Brevibacterium litoralis sp. nov., a cellulose-degrading strain isolated from marine surface sediment

Brevibacterium litoralis sp. nov., a cellulose-degrading strain isolated from marine surface sediment | 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 Brevibacterium litoralis sp. nov., a cellulose-degrading strain isolated from marine surface sediment Quan Yang, Aolin Zhao, Haifei Liu, Jiawei Li, Shujing Wu, Ying Huang, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4724416/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 14 Oct, 2024 Read the published version in Antonie van Leeuwenhoek → Version 1 posted 4 You are reading this latest preprint version Abstract A Gram stain-positive, non-spore-forming, non-motile, short-rod actinomyces strain GXQ1321 T was isolated from maritime surface sediments in Beihai(11°46′21.11″N, 109°62′56.25″E), Guangxi Zhuang Autonomous Region, and a number of categorization studies were performed. Following a period of 72 hours of incubation at a temperature of 30°C within an actinomycetes culture medium, the colony was yellow, circular, smooth, central bulge, convex, opaque, with a 1.8-3.0 mm diameter. Strain GXQ1321 T has the ability to produce amylase and cellulase. Chemotaxonomic studies revealed that the major menaquinone in strain GXQ1321 T is MK-8. The most prevalent cellular fatty acids were anteiso -C 19:0 (27.28%), anteiso -C 15:0 (18.97%), anteiso -C 17:0 (15.95%), and iso -C 16:0 (12.21%). The whole-cell sugars of the strain GXQ1321 T identified were rhamnose, xylose and glucose. Strain GXQ1321 T exhibited the presence of meso-diaminopimelic acid (m-DAP) as a distinctive cell-wall diamino acid, and the polar lipids were identified as diphosphatidylglycerol (DPG), three phosphoglycolipid (PGL), phosphatidylglycerol (PG) and two unknown glycolipid (UG). This strain had 69.6% DNA G + C content. Strain GXQ1321 T is classified as Brevibacterium based on its 16S rRNA gene sequence. It is closely related to Brevibacterium samyangense SST-8 T (96.77%) and Brevibacterium rongguiense 5221 T (96.32%). The results showed that the average nucleotide identity (ANI) values of GXQ1321 T and the above two strain tyoes were 73.91–77.14%, and the digital DNA-DNA hybridisation (dDDH) values were 15.3–21.1%. Based on the phylogenetic, chemotaxonomi and physiologicalc data, strain GXQ1321 T was considered to be a new species of the genus Brevibacterium , named Brevibacterium litoralis sp. nov, with the type strain GXQ1321 T (= MCCC 1K08964 T = KCTC 59167 T ). Surface sediments Marine actinomycetes Novel species Species identification Figures Figure 1 Figure 2 Introduction Breed established the genus Brevibacterium in 1953, and its type species is Brevibacterium linens (Breed 1953). There are currently 57 recognised species in the genus Brevibacterium . The genus is mainly isolated from soil (Tang et al. 2008; Jung et al. 2018), sediment (Pei et al. 2021; Bhadra et al. 2008; Deng et al. 2020; Pei et al. 2020), insects (Kati et al. 2010), marine animals (Zhang et al. 2023), wall (Kampfer et al. 2010), clinical specimens (Wauters et al. 2001; Mages et al. 2008), and human samples (McBride et al. 1993; Pascual et al. 1996). Brevibacterium species are rod-shaped, nonspore-forming, Gram-positive bacteria (Bernard et al. 2010). Brevibacterium has anteiso -C 15:0 and anteiso -C 17:0 as its major cellular fatty acids, while their primary methylnaphthoquinone is MK-8 (H2) (Bernard et al. 2010). In addition, phosphatidylglycerol is often detected as a major lipid (Pascual et al. 1999; Komagata et al. 1964; Choi et al. 2013). Brevibacterium strains have many industrial applications. Studies have shown that Brevibacterium can use extracellular proteases and lipases to degrade lipids and proteins (such as casein) (Ozturkoglu-Budak S et al. 2013). At the same time, many Brevibacterium strains also have the ability to modify sulphur-containing amino acids to produce volatile sulphides, which is useful in the production of volatile flavours (Amarita F et al. 2004). In addition, certain strains of the genus Brevibacterium are used to recover phosphorus-rich products by growing in mixed culture wastewater streams (Simoes, F et al. 2018). Due to the remarkable achievements in the application of Brevibacterium in industrial cheese production, biodegradation and wastewater treatment, and other biotechnologies, more and more Brevibacterium strains have been unearthed, laying the foundation for future industrial development. After being separated from the seawater's surface sediment, the strain GXQ1321 T was discovered to be a novel species of Brevibacterium . This study was undertaken to define the strain's classification status and to provide a foundation for further exploration and development. Materials and Methods Strain Isolation and Culture Conditions The strain GXQ1321 T was isolated from marine surface sediments in Beihai (11°46′21.11″N, 109°62′56.25″E), Guangxi, Guangxi, China. A modified actinomycetes culture medium (0.15 g K 2 HPO 4 , 0.15 g NaCl, 0.30 g KNO 3 , 0.15 g MgSO 4 , 0.003 g FeSO 4 , 6.0 g soluble starch, 4.5 g agar, 300 mL sterile seawater, pH = 7.5) was used as the culture media for the separation. The surface sediment is naturally air dried, then ground to a powder with a sterile grinder and filtered. 30 mL of sterile seawater were mixed with 3 g of the sediment powder samples, and the combination was agitated for 30 minutes at 30°C. It was then diluted in gradient with sterile seawater to 10 − 4 , 10 − 5 , 10 − 6 . Diluents of different concentrations were coated with actinomycetes culture medium and cultured at 30 ℃ for 5 days. To obtain pure culture, strains isolated from the plate were cultivated under the same conditions and purified in actinomycetes culture medium. Store the pure culture solution in glycerol (40%, v/v) at -80°C. 16S rRNA Gene Sequencing and Phylogenetic Analysis DNA extraction was with TIANamp Bacteria DNA Kit (Lamballerie et al. 1992). The 16S rRNA gene was amplified using PCR in accordance with Chun et al. (1995) and Edwards et al. (1989). The particular primers 27F and 1492R were used to get the sequencing of the 16S rRNA gene (Lane et al. 1991), determined by AuGCT Biotech (Wu Han, China) and analysed in EzBioCloud ( http://ezbiocloud.net/ ) (Yoon et al. 2017). Evolutionary analysis was performed using MEGA 11 as previously described by Tamura et al. (2021). Phylogenetic tree models are constructed using the following three algorithms: maximum likelihood (ML) (Felsenstein 1981), maximum parsimony (MP) (Fitch 1971), and neighbour-joining (NJ) (Saitou and Nei 1987). The bootstrap method was used to test the stability of the phylogenetic tree topology, using 1000 replicates overall. Utilizing the Kimura two-parameter method, the evolutionary distance was determined (Kimura 1980; Tamura et al. 2013). Genomic Analysis The whole genome of strain GXQ1321 T was sequenced on Illumina Hiseq 4000 (Shanghai Magi Biomedical Technology Co., LTD., Shanghai, China) and calibrated using SOAP denovo 2.04 and SOAP aligner 2.21 (Li et al. 2008). Draft genomes of 20 Brevibacterium strains were collected from the NCBI ( http://www.ncbi.nlm.nih.gov/ ) database species in order to construct the phylogenetic tree. Pseudoclavibacter helvolus DSM 20419 T (JACHWJ000000000) was used as an outgroup. Genome sequence analysis was performed using a rapid annotation technique based on Subsystem Technology 2.0 ( https://rast.nmpdr.org/ ) (Aziz et al. 2008). Utilizing the ChunLab online ANI calculator from EzBioCloud ( https://www.ezbiocloud.net/ ), strains closely related to GXQ1321 T were analyzed for average nucleotide identity (ANI). Digital DNA-DNA hybridization (dDDH) values between GXQ1321 T and related Brevibacterium members were computed using the genome-genome distance calculator ( https://ggdc.dsmz.de/ggdc.php ) (Meier-Kolthoff et al. 2013). Using the Majorbio amino acids identified online calculator ( https://www.majorbio.com/ ) is closely related to the GXQ1321 T short bacillus strains of amino acids identified (AAI) on average. The antismash.secondarymetabolites.org/ website was used in conjunction with the antiSMASH v. 7.1 analysis tools to rapidly identify and analyse clusters of secondary metabolite biosynthesis genes in the strain genome (Blin et al. 2021). Phylogenetic tree of the genome was reconstructed using bacterial core genes (UBCG) (Na, S. I. et al. 2018), online KEGG metabolic pathway analysis of the strain GXQ1321 T was performed using the KEGG annotation website ( https://www.genome.jp/kegg/kaas/ ) and the egggnog-mapper website ( http://eggnog-mapper.embl.de ) was used to perform the COG categorisation analysis of the encoded proteins (Carlos et al. 2021). Ortho Venn3 software ( https://orthovenn3 . bioinfotoo lkits. net/ home) was used to generate ortho analysis Venn diagrams and pairwise ortho analysis heatmap between the three genomes (Sun et al. 2003). Physiological and biochemical characteristics Scanning electron microscopy and transmission electron microscopy (Talos F200C) were used to observe the morphology and flagellum of the strain. The test strains were stained using the Hopebiol (Qing Dao) Gram Stain Kit's operating methods. The strains were inoculated on actinomycetes culture medium and cultured at different temperatures (0, 4, 10, 20, 30, 37, 40, 50℃) for 4 weeks. The strain's growth was assessed using OD 600 , and pH tolerance (pH 4.0–10.0) was determined using actinomycetes media. Tolerance to 0–20% w/v strains was assessed liquid using actinomycetes culture medium devoid of NaCl. The strain's activity was assessed using the turbidity of a semi-solid medium containing 0.4% agar (Zhao et al. 2024). Catalase activity was measured using 3% (v/v) H 2 O 2 . Additional physiological tests, such as absorption, acid generation, and enzymology, were performed with API 20NE, API 50CH, API ZYM, and Biolog Gen III test strips (bioMerieux). The carbon utilisation of the strains was tested using Biolog's Gen III eco-plate. Follow the directions in the manual. The strains were tested for antibiotic susceptibility using multi-antibiotic susceptibility test tablets (Zhao et al. 2024). In order to determine the enzyme production of strain GXQ1321 T , eight enzyme production function plates (cellulase production, protease production, amylase production, urease production, inorganic phosphorus utilization, organophosphorus utilization, CAS, nitrogen fixation function) were selected for detection. Whether strain GXQ1321T has enzyme production function was preliminarily determined by the presence or absence of the transparent ring. Chemotaxonomic characteristics Bacteria were collected after 4 days of cultivation at 30℃ on actinomycetes culture medium and washed three times with 0.9% NaCl solution. The obtained sediments were freeze-dried by vacuum freeze dryer for 3 days for chemical experiments. The Sherlock Microbial Identification System (MIDI Inc., Newark, USA) operational parameters were followed for the extraction of fatty acid samples. Prepared and analysed according to Sherlock Microbial Identification System instructions (Whittaker 2011). The procedure for extraction of polar lipids was followed as described in detail by Song Y et al. (2000). Methylnaphthoquinone drugs were extracted utilizing the technique of Collins et al. (1979) and high performance liquid chromatography (HPLC) was used to identify them (Kroppenstedt and Reiner 1982). The instructions of Schleifer et al. (1972) for the preparation and analysis of cell wall peptidoglycan were followed. Hasegawa et al.'s (1983) methodology for analyzing whole cell sugars was used. Results and Discussion Phylogenetic characterization The complete 16S rRNA gene sequence of strain GXQ1321 T (GenBank/EMBL/DDBJ accession number OR661284) was found with a total length of 1459 bp. 16S rRNA sequencing showed that the strain belonged to Brevibacterium . Using the EZBiocloud database comparison, the 16S rRNA with the highest similarity to GXQ1321 T were B. samyangense SST-8 T (96.77%) and B. rongguiense 5221 T (96.32%). Strain GXQ1321 T developed an isolated subbranch structure within a broader clade encompassing B. samyangense SST-8 T (96.77%) and B. rongguiense 5221 T (96.32%), according to an adjacency tree analysis based on 16S rRNA gene sequences. Maximum parsimonious and maximum likelihood phylogenetic trees also corroborate this relationship. Maximum likelihood and maximum parsimonious phylogenetic trees also corroborate this relationship. Fig. S1 and Fig. S2 show the results of the maximum likelihood and maximum parsimony calculations. It unstable tree topologies indicate that GXQ1321 T does not belong to any of the Brevibacterium strains that have been successfully characterised. . Genomic analysis The strain GXQ1321 T (GenBank accession number JBDIUY000000000) has a genomic length of 3347008 bp and 69.6% G + C. The genome contains 26 overlapping groups with maximum and minimum contigs lengths of 315,757 and 1251bp, N50 values of 227,695 bp, N90 values of 94,674 bp, and L50 values of 6, respectively. With maximum and minimum contig lengths of 315,757 and 1251 bp, N50 values of 227,695 bp, N90 values of 94,674 bp, and L50 values of 6, the genome is composed of 26 overlapping groups. Genome-wide information is used to construct the phylogenetic tree. Strain GXQ1321 T and reference strain SST-8 T clustered together to form a branch, which proved that strain GXQ1321 T was the closest relative to reference strain B. samyangense SST-8 T (Fig. 2). Table 1 shows that the ANI values of B. rongguiense 5221 T , B. samyangense SST-8 T (96.77%), and GXQ1321 T were all below the species criterion (ANI 95%). dDDH calculation showed that the DNA affinity of GXQ1321 T and B. samyangense SST-8 T (96.77%) was 21.2%. DNA affinity with B. rongguiense 5221 T (96.32%) was 15.3%. The results were below the 70% threshold required to discriminate between species. The AAI values of GXQ1321 T , B. samyangense SST-8 T (96.77%) and B. rongguiense 5221 T were also below the species threshold (AAI 95%) (Fig. S3). The results showed that GXQ1321 T belongs tothe genus