Alcanivorax beigongshangi sp. nov., isolated from the fermented grains of Chinese baijiu
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Abstract A bacterial strain, REN37T, was isolated from fermented grains of Baijiu samples collected from Sichuan, PR China. The cells of strain REN37T was Gram-negative and aerobic. The cellular morphology exhibited rod-shaped cells without flagellum, displaying non-motility. The optimal growth condition was at 32–37 oC, pH 6.0–7.0, and with a NaCl concentration of 1%-2% (w/v). Strain REN37T was positive for amylase, catalase and oxidase activities and aesculin, casein, starch hydrolysis. Based on the analysis of 16S rRNA gene sequence, strains REN37T was identified to belong to the genus of Alcanivorax. Its closest species was Alcanivorax pacificus W11-5T (96.1%). The polar lipids were identified to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, unidentified glycolipid, two unidentified phospholipids. The predominant menaquinone was MK-12. The predominant fatty acids were C15:0 anteiso (49.4%), C16:0 iso (18.1%), C17:0 anteiso (16.5%), C15:0 iso (9.5%). The digital DNA-DNA hybridization (dDDH), average nucleotide identity (OrthoANI) and average amino acid identify (AAI) values between strain REN37T and its most similar species were 19.2%, 74.0% and 92.0%, respectively. The DNA G + C content of the strain REN37T was 63.0 mol%. Based on the results, REN37T represents a novel strain, and the name Alcanivorax beigongshangi sp. nov. was proposed. The type strain is REN37T (= GDMCC 1.3120T = JCM 35319T).
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Alcanivorax beigongshangi sp. nov., isolated from the fermented grains of Chinese baijiu | 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 Alcanivorax beigongshangi sp. nov., isolated from the fermented grains of Chinese baijiu Tong-Xi Zhang, Yi-Ming Li, Hao-Yue Gu, Ru Zhang, Zhan-Bin Sun, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5108599/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 28 Nov, 2024 Read the published version in Antonie van Leeuwenhoek → Version 1 posted 4 You are reading this latest preprint version Abstract A bacterial strain, REN37 T , was isolated from fermented grains of Baijiu samples collected from Sichuan, PR China. The cells of strain REN37 T was Gram-negative and aerobic. The cellular morphology exhibited rod-shaped cells without flagellum, displaying non-motility. The optimal growth condition was at 32–37 o C, pH 6.0–7.0, and with a NaCl concentration of 1%-2% (w/v). Strain REN37 T was positive for amylase, catalase and oxidase activities and aesculin, casein, starch hydrolysis. Based on the analysis of 16S rRNA gene sequence, strains REN37 T was identified to belong to the genus of Alcanivorax. Its closest species was Alcanivorax pacificus W11-5 T (96.1%). The polar lipids were identified to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, unidentified glycolipid, two unidentified phospholipids. The predominant menaquinone was MK-12. The predominant fatty acids were C 15:0 anteiso (49.4%), C 16:0 iso (18.1%), C 17:0 anteiso (16.5%), C 15:0 iso (9.5%). The digital DNA-DNA hybridization (dDDH), average nucleotide identity (OrthoANI) and average amino acid identify (AAI) values between strain REN37 T and its most similar species were 19.2%, 74.0% and 92.0%, respectively. The DNA G + C content of the strain REN37 T was 63.0 mol%. Based on the results, REN37 T represents a novel strain, and the name Alcanivorax beigongshangi sp. nov. was proposed. The type strain is REN37 T (= GDMCC 1.3120 T = JCM 35319 T ). Alcanivorax beigongshangi baijiu fermented grains polyphasic taxonomy Figures Figure 1 Figure 2 INTRODUCTION Strong-flavor baijiu is a traditional Chinese fermented beverage produced from cereals through solid fermentation (Liu and Sun 2018 ). During the fermentation process, a variety of flavor compounds, including ethanol, aldehydes, esters and organic acids, are predominantly generated through microbial metabolism to endow baijiu with distinct flavors (Hu et al. 2021 ; Wei et al. 2022 ). The baijiu brewing process involves a complex interplay of diverse microorganisms. Research on microbial carriers of baijiu primarily focuses on pit mud, daqu and fermented grains (Zhao et al. 2023 ), with the latter serving as the ultimate environment where various microorganisms interact. The genus Alcanivorax , belonging to the family Alcanivoracaceae within the order Oceanospirillales and Class Gammaproteobacteria, currently encompass 18 species published ( http://www.bacterio.net/a/alcanivorax.html ) (Rivas et al. 2007 ; Yakimov et al. 1998 ; Zhu et al. 2021 ). Alcanivorax borkumensis , the first discovered strain in the genus of Alcanivorax , was initially isolated from seawater and sediment samples collected near Borkum Island in the North Sea (Yakimov et al. 1998 ). The description of the genus was subsequently revised by Ferna´ndez-Martı´nez et al. (2003). The Alcanivorax species, which primarily utilizes petroleum hydrocarbons as its major source of carbon and energy, has been employed in bioremediation interventions within contaminated marine and coastal systems (Meier-Kolthoff et al. 2013 ). Furthermore, this species holds potential for the production of biocatalysts in environmentally friendly industrial processes and for use as biosensors to enable in situ monitoring of aromatic or aliphatic compounds (Golyshin et al. 2003 ). In this study, a novel strain was isolated from the baijiu fermented grains, which was designated REN37 T . The classification of REN37 T was investigated through polyphasic taxonomy approach, which encompassed the analysis of morphological, physiological, biochemical chemotaxonomic and molecular biological characteristics. MATERIALS AND METHODS Isolation and culture conditions REN37 T was isolated from baijiu fermented grains collected from Sichuan Province, China (N 30 o 67'; E 104 o 06'). For strain isolating, 10 g sample was dispersed in 200 ml of sterile NaCl solution (0.85%, w/v), thoroughly mixed and then 100 µL was spread onto TSA (Trypticase Soy Agar) medium with a pH of 7.3 ± 0.2. The culture plates were aerobically incubated at 37 o C. Then, single colonies were picked and streaked three times in order to obtain pure strain. The isolated strain was preserved in 30% (v/v) glycerin and stored at -80 o C. Morphological, physiological, and biochemical characterisation After aerobically cultured on TSA medium at 37 o C for 48 h, cell morphological characteristics of strain REN37 T were observed using transmission electron microscopy (HT7700, Hitachi). The Gram’s staining was carried out according to the method described by Buck ( 1982 ). The pH tolerance (pH 5, 6, 7, 8, 9, 10 and 11), NaCl tolerance (concentrations of 0, 0.5, 1, 2, 2.5, 3, 1.5, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 17 and 19%, w/v) and temperature range (4, 15, 20, 25, 32, 37, 45 and 55 o C) were determined through measuring OD 600 values in TSB medium. The growth ability of REN37 T was determined in different media, including MB (Marine Broth), LB (Luria-Bertani Broth), NB (Nutrient Broth), TSB (trypsin Soy Broth), and R 2 A (Reasoner's 2A) medium, also through OD 600 measuring. Hydrolysis of starch, catalase and oxidase activities, casein utilisation, as well as the utilization of Tween 40 and Tween 80 were evaluated using established methodologies (Xu et al. 2005 ). Biochemical assays for assimilation, enzyme activity and antibiotic susceptibility were examined by inoculation with API 20NE, API ZYM strips (bioMerieux) according to the manufacturer's instructions. Chemotaxonomic characterisation REN37 T was cultured in TSB medium at 37 o C for 48 h for chemotaxonomic characterisation tests. Fatty acids of strain REN37 T were extracted and analysed through the Sherlock Microbial Identification System (MIDI) (Sasser 1990 ). The total polar lipids were stained with molybdenum blue, ninhydrin, phosphomolybdate, D reagent, 1-methylnaphthol reagents, respectively, and further separated via two-dimensional thin layer chromatography (TLC) (Minnikin et al. 1984 ). After the cells of REN37 T were lyophilised, cellular respiratory quinones were extracted, purified, and subsequently subjected to analysis using high performance liquid chromatography (HPLC) (Hu et al. 2001 ). Phylogenetic analysis based on 16S rRNA gene sequence and whole genome sequences Strain REN37 T was incubated on TSA plate at 37 o C for 48 h before DNA extraction. Genomic DNA of REN37 T was conducted by using a commercial bacterial DNA extraction kit (Tiangen, China) (Dai et al. 2015 ). The 16S rRNA gene was amplified using the primers of 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3'), followed by the sequencing at Sangon Biotech Co., Ltd. (Shanghai, China). The 16S rRNA of REN37 T was compared with its closely related species through the NCBI-BLAST algorithm ( https://blast.ncbi.nlm.nih.gov/Blast.cgi ). The 16S rRNA gene similarity between these strains was calculated by the EzBioCloud ( www.ezbiocloud.net/identify ). Then, REN37 T and species with high similarity were selected for phylogenetic tree construction with MEGA software (version 11.0) (Altschul et al. 1990 ; Yoon et al. 2017 ). For whole genome sequencing, the genomic DNA was quantified, purified