 6(1):39403\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eŠuransk\u0026aacute; H, Vr\u0026aacute;nov\u0026aacute; D, Omelkov\u0026aacute; J (2016) Isolation, identification and characterization of regional indigenous Saccharomyces cerevisiae strains. Brazilian J Microbiol 47(1):181\u0026ndash;190\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGientka I, Kieliszek M, Jermacz K, Błażejak S (2017) Identification and characterization of oleaginous yeast isolated from kefir and its ability to accumulate intracellular fats in deproteinated potato wastewater with different carbon sources. Biomed Res Int 2017(1):6061042\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOkerentugba PO, Ataikiru TL, Ichor T (2016) Isolation and characterization of hydrocarbon utilizing yeast (HUY) isolates from palm wine. Am J Mol Biology 6(2):63\u0026ndash;70\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHern\u0026aacute;ndez A, Mart\u0026iacute;n A, Aranda E, P\u0026eacute;rez-Nevado F, C\u0026oacute;rdoba MG (2007) Identification and characterization of yeast isolated from the elaboration of seasoned green table olives. Food Microbiol 24(4):346\u0026ndash;351\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlves M, Gon\u0026ccedil;alves T, Quintas C (2012) Microbial quality and yeast population dynamics in cracked green table olives\u0026rsquo; fermentations. Food Control 23(2):363\u0026ndash;368\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKarimi M, Hassanshahian M (2016) Isolation and characterization of phenol degrading yeasts from wastewater in the coking plant of Zarand, Kerman. Brazilian J Microbiol 47(1):18\u0026ndash;24\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZaky AS, Tucker GA, Daw ZY, Du C (2014) Marine yeast isolation and industrial application. FEMS Yeast Res 14(6):813\u0026ndash;825\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePadilla B, Gil JV, Manzanares P (2016) Past and future of non-Saccharomyces yeasts: From spoilage microorganisms to biotechnological tools for improving wine aroma complexity. Front Microbiol 7:411\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSteensels J, Snoek T, Meersman E, Nicolino MP, Voordeckers K, Verstrepen KJ (2014) Improving industrial yeast strains: exploiting natural and artificial diversity. FEMS Microbiol Rev 38(5):947\u0026ndash;995\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarrasco M, Rozas JM, Barahona S, Alca\u0026iacute;no J, Cifuentes V, Baeza M (2012) Diversity and extracellular enzymatic activities of yeasts isolated from King George Island, the sub-Antarctic region. BMC Microbiol 12(1):251\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaxby D, Blundell N, Hart CA (1984) The development and performance of a simple, sensitive method for the detection of Cryptosporidium oocysts in faces. Epidemiol Infect 93(2):317\u0026ndash;323\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHiMedia Laboratories Pvt Ltd. A-516, Swastik Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6147\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBertolini MC, Ernandes JR, Laluce C (1991) New yeast strains for alcoholic fermentation at higher sugar concentrations. Biotechnol Lett 13(3):197\u0026ndash;202\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRoostita R (1993) Occurrence, growth and biochemical properties of yeasts in cheese and milk (Doctoral dissertation, UNSW Sydney)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRoostita LB, Fleet GH, Wendry SP, Apon ZM, Gemilang LU (2011) Determination of yeasts antimicrobial activity in milk and meat products. Adv J Food Sci Technol 3(6):442\u0026ndash;445\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYounis G, Awad A, Dawod RE, Yousef NE (2017) Antimicrobial activity of yeasts against some pathogenic bacteria. Veterinary World 10(8):979\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThe Shroomery (2005) \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.shroomery.org/images/23418/green5.jpg\u003c/span\u003e\u003cspan address=\"http://www.shroomery.org/images/23418/green5.jpg\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e, accessed January 14\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMartini A (1993) Origin and domestication of the wine yeast Saccharomyces cerevisiae. J Wine Res 4(3):165\u0026ndash;176\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eReis VR, Bassi APG, Silva JCGD, Ceccato-Antonini SR (2013) Characteristics of Saccharomyces cerevisiae yeasts exhibiting rough colonies and pseudo hyphal morphology with respect to alcoholic fermentation. Brazilian J Microbiol 44:1121\u0026ndash;1131\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCaputi A, Ueda M, Brown T (1968) Spectrophotometric determination of ethanol in wine. Am J Enol Viticult 19(3):160\u0026ndash;165\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBkyeast Dyeing Yeast Cells: Life vs Death (2013) \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://bkyeast.wordpress.com/2013/01/26/dyeing-yeast-cells-life-vs-death/\u003c/span\u003e\u003cspan