Brevibacterium . Table 1 ANI and dDDH values between strain GXQ1321 T and other closely related strains. Strain 1 Strain 2 ANI (%) dDDH (%) 16S rRNA gene identity (%) GXQ1321 T B. Rongguiense 5221 T 73.91 15.3 96.32 B.samyangense SST-8 T 77.14 21.1 96.77 Several features of the genome of strain GXQ1321 T distinguished it from other Brevibacterium strains by annotation and analysis of the genome (Table 2 and Table S1 ). The annotation results showed that GXQ1321 T , B. samyangense SST-8 T and B. rongguiense 5221 T all contained terpene and NAPPA genes. Annotation results from the antiSMASH 7.1 database showed that GXQ1321 T contained a terpene gene cluster with 33% similarity to carotenoids. The yellow-green colour of the GXQ1321 T colony was also consistent with the phenotypic characteristics. In addition, GXQ1321 T also contains 75% ectoine, whereas the B. samyangense SST-8 T and B. rongguiense 5221 T strains did not contain the ectoine gene (Table S2). Ectoine helps organisms withstand high osmotic pressure environments. Ectoine protects enzymes, membranes and whole cells from the stresses of salt, heating, freezing and drying. The ability of strain GXQ1321 T to endure in marine sediments was also verified by this outcome. This gene cluster is present in the genus Brevibacterium , which was isolated from a harsh environment (Pei S et al. 2020). This result also proved that strain GXQ1321 T belonged to the genus Brevibacterium . Table 2 Comparison of the genomic characteristics of GXQ1321 T and related members of the genus Brevibacterium . +, positive; –, negative. Genes putatively encoding GXQ1321 T B. samyangense SST-8 T B. Rongguiense 5221 T GenBank accession number JAOEFD000000000 BAAANO000000000 JACCFQ010000000 Genome size (bp) 3347008 3394428 3117581 DNA G + C content (mol%) 69.6 68.95 72.4 N50 value (bp) 139003 41933 42508 L50 value (bp) 9 8 23 Number of coding sequences 3088 3197 2904 features in the draft genome 3147 3252 2952 Number of rRNAs 59 55 48 Number of Subsystems 235 238 236 The results of the pan-genomic comparison between strain GXQ1321 T and its reference strains were OrthoVenn3 (Fig. S4). The strain GXQ1321 T had a total of 2243 gene clusters, while the other two reference strains had 2298 and 1790 gene clusters, respectively. The results showed that strains GXQ1321 T and B. samyangense SST-8 T had the highest similarity, with a total of 594 gene clusters, which exceeded the 86 gene clusters of strains GXQ1321 T and B. rongguiense 5221 T . Analysis of the bacteria revealed that the three strains share a "core" genome consisting of 1,548 homologous gene clusters, most of which code for proteins with functions related to cellular metabolism and specialised membrane exchange systems. Strain GXQ1321 T and strain B. samyangense SST-8 T showed the deepest colour representation in the pin-to-pair orthogonal heat maps of the three genomes, indicating a close relationship between them (Fig. S5). Based on the above analysis, strain GXQ1321 T and reference strain SST-8 T were more closely related, which was also confirmed by the genome-wide evolutionary tree. A total of 2,061 genes involved in six pathways were detected in the KEGG database. Of these, metabolism was the most common, including amino acid metabolism, carbohydrate metabolism, cofactor and vitamin metabolism, and energy metabolism (Fig. S6). We discovered that "amino acid transport and metabolism" was the largest of the known coding proteins of strain GXQ1321 T based on a COG classification analysis (Table S3). Since strain GXQ1321 T was isolated from marine surface sediments, we hypothesised that ectoine secreted by GXQ1321 T may affect the growth and development and stress resistance of marine sediment plants. The characteristics of nitrogen and sulphur metabolism in subsystems were compared and analysed. Strains B. samyangense SST-8 T and B. rongguiense 5221 T lacked genes related to cyanate hydrolysis, nitrate and nitrite antimonization, and only ammonia assimilation genes. On the contrary, strain GXQ1321 T contained genes related to cyanate hydrolysis, nitrate and nitrite antimonization and ammonia assimilation. In the characteristics of sulfur metabolism, both strain GXQ1321 T and the reference strains B. samyangense SST-8 T and B. rongguiense 5221 T contained thioredoxin-disulfide reductase, but the reference strain B. samyangense SST-8 T contained genes related to organic sulfur assimilation. Therefore, B. samyangense SST-8 T had the highest abundance, with a total of 11 genes related to sulfur metabolism (Table S4). According to the analysis of the NCBI database, we found that strain GXQ1321 T contained the transporters ABC(WP_349829439.1), BCCT(WP_349828541.1), MFS(WP_349829450.1), NCS2(WP_349828315.1), ACR3(WP_349826989.1), etc., which are basically the same as the transporters contained in Brevibacterium isolated from the sea, which further confirmed that strain GXQ1321 T belongs to a genus of Brevibacterium . Among them, BCCT contributes to the accumulation of compatible solutes betaine, carnitine and choline. This is not only beneficial for maintaining the osmotic balance of the cell, but also acts as a stabiliser for proteins and cellular components, preventing denaturation and resisting the effects of high ionic strength. ABC transporters may be related to their ability to transport and metabolise various toxic substances and chitin, amylopectin, cellulose and starch. The Rast analysis showed that 235 subsystems existed in strain GXQ1321 T , and 15 glycoside hydrolases (GH), 26 glycosyltransferases (GTs), 1 carbohydrate binding module (CBM), 14 carbohydrate esterases (CEs) and 8 auxiliary activities (AAs) were identified. Since lignocellulose is abundant in plants growing in Marine surface sediments, the coexistence of these genes suggests that they play an important role in the breakdown and modification of carbohydrates in Marine surface sediments. This was consistent with our subsequent functional assay finding that strain GXQ1321 T had the ability to produce amylase and cellulase. MFS promotes the transmembrane transport of solutes such as sugars, drug molecules, peptides, tricarboxylic acid cycle metabolites, organic anions and inorganic anions under electrochemical gradients. This corresponds to the maximum "amino acid transport and metabolism" shown in COG. At the same time, KEGG metabolic pathway analysis revealed that strain GXQ1321 T contained the SecE gene. The Sec pathway is the first secretory pathway discovered in bacteria to export proteins across the plasma membrane to the periplasm and outer membrane of bacteria. The SecE gene is a gene associated with extracellular transport of cells, possibly because most marine Brevibacterium release a class of macromolecular protein heteropolysaccharide extracellular polymers into the environment to prevent ice crystal formation and adapt to low temperatures. We also identified genes related to salt tolerance and alkaline resistance in the gene annotation results of strain GXQ1321T, including the Na+/H + reverse transporters NhaC and NhaD, and the Na+-driven multidrug efflux pump, the DinF/NorM/MATE family. The results are also consistent with previous physiological experiments testing the bacteria's tolerance to saline alkaline conditions. Morphological, physiological, and biochemical characteristics The GXQ1321 T strain's colony was circular, convex, golden, and free of diffuse pigment. Its margins were unbroken. Following 48–72 hours of cultivation at 30 ℃, the strain's cells had a short rod-like morphology (0.51–0.59 × 1.1–1.6 µm) (Fig. S7), with a colony diameter of 1.2–2.3 mm. Strain GXQ1321 T was positive for Gram staining and aerobic. Transmission electron microscopy revealed that strain GXQ1321 T was flagellated and nonmotile (Fig. S8). This is similar to the reference strains, the reference strains B. samyangense SST-8 T and B. rongguiense 5221 T have no flagellum and are non-motile, while most strains of the genus Brevibacterium have no flagellum. The strain was grown on actinomycetes culture media for three days at 30°C. The colonies were 1.1–1.8 mm in diameter and yellow in colour. After one month of observation, strain GXQ1321 T grew at 4–50℃ (optimal 30 ℃). The concentration of NaCl tolerance ranged from 0 to 20% (4% was optimal). Catalase detection results of strain GXQ1321 T were negative. The pH range in which the strains can grow is 4.0–10 (7.0 is optimal). Table 3 illustrates the observed variations in physiological characteristics between GXQ1321 T and the reference strains B. samyangense SST-8 T and B. rongguiense 5221 T . Strain GXQ1321 T and the reference strains B. samyangense SST-8 T , B. rongguiense 5221 T were sensitive to amoxicillin, streptomycin, erythromycin, rifampicin, neomycin and chloramphenicol. Compared to the reference strain B. rongguiense 5221 T , strain GXQ1321 T and reference strain B. samyangense SST-8 T were not resistant to neomycin and gentamicin. Table 3 displays the differences in API 20NE, API 50CH, API ZYM, and Biolog GEN III as well as the physiological and biochemical features of strain GXQ1321 T compared to reference strains B. samyangense SST-8 T and B. rongguiense 5221 T . According to the appearance of the transparent circle, we found that strain GXQ1321 T had the function of producing amylase, amylase and ferriferite. Table 3 Differential physiological characteristics of GXQ1321 T from the closely related species B.samyangense SST-8 T and B. Rongguiense 5221 T . All data presented are from this study. +, positive; –, negative;W, weak reaction. Characteristic GXQ1321 T B.samyangense SST-8 T B. Rongguiense 5221 T Colony color Light-yellow Bright-yellow Milky - white Motility - + - pH(optimum) 4.0–10.0(7.0) 6.0-11.5(10.0) 5.0-9.5(7.5) Temperature (optimum) (°C) 4–50( 30 ) 10–45( 30 ) 10–40( 35 ) NaCl (optimum) (%, w/v) (0–20)4.0% 0–15(7.5) ( 1 – 12 )4% Assimilation of (API 20NE): Urea w - + Maltose - w + Gelatin + - - Esculin + - - Acid production from (API 50CH): D-Mannose - + + D-GALactose + - - D-MELibiose + - - L-RHamnose - - + D-Fruetose - - + Arbutin + w - Carbon source utilization (Biolog GEN III): α-D-Glucose + - - D-Salicin - - w Dextrin + + - Gentiobiose + - - D-Turanose + + - α-D-lactose + + - L-Fucose + + - Inosine + + - D-Mannitol + + - D-arabitol + + - L-Aspartic acid + - - Gelatin + - - Pectin + - - L-Alanine - + + Sodium citrate W - + Sodium formate W - + Disodium-D,L-malate - - + Inulin - + - Enzymology (API ZYM): + Acid phosphatase - + Valine arylamidase + - - Cysteine arylamidase + W - Trypsin - W + β-glucosidase - - + Antibiotic tolerance: - Gentamicin - + Neomycin - - + The peptidoglycan of strain GXQ1321 T is consistent with the peptidoglycan of the reference strains B. samyangense SST-8 T and B. rongguiense 5221 T , whole cell hydrolysates containing meso-diaminopimelic acid (Fig. S9). Anteiso -C 19:0 (27.28%), anteiso -C 15:0 (18.97%), anteiso -C 17:0 (15.95%), and iso -C 16:0 (12.21%) were the major fatty acids (> 10%) of strain GXQ1321 T . The major fatty acids (> 10%) of B. samyangense SST-8 T were anteiso -C 17:0 (35.3%), anteiso -C 15:0 (29.9%) and iso -C 15:0 (15.5%). The major fatty acids (> 10%) of B. rongguiense 5221 T were anteiso -C 15:0 (46.2%), anteiso -C 17:0 (39.3%) and C 16:0 (11.9%) (Table S5). The main fatty acids of strain GXQ1321 T were different from those of B. samyangense SST-8 T and B. rongguiense 5221 T . Phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), three phosphoglycolipids (PGL) and two unknown glycolipid (UG) were the major polar lipids of strain GXQ1321 T (Fig. S10). Compared to GXQ1321 T , the major polar lipid of B. samyangense SST-8 T was phosphatidylglycerol (PG), and the major polar lipids of B. rongguiense 5221 T were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), and three types of phosphoglycolipids (PGL). The respiratory quinones of strain GXQ1321 T were mainly MK-8 (90%). The reference strains B. samyangense SST-8 T and B. rongguiense 5221 T breathed predominantly quinones as MK-8, consistent with most of the genus Brevibacterium . The results of physicochemical experiments demonstrated that strain GXQ1321 T could be distinguished from the reference strains B. samyangense SST-8 T and B. rongguiense 5221 T . The growth rate of strain GXQ1321 T on actinomycetes culture medium was higher than the reference strains B. samyangense SST-8 T and B. rongguiense 5221 T . The colony colour and moisture of strain GXQ1321 T were also different from the reference strains B. samyangense SST-8 T and B. rongguiense 5221 T . Strain GXQ1321 T and the reference strains B. samyangense SST-8 T , B. rongguiense 5221 T were sensitive to amoxicillin (10 µg/tablet), streptomycin (10 µg/tablet), erythromycin (15 µg/tablet), rifampicin (5 µg/tablet), neomycin (30 µg/tablet) and chloramphenicol (30 µg/tablet). Strain GXQ1321 T and the reference strains B. samyangense SST-8 T were resistant to neomycin (30 µg/tablet) and gentamicin (10 µg/tablet), while B. rongguiense 5221 T was resistant to neomycin and gentamicin. Following an analysis of the strain's physiology, chemistry, and phylogeny, it was concluded that strain GXQ1321 T represents a novel species within the Brevibacterium genus, designated as Brevibacterium litoralis sp. nov. Description of Brevibacterium litoralis sp. nov Brevibacterium litoralis sp. nov (li.to.ra′lis. L. masc. adj. litoralis of or belonging to the sea shore). The strain is an aerobic, Gram-positive actinomycetes that is non-motile and does not generate spores. The cells measured 0.51–0.59 × 1.1–1.6 µm and had a short rod shape. The colony is yellow, round, convex, with intact margins and does not produce diffuse pigments. Negative for catalase. The optimal growth conditions for the organism in question are 4–50 ℃, pH 4.0–10.0, and NaCl concentration ranging between 0–20% w/v (optimum 4.0%). Maltose could not be hydrolyzed by the strain, while gelatin and aesculin could. Arbutin, D-galactose, and D-meliose were converted to