and cut into 400–500 bp fragments. Then, sequencing libraries were prepared in cleaved fragments using the NEXTflex Rapid DNA-Seq kit, followed by the sequencing through the Illumina NovaSeq6000 sequencing platform at Majorbio Biopharm Technology Co., Ltd. (Shanghai, China). The raw reads obtained after sequencing were screened by fastp software (version 0.19.6) (Chen et al. 2018 ) and used for assembly by SOAPdenovo version 2.0 (Saitou and Nei 1987 ). Average nucleotide identity by orthology (OrthoANI) values between strain REN37 T and its similar strains were calculated using the OrthoANI tool (version 0.93.1). Gene annotation was performed via the Rapid Annotation using Subsystem Technology (RAST) server ( https://rast.nmpdr.org/ ; the Genetic Codes version uploaded on). The average amino acid identity (AAI) values were calculated using the AAI calculator ( http://enve-omics.ce.gatech.edu/aai/ ) (Rodriguezr and Konstantinidis 2016 ). Digital DNA-DNA hybridisation (dDDH) values were estimated using the Genome-to-Genome Distance Calculator (GGDC, version 3.0; http://ggdc.dsmz.de/ggdc.php ) with formula 2 (Meier-Kolthoff et al. 2013 ; Meier-Kolthoff et al. 2022 ; Meier-Kolthoff and Göker 2019 ). The whole Genome phylogenetic tree was constructed using the Type (Strain) Genome Server ( https://tygs.dsmz.de/ ) (Felsenstein 1981 ) and MEGA11 (Tamura et al. 2021 ) to assess the genome-based phylogeny. RESULTS Phylogenetic analysis based on 16S rRNA gene sequence After PCR amplification, the 16S rRNA gene sequence (1409 bp) was obtained and deposited in GenBank with the accession number of OR405985. Then, the 16S rRNA of REN37 T and its closely related species were compared by the NCBI-BLAST algorithm. Alcanivorax pacificus W11-5 T (96.10%) was found to share the highest similarity with REN37 T , followed by Alcanivorax indicus SW127 T (94.39%), Alcanivorax profundi MTEO17 T (93.19%), Alcanivorax mobilis MT13131 T (92.83%), Alcanivorax sediminis PA15-N-34 T (92.83%), Alcanivorax nanhaiticus 19-m-6 T (92.76%), Alcanivorax xenomutans JC109 T (92.71%), Alcanivorax dieselolei B-5 T (92.69%) and Alcanivorax venustensis ISO4 T (91.19%). The 16S rRNA gene similarity between strain REN37 T and its similar species were compared (Tables S2) and the phylogenetic tree based on their16S rRNA gene sequences were constructed (Fig. 1 ). Results showed that REN37 T formed a distinct cluster within the genus of Alcanivorax (Fig. 1 ). Therefore, above results indicated that the strain REN37 T belonged to Alcanivorax genus. Morphological, physiological, and biochemical characterisation The bacterial colonies of strain REN37 T exhibited a round and milky white appearance after 48 h of cultivation on TSA medium at 37 o C. Strain REN37 T was Gram-negative and aerobic. The cellular morphology exhibited rod-shaped cells lacking flagella, displaying non-motility, with a cell length and width of approximately 1.2–1.4 µm and 0.3–0.4 µm, respectively (Fig. S1 ). Similar species in Alcanivorax sp. also exhibit rod-shaped cell morphology, however, A. venustensis ISO4 T , A. dieselolei B-5 T , A. indicus SW127 T , A. mobilis MT13131 T and A. xenomutans JC109 T displayed motility (Fernández-Martínez et al. 2003 ; Liu and Shao 2005 ; Rahul et al. 2014 ; Song et al. 2018 ; Yang et al. 2018 ). Strain REN37 T could grow within the temperature range of 25–45 o C, pH range of 5.0–10.0, NaCl concentration range of 1.0%-5.0% (w/v). REN37 T was able to grow in MB, LB, NB, TSB and R 2 A medium, among which TSB was the optimal medium. The optimal growth condition was at 32–37 o C, in pH 6.0–7.0, and 1.0%-2.0% (w/v) NaCl. REN37 T was positive for catalase and oxidase tests, which was similar to A. dieselolei B-5 T , A. indicus SW127 T , A. venustensis ISO4 T , A. mobilis MT13131 T , A. nanhaiticus 19-m-6 T , A. pacificus W11-5 T and A. sediminis PA15-N-34 T (Fernández-Martínez et al. 2003 ; Lai et al. 2011 ; Lai et al. 2016 ; Liao et al. 2020 ; Liu and Shao 2005 ; Song et al. 2018 ; Yang et al. 2018 ). Besides, the positive reaction for amylase test was different from the reference strains of A. dieselolei B-5 T (Liu and Shao 2005 ), A. mobilis MT13131 T (Yang et al. 2018 ), A. sediminis PA15-N-34 T (Liao et al. 2020 )d xenomutans JC109 T (Rahul et al. 2014 ). Strain REN37 T exhibited the ability to hydrolyse aesculin, casein and starch, while showing no activity towards gelatin, Tween 40 and Tween 80. The results were consistent with the species of A. nanhaiticus 19-m-6 T (Lai et al. 2016 ) but different from A. sediminis PA15-N-34 T (Liao et al. 2020 ). REN37 T could assimilate trisodium CITrate, which was the same as A. xenomutans JC109 T (Rahul et al. 2014 ). Based on the API ZYM results, REN37 T produced L-cystyl-2-naphthylamide, 2-naphthyl myristate and L-valyl-2-naphthylamide, but did not produce Naphthol-AS-Bl-phosphate. In contrast, among its closely related species, A. dieselolei B-5 T (Liu and Shao 2005 ), A. nanhaiticus 19-m-6 T (Lai et al. 2016 )d sediminis PA15-N-34 T (Liao et al. 2020 ) could produce Naphthol-AS-Bl-phosphate. Chemotaxonomic characteristics C 15:0 anteiso (49.4%) is the major cellular fatty acid of strain REN37 T , followed by C 16:0 iso (18.1%), C 17:0 anteiso (16.5%) and C 15:0 iso (9.5%) (Tables S1). The polar lipids of strain REN37 T were identified to be diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylcholine (PC), unidentified glycolipid (GL), two unidentified phospholipids (PL1-2) (Fig. S2). Consistently, PG was existed in A. mobilis MT13131 T , A. nanhaiticus 19-m-6 T , A. profundi MTEO17 T (Liu et al. 2019 ), A. sediminis PA15-N-34 T , A. xenomutans JC109 T and DPG was existed in A. indicus SW127 T , A. xenomutans JC109 T (Table 1 ). The predominant respiratory quinone of strain REN37 T was MK-12. Table 1. Characteristics of REN37 T and other closely related species in the genus Alcanivorax . Characteristic 1 2 3 4 5 6 7 8 9 10 Isolated from fermented grains oil-contaminated sea water seawater filtered sea water deep-sea sediment deep sea sediment deep-sea sediment deep seawater deep-sea sediment shrimp cultivation pond sediment Gram staining - - - - - - - - + - Motility - + + + + - - - - + Cell shape rods rods rods rods rods short rods short rods rods rods short rods Optimal medium TSB HLB 216L agar MA MA 216L agar MA MA 216L agar NA Temperatures range( o C) 25-45 15-45 8-42 4-40 10-42 4-41 10-42 10-45 10-45 25-40 Temperatures preference(°C) 32-37 28-35 30.0 22-25 28 28 25-28 37.0 28-30 30 pH range 5-10 ND 5.5-9 ND 6-10 ND ND 6-10 6-10 4-9 pH preference 6-7 ND 7.5 ND 7-8 ND ND 7-8 7-8 6 NaCl tolerance range for growth (w/v%) 1.0-5.0 1.0-15.0 0.5-11.0 ND 1.0-12.0 0.5-15.0 0.5-12.0 0.0-12.0 0.0-15.0 0.5-20.0 Optimum NaCl concentration(w/v%) 1.0-2.0 3.0-7.5 3.0-4.0 10.0 3.0-4.0 3.0 3.0-5.0 3.0 3.0-5.0 2.0-5.0 NO 3 - reduction - + + - ND - + ND - - Tween 40 hydrolysis - + + + + ND + + + ND Tween 80 hydrolysis - + + + + - + + + + Gelatin hydrolysis - - + - - - - + + - Aesculin hydrolysis + + + ND - ND + + - - Casein hydrolysis + ND + - ND ND ND - ND + Starch hydrolysis + - - - - ND - - ND - Catalase + + + + + + + - + + Oxidase + + + + + + + + + w Amylase + - ND ND - ND ND ND - - CAPric acid assimilation - ND ND ND ND + - - ND ND ADIpic acid assimilation - ND ND ND + ND w ND ND ND MaLaTe assimilation - ND ND ND ND ND + ND ND ND trisodium CITrate assimilation + ND ND ND ND ND - ND ND + API ZYM L-cystyl-2-naphthylamide + ND ND ND ND ND ND ND ND ND 2-naphthyl myristate + ND ND ND ND ND ND ND ND ND L-valyl-2-naphthylamide + ND ND ND ND ND ND ND ND ND Naphthol-AS-Bl-phosphate - + ND ND ND + w ND + ND Polar lipids DPG, PG ND DPG, PE ND PG, PE PG, PE ND PE2, PG PE, PG DPG, PG DNA G+C content (mol%) 63.0 62.1-62.5 62.8 66.4 64.2 56.4 60.8 57.5 57.2 54.5 Strains: 1. REN37 T ; 2. A. dieselolei B-5 T (Liu and Shao 2005); 3. A. indicus SW127 T (Song et al. 2018); 4. A. venustensis ISO4 T (Fernández-Martínez et al. 2003); 5. A. mobilis MT13131 T (Yang et al. 2018); 6. A. nanhaiticus 19-m-6 T (Lai et al. 2016); 7. A. pacificus W11-5 T (Lai et al. 2011); 8. A. profundi MTEO17 T (Liu et al. 2019); 9. A. sediminis PA15-N-34 T (Liao et al. 2020); 10. A. xenomutans JC109 T (Rahul et al. 2014) . +, positive reaction or growth; -, no reaction or growth; w, weak reaction; ND, no data. TSB, trypsin Soy Broth medium; HLB, HLB was modified from Luria–Bertani (LB) medium; 216L agar, 216L marine agar medium; MA, marine agar medium; NA, Nutrient Agar medium. DPG, diphosphatidylglycerol; PG, phosphatidylglycerol; PE, phosphatidylethanolamin Genomic features and phylogenetic analysis based on whole genome sequences The whole genome sequence was submitted to GenBank with the accession number of GCA 041102775.1. Based on the genomic information analysis, the genome of REN37 T was characterised by a size of 3.02 Mbp and a DNA G + C content of 63.0 mol%. The OrthoANI, dDDH, and AAI values between REN37 T and its most similar strain of Alcanivorax pacificus W11-5 T were 74.0%, 19.2% and 92.0%, respectively. The values were lower than the threshold values of 95% (OrthoANI), 70% (dDDH), and 95% (AAI), indicating that strain REN37 T represented a novel species in the genus of Alcanivorax (Felsenstein 1981 ; Goris et al. 2007 ; Kim et al. 2014 ; Meier-Kolthoff et al. 2013 ; Richter and Rosselló-Móra 2009 ). Furthermore, Alcanivorax