address=\"https://bkyeast.wordpress.com/2013/01/26/dyeing-yeast-cells-life-vs-death/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTankeshwar A (2015) \u003cem\u003eZiehl-Neelsen technique (AFB Staining): Principle, Procedure and Reporting\u003c/em\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHahn-H\u0026auml;gerdal B, Karhumaa K, Fonseca C, Spencer-Martins I, Gorwa-Grauslund MF (2007) Towards industrial pentose-fermenting yeast strains. Appl Microbiol Biotechnol 74(5):937\u0026ndash;953\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuimar\u0026atilde;es TM, Moriel DG, Machado IP, Picheth CM, Bonfim T (2006) Isolation and characterization of Saccharomyces cerevisiae strains of winery interest. Revista Brasileira de Ci\u0026ecirc;ncias Farmac\u0026ecirc;uticas 42:119\u0026ndash;126\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGraeme MW (2005) In: Kevin K (ed) and A.W.Nia, Introduction to Fungal Physiology. Biology and Application, John Wiley and Sons, Ltd, Fungi, pp 1\u0026ndash;34\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAzhar SHM, Abdulla R, Jambo SA, Marbawi H, Gansau JA, Faik AAM, Rodrigues KF (2017) Yeasts in sustainable bioethanol production: A review. Biochem Biophys Rep 10:52\u0026ndash;61\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCharoenchai C, Fleet GH, Henschke PA (1998) Effects of temperature, pH, and sugar concentration on the growth rates and cell biomass of wine yeasts. Am J Enol Viticult 49(3):283\u0026ndash;288\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFleet GH (1993) Wine microbiology and biotechnology. CRC\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLin Y, Zhang W, Li C, Sakakibara K, Tanaka S, Kong H (2012) Factors affecting ethanol fermentation using Saccharomyces cerevisiae BY4742. Biomass Bioenergy 47:395\u0026ndash;401\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZabed H, Faruq G, Sahu JN, Azirun MS, Hashim R, Boyce N, A (2014) Bioethanol production from fermentable sugar juice. Sci world J 2014(1):957102\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTikka, C., Osuru, H. P., Atluri, N., Raghavulu, P. C. V., Mannur, I. S., Prasad, U.V., \u0026hellip; Bhaskar, M. (2013). Isolation and characterization of ethanol tolerant yeast strains. Bioinformation, 9(8), 421.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBen\u0026iacute;tez T, del Castillo L, Aguilera A, Conde J, Cerd\u0026aacute;olmedo E (1983) Selection of wine yeasts for growth and fermentation in the presence of ethanol and sucrose. Appl Environ Microbiol 45(5):1429\u0026ndash;1436\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Mekelle University","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"bioethanol, fermentation, local drinks, molasses, yeasts","lastPublishedDoi":"10.21203/rs.3.rs-9532904/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9532904/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe depletion of fossil fuels forced researchers to find alternative, cheap, locally variable, and renewable sources of energy. Molasses is among the promised sources of fuel-grade ethanol. However, yeast strains that can produce large amounts of ethanol while resisting high sugar concentrations are difficult. Therefore, this study aims to investigate the potential of yeasts from Tigray\u0026rsquo;s local drinks and their potential for bioethanol production. Accordingly, yeasts were isolated from \u003cem\u003eKorefe, Areki, teff\u003c/em\u003e dough, and \u003cem\u003eTej\u003c/em\u003e. Isolation, morphological characterization, and biochemical tests were carried out to identify potential yeasts. Then the yeasts were applied in molasses fermentation to examine their potential for bioethanol production. Bioethanol was estimated using the estimation of ethanol content using potassium dichromate solution and a standard curve-generated linear equation (absorbance = -0.09700\u0026thinsp;+\u0026thinsp;0.3691x). Results showed that the yeasts have the capability of osmotolerance, and the yeasts isolated from \u003cem\u003eAreki\u003c/em\u003e (A-pen-3) produce a high amount of ethanol (8.5%) from molasses. Hence, production of ethanol from molasses using current yeasts has moderate potential and contributes to the effort of creating a sustainable and green environment to make the earth suitable for living. This study provides a good insight into using wild yeasts, and conducting further comprehensive research is appreciated.\u003c/p\u003e","manuscriptTitle":"Evaluation of Indigenous Yeasts from Local Drinks (Korefe, Areki and Tej) and Dough as Potential Fermenters for Bioethanol Production","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-28 09:21:29","doi":"10.21203/rs.3.rs-9532904/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6b0ab3e4-5aa0-4c55-bf6d-b25016061468","owner":[],"postedDate":"April 28th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-28T09:21:29+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-28 09:21:29","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9532904","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9532904","identity":"rs-9532904","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}