acid. Dextrin, L-fructose, L-rhamnose, D-maltose, D-trehalose, myo-inositol, D-cellulobiose, sucrose, D-terabiose, stachyose, α-D-lactose,glycerol, D-glucose-6-phosphate, D-fructose-6-phosphate, β-methyl-D-glucoside, D-salicin, N-acetyl-D-glucosamine, alpha-D-glucose, N-acetyl-β-D-mannosamine, D-mannose, gelatine, D-fructose, sodium lactate, D-sorbitol, inosine, D-mannitol, D-arabitol, D-aspartate, L-alanine, gentiobiose, L-aspartic acid, L-glutamic acid, L-histidine, L-pyroglutamic acid, L-serine, pectin, D-galacturonic acid, L-galactonic acid lactone, D-glucuronic acid, D-saccharic acid, L-lactic acid, α-ketoglutaric acid, D-malic acid, D-lactic acid methyl ester, L-malic acid, Tween 40, propionic acid, acetic acid, formic acid can be used as a sources of carbon. In accordance with the API ZYM test paper, alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, valine arylamidase, cystine arylamidase, trypsin, acid phosphatase, naphtho-AS-BI-phosphohydrolase were all positive. Lipase (C14), chymotrypsin, β-fucosidase, alpha-mannosidase, α-galactosidase, β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-glucosaminase were negative. The enzymes produced by this strain were esterase, amylase and cellulase. The strain was resistant to neomycin, amoxicillin, streptomycin, chloramphenicol, rifampicin, novobiocin, gentamicin and erythromycin. The main respiratory quinone was MK-8 (90%). The main polar lipids of strain GXQ1321 T were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), three phosphoglycolipids (PGL) and two unknown glycolipid (UG). The whole cell hydrolysates containing meso-diaminopimelic acid. Anteiso -C 19:0 (27.28%), anteiso -C 15:0 (18.97%), anteiso -C 17:0 (15.95%), and iso -C 16:0 (12.21%) were the major fatty acids (> 10%) of strain GXQ1321 T . Strain GXQ1321 T (= MCCC 1K08964 T = KCTC 59167 T ) was the first Brevibacterium isolated from surface sediments in Beihai, Guangxi. The 16S rRNA gene for the strain GXQ1321 T has been assigned the GenBank accession number OR661284. The Whole Genome Shotgun project has been deposited at DDBJ/ENA/GenBank under the accession JBDIUY000000000. Declarations Author contributions Mingguo Jiang, conceptualisation and project administration. Contributed to the conception of the study, formulation or evolution of overarching research goals and aims. Yi Jiang, resources. Provision of some study materials. Quan Yang, investigation and writing-original draft preparation, performing all of the experiments. Aolin Zhao, review and editing, performing part of the experiments. Haifei Liu, performed research; Jiawei Li, performed research; Shujing Wu, performed research; Ying Huang, analyzed data. Jie Weng, analyzed data. Funding information This study was funded by the Science and Technology Major Project of Guangxi (AA18242026), recipient: Mingguo Jiang. Compliance with Ethical Standards Conflict of interest All authors confirm that no competing interests, both financial and personal, are with this manuscript. Data The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene and the draft genome sequences of strain GXQ1321 T are OR661284 and JBDIUY000000000, respectively. References Breed, R. (1953) The Brevibacteriaceae fam. nov. of order Eubacteriales. Rias Commun VI Congr Int Microbiol Roma 1, 13–14. https://doi.org/10.1007/978-3-642-30138-4_169 Tang, S. K., Wang, Y., Schumann, P., Stackebrandt, E., Lou, K., Jiang, C. L., Xu, L. H., & Li, W. J. (2008) Brevibacterium album sp. nov., a novel actinobacterium isolated from a saline soil in China. International journal of systematic and evolutionary microbiology, 58(Pt 3), 574–577. https://doi.org/10.1099/ijs.0.65183-0 Jung MS, Quan XT, Siddiqi MZ, Liu Q, Kim SY, Wee JH, Im WT. (2018) Brevibacterium anseongense sp. nov., isolated from soil of ginseng field. J Microbiol 56:706–712. https://doi.org/10.1007/s12275-018-8181-5 Pei S, Niu S, Xie F, Wang W, Zhang S, Zhang G. (2021) Brevibacterium limosum sp. nov., Brevibacterium pigmenatum sp. nov., and Brevibacterium atlanticum sp. nov., three novel dye decolorizing actinobacteria isolated from ocean sediments. J Microbiol 59:898–910. https://doi.org/10.1007/s12275-021-1235-0 Bhadra B, Raghukumar C, Pindi PK, Shivaji S. (2008) Brevibacterium oceani sp. nov., isolated from deep-sea sediment of the Chagos Trench, Indian Ocean. Int J Syst Evol Microbiol 58:57–60. https://doi.org/10.1099/ijs.0.64869-0 Deng T, Lu H, Qian Y, Chen X, Yang X, Guo J, Sun G, Xu M. (2020) Brevibacterium rongguiense sp. nov., isolated from freshwater sediment. Int J Syst Evol Microbiol 70:5205–5210. https://doi.org/10.1099/ijsem.0.004379 Pei S, Xie F, Niu S, Ma L, Zhang R, Zhang G. (2020) Brevibacterium profundi sp. nov., isolated from deep-sea sediment of the Western Pacific Ocean. Int J Syst Evol Microbiol 70:5818–5823. https://doi.org/10.1099/ijsem.0.004483 Kati H, Ince IA, Demir I, Demirbag Z. (2010) Brevibacterium pityocampae sp. nov., isolated from caterpillars of Thaumetopoea pityocampa (Lepidoptera, Thaumetopoeidae). Int J Syst Evol Microbiol 60:312–316. https://doi.org/10.1099/ijs.0.006692-0 Zhang M, Song Q, Sang J, Li Z. (2023) Brevibacterium spongiae sp. nov., isolated from marine sponge Hymeniacidon sp. Int J Syst Evol Microbiol 2023; 73:5869. https://doi.org/10.1099/ijsem.0.005869 Kampfer P, Schafer J, Lodders N, Busse HJ. (2010) Brevibacterium sandarakinum sp. nov., isolated from a wall of an indoor environment. Int J Syst Evol Microbiol 60:909–913. https://doi.org/10.1099/ijs.0.014100-0 Wauters G, Charlier J, Janssens M, Delmee M. (2001) Brevibacterium paucivorans sp. nov., from human clinical specimens. Int J Syst Evol Microbiol 51:1703–1707. https://doi.org/10.1099/00207713-51-5-1703 Mages IS, Frodl R, Bernard KA, Funke G. (2008) Identities of Arthrobacter spp. and Arthrobacter-like bacteria encountered in human clinical specimens. J Clin Microbiol 46:2980–2986. https://doi.org/10.1128/jcm.00658-08 McBride ME, Ellner KM, Black HS, Clarridge JE, Wolf JE. (1993) A new Brevibacterium sp. isolated from infected genital hair of patients with white piedra. J Med Microbiol 39:255–261. https://doi.org/10.1099/00222615-39-4-255 Pascual C, Collins MD, Funke G, Pitcher DG. (1996) Phenotypic and genotypic characterization of two Brevibacterium strains from the human ear: description of Brevibacterium otitidis sp. nov. Med. Microbiol. Lett. 5:113–123. http://dx.doi.org/10.1099/jmm.0.043109-0 Bernard, K. A., Wiebe, D., Burdz, T., Reimer, A., Ng, B., Singh, C., Schindle, S., & Pacheco, A. L. (2010) Assignment of Brevibacterium stationis (ZoBell and Upham 1944) Breed 1953 to the genus Corynebacterium, as Corynebacterium stationis comb. nov., and emended description of the genus Corynebacterium to include isolates that can alkalinize citrate. International journal of systematic and evolutionary microbiology, 60(Pt 4), 874–879. https://doi.org/10.1099/ijs.0.012641-0 Pascual C, Collins MD. (1999) Brevibacterium avium sp. nov., isolated from poultry. Int J Syst Bacteriol 9:1527–1530. https://doi.org/10.1099/00207713-49-4-1527 Komagata, K., & Iizuka, H. (1964) New species of Brevibacterium Isolated from Rice. Bulletin of the Agricultural Chemical Society of Japan, 38, 496–502. https://doi.org/10.1271/nogeikagaku1924.38.496 Choi EJ, Lee SH, Jung JY, Jeon CO. (2013) Brevibacterium jeotgali sp. nov., isolated from jeotgal, a traditional Korean fermented seafood. Int J Syst Evol Microbiol 63:3430–3436. https://doi.org/10.1099/ijs.0.049197-0 Ozturkoglu-Budak S, Wiebenga A, Bron PA, de Vries RP. (2016) Protease and lipase activities of fungal and bacterial strains derived from an artisanal raw ewe's milk cheese, International Journal of Food Microbiology, 237: 17–27. https://doi.org/10.1016/j.ijfoodmicro.2016.08.007 Amarita F, Nardi M, Chambellon E, Delettre J, Bonnarme P. (2004). Identification and Functional Analysis of the Gene Encoding Methionine-γ-Lyase in Brevibacterium linens. Appl Environ Microbiol 70(12):7348–7354. https://doi.org/10.1128/AEM.70.12.7348-7354.2004 Simoes, F., Vale, P., Stephenson, T., & Soares, A. (2018). Understanding the growth of the bio-struvite production Brevibacterium antiquum in sludge liquors. Environmental technology, 39(17), 2278–2287. https://doi.org/10.1080/09593330.2017.1411399 Lamballerie de, Zandotti C, Vignoli C, Bollet C, de Micco P. A onestep microbial DNA extraction method using “Chelex 100” suitable for gene amplification. Res Microbiol 1992; 143:785–790. https://doi.org/10.1016/0923-2508(92)90107-y Chun J, Goodfellow M. (1995) A phylogenetic analysis of the genus Nocardia with 16S rRNA gene sequences. Int J Syst Bacteriol 45:240–245. https://doi.org/10.1099/00207713-45-2-240 Edwards U, Rogall T, Blöcker H, Emde M, Böttger EC. (1989) Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 17:7843–7853. https://doi.org/10.1093/nar/17.19.7843 Lane DJ. (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, Chichester, pp 115–175. Yoon SH, SM H, Kwon S, Lim J, Kim Y. (2017) Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol; 67:1613–1617. https://doi.org/10.1099/ijsem.0.001755 Koichiro Tamura, Glen Stecher, Sudhir Kumar. MEGA11: Molecular Evolutionary Genetics Analysis version 11. Molecular Biology and Evolution, Volume 38, Issue 7, July 2021, Pages 3022–3027. https://doi.org/10.1093/molbev/msab120 Felsenstein J. (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 17:368–376. https://doi.org/10.1007/BF01734359 Fitch WM. (1971) Toward defining the course of evolution: minimum change for a specific tree topology. Syst Zool 20:406–416. https://doi.org/10.1093/sysbio/20.4.406 Saitou N, Nei M. (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454 Kimura M. (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol; 16:111–120. https://doi.org/10.1007/BF01731581 Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 2013; 30:2725–2729. https://doi.org/10.1093/molbev/mst197 Li R, Li Y, Kristiansen K, Wang J. (2008) SOAP: short oligonucleotide alignment program. Bioinformatics 24:713–714. https://doi.org/10.1093/bioinformatics/btn025 Aziz RK, Bartels D, Best AA, DeJongh M, Disz T, Edwards RA, Formsma K, Gerdes S, Glass EM, Kubal M, Meyer F, Olsen GJ, Olson R, Osterman AL, Overbeek RA, McNeil LK, Paarmann D, Paczian T, Parrello B, Pusch GD, Reich C, Stevens R, Vassieva O, Vonstein V, Wilke A, Zagnitko O. (2008) The RAST server: rapid annotations using subsystems technology. BMC Genomics 9:75. https://doi.org/10.1186/1471-2164-9-75 Meier-Kolthoff JP, Auch AF, Klenk HP, Göker M. (2013) Genome sequencebased species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 14:60. https://doi.org/10.1186/1471-2105-14-60 Blin K, Shaw S, Kloosterman AM et al. (2021) antiSMASH 6.0: improving cluster detection and comparison capabilities. Nucl Acids Res 49(W1): W29-W35. https://doi.org/10.1093/nar/gkab335 Na, S. I., Kim, Y. O., Yoon, S. H., Ha, S. M., Baek, I., & Chun, J. (2018). UBCG: Up-to-date bacterial core gene set and pipeline for phylogenomic tree reconstruction. Journal of microbiology (Seoul, Korea), 56(4), 280–285. https://doi.org/10.1007/s12275-018-8014-6 Wattam AR, Davis JJ, Assaf R, Boisvert S, Brettin T, et al. (2017) Improvements to PATRIC, the all-bacterial bioinformatics database and analysis resource center. Nucleic Acids Res 45: D535–D542. https://doi.org/10.1093/nar/gkw1017 Carlos P. Cantalapiedra, Ana Hernandez-Plaza, Ivica Letunic, Peer Bork, and Jaime Huerta -Cepas. eggNOG-mapper v2: functional annotation, orthology assignments, and domain prediction at the metagenomic scale. Mol. Biol. Evol. 38(12):5825–5829. https://doi.org/10.1093/molbev/msab293 Sun, J., Lu, F., Luo, Y., Bie, L., Xu, L., & Wang, Y. (2023). OrthoVenn3: an integrated platform for exploring and visualizing orthologous data across genomes. Nucleic acids research, 51(W1), W397–W403. https://doi.org/10.1093/nar/gkad313 Zhao, A., Cai, H., Huang, Y. et al. (2024) Nesterenkonia marinintestina sp. nov., isolated from the fish intestine. Arch Microbiol 206, 110. https://doi.org/10.1007/s00203-023-03825-0 Whittaker P. (2011) Identification of six species in the new genus Cronobacter (Enterobacter sakazakii) from culture using gas chromatographic analysis of fatty acid methyl esters. Journal of AOAC International. 