Among the annotated genes, 2.34 Mbp (77.48%) were assigned to functional categories. Rast server analysis showed that amino acids and derivatives (170) had the highest counts, followed by protein metabolism (168) and Cofactors, Vitamins, Prosthetic Groups, Pigments (146) (Fig. S3). The whole genome sequence of strain REN37 T was compared with its similar strains in the corresponding genus (Table S3), and phylogenetic analysis was performed. Results showed that strain REN37 T also formed a separate clade within the genus of Alcanivorax (Fig. 2 ). Description of Alcanivorax beigongshangi sp. nov. Alcanivorax beigongshangi (bei. gong. shang'i. N.L. gen. n. beigongshangi , as Beijing Technology and Business University, the strain was cultured and isolated in Beijing Technology and Business University). The strain is Gram-negative, aerobic. Cells of REN37 T are rod-shaped (1.2–1.4 µm in length and 0.3–0.4 µm in width), lacking flagella, and thus displaying non-motility. It can grow under 25–45 o C, pH5-10, 1%-5% (w/v) NaCl and in MB, LB, NB, TSB and R 2 A medium. The optimal growth condition is 32–37 o C, pH 6–7, NaCl concentration 1%-2% (w/v) in TSB medium. Based on the API ZYM results, REN37 T produces 2-naphthyl myristate, L-valyl-2-naphthylamide, L-cystyl-2-naphthylamide, N-benzoyl-DL-arginine-2-naphthylamide, N-glutaryl-phenylalanine-2-naphthylamide, 6-Br-2-naphthyl-αD-galactopyranoside, 2-naphthyl-αD-galactopyranoside, Naphthol-AS-Bl-SD-glucuronide, 2-naphthyl-αD-glucopyranoside, 6-Br-2-naphthyl-BD-glucopyranoside, 1-naphthyl-N-acetyl-BD-glucosaminide, 6-Br-2-naphthyl-αD-mannopyranoside, 2-naphthyl-aL-fucopyranoside. Positive for amylase, catalase, oxidase and hydrolysis aesculin, casein, starch. Assimilation trisodium CITrate. Major fatty acids are C 14:0 iso, C 14:0 anteiso, C 14:0 , C 15:1 anteiso A, C 15:0 iso, C 15:0 anteiso, C 16:0 iso, C 16:0 , C 15:0 3OH, C 17:0 iso, C 17:0 anteiso, C 17:1 w5c, C 17:0 , C 18:0 , C 17:0 3OH, C 19:0 anteiso. The major polar lipids are DPG, PL1-2, PG, GL and PC. The major respiratory quinone of strain REN37 T is MK-12 (100%). The genomic DNA G + C content is 63.0 mol%. The type strain, REN37 T (= GDMCC 1.3120 T = JCM 35319 T ), was isolated from Baijiu fermented grains from Sichuan, PR China. The GenBank accession numbers for the 16S rRNA gene sequence and the whole genome sequence is OR405985 and GCA 041102775.1, respectively. Abbreviations OrthoANI , average nucleotide identity by Orthology; dDDH , digital DNA-DNA hybridization; AAI , average amino acid identity; TSB , trypsin Soy Broth medium; LB, Luria-Bertani Broth; NB, Nutrient Broth; R 2 A , Reasoner's 2A; HLB , HLB was modified from Luria-Bertani ; 216L agar , 216L marine agar; MA , marine agar; NA , Nutrient Agar; DPG , diphosphatidylglycerol; PG , phosphatidylglycerol; PE , phosphatidylethanolamine; PC , phosphatidylcholine; GL , unidentified glycolipid; PL , phospholipid. Declarations Funding: the authors declare that no funds, grants, or other support were received during the preparation of this manuscript. Competing interests : the authors declare that there are no conflicts of interest. Author Contributions Qing Ren and Han-Xu Pan contributed to the study conception and design. Material preparation, data collection and analysis were performed by Yi-Ming Li, Hao-Yue Gu, Ru Zhang and Zhan-Bin Sun. The first draft of the manuscript was written by Tong-xi Zhang and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Data Availability : all data have been made fully available to the public. The GenBank accession numbers for the 16S rRNA sequence and the full genome sequence of Alcanivorax beigongshangi REN37 T are OR405985 and GCA 041102775.1, respectively. Ethical approval: this article does not contain any studies with human participants or animals performed by any of the authors. Consent to participate : all authors gave their consent to participate in this study. 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Int J Syst Evol Microbiol 70:4280-4284. https://doi.org/10.1099/ijsem.0.004285 Liu C, Shao Z (2005) Alcanivorax dieselolei sp. Nov., A novel alkane-degrading bacterium isolated from sea water and deep-sea sediment. Int J Syst Evol Microbiol 55:1181-1186. https://doi.org/10.1099/ijs.0.63443-0 Liu H, Sun B (2018) Effect of Fermentation Processing on the Flavor of Baijiu. J Agric Food Chem 66:5425-5432. https://doi.org/10.1021/acs.jafc.8b00692 Liu J, Ren Q, Zhang Y, Li Y, Tian X, Wu Y, Tian J, Zhang XH (2019) Alcanivorax profundi sp. Nov., Isolated from deep seawater of the Mariana Trench. Int J Syst Evol Microbiol 69:371-376. https://doi.org/10.1099/ijsem.0.003145 Meier-Kolthoff JP, Auch AF, Klenk HP, Göker M (2013) Genome sequence-based species delimitation with confidence intervals and improved distance functions. Bmc Bioinformatics 14: 60.https:// doi.org/ 10.1186/1471-2105-14-60 Meier-Kolthoff JP, Carbasse JS, Peinado-Olarte RL, Göker M (2022) TYGS and LPSN: a database tandem for fast and reliable genome-based classification and nomenclature of prokaryotes. Nucleic Acids Res 50:D801-D807. https://doi.org/10.1093/nar/gkab902 Meier-Kolthoff JP, Göker M (2019) TYGS is an automated high-throughput platform for state-of-the-art genome-based taxonomy. Nat Commun 10:2182. https://doi.org/10.1038/s41467-019-10210-3 Minnikin DE, O'Donnell AG, Goodfellow M, Alderson G, Athalye M, Schaal A, Parlett JH (1984) An integrated procedure for the extraction of bacterial isoprenoid quinones and polar lipids. J Microbiol Methods 2:233-241. https://doi.org/https://doi.org/10.1016/0167-7012(84)90018-6 Rahul K, Sasikala C, Tushar L, Debadrita R, Ramana CV (2014) Alcanivorax xenomutans sp. Nov., A hydrocarbonoclastic bacterium isolated from a shrimp cultivation pond. Int J Syst Evol Microbiol 64:3553-3558. https://doi.org/10.1099/ijs.0.061168-0 Richter M, Rosselló-Móra R (2009) Shifting the genomic gold standard for the prokaryotic species definition. Proc Natl Acad Sci U S A 106:19126-19131. https://doi.org/10.1073/pnas.0906412106 Rivas R, García-Fraile P, Peix A, Mateos PF, Martínez-Molina E, Velázquez E (2007) Alcanivorax balearicus sp. Nov., Isolated from Lake Martel. Int J Syst Evol Microbiol 57:1331-1335. https://doi.org/10.1099/ijs.0.64912-0 Rodriguezr LM, Konstantinidis KT (2016) The enveomics collection: a toolbox for specialized analyses of microbial genomes and metagenomes Saitou NNM, Nei MC (1987) Saitou n, nei m.. The Neighbor-Joining Method-a New Method for Reconstructing Phylogenetic Trees. Mol biol evol 4:406-425. Mol Biol Evol 4:406-425 Sasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids. Usfcc Newsl Song L, Liu H, Cai S, Huang Y, Dai X, Zhou Y (2018) Alcanivorax indicus sp. Nov., Isolated from seawater. Int J Syst Evol Microbiol 68:3785-3789. https://doi.org/10.1099/ijsem.0.003058 Tamura K, Stecher G, Kumar S (2021) MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol 38:3022-3027. https://doi.org/10.1093/molbev/msab120 Wei CH, Zheng ZQ, Li H (2022) Spatial and temporal differences of flavor substances during fermentation of Luzhou-flavor liquor. Food and Fermentation Industry 48:240-246. https://doi.org/10.13995/j.cnki.11-1802/ts.027617 Xu P, Li WJ, Tang SK, Zhang YQ, Chen GZ, Chen HH, Xu LH, Jiang CL (2005) Naxibacter alkalitolerans gen. Nov., Sp. Nov., A novel member of the family 'Oxalobacteraceae' isolated from China. Int J Syst Evol Microbiol 55:1149-1153. https://doi.org/10.1099/ijs.0.63407-0 Yakimov MM, Golyshin PN, Lang S, Moore ER, Abraham WR, Lünsdorf H, Timmis KN (1998) Alcanivorax borkumensis gen. Nov., Sp. Nov., A new, hydrocarbon-degrading and surfactant-producing marine bacterium. Int J Syst Bacteriol 48 Pt 2:339-348. https://doi.org/10.1099/00207713-48-2-339 Yang S, Li M, Lai Q, Li G, Shao Z (2018) Alcanivorax mobilis sp. Nov., A new hydrocarbon-degrading bacterium isolated from deep-sea sediment. Int J Syst Evol Microbiol 68:1639-1643. https://doi.org/10.1099/ijsem.0.002612 Yoon SH, Ha SM, Kwon S, Lim J, Kim Y, Seo H, Chun J (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 Zhao QZ, Zhang MM, Miao KC (2023) Research status and visual analysis of microorganism in liquor brewing based on bibliometrics. Food Industry Science and Technology 44:492-500. https://doi.org/10.13386/j.issn1002-0306.2022120042 Zhu L, Wang Y, Ding Y, Luo K, Yang B, Yang S, Liu S, Cui H, Wei W (2021) Alcanivorax limicola sp. Nov., Isolated from a soda alkali-saline soil. Arch Microbiol 204:106. https://doi.org/10.1007/s00203-021-02638-3 Additional Declarations No competing interests reported. Supplementary Files SupplementalMaterials.docx Cite Share Download PDF Status: Published Journal Publication published 28 Nov, 2024 Read the published version in Antonie van Leeuwenhoek → Version 1 posted Editorial decision: Revision requested 21 Sep, 2024 Editor assigned by journal 20 Sep, 2024 Submission checks completed at journal 20 Sep, 2024 First submitted to journal 18 Sep, 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. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5108599","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":357174498,"identity":"9f08fd12-ec46-4eac-b019-1b09c308db3f","order_by":0,"name":"Tong-Xi Zhang","email":"","orcid":"","institution":"Beijing Technology and Business University","correspondingAuthor":false,"prefix":"","firstName":"Tong-Xi","middleName":"","lastName":"Zhang","suffix":""},{"id":357174499,"identity":"d7aa2d10-47aa-40a3-8e1d-96ef46fb94fd","order_by":1,"name":"Yi-Ming Li","email":"","orcid":"","institution":"Beijing Technology and Business University","correspondingAuthor":false,"prefix":"","firstName":"Yi-Ming","middleName":"","lastName":"Li","suffix":""},{"id":357174501,"identity":"6f9d25a6-d6ea-4492-9312-647d1ecbe4e5","order_by":2,"name":"Hao-Yue Gu","email":"","orcid":"","institution":"Beijing Technology and Business University","correspondingAuthor":false,"prefix":"","firstName":"Hao-Yue","middleName":"","lastName":"Gu","suffix":""},{"id":357174504,"identity":"65a028ea-03d3-4d24-b390-37ffe2916b4f","order_by":3,"name":"Ru Zhang","email":"","orcid":"","institution":"Beijing Technology and Business University","correspondingAuthor":false,"prefix":"","firstName":"Ru","middleName":"","lastName":"Zhang","suffix":""},{"id":357174505,"identity":"794bce0f-d743-49c3-9eed-29508fd54f96","order_by":4,"name":"Zhan-Bin Sun","email":"","orcid":"","institution":"Beijing Technology and Business University","correspondingAuthor":false,"prefix":"","firstName":"Zhan-Bin","middleName":"","lastName":"Sun","suffix":""},{"id":357174506,"identity":"e794470b-1e92-43d9-b326-d4d8f6b05df2","order_by":5,"name":"Han-Xu Pan","email":"","orcid":"","institution":"Beijing Technology and Business University","correspondingAuthor":false,"prefix":"","firstName":"Han-Xu","middleName":"","lastName":"Pan","suffix":""},{"id":357174507,"identity":"e92792aa-291a-4c66-a9d1-4fa5b0d1a72a","order_by":6,"name":"Qing Ren","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6ElEQVRIie3RuwrCMBSA4VOEdgm4RhR9hYiggoOv0tDBpYKroNIpXdLd13DrZkOhLtG5borg5OLYzbZeNtuOgvmHEwL5OEMAVKpfzQRoAwYdNOd1rUJ6KCOiMkmjvDIZuJ6Iz/5ysm06+uXuh1A3bAKJ/5205MEaUrmb8lZgECFDaPAb0Tz5nWBs9zFl0ZRjU8eChUBim9Q0VkA6t5xM0JuMSwlGGVmYH0JwGUF2b0hZ0OUxdcleptvkdSa8ImLI7jFhq46xtqLT3B+16661OSUFJGcAYXo8vx5lIygGOVmVvVGpVKp/7gGBgVBdMrUP5gAAAABJRU5ErkJggg==","orcid":"","institution":"Beijing Technology and Business University","correspondingAuthor":true,"prefix":"","firstName":"Qing","middleName":"","lastName":"Ren","suffix":""}],"badges":[],"createdAt":"2024-09-18 09:07:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5108599/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5108599/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10482-024-02043-y","type":"published","date":"2024-11-28T15:57:17+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":69002982,"identity":"dc60dfca-fcf5-41ac-ba28-84e2670c3311","added_by":"auto","created_at":"2024-11-14 12:01:33","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":3156998,"visible":true,"origin":"","legend":"\u003cp\u003eThe relationship between \u003cem\u003eAlcanivorax beigongshangi\u003c/em\u003e (REN37\u003csup\u003eT\u003c/sup\u003e) and related types of strains of the genus \u003cem\u003eAlcanivorax\u003c/em\u003e was constructed based on the adjacent phylogenetic tree of 16S rRNA gene sequences. \u003cem\u003eRhodospirillum centenum\u003c/em\u003e DSM 8284\u003csup\u003eT\u003c/sup\u003e (D12701.1) was used as the outgroup. Bootstrap values above 50% are represented on the branch node. The GenBank accession number for the 16S rRNA sequences of the species is provided in parentheses. Bar, 0.01, substitutions per nucleotide position.\u003c/p\u003e","description":"","filename":"Fig.1.png","url":"https://assets-eu.researchsquare.com/files/rs-5108599/v1/8842c80fdcc9df2ba5c6d021.png"},{"id":69002984,"identity":"271a7dbb-153d-4357-b5c2-c9b65005f43f","added_by":"auto","created_at":"2024-11-14 12:01:33","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":6166615,"visible":true,"origin":"","legend":"\u003cp\u003eThe relationship between \u003cem\u003eAlcanivorax beigongshangi\u003c/em\u003e (REN37\u003csup\u003eT\u003c/sup\u003e) and related types of strains of the genus \u003cem\u003eAlcanivorax\u003c/em\u003e was constructed based on the adjacent phylogenetic tree of whole genome sequences. The distance to GBDP was calculated based on the genome sequence and the tree was inferred by FastME 2.1.6.1 (Kreft et al. 2017), and the length of the branches was scaled according to the formula d5. \u003cem\u003eRhodospirillum centenum\u003c/em\u003e DSM 8284\u003csup\u003eT\u003c/sup\u003e (GCA 000016185.1) was used as the outgroup. Bootstrap values above 50% are represented on the branch node. The GenBank accession number for the whole genome sequences of the species is provided in parentheses. Bar, 0.02, substitutions per nucleotide position.\u003c/p\u003e","description":"","filename":"Fig.2.png","url":"https://assets-eu.researchsquare.com/files/rs-5108599/v1/1689d3760e5f6f75c238071e.png"},{"id":70382159,"identity":"22ca4919-e10c-4073-b46f-dc44967227ba","added_by":"auto","created_at":"2024-12-02 16:24:49","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8182375,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5108599/v1/1b7f00e5-81e8-42d2-872f-1c291bd851e5.pdf"},{"id":69002985,"identity":"9bbe978c-f9fd-4106-acd2-10a1a6befcba","added_by":"auto","created_at":"2024-11-14 12:01:33","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2729170,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementalMaterials.docx","url":"https://assets-eu.researchsquare.com/files/rs-5108599/v1/7da86baff918273309d74183.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Alcanivorax beigongshangi sp. nov., isolated from the fermented grains of Chinese baijiu","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eStrong-flavor baijiu is a traditional Chinese fermented beverage produced from cereals through solid fermentation (Liu and Sun \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). During the fermentation process, a variety of flavor compounds, including ethanol, aldehydes, esters and organic acids, are predominantly generated through microbial metabolism to endow baijiu with distinct flavors (Hu et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The baijiu brewing process involves a complex interplay of diverse microorganisms. Research on microbial carriers of baijiu primarily focuses on pit mud, daqu and fermented grains (Zhao et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), with the latter serving as the ultimate environment where various microorganisms interact. The genus \u003cem\u003eAlcanivorax\u003c/em\u003e, belonging to the family Alcanivoracaceae within the order Oceanospirillales and Class Gammaproteobacteria, currently encompass 18 species published (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.bacterio.net/a/alcanivorax.html\u003c/span\u003e\u003cspan address=\"http://www.bacterio.net/a/alcanivorax.html\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) (Rivas et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Yakimov et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Zhu et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). \u003cem\u003eAlcanivorax borkumensis\u003c/em\u003e, the first discovered strain in the genus of \u003cem\u003eAlcanivorax\u003c/em\u003e, was initially isolated from seawater and sediment samples collected near Borkum Island in the North Sea (Yakimov et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). The description of the genus was subsequently revised by Ferna\u0026acute;ndez-Martı\u0026acute;nez et al. (2003). The \u003cem\u003eAlcanivorax\u003c/em\u003e species, which primarily utilizes petroleum hydrocarbons as its major source of carbon and energy, has been employed in bioremediation interventions within contaminated marine and coastal systems (Meier-Kolthoff et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Furthermore, this species holds potential for the production of biocatalysts in environmentally friendly industrial processes and for use as biosensors to enable \u003cem\u003ein situ\u003c/em\u003e monitoring of aromatic or aliphatic compounds (Golyshin et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). In this study, a novel strain was isolated from the baijiu fermented grains, which was designated REN37\u003csup\u003eT\u003c/sup\u003e. The classification of REN37\u003csup\u003eT\u003c/sup\u003e was investigated through polyphasic taxonomy approach, which encompassed the analysis of morphological, physiological, biochemical chemotaxonomic and molecular biological characteristics.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eIsolation and culture conditions\u003c/h2\u003e\n \u003cp\u003eREN37\u003csup\u003eT\u003c/sup\u003e was isolated from baijiu fermented grains collected from Sichuan Province, China (N 30\u003csup\u003eo\u003c/sup\u003e67\u0026apos;; E 104\u003csup\u003eo\u003c/sup\u003e 06\u0026apos;). For strain isolating, 10 g sample was dispersed in 200 ml of sterile NaCl solution (0.85%, w/v), thoroughly mixed and then 100 \u0026micro;L was spread onto TSA (Trypticase Soy Agar) medium with a pH of 7.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2. The culture plates were aerobically incubated at 37\u003csup\u003eo\u003c/sup\u003eC. Then, single colonies were picked and streaked three times in order to obtain pure strain. The isolated strain was preserved in 30% (v/v) glycerin and stored at -80 \u003csup\u003eo\u003c/sup\u003eC.\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eMorphological, physiological, and biochemical characterisation \u003c/h3\u003e\n\u003cp\u003eAfter aerobically cultured on TSA medium at 37 \u003csup\u003eo\u003c/sup\u003eC for 48 h, cell morphological characteristics of strain REN37\u003csup\u003eT\u003c/sup\u003e were observed using transmission electron microscopy (HT7700, Hitachi). The Gram\u0026rsquo;s staining was carried out according to the method described by Buck (\u003cspan class=\"CitationRef\"\u003e1982\u003c/span\u003e). The pH tolerance (pH 5, 6, 7, 8, 9, 10 and 11), NaCl tolerance (concentrations of 0, 0.5, 1, 2, 2.5, 3, 1.5, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 17 and 19%, w/v) and temperature range (4, 15, 20, 25, 32, 37, 45 and 55 \u003csup\u003eo\u003c/sup\u003eC) were determined through measuring OD\u003csub\u003e600\u003c/sub\u003e values in TSB medium. The growth ability of REN37\u003csup\u003eT\u003c/sup\u003e was determined in different media, including MB (Marine Broth), LB (Luria-Bertani Broth), NB (Nutrient Broth), TSB (trypsin Soy Broth), and R\u003csub\u003e2\u003c/sub\u003eA (Reasoner\u0026apos;s 2A) medium, also through OD\u003csub\u003e600\u003c/sub\u003e measuring. Hydrolysis of starch, catalase and oxidase activities, casein utilisation, as well as the utilization of Tween 40 and Tween 80 were evaluated using established methodologies (Xu et al. \u003cspan class=\"CitationRef\"\u003e2005\u003c/span\u003e). Biochemical assays for assimilation, enzyme activity and antibiotic susceptibility were examined by inoculation with API 20NE, API ZYM strips (bioMerieux) according to the manufacturer\u0026apos;s instructions.\u003c/p\u003e\n\u003ch3\u003eChemotaxonomic characterisation\u003c/h3\u003e\n\u003cp\u003eREN37\u003csup\u003eT\u003c/sup\u003e was cultured in TSB medium at 37 \u003csup\u003eo\u003c/sup\u003eC for 48 h for chemotaxonomic characterisation tests. Fatty acids of strain REN37\u003csup\u003eT\u003c/sup\u003e were extracted and analysed through the Sherlock Microbial Identification System (MIDI) (Sasser \u003cspan class=\"CitationRef\"\u003e1990\u003c/span\u003e). The total polar lipids were stained with molybdenum blue, ninhydrin, phosphomolybdate, D reagent, 1-methylnaphthol reagents, respectively, and further separated via two-dimensional thin layer chromatography (TLC) (Minnikin et al. \u003cspan class=\"CitationRef\"\u003e1984\u003c/span\u003e). After the cells of REN37\u003csup\u003eT\u003c/sup\u003e were lyophilised, cellular respiratory quinones were extracted, purified, and subsequently subjected to analysis using high performance liquid chromatography (HPLC) (Hu et al. \u003cspan class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003ePhylogenetic analysis based on 16S rRNA gene sequence and whole genome sequences\u003c/h3\u003e\n\u003cp\u003eStrain REN37\u003csup\u003eT\u003c/sup\u003e was incubated on TSA plate at 37 \u003csup\u003eo\u003c/sup\u003eC for 48 h before DNA extraction. Genomic DNA of REN37\u003csup\u003eT\u003c/sup\u003e was conducted by using a commercial bacterial DNA extraction kit (Tiangen, China) (Dai et al. \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e). The 16S rRNA gene was amplified using the primers of 27F (5\u0026apos;-AGAGTTTGATCCTGGCTCAG-3\u0026apos;) and 1492R (5\u0026apos;-TACGGCTACCTTGTTACGACTT-3\u0026apos;), followed by the sequencing at Sangon Biotech Co., Ltd. (Shanghai, China). The 16S rRNA of REN37\u003csup\u003eT\u003c/sup\u003e was compared with its closely related species through the NCBI-BLAST algorithm (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://blast.ncbi.nlm.nih.gov/Blast.cgi\u003c/span\u003e\u003c/span\u003e). The 16S rRNA gene similarity between these strains was calculated by the EzBioCloud (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ewww.ezbiocloud.net/identify\u003c/span\u003e\u003c/span\u003e). Then, REN37\u003csup\u003eT\u003c/sup\u003e and species with high similarity were selected for phylogenetic tree construction with MEGA software (version 11.0) (Altschul et al. \u003cspan class=\"CitationRef\"\u003e1990\u003c/span\u003e; Yoon et al. \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eFor whole genome sequencing, the genomic DNA was quantified, purified and cut into 400\u0026ndash;500 bp fragments. Then, sequencing libraries were prepared in cleaved fragments using the NEXTflex Rapid DNA-Seq kit, followed by the sequencing through the Illumina NovaSeq6000 sequencing platform at Majorbio Biopharm Technology Co., Ltd. (Shanghai, China). The raw reads obtained after sequencing were screened by fastp software (version 0.19.6) (Chen et al. \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e) and used for assembly by SOAPdenovo version 2.0 (Saitou and Nei \u003cspan class=\"CitationRef\"\u003e1987\u003c/span\u003e). Average nucleotide identity by orthology (OrthoANI) values between strain REN37\u003csup\u003eT\u003c/sup\u003e and its similar strains were calculated using the OrthoANI tool (version 0.93.1). Gene annotation was performed via the Rapid Annotation using Subsystem Technology (RAST) server (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://rast.nmpdr.org/\u003c/span\u003e\u003c/span\u003e; the Genetic Codes version uploaded on). The average amino acid identity (AAI) values were calculated using the AAI calculator (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://enve-omics.ce.gatech.edu/aai/\u003c/span\u003e\u003c/span\u003e) (Rodriguezr and Konstantinidis \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). Digital DNA-DNA hybridisation (dDDH) values were estimated using the Genome-to-Genome Distance Calculator (GGDC, version 3.0; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://ggdc.dsmz.de/ggdc.php\u003c/span\u003e\u003c/span\u003e) with formula 2 (Meier-Kolthoff et al. \u003cspan class=\"CitationRef\"\u003e2013\u003c/span\u003e; Meier-Kolthoff et al. \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e; Meier-Kolthoff and G\u0026ouml;ker \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). The whole Genome phylogenetic tree was constructed using the Type (Strain) Genome Server (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://tygs.dsmz.de/\u003c/span\u003e\u003c/span\u003e) (Felsenstein \u003cspan class=\"CitationRef\"\u003e1981\u003c/span\u003e) and MEGA11 (Tamura et al. \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e) to assess the genome-based phylogeny.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec8\"\u003e\n \u003ch2\u003ePhylogenetic analysis based on 16S rRNA gene sequence\u003c/h2\u003e\n \u003cp\u003eAfter PCR amplification, the 16S rRNA gene sequence (1409 bp) was obtained and deposited in GenBank with the accession number of OR405985. Then, the 16S rRNA of REN37\u003csup\u003eT\u003c/sup\u003e and its closely related species were compared by the NCBI-BLAST algorithm. \u003cem\u003eAlcanivorax pacificus\u003c/em\u003e W11-5\u003csup\u003eT\u003c/sup\u003e (96.10%) was found to share the highest similarity with REN37\u003csup\u003eT\u003c/sup\u003e, followed by \u003cem\u003eAlcanivorax indicus\u003c/em\u003e SW127\u003csup\u003eT\u003c/sup\u003e (94.39%), \u003cem\u003eAlcanivorax profundi\u003c/em\u003e MTEO17\u003csup\u003eT\u003c/sup\u003e (93.19%), \u003cem\u003eAlcanivorax mobilis\u003c/em\u003e MT13131\u003csup\u003eT\u003c/sup\u003e (92.83%), \u003cem\u003eAlcanivorax sediminis\u003c/em\u003e PA15-N-34\u003csup\u003eT\u003c/sup\u003e (92.83%), \u003cem\u003eAlcanivorax nanhaiticus\u003c/em\u003e 19-m-6\u003csup\u003eT\u003c/sup\u003e (92.76%), \u003cem\u003eAlcanivorax xenomutans\u003c/em\u003e JC109\u003csup\u003eT\u003c/sup\u003e (92.71%), \u003cem\u003eAlcanivorax dieselolei\u003c/em\u003e B-5\u003csup\u003eT\u003c/sup\u003e (92.69%) and \u003cem\u003eAlcanivorax venustensis\u003c/em\u003e ISO4\u003csup\u003eT\u003c/sup\u003e (91.19%). The 16S rRNA gene similarity between strain REN37\u003csup\u003eT\u003c/sup\u003e and its similar species were compared (Tables S2) and the phylogenetic tree based on their16S rRNA gene sequences were constructed (Fig.\u0026nbsp;\u003cspan\u003e1\u003c/span\u003e). Results showed that REN37\u003csup\u003eT\u003c/sup\u003e formed a distinct cluster within the genus of \u003cem\u003eAlcanivorax\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan\u003e1\u003c/span\u003e). Therefore, above results indicated that the strain REN37\u003csup\u003eT\u003c/sup\u003e belonged to \u003cem\u003eAlcanivorax\u003c/em\u003e genus.