94(5):1581–1584. https://doi.org/10.5740/jaoacint.10-451 Song Y, Yang R, Guo Z, Zhang M, Wang X et al. (2000) Distinctness of spore and vegetative cellular fatty acid profiles of some aerobic endospore-forming bacilli. J Microbiol Methods 39:225–241. https://doi.org/10.5740/jaoacint.10-451 Collins, M. D., Goodfellow, M. & Minnikin, D. E. (1979) Isoprenoid quinones in the classification of coryneform and related bacteria. J Gen Microbiol 110, 127–136. https://doi.org/10.1099/00221287-110-1-127 Kroppenstedt, Reiner M. (1982) Separation of bacterial menaquinones by HPLC using reverse phase (RP18) and a silver loaded ion exchanger as stationary phases. J Liq Chromatogr 5:2359–2367. https://doi.org/10.1080/01483918208067640 Schleifer, K. H. & Kandler, O. (1972) Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36, 407–477. https://doi.org/10.1128/br.36.4.407-477.1972 Hasegawa, T., Takizawa, M., & Tanida, S. (1983) A rapid analysis for chemical grouping of aerobic actinomycetes. Journal of General and Applied Microbiology, 29, 319–322. https://doi.org/10.2323/jgam.29.319 Additional Declarations No competing interests reported. Supplementary Files Supplementarymaterial.docx Cite Share Download PDF Status: Published Journal Publication published 14 Oct, 2024 Read the published version in Antonie van Leeuwenhoek → Version 1 posted Editorial decision: Revision requested 15 Jul, 2024 Editor assigned by journal 12 Jul, 2024 Submission checks completed at journal 12 Jul, 2024 First submitted to journal 11 Jul, 2024 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-4724416","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":327098373,"identity":"bae2018f-9335-4947-8619-7aed099e90d3","order_by":0,"name":"Quan Yang","email":"","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":false,"prefix":"","firstName":"Quan","middleName":"","lastName":"Yang","suffix":""},{"id":327098374,"identity":"2a5cbbc7-1e2e-412c-8a67-4a885e4af37b","order_by":1,"name":"Aolin Zhao","email":"","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":false,"prefix":"","firstName":"Aolin","middleName":"","lastName":"Zhao","suffix":""},{"id":327098375,"identity":"90fedf1f-5535-4ef6-9976-b3f7b83f2a29","order_by":2,"name":"Haifei Liu","email":"","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":false,"prefix":"","firstName":"Haifei","middleName":"","lastName":"Liu","suffix":""},{"id":327098376,"identity":"ad801c13-a5b6-4738-b536-ed550a7fcde3","order_by":3,"name":"Jiawei Li","email":"","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":false,"prefix":"","firstName":"Jiawei","middleName":"","lastName":"Li","suffix":""},{"id":327098378,"identity":"d7dadf4b-1a27-4176-aa89-869bb8612cbf","order_by":4,"name":"Shujing Wu","email":"","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":false,"prefix":"","firstName":"Shujing","middleName":"","lastName":"Wu","suffix":""},{"id":327098380,"identity":"2d6d4784-09a5-4c6b-83a5-76f6330aa43c","order_by":5,"name":"Ying Huang","email":"","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":false,"prefix":"","firstName":"Ying","middleName":"","lastName":"Huang","suffix":""},{"id":327098383,"identity":"f9d0bb26-7f89-4c09-9fe6-0dfc2379e755","order_by":6,"name":"Jie Weng","email":"","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":false,"prefix":"","firstName":"Jie","middleName":"","lastName":"Weng","suffix":""},{"id":327098385,"identity":"4d974f9f-625d-4c8d-b3e2-3734f20a1031","order_by":7,"name":"Mingguo Jiang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAy0lEQVRIiWNgGAWjYLACCQMgwd7Y+PADaVp4DjcbS5BoVXqbAA8xCnX7z5hJWBTY5clHPmxjkGCwk9NtIKDF7MCxNAkJg+Riw9uJbQ8KGJKNzQ4Q0nKw+RhQC3PixtmJ7QYSDAcStxHUcpixDailPnHjzINtEjxEaTnGDLLlcOJ8CUZitZxhS7aQMDieuIEnERjIBsT45fwZw9sSf6oT57cff/jwQ4WdHEEtQMAiDYpBA7BKA8LKQYD5IyidyDcQp3oUjIJRMApGIAAASO9BqW8u554AAAAASUVORK5CYII=","orcid":"","institution":"Guangxi Minzu Unversity","correspondingAuthor":true,"prefix":"","firstName":"Mingguo","middleName":"","lastName":"Jiang","suffix":""},{"id":327098386,"identity":"f546e281-a694-46b9-bf0d-6cd617a3f0b7","order_by":8,"name":"Yi Jiang","email":"","orcid":"","institution":"Yunnan University","correspondingAuthor":false,"prefix":"","firstName":"Yi","middleName":"","lastName":"Jiang","suffix":""}],"badges":[],"createdAt":"2024-07-11 13:21:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4724416/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4724416/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10482-024-02031-2","type":"published","date":"2024-10-14T15:56:54+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":61868092,"identity":"8bbfb05e-1026-4377-b1e0-72be7aaac8d7","added_by":"auto","created_at":"2024-08-06 12:34:21","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":436473,"visible":true,"origin":"","legend":"\u003cp\u003eThe neighbor-joining tree based on the 16S rRNA gene sequences showing the phylogenetic relationship of GXQ1321\u003csup\u003eT\u003c/sup\u003e with related taxa. The evolutionary distances were computed using the Kimura 2-parameter method. The sequence of \u003cem\u003eConexibacter arvalis\u003c/em\u003e KV-962\u003csup\u003eT\u003c/sup\u003e(AB597950) was used as an outgroup. Bootstrap support values were calculated from 1000 replicates.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4724416/v1/5d4032ccfd0ecfcda8bdf7a8.jpeg"},{"id":61868093,"identity":"158a91ff-a02b-4a41-a344-33a1ee081250","added_by":"auto","created_at":"2024-08-06 12:34:21","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":594950,"visible":true,"origin":"","legend":"\u003cp\u003eLegend not included with this version\u003c/p\u003e","description":"","filename":"Fig.2.png","url":"https://assets-eu.researchsquare.com/files/rs-4724416/v1/0c1e7833c59ba00537cc6821.png"},{"id":67148989,"identity":"ec58c83b-9053-4e86-85e8-5e6eef1917fa","added_by":"auto","created_at":"2024-10-21 16:10:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1827015,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4724416/v1/ce62927f-1c89-480b-9cb9-7f1aca102de1.pdf"},{"id":61868095,"identity":"e7ac1b7c-18fa-4b31-b05b-b7b7ccb748ce","added_by":"auto","created_at":"2024-08-06 12:34:22","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":3150396,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-4724416/v1/fd049969ed8e6366ed30b2dd.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Brevibacterium litoralis sp. nov., a cellulose-degrading strain isolated from marine surface sediment","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBreed established the genus \u003cem\u003eBrevibacterium\u003c/em\u003e in 1953, and its type species is \u003cem\u003eBrevibacterium\u003c/em\u003e linens (Breed 1953). There are currently 57 recognised species in the genus \u003cem\u003eBrevibacterium\u003c/em\u003e. The genus is mainly isolated from soil (Tang et al. 2008; Jung et al. 2018), sediment (Pei et al. 2021; Bhadra et al. 2008; Deng et al. 2020; Pei et al. 2020), insects (Kati et al. 2010), marine animals (Zhang et al. 2023), wall (Kampfer et al. 2010), clinical specimens (Wauters et al. 2001; Mages et al. 2008), and human samples (McBride et al. 1993; Pascual et al. 1996). \u003cem\u003eBrevibacterium\u003c/em\u003e species are rod-shaped, nonspore-forming, Gram-positive bacteria (Bernard et al. 2010). \u003cem\u003eBrevibacterium\u003c/em\u003e has \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e and \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e as its major cellular fatty acids, while their primary methylnaphthoquinone is MK-8 (H2) (Bernard et al. 2010). In addition, phosphatidylglycerol is often detected as a major lipid (Pascual et al. 1999; Komagata et al. 1964; Choi et al. 2013).\u003c/p\u003e \u003cp\u003e \u003cem\u003eBrevibacterium\u003c/em\u003e strains have many industrial applications. Studies have shown that \u003cem\u003eBrevibacterium\u003c/em\u003e can use extracellular proteases and lipases to degrade lipids and proteins (such as casein) (Ozturkoglu-Budak S et al. 2013). At the same time, many \u003cem\u003eBrevibacterium\u003c/em\u003e strains also have the ability to modify sulphur-containing amino acids to produce volatile sulphides, which is useful in the production of volatile flavours (Amarita F et al. 2004). In addition, certain strains of the genus \u003cem\u003eBrevibacterium\u003c/em\u003e are used to recover phosphorus-rich products by growing in mixed culture wastewater streams (Simoes, F et al. 2018). Due to the remarkable achievements in the application of \u003cem\u003eBrevibacterium\u003c/em\u003e in industrial cheese production, biodegradation and wastewater treatment, and other biotechnologies, more and more \u003cem\u003eBrevibacterium\u003c/em\u003e strains have been unearthed, laying the foundation for future industrial development. After being separated from the seawater's surface sediment, the strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was discovered to be a novel species of \u003cem\u003eBrevibacterium\u003c/em\u003e. This study was undertaken to define the strain's classification status and to provide a foundation for further exploration and development.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStrain Isolation and Culture Conditions\u003c/h2\u003e \u003cp\u003eThe strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was isolated from marine surface sediments in Beihai (11\u0026deg;46\u0026prime;21.11\u0026Prime;N, 109\u0026deg;62\u0026prime;56.25\u0026Prime;E), Guangxi, Guangxi, China. A modified actinomycetes culture medium (0.15 g K\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e, 0.15 g NaCl, 0.30 g KNO\u003csub\u003e3\u003c/sub\u003e, 0.15 g MgSO\u003csub\u003e4\u003c/sub\u003e, 0.003 g FeSO\u003csub\u003e4\u003c/sub\u003e, 6.0 g soluble starch, 4.5 g agar, 300 mL sterile seawater, pH\u0026thinsp;=\u0026thinsp;7.5) was used as the culture media for the separation. The surface sediment is naturally air dried, then ground to a powder with a sterile grinder and filtered. 30 mL of sterile seawater were mixed with 3 g of the sediment powder samples, and the combination was agitated for 30 minutes at 30\u0026deg;C. It was then diluted in gradient with sterile seawater to 10\u003csup\u003e\u0026minus;\u0026thinsp;4\u003c/sup\u003e, 10\u003csup\u003e\u0026minus;\u0026thinsp;5\u003c/sup\u003e, 10\u003csup\u003e\u0026minus;\u0026thinsp;6\u003c/sup\u003e. Diluents of different concentrations were coated with actinomycetes culture medium and cultured at 30 ℃ for 5 days. To obtain pure culture, strains isolated from the plate were cultivated under the same conditions and purified in actinomycetes culture medium. Store the pure culture solution in glycerol (40%, v/v) at -80\u0026deg;C.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e16S rRNA Gene Sequencing and Phylogenetic Analysis\u003c/h2\u003e \u003cp\u003eDNA extraction was with TIANamp Bacteria DNA Kit (Lamballerie et al. 1992). The 16S rRNA gene was amplified using PCR in accordance with Chun et al. (1995) and Edwards et al. (1989). The particular primers 27F and 1492R were used to get the sequencing of the 16S rRNA gene (Lane et al. 1991), determined by AuGCT Biotech (Wu Han, China) and analysed in EzBioCloud (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://ezbiocloud.net/\u003c/span\u003e\u003cspan address=\"http://ezbiocloud.net/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) (Yoon et al. 2017). Evolutionary analysis was performed using MEGA 11 as previously described by Tamura et al. (2021). Phylogenetic tree models are constructed using the following three algorithms: maximum likelihood (ML) (Felsenstein 1981), maximum parsimony (MP) (Fitch 1971), and neighbour-joining (NJ) (Saitou and Nei 1987). The bootstrap method was used to test the stability of the phylogenetic tree topology, using 1000 replicates overall. Utilizing the Kimura two-parameter method, the evolutionary distance was determined (Kimura 1980; Tamura et al. 2013).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eGenomic Analysis\u003c/h2\u003e \u003cp\u003eThe whole genome of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was sequenced on Illumina Hiseq 4000 (Shanghai Magi Biomedical Technology Co., LTD., Shanghai, China) and calibrated using SOAP denovo 2.04 and SOAP aligner 2.21 (Li et al. 2008). Draft genomes of 20 \u003cem\u003eBrevibacterium\u003c/em\u003e strains were collected from the NCBI (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.ncbi.nlm.nih.gov/\u003c/span\u003e\u003cspan address=\"http://www.ncbi.nlm.nih.gov/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) database species in order to construct the phylogenetic tree. \u003cem\u003ePseudoclavibacter helvolus\u003c/em\u003e DSM 20419\u003csup\u003eT\u003c/sup\u003e (JACHWJ000000000) was used as an outgroup. Genome sequence analysis was performed using a rapid annotation technique based on Subsystem Technology 2.0 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://rast.nmpdr.org/\u003c/span\u003e\u003cspan address=\"https://rast.nmpdr.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) (Aziz et al. 2008). Utilizing the ChunLab online ANI calculator from EzBioCloud (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ezbiocloud.net/\u003c/span\u003e\u003cspan address=\"https://www.ezbiocloud.net/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), strains closely related to GXQ1321\u003csup\u003eT\u003c/sup\u003e were analyzed for average nucleotide identity (ANI). Digital DNA-DNA hybridization (dDDH) values between GXQ1321\u003csup\u003eT\u003c/sup\u003e and related \u003cem\u003eBrevibacterium\u003c/em\u003e members were computed using the genome-genome distance calculator (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://ggdc.dsmz.de/ggdc.php\u003c/span\u003e\u003cspan address=\"https://ggdc.dsmz.de/ggdc.php\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) (Meier-Kolthoff et al. 2013). Using the Majorbio amino acids identified online calculator (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.majorbio.com/\u003c/span\u003e\u003cspan address=\"https://www.majorbio.com/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) is closely related to the GXQ1321\u003csup\u003eT\u003c/sup\u003e short bacillus strains of amino acids identified (AAI) on average. The antismash.secondarymetabolites.org/ website was used in conjunction with the antiSMASH v. 7.1 analysis tools to rapidly identify and analyse clusters of secondary metabolite biosynthesis genes in the strain genome (Blin et al. 2021). Phylogenetic tree of the genome was reconstructed using bacterial core genes (UBCG) (Na, S. I. et al. 2018), online KEGG metabolic pathway analysis of the strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was performed using the KEGG annotation website (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.genome.jp/kegg/kaas/\u003c/span\u003e\u003cspan address=\"https://www.genome.jp/kegg/kaas/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and the egggnog-mapper website (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://eggnog-mapper.embl.de\u003c/span\u003e\u003cspan address=\"http://eggnog-mapper.embl.de\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) was used to perform the COG categorisation analysis of the encoded proteins (Carlos et al. 2021). Ortho Venn3 software (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://orthovenn3\u003c/span\u003e\u003cspan address=\"https://orthovenn3\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. bioinfotoo lkits. net/ home) was used to generate ortho analysis Venn diagrams and pairwise ortho analysis heatmap between the three genomes (Sun et al. 2003).