\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eMorphological, physiological, and biochemical characterisation\u003c/h3\u003e\n\u003cp\u003eThe bacterial colonies of strain REN37\u003csup\u003eT\u003c/sup\u003e exhibited a round and milky white appearance after 48 h of cultivation on TSA medium at 37 \u003csup\u003eo\u003c/sup\u003eC. Strain REN37\u003csup\u003eT\u003c/sup\u003e was Gram-negative and aerobic. The cellular morphology exhibited rod-shaped cells lacking flagella, displaying non-motility, with a cell length and width of approximately 1.2\u0026ndash;1.4 \u0026micro;m and 0.3\u0026ndash;0.4 \u0026micro;m, respectively (Fig. \u003cspan\u003eS1\u003c/span\u003e). Similar species in \u003cem\u003eAlcanivorax\u003c/em\u003e sp. also exhibit rod-shaped cell morphology, however, \u003cem\u003eA. venustensis\u003c/em\u003e ISO4\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. dieselolei\u003c/em\u003e B-5\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. indicus\u003c/em\u003e SW127\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. mobilis\u003c/em\u003e MT13131\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eA. xenomutans\u003c/em\u003e JC109\u003csup\u003eT\u003c/sup\u003e displayed motility (Fern\u0026aacute;ndez-Mart\u0026iacute;nez et al. \u003cspan\u003e2003\u003c/span\u003e; Liu and Shao \u003cspan\u003e2005\u003c/span\u003e; Rahul et al. \u003cspan\u003e2014\u003c/span\u003e; Song et al. \u003cspan\u003e2018\u003c/span\u003e; Yang et al. \u003cspan\u003e2018\u003c/span\u003e). Strain REN37\u003csup\u003eT\u003c/sup\u003e could grow within the temperature range of 25\u0026ndash;45 \u003csup\u003eo\u003c/sup\u003eC, pH range of 5.0\u0026ndash;10.0, NaCl concentration range of 1.0%-5.0% (w/v). REN37\u003csup\u003eT\u003c/sup\u003e was able to grow in MB, LB, NB, TSB and R\u003csub\u003e2\u003c/sub\u003eA medium, among which TSB was the optimal medium. The optimal growth condition was at 32\u0026ndash;37 \u003csup\u003eo\u003c/sup\u003eC, in pH 6.0\u0026ndash;7.0, and 1.0%-2.0% (w/v) NaCl. REN37\u003csup\u003eT\u003c/sup\u003e was positive for catalase and oxidase tests, which was similar to \u003cem\u003eA. dieselolei\u003c/em\u003e B-5\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. indicus\u003c/em\u003e SW127\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. venustensis\u003c/em\u003e ISO4\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. mobilis\u003c/em\u003e MT13131\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. nanhaiticus\u003c/em\u003e 19-m-6\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. pacificus\u003c/em\u003e W11-5\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eA. sediminis\u003c/em\u003e PA15-N-34\u003csup\u003eT\u003c/sup\u003e (Fern\u0026aacute;ndez-Mart\u0026iacute;nez et al. \u003cspan\u003e2003\u003c/span\u003e; Lai et al. \u003cspan\u003e2011\u003c/span\u003e; Lai et al. \u003cspan\u003e2016\u003c/span\u003e; Liao et al. \u003cspan\u003e2020\u003c/span\u003e; Liu and Shao \u003cspan\u003e2005\u003c/span\u003e; Song et al. \u003cspan\u003e2018\u003c/span\u003e; Yang et al. \u003cspan\u003e2018\u003c/span\u003e). Besides, the positive reaction for amylase test was different from the reference strains of \u003cem\u003eA. dieselolei\u003c/em\u003e B-5\u003csup\u003eT\u003c/sup\u003e (Liu and Shao \u003cspan\u003e2005\u003c/span\u003e), \u003cem\u003eA. mobilis\u003c/em\u003e MT13131\u003csup\u003eT\u003c/sup\u003e (Yang et al. \u003cspan\u003e2018\u003c/span\u003e), A. \u003cem\u003esediminis\u003c/em\u003e PA15-N-34\u003csup\u003eT\u003c/sup\u003e (Liao et al. \u003cspan\u003e2020\u003c/span\u003e)d \u003cem\u003exenomutans\u003c/em\u003e JC109\u003csup\u003eT\u003c/sup\u003e (Rahul et al. \u003cspan\u003e2014\u003c/span\u003e). Strain REN37\u003csup\u003eT\u003c/sup\u003e exhibited the ability to hydrolyse aesculin, casein and starch, while showing no activity towards gelatin, Tween 40 and Tween 80. The results were consistent with the species of \u003cem\u003eA. nanhaiticus\u003c/em\u003e 19-m-6\u003csup\u003eT\u003c/sup\u003e (Lai et al. \u003cspan\u003e2016\u003c/span\u003e) but different from \u003cem\u003eA. sediminis\u003c/em\u003e PA15-N-34\u003csup\u003eT\u003c/sup\u003e (Liao et al. \u003cspan\u003e2020\u003c/span\u003e). REN37\u003csup\u003eT\u003c/sup\u003e could assimilate trisodium CITrate, which was the same as \u003cem\u003eA. xenomutans\u003c/em\u003e JC109\u003csup\u003eT\u003c/sup\u003e (Rahul et al. \u003cspan\u003e2014\u003c/span\u003e). Based on the API ZYM results, REN37\u003csup\u003eT\u003c/sup\u003e produced L-cystyl-2-naphthylamide, 2-naphthyl myristate and L-valyl-2-naphthylamide, but did not produce Naphthol-AS-Bl-phosphate. In contrast, among its closely related species, \u003cem\u003eA. dieselolei\u003c/em\u003e B-5\u003csup\u003eT\u003c/sup\u003e (Liu and Shao \u003cspan\u003e2005\u003c/span\u003e), \u003cem\u003eA. nanhaiticus\u003c/em\u003e 19-m-6\u003csup\u003eT\u003c/sup\u003e (Lai et al. \u003cspan\u003e2016\u003c/span\u003e)d \u003cem\u003esediminis\u003c/em\u003e PA15-N-34\u003csup\u003eT\u003c/sup\u003e (Liao et al. \u003cspan\u003e2020\u003c/span\u003e) could produce Naphthol-AS-Bl-phosphate.\u003c/p\u003e\n\u003ch3\u003eChemotaxonomic characteristics\u003c/h3\u003e\n\u003cp\u003eC\u003csub\u003e15:0\u003c/sub\u003e anteiso (49.4%) is the major cellular fatty acid of strain REN37\u003csup\u003eT\u003c/sup\u003e, followed by C\u003csub\u003e16:0\u003c/sub\u003e iso (18.1%), C\u003csub\u003e17:0\u003c/sub\u003e anteiso (16.5%) and C\u003csub\u003e15:0\u003c/sub\u003e iso (9.5%) (Tables S1). The polar lipids of strain REN37\u003csup\u003eT\u003c/sup\u003e were identified to be diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylcholine (PC), unidentified glycolipid (GL), two unidentified phospholipids (PL1-2) (Fig. S2). Consistently, PG was existed in \u003cem\u003eA. mobilis\u003c/em\u003e MT13131\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. nanhaiticus\u003c/em\u003e 19-m-6\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. profundi\u003c/em\u003e MTEO17\u003csup\u003eT\u003c/sup\u003e(Liu et al. \u003cspan\u003e2019\u003c/span\u003e), A. \u003cem\u003esediminis\u003c/em\u003e PA15-N-34\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. xenomutans\u003c/em\u003e JC109\u003csup\u003eT\u003c/sup\u003e and DPG was existed in \u003cem\u003eA. indicus\u003c/em\u003e SW127\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eA. xenomutans\u003c/em\u003e JC109\u003csup\u003eT\u003c/sup\u003e (Table\u0026nbsp;\u003cspan\u003e1\u003c/span\u003e). The predominant respiratory quinone of strain REN37\u003csup\u003eT\u003c/sup\u003e was MK-12.\u003c/p\u003e\n\u003cdiv\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1.\u003c/strong\u003e Characteristics of REN37\u003csup\u003eT\u003c/sup\u003e and other closely related species in the genus\u0026nbsp;\u003cem\u003eAlcanivorax\u003c/em\u003e.\u003c/p\u003e\n \u003cdiv align=\"center\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"933\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCharacteristic\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eIsolated from\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003efermented grains\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eoil-contaminated sea water\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eseawater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003efiltered sea water\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003edeep-sea sediment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003edeep sea sediment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003edeep-sea sediment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003edeep seawater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003edeep-sea sediment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eshrimp cultivation pond\u0026nbsp;sediment\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGram staining\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eMotility\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCell shape\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003erods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003erods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003erods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003erods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003erods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eshort rods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eshort rods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003erods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003erods\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eshort rods\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eOptimal medium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eTSB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHLB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e216L agar\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eMA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eMA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e216L agar\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eMA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eMA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e216L agar\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTemperatures range(\u003csup\u003eo\u003c/sup\u003eC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e25-45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e15-45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e8-42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4-40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e10-42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4-41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e10-42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e10-45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e10-45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e25-40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTemperatures preference(\u0026deg;C)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e32-37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e28-35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e30.0\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e22-25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e25-28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e37.0\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e28-30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e30\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003epH range\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5-10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5.5-9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6-10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6-10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6-10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4-9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003epH preference\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6-7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7.5\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7-8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7-8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7-8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eNaCl tolerance range for growth (w/v%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.0-5.