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003ePhysiological and biochemical characteristics\u003c/h2\u003e \u003cp\u003eScanning electron microscopy and transmission electron microscopy (Talos F200C) were used to observe the morphology and flagellum of the strain. The test strains were stained using the Hopebiol (Qing Dao) Gram Stain Kit's operating methods. The strains were inoculated on actinomycetes culture medium and cultured at different temperatures (0, 4, 10, 20, 30, 37, 40, 50℃) for 4 weeks. The strain's growth was assessed using OD\u003csub\u003e600\u003c/sub\u003e, and pH tolerance (pH 4.0\u0026ndash;10.0) was determined using actinomycetes media. Tolerance to 0\u0026ndash;20% w/v strains was assessed liquid using actinomycetes culture medium devoid of NaCl. The strain's activity was assessed using the turbidity of a semi-solid medium containing 0.4% agar (Zhao et al. 2024). Catalase activity was measured using 3% (v/v) H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e. Additional physiological tests, such as absorption, acid generation, and enzymology, were performed with API 20NE, API 50CH, API ZYM, and Biolog Gen III test strips (bioMerieux). The carbon utilisation of the strains was tested using Biolog's Gen III eco-plate. Follow the directions in the manual. The strains were tested for antibiotic susceptibility using multi-antibiotic susceptibility test tablets (Zhao et al. 2024). In order to determine the enzyme production of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e, eight enzyme production function plates (cellulase production, protease production, amylase production, urease production, inorganic phosphorus utilization, organophosphorus utilization, CAS, nitrogen fixation function) were selected for detection. Whether strain GXQ1321T has enzyme production function was preliminarily determined by the presence or absence of the transparent ring.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eChemotaxonomic characteristics\u003c/h2\u003e \u003cp\u003eBacteria were collected after 4 days of cultivation at 30℃ on actinomycetes culture medium and washed three times with 0.9% NaCl solution. The obtained sediments were freeze-dried by vacuum freeze dryer for 3 days for chemical experiments. The Sherlock Microbial Identification System (MIDI Inc., Newark, USA) operational parameters were followed for the extraction of fatty acid samples. Prepared and analysed according to Sherlock Microbial Identification System instructions (Whittaker 2011). The procedure for extraction of polar lipids was followed as described in detail by Song Y et al. (2000). Methylnaphthoquinone drugs were extracted utilizing the technique of Collins et al. (1979) and high performance liquid chromatography (HPLC) was used to identify them (Kroppenstedt and Reiner 1982). The instructions of Schleifer et al. (1972) for the preparation and analysis of cell wall peptidoglycan were followed. Hasegawa et al.'s (1983) methodology for analyzing whole cell sugars was used.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic characterization\u003c/h2\u003e \u003cp\u003eThe complete 16S rRNA gene sequence of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e (GenBank/EMBL/DDBJ accession number OR661284) was found with a total length of 1459 bp. 16S rRNA sequencing showed that the strain belonged to \u003cem\u003eBrevibacterium\u003c/em\u003e. Using the EZBiocloud database comparison, the 16S rRNA with the highest similarity to GXQ1321\u003csup\u003eT\u003c/sup\u003e were \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e (96.77%) and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e (96.32%). Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e developed an isolated subbranch structure within a broader clade encompassing \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e (96.77%) and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e (96.32%), according to an adjacency tree analysis based on 16S rRNA gene sequences. Maximum parsimonious and maximum likelihood phylogenetic trees also corroborate this relationship. Maximum likelihood and maximum parsimonious phylogenetic trees also corroborate this relationship. Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e and Fig. S2 show the results of the maximum likelihood and maximum parsimony calculations. It unstable tree topologies indicate that GXQ1321\u003csup\u003eT\u003c/sup\u003e does not belong to any of the Brevibacterium strains that have been successfully characterised. .\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eGenomic analysis\u003c/h2\u003e \u003cp\u003eThe strain GXQ1321\u003csup\u003eT\u003c/sup\u003e (GenBank accession number JBDIUY000000000) has a genomic length of 3347008 bp and 69.6% G\u0026thinsp;+\u0026thinsp;C. The genome contains 26 overlapping groups with maximum and minimum contigs lengths of 315,757 and 1251bp, N50 values of 227,695 bp, N90 values of 94,674 bp, and L50 values of 6, respectively. With maximum and minimum contig lengths of 315,757 and 1251 bp, N50 values of 227,695 bp, N90 values of 94,674 bp, and L50 values of 6, the genome is composed of 26 overlapping groups. Genome-wide information is used to construct the phylogenetic tree. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and reference strain SST-8\u003csup\u003eT\u003c/sup\u003e clustered together to form a branch, which proved that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was the closest relative to reference strain \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e (Fig.\u0026nbsp;2). Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows that the ANI values of \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e (96.77%), and GXQ1321\u003csup\u003eT\u003c/sup\u003e were all below the species criterion (ANI 95%). dDDH calculation showed that the DNA affinity of GXQ1321\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e (96.77%) was 21.2%. DNA affinity with \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e (96.32%) was 15.3%. The results were below the 70% threshold required to discriminate between species. The AAI values of GXQ1321\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e (96.77%) and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e were also below the species threshold (AAI 95%) (Fig. S3). The results showed that GXQ1321\u003csup\u003eT\u003c/sup\u003e belongs tothe genus \u003cem\u003eBrevibacterium\u003c/em\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\u003eANI and dDDH values between strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and other closely related strains.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStrain 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStrain 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eANI (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003edDDH (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16S rRNA gene identity (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGXQ1321\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eB. Rongguiense\u003c/em\u003e\u003c/p\u003e \u003cp\u003e5221\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e73.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e15.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96.32\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eB.samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e77.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e21.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96.77\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\u003eSeveral features of the genome of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e distinguished it from other \u003cem\u003eBrevibacterium\u003c/em\u003e strains by annotation and analysis of the genome (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The annotation results showed that GXQ1321\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e all contained terpene and NAPPA genes. Annotation results from the antiSMASH 7.1 database showed that GXQ1321\u003csup\u003eT\u003c/sup\u003e contained a terpene gene cluster with 33% similarity to carotenoids. The yellow-green colour of the GXQ1321\u003csup\u003eT\u003c/sup\u003e colony was also consistent with the phenotypic characteristics. In addition, GXQ1321\u003csup\u003eT\u003c/sup\u003e also contains 75% ectoine, whereas the \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e strains did not contain the ectoine gene (Table S2). Ectoine helps organisms withstand high osmotic pressure environments. Ectoine protects enzymes, membranes and whole cells from the stresses of salt, heating, freezing and drying. The ability of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e to endure in marine sediments was also verified by this outcome. This gene cluster is present in the genus \u003cem\u003eBrevibacterium\u003c/em\u003e, which was isolated from a harsh environment (Pei S et al. 2020). This result also proved that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e belonged to the genus \u003cem\u003eBrevibacterium\u003c/em\u003e.\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\u003eComparison of the genomic characteristics of GXQ1321\u003csup\u003eT\u003c/sup\u003e and related members of the genus \u003cem\u003eBrevibacterium\u003c/em\u003e. +, positive; \u0026ndash;, negative.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGenes putatively encoding\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGXQ1321\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eB. \u003cem\u003esamyangense\u003c/em\u003e\u003c/p\u003e \u003cp\u003eSST-8\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eB. Rongguiense\u003c/em\u003e\u003c/p\u003e \u003cp\u003e5221\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGenBank accession number\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eJAOEFD000000000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBAAANO000000000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eJACCFQ010000000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGenome size (bp)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3347008\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3394428\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3117581\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDNA G\u0026thinsp;+\u0026thinsp;C content (mol%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e68.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e72.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN50 value (bp)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e139003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e41933\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e42508\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL50 value (bp)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNumber of coding sequences\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3088\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3197\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2904\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003efeatures in the draft genome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3147\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3252\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2952\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNumber of rRNAs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNumber of Subsystems\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e235\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e238\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e236\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\u003eThe results of the pan-genomic comparison between strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and its reference strains were OrthoVenn3 (Fig. S4). The strain GXQ1321\u003csup\u003eT\u003c/sup\u003e had a total of 2243 gene clusters, while the other two reference strains had 2298 and 1790 gene clusters, respectively. The results showed that strains GXQ1321\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e had the highest similarity, with a total of 594 gene clusters, which exceeded the 86 gene clusters of strains GXQ1321\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. Analysis of the bacteria revealed that the three strains share a \"core\" genome consisting of 1,548 homologous gene clusters, most of which code for proteins with functions related to cellular metabolism and specialised membrane exchange systems. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and strain \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e showed the deepest colour representation in the pin-to-pair orthogonal heat maps of the three genomes, indicating a close relationship between them (Fig. S5). Based on the above analysis, strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and reference strain SST-8\u003csup\u003eT\u003c/sup\u003e were more closely related, which was also confirmed by the genome-wide evolutionary tree.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eA total of 2,061 genes involved in six pathways were detected in the KEGG database. Of these, metabolism was the most common, including amino acid metabolism, carbohydrate metabolism, cofactor and vitamin metabolism, and energy metabolism (Fig. S6). We discovered that \"amino acid transport and metabolism\" was the largest of the known coding proteins of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e based on a COG classification analysis (Table S3). Since strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was isolated from marine surface sediments, we hypothesised that ectoine secreted by GXQ1321\u003csup\u003eT\u003c/sup\u003e may affect the growth and development and stress resistance of marine sediment plants.\u003c/p\u003e \u003cp\u003eThe characteristics of nitrogen and sulphur metabolism in subsystems were compared and analysed. Strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e lacked genes related to cyanate hydrolysis, nitrate and nitrite antimonization, and only ammonia assimilation genes. On the contrary, strain GXQ1321\u003csup\u003eT\u003c/sup\u003e contained genes related to cyanate hydrolysis, nitrate and nitrite antimonization and ammonia assimilation. In the characteristics of sulfur metabolism, both strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e contained thioredoxin-disulfide reductase, but the reference strain \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e contained genes related to organic sulfur assimilation. Therefore, \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e had the highest abundance, with a total of 11 genes related to sulfur metabolism (Table S4).