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.0-15.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.5-11.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.0-12.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.5-15.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.5-12.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0-12.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0-15.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.5-20.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eOptimum NaCl concentration(w/v%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.0-2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.0-7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.0-4.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e10.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.0-4.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.0-5.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.0\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.0-5.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.0-5.0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eNO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e reduction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTween 40 hydrolysis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTween 80 hydrolysis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGelatin hydrolysis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAesculin hydrolysis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCasein hydrolysis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eStarch hydrolysis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCatalase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eOxidase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ew\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAmylase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCAPric acid assimilation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eADIpic acid assimilation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ew\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eMaLaTe assimilation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003etrisodium CITrate\u0026nbsp;assimilation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eAPI ZYM\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eL-cystyl-2-naphthylamide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e2-naphthyl myristate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u0026nbsp;L-valyl-2-naphthylamide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eNaphthol-AS-Bl-phosphate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ew\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePolar lipids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eDPG, PG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eDPG, PE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePG, PE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePG, PE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePE2, PG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePE, PG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eDPG, PG\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eDNA G+C content (mol%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e63.0\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e62.1-62.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e62.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e66.4\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e64.2\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e56.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e60.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e57.5\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e57.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e54.5\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eStrains: 1. REN37\u003csup\u003eT\u003c/sup\u003e; 2. \u003cem\u003eA. dieselolei\u003c/em\u003e B-5\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Liu and Shao 2005); 3. \u003cem\u003eA. indicus\u0026nbsp;\u003c/em\u003eSW127\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Song et al. 2018); 4. \u003cem\u003eA.\u003c/em\u003e \u003cem\u003evenustensis\u003c/em\u003e ISO4\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Fern\u0026aacute;ndez-Mart\u0026iacute;nez et al. 2003); 5. \u003cem\u003eA. mobilis\u003c/em\u003e MT13131\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Yang et al. 2018); 6. \u003cem\u003eA. nanhaiticus\u003c/em\u003e 19-m-6\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Lai et al. 2016); 7. \u003cem\u003eA. pacificus\u003c/em\u003e W11-5\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Lai et al. 2011); 8. \u003cem\u003eA. profundi\u003c/em\u003e MTEO17\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Liu et al. 2019); 9. \u003cem\u003eA.\u003c/em\u003e \u003cem\u003esediminis\u0026nbsp;\u003c/em\u003ePA15-N-34\u003csup\u003eT\u003c/sup\u003e\u003csup\u003e\u0026nbsp;\u003c/sup\u003e(Liao et al. 2020); 10. \u003cem\u003eA. xenomutans\u003c/em\u003e JC109\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Rahul et al. 2014)\u003cem\u003e.\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e+, positive reaction or growth; -, no reaction or growth; w, weak reaction; ND, no data.\u003c/p\u003e\n \u003cp\u003eTSB, trypsin Soy Broth\u0026nbsp;medium; HLB, HLB was modified from Luria\u0026ndash;Bertani (LB) medium;\u0026nbsp;216L agar, 216L marine agar medium;\u0026nbsp;MA, marine agar medium; NA,\u0026nbsp;Nutrient Agar\u0026nbsp;medium.\u003c/p\u003e\n \u003cp\u003eDPG,\u0026nbsp;diphosphatidylglycerol;\u0026nbsp;PG,\u0026nbsp;phosphatidylglycerol; PE, phosphatidylethanolamin\u003c/p\u003e\u0026nbsp;\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\"\u003e\n \u003ch2\u003eGenomic features and phylogenetic analysis based on whole genome sequences\u003c/h2\u003e\n \u003cp\u003eThe whole genome sequence was submitted to GenBank with the accession number of GCA 041102775.1. Based on the genomic information analysis, the genome of REN37\u003csup\u003eT\u003c/sup\u003e was characterised by a size of 3.02 Mbp and a DNA G\u0026thinsp;+\u0026thinsp;C content of 63.0 mol%. The OrthoANI, dDDH, and AAI values between REN37\u003csup\u003eT\u003c/sup\u003e and its most similar strain of \u003cem\u003eAlcanivorax pacificus\u003c/em\u003e W11-5\u003csup\u003eT\u003c/sup\u003e were 74.0%, 19.2% and 92.0%, respectively. The values were lower than the threshold values of 95% (OrthoANI), 70% (dDDH), and 95% (AAI), indicating that strain REN37\u003csup\u003eT\u003c/sup\u003e represented a novel species in the genus of \u003cem\u003eAlcanivorax\u003c/em\u003e (Felsenstein \u003cspan\u003e1981\u003c/span\u003e; Goris et al. \u003cspan\u003e2007\u003c/span\u003e; Kim et al. \u003cspan\u003e2014\u003c/span\u003e; Meier-Kolthoff et al. \u003cspan\u003e2013\u003c/span\u003e; Richter and Rossell\u0026oacute;-M\u0026oacute;ra \u003cspan\u003e2009\u003c/span\u003e). Furthermore, \u003cem\u003eAlcanivorax\u003c/em\u003e Among the annotated genes, 2.34 Mbp (77.48%) were assigned to functional categories. Rast server analysis showed that amino acids and derivatives (170) had the highest counts, followed by protein metabolism (168) and Cofactors, Vitamins, Prosthetic Groups, Pigments (146) (Fig. S3).\u003c/p\u003e\n \u003cp\u003eThe whole genome sequence of strain REN37\u003csup\u003eT\u003c/sup\u003e was compared with its similar strains in the corresponding genus (Table S3), and phylogenetic analysis was performed. Results showed that strain REN37\u003csup\u003eT\u003c/sup\u003e also formed a separate clade within the genus of \u003cem\u003eAlcanivorax\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan\u003e2\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eDescription of\u003c/strong\u003e \u003cstrong\u003eAlcanivorax beigongshangi\u003c/strong\u003e \u003cstrong\u003esp. nov.\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eAlcanivorax beigongshangi\u003c/em\u003e (bei. gong. shang\u0026apos;i. N.L. gen. n. \u003cem\u003ebeigongshangi\u003c/em\u003e, as Beijing Technology and Business University, the strain was cultured and isolated in Beijing Technology and Business University).\u003c/p\u003e\n \u003cp\u003eThe strain is Gram-negative, aerobic. Cells of REN37\u003csup\u003eT\u003c/sup\u003e are rod-shaped (1.2\u0026ndash;1.4 \u0026micro;m in length and 0.3\u0026ndash;0.4 \u0026micro;m in width), lacking flagella, and thus displaying non-motility. It can grow under 25\u0026ndash;45 \u003csup\u003eo\u003c/sup\u003eC, pH5-10, 1%-5% (w/v) NaCl and in MB, LB, NB, TSB and R\u003csub\u003e2\u003c/sub\u003eA medium. The optimal growth condition is 32\u0026ndash;37 \u003csup\u003eo\u003c/sup\u003eC, pH 6\u0026ndash;7, NaCl concentration 1%-2% (w/v) in TSB medium. Based on the API ZYM results, REN37\u003csup\u003eT\u003c/sup\u003e produces 2-naphthyl myristate, L-valyl-2-naphthylamide, L-cystyl-2-naphthylamide, N-benzoyl-DL-arginine-2-naphthylamide, N-glutaryl-phenylalanine-2-naphthylamide, 6-Br-2-naphthyl-\u0026alpha;D-galactopyranoside, 2-naphthyl-\u0026alpha;D-galactopyranoside, Naphthol-AS-Bl-SD-glucuronide, 2-naphthyl-\u0026alpha;D-glucopyranoside, 6-Br-2-naphthyl-BD-glucopyranoside, 1-naphthyl-N-acetyl-BD-glucosaminide, 6-Br-2-naphthyl-\u0026alpha;D-mannopyranoside, 2-naphthyl-aL-fucopyranoside. Positive for amylase, catalase, oxidase and hydrolysis aesculin, casein, starch. Assimilation trisodium CITrate. Major fatty acids are C\u003csub\u003e14:0\u003c/sub\u003e iso, C\u003csub\u003e14:0\u003c/sub\u003e anteiso, C\u003csub\u003e14:0\u003c/sub\u003e, C\u003csub\u003e15:1\u003c/sub\u003e anteiso A, C\u003csub\u003e15:0\u003c/sub\u003e iso, C\u003csub\u003e15:0\u003c/sub\u003e anteiso, C\u003csub\u003e16:0\u003c/sub\u003e iso, C\u003csub\u003e16:0\u003c/sub\u003e, C\u003csub\u003e15:0\u003c/sub\u003e 3OH, C\u003csub\u003e17:0\u003c/sub\u003e iso, C\u003csub\u003e17:0\u003c/sub\u003e anteiso, C\u003csub\u003e17:1\u003c/sub\u003e w5c, C\u003csub\u003e17:0\u003c/sub\u003e, C\u003csub\u003e18:0\u003c/sub\u003e, C\u003csub\u003e17:0\u003c/sub\u003e 3OH, C\u003csub\u003e19:0\u003c/sub\u003e anteiso. The major polar lipids are DPG, PL1-2, PG, GL and PC. The major respiratory quinone of strain REN37\u003csup\u003eT\u003c/sup\u003e is MK-12 (100%). The genomic DNA G\u0026thinsp;+\u0026thinsp;C content is 63.0 mol%. The type strain, REN37\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;GDMCC 1.3120\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;JCM 35319\u003csup\u003eT\u003c/sup\u003e), was isolated from Baijiu fermented grains from Sichuan, PR China. The GenBank accession numbers for the 16S rRNA gene sequence and the whole genome sequence is OR405985 and GCA 041102775.1, respectively.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Abbreviations","content":"\u003cp\u003e\u003cstrong\u003eOrthoANI\u003c/strong\u003e, average nucleotide identity by Orthology; \u003cstrong\u003edDDH\u003c/strong\u003e, digital DNA-DNA hybridization; \u003cstrong\u003eAAI\u003c/strong\u003e, average amino acid identity; \u003cstrong\u003eTSB\u003c/strong\u003e, trypsin Soy Broth medium; \u003cstrong\u003eLB,\u0026nbsp;\u003c/strong\u003eLuria-Bertani Broth; \u003cstrong\u003eNB,\u003c/strong\u003e Nutrient Broth; \u003cstrong\u003eR\u003csub\u003e2\u003c/sub\u003eA\u003c/strong\u003e, Reasoner\u0026apos;s 2A; \u003cstrong\u003eHLB\u003c/strong\u003e, HLB was modified from Luria-Bertani ; \u003cstrong\u003e216L agar\u003c/strong\u003e, 216L marine agar; \u003cstrong\u003eMA\u003c/strong\u003e, marine agar; \u003cstrong\u003eNA\u003c/strong\u003e, Nutrient Agar; \u003cstrong\u003eDPG\u003c/strong\u003e, diphosphatidylglycerol; \u003cstrong\u003ePG\u003c/strong\u003e, phosphatidylglycerol; \u003cstrong\u003ePE\u003c/strong\u003e, phosphatidylethanolamine; \u003cstrong\u003ePC\u003c/strong\u003e, phosphatidylcholine; \u003cstrong\u003eGL\u003c/strong\u003e, unidentified glycolipid; \u003cstrong\u003ePL\u003c/strong\u003e, phospholipid.\u003c/p\u003e\n"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003ethe authors declare that no funds, grants, or other support were received during the preparation of this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e: the authors declare that there are no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u0026nbsp;\u003c/strong\u003eQing Ren and Han-Xu Pan contributed to the study conception and design. Material preparation, data collection and analysis were performed by Yi-Ming Li, Hao-Yue Gu, Ru Zhang and Zhan-Bin Sun. The first draft of the manuscript was written by Tong-xi Zhang and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e: all data have been made fully available to the public. The GenBank accession numbers for the 16S rRNA sequence and the full genome sequence of \u003cem\u003eAlcanivorax beigongshangi\u003c/em\u003e REN37\u003csup\u003eT\u003c/sup\u003e are OR405985 and GCA 041102775.1, respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval:\u0026nbsp;\u003c/strong\u003ethis article does not contain any studies with human participants or animals performed by any of the authors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e: all authors gave their consent to participate in this study.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAltschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. 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Arch Microbiol 204:106. https://doi.org/10.1007/s00203-021-02638-3\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":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":"Alcanivorax beigongshangi, baijiu, fermented grains, polyphasic taxonomy","lastPublishedDoi":"10.21203/rs.3.rs-5108599/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5108599/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA bacterial strain, REN37\u003csup\u003eT\u003c/sup\u003e, was isolated from fermented grains of Baijiu samples collected from Sichuan, PR China. The cells of strain REN37\u003csup\u003eT\u003c/sup\u003e was Gram-negative and aerobic. The cellular morphology exhibited rod-shaped cells without flagellum, displaying non-motility. The optimal growth condition was at 32\u0026ndash;37 \u003csup\u003eo\u003c/sup\u003eC, pH 6.0\u0026ndash;7.0, and with a NaCl concentration of 1%-2% (w/v). Strain REN37\u003csup\u003eT\u003c/sup\u003e was positive for amylase, catalase and oxidase activities and aesculin, casein, starch hydrolysis. Based on the analysis of 16S rRNA gene sequence, strains REN37\u003csup\u003eT\u003c/sup\u003e was identified to belong to the genus of \u003cem\u003eAlcanivorax.\u003c/em\u003e Its closest species was \u003cem\u003eAlcanivorax pacificus\u003c/em\u003e W11-5\u003csup\u003eT\u003c/sup\u003e (96.1%). The polar lipids were identified to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, unidentified glycolipid, two unidentified phospholipids. The predominant menaquinone was MK-12. The predominant fatty acids were C\u003csub\u003e15:0\u003c/sub\u003e anteiso (49.4%), C\u003csub\u003e16:0\u003c/sub\u003e iso (18.1%), C\u003csub\u003e17:0\u003c/sub\u003e anteiso (16.5%), C\u003csub\u003e15:0\u003c/sub\u003e iso (9.5%). The digital DNA-DNA hybridization (dDDH), average nucleotide identity (OrthoANI) and average amino acid identify (AAI) values between strain REN37\u003csup\u003eT\u003c/sup\u003e and its most similar species were 19.2%, 74.0% and 92.0%, respectively. The DNA G\u0026thinsp;+\u0026thinsp;C content of the strain REN37\u003csup\u003eT\u003c/sup\u003e was 63.0 mol%. Based on the results, REN37\u003csup\u003eT\u003c/sup\u003e represents a novel strain, and the name \u003cem\u003eAlcanivorax beigongshangi\u003c/em\u003e sp. nov. was proposed. The type strain is REN37\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;GDMCC 1.3120\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;JCM 35319\u003csup\u003eT\u003c/sup\u003e).\u003c/p\u003e","manuscriptTitle":"Alcanivorax beigongshangi sp. nov., isolated from the fermented grains of Chinese baijiu","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-14 12:01:28","doi":"10.21203/rs.3.rs-5108599/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-09-21T10:12:19+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-09-20T13:54:56+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-09-20T13:45:51+00:00","index":"","fulltext":""},{"type":"submitted","content":"Antonie van Leeuwenhoek","date":"2024-09-18T09:05:52+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":"abcf217e-46f1-4a58-8415-079c31492924","owner":[],"postedDate":"November 14th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-12-02T16:00:58+00:00","versionOfRecord":{"articleIdentity":"rs-5108599","link":"https://doi.org/10.1007/s10482-024-02043-y","journal":{"identity":"antonie-van-leeuwenhoek","isVorOnly":false,"title":"Antonie van Leeuwenhoek"},"publishedOn":"2024-11-28 15:57:17","publishedOnDateReadable":"November 28th, 2024"},"versionCreatedAt":"2024-11-14 12:01:28","video":"","vorDoi":"10.1007/s10482-024-02043-y","vorDoiUrl":"https://doi.org/10.1007/s10482-024-02043-y","workflowStages":[]},"version":"v1","identity":"rs-5108599","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5108599","identity":"rs-5108599","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}
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