\u003c/p\u003e \u003cp\u003eAccording to the analysis of the NCBI database, we found that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e contained the transporters ABC(WP_349829439.1), BCCT(WP_349828541.1), MFS(WP_349829450.1), NCS2(WP_349828315.1), ACR3(WP_349826989.1), etc., which are basically the same as the transporters contained in \u003cem\u003eBrevibacterium\u003c/em\u003e isolated from the sea, which further confirmed that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e belongs to a genus of \u003cem\u003eBrevibacterium\u003c/em\u003e. Among them, BCCT contributes to the accumulation of compatible solutes betaine, carnitine and choline. This is not only beneficial for maintaining the osmotic balance of the cell, but also acts as a stabiliser for proteins and cellular components, preventing denaturation and resisting the effects of high ionic strength. ABC transporters may be related to their ability to transport and metabolise various toxic substances and chitin, amylopectin, cellulose and starch. The Rast analysis showed that 235 subsystems existed in strain GXQ1321\u003csup\u003eT\u003c/sup\u003e, and 15 glycoside hydrolases (GH), 26 glycosyltransferases (GTs), 1 carbohydrate binding module (CBM), 14 carbohydrate esterases (CEs) and 8 auxiliary activities (AAs) were identified. Since lignocellulose is abundant in plants growing in Marine surface sediments, the coexistence of these genes suggests that they play an important role in the breakdown and modification of carbohydrates in Marine surface sediments. This was consistent with our subsequent functional assay finding that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e had the ability to produce amylase and cellulase.\u003c/p\u003e \u003cp\u003eMFS promotes the transmembrane transport of solutes such as sugars, drug molecules, peptides, tricarboxylic acid cycle metabolites, organic anions and inorganic anions under electrochemical gradients. This corresponds to the maximum \"amino acid transport and metabolism\" shown in COG. At the same time, KEGG metabolic pathway analysis revealed that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e contained the SecE gene. The Sec pathway is the first secretory pathway discovered in bacteria to export proteins across the plasma membrane to the periplasm and outer membrane of bacteria. The SecE gene is a gene associated with extracellular transport of cells, possibly because most marine \u003cem\u003eBrevibacterium\u003c/em\u003e release a class of macromolecular protein heteropolysaccharide extracellular polymers into the environment to prevent ice crystal formation and adapt to low temperatures. We also identified genes related to salt tolerance and alkaline resistance in the gene annotation results of strain GXQ1321T, including the Na+/H\u0026thinsp;+\u0026thinsp;reverse transporters NhaC and NhaD, and the Na+-driven multidrug efflux pump, the DinF/NorM/MATE family. The results are also consistent with previous physiological experiments testing the bacteria's tolerance to saline alkaline conditions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eMorphological, physiological, and biochemical characteristics\u003c/h2\u003e \u003cp\u003eThe GXQ1321\u003csup\u003eT\u003c/sup\u003e strain's colony was circular, convex, golden, and free of diffuse pigment. Its margins were unbroken. Following 48\u0026ndash;72 hours of cultivation at 30 ℃, the strain's cells had a short rod-like morphology (0.51\u0026ndash;0.59 \u0026times; 1.1\u0026ndash;1.6 \u0026micro;m) (Fig. S7), with a colony diameter of 1.2\u0026ndash;2.3 mm. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was positive for Gram staining and aerobic. Transmission electron microscopy revealed that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was flagellated and nonmotile (Fig. S8). This is similar to the reference strains, the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e have no flagellum and are non-motile, while most strains of the genus \u003cem\u003eBrevibacterium\u003c/em\u003e have no flagellum. The strain was grown on actinomycetes culture media for three days at 30\u0026deg;C. The colonies were 1.1\u0026ndash;1.8 mm in diameter and yellow in colour. After one month of observation, strain GXQ1321\u003csup\u003eT\u003c/sup\u003e grew at 4\u0026ndash;50℃ (optimal 30 ℃). The concentration of NaCl tolerance ranged from 0 to 20% (4% was optimal). Catalase detection results of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e were negative. The pH range in which the strains can grow is 4.0\u0026ndash;10 (7.0 is optimal). Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e illustrates the observed variations in physiological characteristics between GXQ1321\u003csup\u003eT\u003c/sup\u003e and the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e were sensitive to amoxicillin, streptomycin, erythromycin, rifampicin, neomycin and chloramphenicol. Compared to the reference strain \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e, strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and reference strain \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e were not resistant to neomycin and gentamicin. Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e displays the differences in API 20NE, API 50CH, API ZYM, and Biolog GEN III as well as the physiological and biochemical features of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e compared to reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. According to the appearance of the transparent circle, we found that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e had the function of producing amylase, amylase and ferriferite.\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\u003eDifferential physiological characteristics of GXQ1321\u003csup\u003eT\u003c/sup\u003e from the closely related species \u003cem\u003eB.samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. Rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. All data presented are from this study. +, positive; \u0026ndash;, negative;W, weak reaction.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCharacteristic\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGXQ1321\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eB.samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eB. Rongguiense\u003c/em\u003e\u003c/p\u003e \u003cp\u003e5221\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eColony color\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLight-yellow\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBright-yellow\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMilky - white\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMotility\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003epH(optimum)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.0\u0026ndash;10.0(7.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.0-11.5(10.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.0-9.5(7.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTemperature (optimum) (\u0026deg;C)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4\u0026ndash;50(\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10\u0026ndash;45(\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10\u0026ndash;40(\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNaCl (optimum) (%, w/v)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0\u0026ndash;20)4.0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026ndash;15(7.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(\u003cspan additionalcitationids=\"CR2 CR3 CR4 CR5 CR6 CR7 CR8 CR9 CR10 CR11\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e)4%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAssimilation of (API 20NE):\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUrea\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ew\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaltose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ew\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGelatin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEsculin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAcid production from (API 50CH):\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-Mannose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-GALactose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-MELibiose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL-RHamnose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-Fruetose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eArbutin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ew\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCarbon source utilization (Biolog GEN III):\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eα-D-Glucose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-Salicin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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\u003ew\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDextrin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGentiobiose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-Turanose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eα-D-lactose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL-Fucose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInosine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-Mannitol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-arabitol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL-Aspartic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGelatin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePectin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL-Alanine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSodium citrate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eW\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSodium formate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eW\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDisodium-D,L-malate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInulin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEnzymology (API ZYM):\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAcid phosphatase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eValine arylamidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCysteine arylamidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrypsin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eβ-glucosidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAntibiotic tolerance:\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGentamicin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNeomycin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\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 \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe peptidoglycan of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e is consistent with the peptidoglycan of the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e, whole cell hydrolysates containing meso-diaminopimelic acid (Fig. S9). \u003cem\u003eAnteiso\u003c/em\u003e-C\u003csub\u003e19:0\u003c/sub\u003e (27.28%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e (18.97%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e (15.95%), and \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e16:0\u003c/sub\u003e (12.21%) were the major fatty acids (\u0026gt;\u0026thinsp;10%) of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e. The major fatty acids (\u0026gt;\u0026thinsp;10%) of \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e were \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e (35.3%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e (29.9%) and \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e (15.5%). The major fatty acids (\u0026gt;\u0026thinsp;10%) of \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e were \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e (46.2%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e (39.3%) and C\u003csub\u003e16:0\u003c/sub\u003e (11.9%) (Table S5). The main fatty acids of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e were different from those of \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. Phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), three phosphoglycolipids (PGL) and two unknown glycolipid (UG) were the major polar lipids of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e (Fig. S10). Compared to GXQ1321\u003csup\u003eT\u003c/sup\u003e, the major polar lipid of \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e was phosphatidylglycerol (PG), and the major polar lipids of \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), and three types of phosphoglycolipids (PGL). The respiratory quinones of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e were mainly MK-8 (90%). The reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e breathed predominantly quinones as MK-8, consistent with most of the genus \u003cem\u003eBrevibacterium\u003c/em\u003e. The results of physicochemical experiments demonstrated that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e could be distinguished from the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. The growth rate of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e on actinomycetes culture medium was higher than the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. The colony colour and moisture of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e were also different from the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e were sensitive to amoxicillin (10 \u0026micro;g/tablet), streptomycin (10 \u0026micro;g/tablet), erythromycin (15 \u0026micro;g/tablet), rifampicin (5 \u0026micro;g/tablet), neomycin (30 \u0026micro;g/tablet) and chloramphenicol (30 \u0026micro;g/tablet). Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e and the reference strains \u003cem\u003eB. samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e were resistant to neomycin (30 \u0026micro;g/tablet) and gentamicin (10 \u0026micro;g/tablet), while \u003cem\u003eB. rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e was resistant to neomycin and gentamicin. Following an analysis of the strain's physiology, chemistry, and phylogeny, it was concluded that strain GXQ1321\u003csup\u003eT\u003c/sup\u003e represents a novel species within the \u003cem\u003eBrevibacterium\u003c/em\u003e genus, designated as \u003cem\u003eBrevibacterium litoralis\u003c/em\u003e sp. nov.\u003c/p\u003e \u003cp\u003e \u003cb\u003eDescription of\u003c/b\u003e \u003cb\u003eBrevibacterium litoralis\u003c/b\u003e \u003cb\u003esp. nov\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eBrevibacterium litoralis\u003c/em\u003e sp. nov (li.to.ra\u0026prime;lis. L. masc. adj. litoralis of or belonging to the sea shore).\u003c/p\u003e \u003cp\u003eThe strain is an aerobic, Gram-positive actinomycetes that is non-motile and does not generate spores. The cells measured 0.51\u0026ndash;0.59 \u0026times; 1.1\u0026ndash;1.6 \u0026micro;m and had a short rod shape. The colony is yellow, round, convex, with intact margins and does not produce diffuse pigments. Negative for catalase. The optimal growth conditions for the organism in question are 4\u0026ndash;50 ℃, pH 4.0\u0026ndash;10.0, and NaCl concentration ranging between 0\u0026ndash;20% w/v (optimum 4.0%). Maltose could not be hydrolyzed by the strain, while gelatin and aesculin could. Arbutin, D-galactose, and D-meliose were converted to acid. Dextrin, L-fructose, L-rhamnose, D-maltose, D-trehalose, myo-inositol, D-cellulobiose, sucrose, D-terabiose, stachyose, α-D-lactose,glycerol, D-glucose-6-phosphate, D-fructose-6-phosphate, β-methyl-D-glucoside, D-salicin, N-acetyl-D-glucosamine, alpha-D-glucose, N-acetyl-β-D-mannosamine, D-mannose, gelatine, D-fructose, sodium lactate, D-sorbitol, inosine, D-mannitol, D-arabitol, D-aspartate, L-alanine, gentiobiose, L-aspartic acid, L-glutamic acid, L-histidine, L-pyroglutamic acid, L-serine, pectin, D-galacturonic acid, L-galactonic acid lactone, D-glucuronic acid, D-saccharic acid, L-lactic acid, α-ketoglutaric acid, D-malic acid, D-lactic acid methyl ester, L-malic acid, Tween 40, propionic acid, acetic acid, formic acid can be used as a sources of carbon. In accordance with the API ZYM test paper, alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, valine arylamidase, cystine arylamidase, trypsin, acid phosphatase, naphtho-AS-BI-phosphohydrolase were all positive. Lipase (C14), chymotrypsin, β-fucosidase, alpha-mannosidase, α-galactosidase, β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-glucosaminase were negative.\u003c/p\u003e \u003cp\u003eThe enzymes produced by this strain were esterase, amylase and cellulase. The strain was resistant to neomycin, amoxicillin, streptomycin, chloramphenicol, rifampicin, novobiocin, gentamicin and erythromycin. The main respiratory quinone was MK-8 (90%). The main polar lipids of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), three phosphoglycolipids (PGL) and two unknown glycolipid (UG). The whole cell hydrolysates containing meso-diaminopimelic acid. \u003cem\u003eAnteiso\u003c/em\u003e-C\u003csub\u003e19:0\u003c/sub\u003e (27.28%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e (18.97%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e (15.95%), and \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e16:0\u003c/sub\u003e (12.21%) were the major fatty acids (\u0026gt;\u0026thinsp;10%) of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eStrain GXQ1321\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;MCCC 1K08964\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;KCTC 59167\u003csup\u003eT\u003c/sup\u003e) was the first \u003cem\u003eBrevibacterium\u003c/em\u003e isolated from surface sediments in Beihai, Guangxi. The 16S rRNA gene for the strain GXQ1321\u003csup\u003eT\u003c/sup\u003e has been assigned the GenBank accession number OR661284. The Whole Genome Shotgun project has been deposited at DDBJ/ENA/GenBank under the accession JBDIUY000000000.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMingguo Jiang, conceptualisation and project administration. Contributed to the conception of the study, formulation or evolution of overarching research goals and aims.\u003c/p\u003e\n\u003cp\u003eYi Jiang, resources. Provision of some study materials.\u003c/p\u003e\n\u003cp\u003eQuan Yang, investigation and writing-original draft preparation, performing all of the experiments.\u003c/p\u003e\n\u003cp\u003eAolin Zhao, review and editing, performing part of the experiments.\u003c/p\u003e\n\u003cp\u003eHaifei Liu, performed research;\u003c/p\u003e\n\u003cp\u003eJiawei Li, performed research;\u003c/p\u003e\n\u003cp\u003eShujing Wu, performed research;\u003c/p\u003e\n\u003cp\u003eYing Huang, analyzed data.\u003c/p\u003e\n\u003cp\u003eJie Weng, analyzed data.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the Science and Technology Major Project of Guangxi\u0026nbsp;(AA18242026), recipient: Mingguo Jiang.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompliance with Ethical Standards\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors confirm that no competing interests, both financial and personal, are with this manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene and the draft genome sequences of strain GXQ1321\u003csup\u003eT\u003c/sup\u003e are OR661284 and JBDIUY000000000, respectively.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBreed, R. (1953) The \u003cem\u003eBrevibacteriaceae fam.\u003c/em\u003e nov. of order Eubacteriales. Rias Commun VI Congr Int Microbiol Roma 1, 13\u0026ndash;14. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-3-642-30138-4_169\u003c/span\u003e\u003cspan address=\"10.1007/978-3-642-30138-4_169\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTang, S. K., Wang, Y., Schumann, P., Stackebrandt, E., Lou, K., Jiang, C. L., Xu, L. H., \u0026amp; Li, W. J. (2008) \u003cem\u003eBrevibacterium album\u003c/em\u003e sp. nov., a novel actinobacterium isolated from a saline soil in China. International journal of systematic and evolutionary microbiology, 58(Pt 3), 574\u0026ndash;577. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.65183-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.65183-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJung MS, Quan XT, Siddiqi MZ, Liu Q, Kim SY, Wee JH, Im WT. (2018) \u003cem\u003eBrevibacterium anseongense\u003c/em\u003e sp. nov., isolated from soil of ginseng field. J Microbiol 56:706\u0026ndash;712. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s12275-018-8181-5\u003c/span\u003e\u003cspan address=\"10.1007/s12275-018-8181-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePei S, Niu S, Xie F, Wang W, Zhang S, Zhang G. (2021) \u003cem\u003eBrevibacterium limosum\u003c/em\u003e sp. nov., \u003cem\u003eBrevibacterium pigmenatum\u003c/em\u003e sp. nov., and \u003cem\u003eBrevibacterium atlanticum\u003c/em\u003e sp. nov., three novel dye decolorizing actinobacteria isolated from ocean sediments. J Microbiol 59:898\u0026ndash;910. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s12275-021-1235-0\u003c/span\u003e\u003cspan address=\"10.1007/s12275-021-1235-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBhadra B, Raghukumar C, Pindi PK, Shivaji S. (2008) \u003cem\u003eBrevibacterium oceani\u003c/em\u003e sp. nov., isolated from deep-sea sediment of the Chagos Trench, Indian Ocean. Int J Syst Evol Microbiol 58:57\u0026ndash;60. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.64869-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.64869-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDeng T, Lu H, Qian Y, Chen X, Yang X, Guo J, Sun G, Xu M. (2020) \u003cem\u003eBrevibacterium rongguiense\u003c/em\u003e sp. nov., isolated from freshwater sediment. Int J Syst Evol Microbiol 70:5205\u0026ndash;5210. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.004379\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.004379\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePei S, Xie F, Niu S, Ma L, Zhang R, Zhang G. (2020) \u003cem\u003eBrevibacterium profundi\u003c/em\u003e sp. nov., isolated from deep-sea sediment of the Western Pacific Ocean. Int J Syst Evol Microbiol 70:5818\u0026ndash;5823. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.004483\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.004483\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKati H, Ince IA, Demir I, Demirbag Z. (2010) \u003cem\u003eBrevibacterium pityocampae\u003c/em\u003e sp. nov., isolated from caterpillars of Thaumetopoea pityocampa (Lepidoptera, Thaumetopoeidae). Int J Syst Evol Microbiol 60:312\u0026ndash;316. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.006692-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.006692-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang M, Song Q, Sang J, Li Z. (2023) \u003cem\u003eBrevibacterium spongiae\u003c/em\u003e sp. nov., isolated from marine sponge Hymeniacidon sp. Int J Syst Evol Microbiol 2023; 73:5869. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.005869\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.005869\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKampfer P, Schafer J, Lodders N, Busse HJ. (2010) \u003cem\u003eBrevibacterium sandarakinum\u003c/em\u003e sp. nov., isolated from a wall of an indoor environment. Int J Syst Evol Microbiol 60:909\u0026ndash;913. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.014100-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.014100-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWauters G, Charlier J, Janssens M, Delmee M. (2001) \u003cem\u003eBrevibacterium paucivorans\u003c/em\u003e sp. nov., from human clinical specimens. Int J Syst Evol Microbiol 51:1703\u0026ndash;1707. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/00207713-51-5-1703\u003c/span\u003e\u003cspan address=\"10.1099/00207713-51-5-1703\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMages IS, Frodl R, Bernard KA, Funke G. (2008) Identities of Arthrobacter spp. and Arthrobacter-like bacteria encountered in human clinical specimens. J Clin Microbiol 46:2980\u0026ndash;2986. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1128/jcm.00658-08\u003c/span\u003e\u003cspan address=\"10.1128/jcm.00658-08\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMcBride ME, Ellner KM, Black HS, Clarridge JE, Wolf JE. (1993) A new \u003cem\u003eBrevibacterium\u003c/em\u003e sp. isolated from infected genital hair of patients with white piedra. J Med Microbiol 39:255\u0026ndash;261. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/00222615-39-4-255\u003c/span\u003e\u003cspan address=\"10.1099/00222615-39-4-255\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePascual C, Collins MD, Funke G, Pitcher DG. (1996) Phenotypic and genotypic characterization of two \u003cem\u003eBrevibacterium\u003c/em\u003e strains from the human ear: description of \u003cem\u003eBrevibacterium otitidis\u003c/em\u003e sp. nov. Med. Microbiol. Lett. 5:113\u0026ndash;123. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.1099/jmm.0.043109-0\u003c/span\u003e\u003cspan address=\"10.1099/jmm.0.043109-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBernard, K. A., Wiebe, D., Burdz, T., Reimer, A., Ng, B., Singh, C., Schindle, S., \u0026amp; Pacheco, A. L. (2010) Assignment of Brevibacterium stationis (ZoBell and Upham 1944) Breed 1953 to the genus Corynebacterium, as Corynebacterium stationis comb. nov., and emended description of the genus Corynebacterium to include isolates that can alkalinize citrate. International journal of systematic and evolutionary microbiology, 60(Pt 4), 874\u0026ndash;879. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.012641-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.012641-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePascual C, Collins MD. (1999) \u003cem\u003eBrevibacterium avium\u003c/em\u003e sp. nov., isolated from poultry. Int J Syst Bacteriol 9:1527\u0026ndash;1530. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/00207713-49-4-1527\u003c/span\u003e\u003cspan address=\"10.1099/00207713-49-4-1527\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKomagata, K., \u0026amp; Iizuka, H. (1964) New species of \u003cem\u003eBrevibacterium\u003c/em\u003e Isolated from Rice. Bulletin of the Agricultural Chemical Society of Japan, 38, 496\u0026ndash;502. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1271/nogeikagaku1924.38.496\u003c/span\u003e\u003cspan address=\"10.1271/nogeikagaku1924.38.496\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoi EJ, Lee SH, Jung JY, Jeon CO. (2013) \u003cem\u003eBrevibacterium jeotgali\u003c/em\u003e sp. nov., isolated from jeotgal, a traditional Korean fermented seafood. Int J Syst Evol Microbiol 63:3430\u0026ndash;3436. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.049197-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.049197-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOzturkoglu-Budak S, Wiebenga A, Bron PA, de Vries RP. (2016) Protease and lipase activities of fungal and bacterial strains derived from an artisanal raw ewe's milk cheese, International Journal of Food Microbiology, 237: 17\u0026ndash;27. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ijfoodmicro.2016.08.007\u003c/span\u003e\u003cspan address=\"10.1016/j.ijfoodmicro.2016.08.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmarita F, Nardi M, Chambellon E, Delettre J, Bonnarme P. (2004). Identification and Functional Analysis of the Gene Encoding Methionine-γ-Lyase in Brevibacterium linens. Appl Environ Microbiol 70(12):7348\u0026ndash;7354. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1128/AEM.70.12.7348-7354.2004\u003c/span\u003e\u003cspan address=\"10.1128/AEM.70.12.7348-7354.2004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSimoes, F., Vale, P., Stephenson, T., \u0026amp; Soares, A. (2018). Understanding the growth of the bio-struvite production Brevibacterium antiquum in sludge liquors. Environmental technology, 39(17), 2278\u0026ndash;2287. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/09593330.2017.1411399\u003c/span\u003e\u003cspan address=\"10.1080/09593330.2017.1411399\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLamballerie de, Zandotti C, Vignoli C, Bollet C, de Micco P. A onestep microbial DNA extraction method using \u0026ldquo;Chelex 100\u0026rdquo; suitable for gene amplification. Res Microbiol 1992; 143:785\u0026ndash;790. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0923-2508(92)90107-y\u003c/span\u003e\u003cspan address=\"10.1016/0923-2508(92)90107-y\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChun J, Goodfellow M. (1995) A phylogenetic analysis of the genus \u003cem\u003eNocardia\u003c/em\u003e with 16S rRNA gene sequences. Int J Syst Bacteriol 45:240\u0026ndash;245. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/00207713-45-2-240\u003c/span\u003e\u003cspan address=\"10.1099/00207713-45-2-240\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEdwards U, Rogall T, Bl\u0026ouml;cker H, Emde M, B\u0026ouml;ttger EC. (1989) Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 17:7843\u0026ndash;7853. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/nar/17.19.7843\u003c/span\u003e\u003cspan address=\"10.1093/nar/17.19.7843\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLane DJ. (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, Chichester, pp 115\u0026ndash;175.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYoon SH, SM H, Kwon S, Lim J, Kim Y. (2017) Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol; 67:1613\u0026ndash;1617. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.001755\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.001755\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoichiro Tamura, Glen Stecher, Sudhir Kumar. MEGA11: Molecular Evolutionary Genetics Analysis version 11. Molecular Biology and Evolution, Volume 38, Issue 7, July 2021, Pages 3022\u0026ndash;3027. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msab120\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msab120\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFelsenstein J. (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 17:368\u0026ndash;376. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01734359\u003c/span\u003e\u003cspan address=\"10.1007/BF01734359\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFitch WM. (1971) Toward defining the course of evolution: minimum change for a specific tree topology. Syst Zool 20:406\u0026ndash;416. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/sysbio/20.4.406\u003c/span\u003e\u003cspan address=\"10.1093/sysbio/20.4.406\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaitou N, Nei M. (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406\u0026ndash;425. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/oxfordjournals.molbev.a040454\u003c/span\u003e\u003cspan address=\"10.1093/oxfordjournals.molbev.a040454\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKimura M. (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol; 16:111\u0026ndash;120. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01731581\u003c/span\u003e\u003cspan address=\"10.1007/BF01731581\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 2013; 30:2725\u0026ndash;2729. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/mst197\u003c/span\u003e\u003cspan address=\"10.1093/molbev/mst197\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi R, Li Y, Kristiansen K, Wang J. (2008) SOAP: short oligonucleotide alignment program. Bioinformatics 24:713\u0026ndash;714. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/bioinformatics/btn025\u003c/span\u003e\u003cspan address=\"10.1093/bioinformatics/btn025\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAziz RK, Bartels D, Best AA, DeJongh M, Disz T, Edwards RA, Formsma K, Gerdes S, Glass EM, Kubal M, Meyer F, Olsen GJ, Olson R, Osterman AL, Overbeek RA, McNeil LK, Paarmann D, Paczian T, Parrello B, Pusch GD, Reich C, Stevens R, Vassieva O, Vonstein V, Wilke A, Zagnitko O. (2008) The RAST server: rapid annotations using subsystems technology. BMC Genomics 9:75. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1471-2164-9-75\u003c/span\u003e\u003cspan address=\"10.1186/1471-2164-9-75\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeier-Kolthoff JP, Auch AF, Klenk HP, G\u0026ouml;ker M. (2013) Genome sequencebased species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 14:60. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1471-2105-14-60\u003c/span\u003e\u003cspan address=\"10.1186/1471-2105-14-60\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBlin K, Shaw S, Kloosterman AM et al. (2021) antiSMASH 6.0: improving cluster detection and comparison capabilities. Nucl Acids Res 49(W1): W29-W35. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/nar/gkab335\u003c/span\u003e\u003cspan address=\"10.1093/nar/gkab335\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNa, S. I., Kim, Y. O., Yoon, S. H., Ha, S. M., Baek, I., \u0026amp; Chun, J. (2018). UBCG: Up-to-date bacterial core gene set and pipeline for phylogenomic tree reconstruction. Journal of microbiology (Seoul, Korea), 56(4), 280\u0026ndash;285. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s12275-018-8014-6\u003c/span\u003e\u003cspan address=\"10.1007/s12275-018-8014-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWattam AR, Davis JJ, Assaf R, Boisvert S, Brettin T, et al. (2017) Improvements to PATRIC, the all-bacterial bioinformatics database and analysis resource center. Nucleic Acids Res 45: D535\u0026ndash;D542. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/nar/gkw1017\u003c/span\u003e\u003cspan address=\"10.1093/nar/gkw1017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarlos P. Cantalapiedra, Ana Hernandez-Plaza, Ivica Letunic, Peer Bork, and Jaime Huerta -Cepas. eggNOG-mapper v2: functional annotation, orthology assignments, and domain prediction at the metagenomic scale. Mol. Biol. Evol. 38(12):5825\u0026ndash;5829. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msab293\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msab293\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSun, J., Lu, F., Luo, Y., Bie, L., Xu, L., \u0026amp; Wang, Y. (2023). OrthoVenn3: an integrated platform for exploring and visualizing orthologous data across genomes. Nucleic acids research, 51(W1), W397\u0026ndash;W403. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/nar/gkad313\u003c/span\u003e\u003cspan address=\"10.1093/nar/gkad313\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhao, A., Cai, H., Huang, Y. et al. (2024) \u003cem\u003eNesterenkonia marinintestina\u003c/em\u003e sp. nov., isolated from the fish intestine. Arch Microbiol 206, 110. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00203-023-03825-0\u003c/span\u003e\u003cspan address=\"10.1007/s00203-023-03825-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWhittaker P. (2011) Identification of six species in the new genus Cronobacter (Enterobacter sakazakii) from culture using gas chromatographic analysis of fatty acid methyl esters. Journal of AOAC International. 94(5):1581\u0026ndash;1584. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5740/jaoacint.10-451\u003c/span\u003e\u003cspan address=\"10.5740/jaoacint.10-451\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSong Y, Yang R, Guo Z, Zhang M, Wang X et al. (2000) Distinctness of spore and vegetative cellular fatty acid profiles of some aerobic endospore-forming bacilli. J Microbiol Methods 39:225\u0026ndash;241. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5740/jaoacint.10-451\u003c/span\u003e\u003cspan address=\"10.5740/jaoacint.10-451\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCollins, M. D., Goodfellow, M. \u0026amp; Minnikin, D. E. (1979) Isoprenoid quinones in the classification of coryneform and related bacteria. J Gen Microbiol 110, 127\u0026ndash;136. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/00221287-110-1-127\u003c/span\u003e\u003cspan address=\"10.1099/00221287-110-1-127\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKroppenstedt, Reiner M. (1982) Separation of bacterial menaquinones by HPLC using reverse phase (RP18) and a silver loaded ion exchanger as stationary phases. J Liq Chromatogr 5:2359\u0026ndash;2367. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/01483918208067640\u003c/span\u003e\u003cspan address=\"10.1080/01483918208067640\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchleifer, K. H. \u0026amp; Kandler, O. (1972) Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36, 407\u0026ndash;477. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1128/br.36.4.407-477.1972\u003c/span\u003e\u003cspan address=\"10.1128/br.36.4.407-477.1972\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHasegawa, T., Takizawa, M., \u0026amp; Tanida, S. (1983) A rapid analysis for chemical grouping of aerobic actinomycetes. Journal of General and Applied Microbiology, 29, 319\u0026ndash;322. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2323/jgam.29.319\u003c/span\u003e\u003cspan address=\"10.2323/jgam.29.319\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"antonie-van-leeuwenhoek","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"anto","sideBox":"Learn more about [Antonie van Leeuwenhoek](https://www.springer.com/journal/10482)","snPcode":"10482","submissionUrl":"https://submission.nature.com/new-submission/10482/3","title":"Antonie van Leeuwenhoek","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Surface sediments, Marine actinomycetes, Novel species, Species identification","lastPublishedDoi":"10.21203/rs.3.rs-4724416/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4724416/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA Gram stain-positive, non-spore-forming, non-motile, short-rod actinomyces strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was isolated from maritime surface sediments in Beihai(11\u0026deg;46\u0026prime;21.11\u0026Prime;N, 109\u0026deg;62\u0026prime;56.25\u0026Prime;E), Guangxi Zhuang Autonomous Region, and a number of categorization studies were performed. Following a period of 72 hours of incubation at a temperature of 30\u0026deg;C within an actinomycetes culture medium, the colony was yellow, circular, smooth, central bulge, convex, opaque, with a 1.8-3.0 mm diameter. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e has the ability to produce amylase and cellulase. Chemotaxonomic studies revealed that the major menaquinone in strain GXQ1321\u003csup\u003eT\u003c/sup\u003e is MK-8. The most prevalent cellular fatty acids were \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e19:0\u003c/sub\u003e (27.28%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e (18.97%), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e (15.95%), and \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e16:0\u003c/sub\u003e (12.21%). The whole-cell sugars of the strain GXQ1321\u003csup\u003eT\u003c/sup\u003e identified were rhamnose, xylose and glucose. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e exhibited the presence of meso-diaminopimelic acid (m-DAP) as a distinctive cell-wall diamino acid, and the polar lipids were identified as diphosphatidylglycerol (DPG), three phosphoglycolipid (PGL), phosphatidylglycerol (PG) and two unknown glycolipid (UG). This strain had 69.6% DNA G\u0026thinsp;+\u0026thinsp;C content. Strain GXQ1321\u003csup\u003eT\u003c/sup\u003e is classified as \u003cem\u003eBrevibacterium\u003c/em\u003e based on its 16S rRNA gene sequence. It is closely related to \u003cem\u003eBrevibacterium samyangense\u003c/em\u003e SST-8\u003csup\u003eT\u003c/sup\u003e (96.77%) and \u003cem\u003eBrevibacterium rongguiense\u003c/em\u003e 5221\u003csup\u003eT\u003c/sup\u003e (96.32%). The results showed that the average nucleotide identity (ANI) values of GXQ1321\u003csup\u003eT\u003c/sup\u003e and the above two strain tyoes were 73.91\u0026ndash;77.14%, and the digital DNA-DNA hybridisation (dDDH) values were 15.3\u0026ndash;21.1%. Based on the phylogenetic, chemotaxonomi and physiologicalc data, strain GXQ1321\u003csup\u003eT\u003c/sup\u003e was considered to be a new species of the genus \u003cem\u003eBrevibacterium\u003c/em\u003e, named \u003cem\u003eBrevibacterium litoralis\u003c/em\u003e sp. nov, with the type strain GXQ1321\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;MCCC 1K08964\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;KCTC 59167\u003csup\u003eT\u003c/sup\u003e).\u003c/p\u003e","manuscriptTitle":"Brevibacterium litoralis sp. nov., a cellulose-degrading strain isolated from marine surface sediment","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-06 12:34:17","doi":"10.21203/rs.3.rs-4724416/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-07-15T11:37:29+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-07-12T11:46:58+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-07-12T11:45:28+00:00","index":"","fulltext":""},{"type":"submitted","content":"Antonie van Leeuwenhoek","date":"2024-07-11T13:19:33+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"antonie-van-leeuwenhoek","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"anto","sideBox":"Learn more about [Antonie van Leeuwenhoek](https://www.springer.com/journal/10482)","snPcode":"10482","submissionUrl":"https://submission.nature.com/new-submission/10482/3","title":"Antonie van Leeuwenhoek","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"4525939d-84de-472b-bbee-553d6cacfcd6","owner":[],"postedDate":"August 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-10-21T16:01:46+00:00","versionOfRecord":{"articleIdentity":"rs-4724416","link":"https://doi.org/10.1007/s10482-024-02031-2","journal":{"identity":"antonie-van-leeuwenhoek","isVorOnly":false,"title":"Antonie van Leeuwenhoek"},"publishedOn":"2024-10-14 15:56:54","publishedOnDateReadable":"October 14th, 2024"},"versionCreatedAt":"2024-08-06 12:34:17","video":"","vorDoi":"10.1007/s10482-024-02031-2","vorDoiUrl":"https://doi.org/10.1007/s10482-024-02031-2","workflowStages":[]},"version":"v1","identity":"rs-4724416","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4724416","identity":"rs-4724416","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}