枣阳酸浆水中乳酸菌的可培养多样性及其分离方式评价
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摘要
采用变形梯度凝胶电泳(DGGE)对浆水样品细菌群落进行分析,使用6种常用的乳酸菌分离方式对酸浆水中的乳酸菌进行了分离与鉴定,并对这些方法的分离效果进行了评价。结果表明,酸浆水样品中乳酸菌的数量约为5×106~2.5×108cfu/m L,不同来源的浆水样品中乳酸菌数量差异较大。通过这6种培养方式共分离得到90株乳酸菌。经过鉴定,它们是德氏乳杆菌(Lactobacillus delbrueckii)、发酵乳杆菌(Lactobacillus fermentium)、副干酪乳杆菌(Lactobacillus paracasei)、植物乳杆菌(Lactobacillus plantarum)、鼠李糖乳杆菌(Lactobacillus rhamnosus)与玉米乳杆菌(Lactobacillus zeae)。在6种乳酸菌分离方式中,非增菌法中厌氧罐-厌氧袋法、厌氧工作站(10%H2,10%CO2,80%N2)及增菌法对酸浆水中的乳酸菌分离效果较好。
DGGE was used to analyze the bacterial community of Suanjiangshui samples,and the microbial compositions in Suanjiangshui were separated and identified by six common lactic acid bacteria separation methods,and the separation effects of these methods were evaluated. The results showed that there were differences in number of lactic acid bacteria among the Suanjiangshui from different sources.The number of lactic acid bacteria were about 5 × 106~ 2.5 × 108cfu/m L. A total of 90 strains of lactic acid bacteria were obtained,which were identified as Lactobacillus delbrueckii,Lactobacillus fermentium,Lactobacillus zeae,Lactobacillus plantarum,Lactobacillus rhamnosus and Lactobacillus paracasei.Among the methods used in this study,anaerobic jar with anaeropack,anaerobic workstation( 10% H2,10% CO2,80% N2) and enumeration by litmus milk medium method were the best methods for LAB isolation.
DGGE was used to analyze the bacterial community of Suanjiangshui samples,and the microbial compositions in Suanjiangshui were separated and identified by six common lactic acid bacteria separation methods,and the separation effects of these methods were evaluated. The results showed that there were differences in number of lactic acid bacteria among the Suanjiangshui from different sources.The number of lactic acid bacteria were about 5 × 106~ 2.5 × 108cfu/m L. A total of 90 strains of lactic acid bacteria were obtained,which were identified as Lactobacillus delbrueckii,Lactobacillus fermentium,Lactobacillus zeae,Lactobacillus plantarum,Lactobacillus rhamnosus and Lactobacillus paracasei.Among the methods used in this study,anaerobic jar with anaeropack,anaerobic workstation( 10% H2,10% CO2,80% N2) and enumeration by litmus milk medium method were the best methods for LAB isolation.
引文
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[24]孟宪刚,李雪萍,李建宏,等.浆水中乳酸菌分离鉴定及其代谢特性的初步研究[J].食品工业科技,2015,36(1):181-186.
[25]Qiu Y L,Hanada S,Ohashi A,et al.Syntrophorhabdus aromaticivorans gen.nov.,sp.nov.,the first cultured anaerobe capable of degrading phenol to acetate in obligate syntrophic assciations with a hydrogenotrophic methanogen[J].Applied and Environmental Microbiology.2008,74(7):2051-2058.
[26]Tohno M,Kitahara M,Irisawa T,et al.Lactobacillus silagei sp.nov.,isolated from orchardgrass silage[J].International Journal of Systematic and Evolutionary Microbiology,2013,63(12):4613-4618.
[27]Roos S,Engstrand L,Jonsson H,et al.Lactobacillus gastricus sp.nov.,Lactobacillus antri sp.nov.,Lactobacillus kalixensis sp.nov.and Lactobacillus ultunensis sp.nov.,isolated from human stomach mucosa[J].International Journal of Systematic and Evolutionary Microbiology,2005,55(1):77-82.
[28]Tohno M,Kitahara M,Inoue H,et al.Weissella oryzae sp.nov.,isolated from fermented rice grains[J].International Journal of Systematic and Evolutionary Microbiology,2013,63(4):1417-1420.
[29]Baele M,Vancannevt M,Devriese L A,et al.Lactobacillus ingluviei sp.nov.,isolated from the intestinal tract of pigeons[J].International Journal of Systematic and Evolutionary Microbiology,2003,53(1):133-136.
[30]Volkhov D V,Amselle M,Beck B J,et al.Lactobacillus brantae sp.nov.,isolated from faeces of Canada geese(Branta canadensis)[J].International Journal of Systematic and Evolutionary Microbiology,2011,62(9):2068-2076.
[2]周书楠,席修璞,董蕴,等.琚湾酸浆面浆水细菌多样性评价[J].2018,37(1):49-53.
[3]Teusink B,Molenaar D.Systems biology of lactic acid bacteria:For food and thought[J].Current Opinion in Systems Biology,2017,6:7-13.
[4]Cappello M S,Zapparoli G,Logrieco A,et al.Linking wine lactic acid bacteria diversity with wine aroma and flavor[J].International Journal of Food Microbiology,2017,243:16-27.
[5]Chen C,Zhao S S,Hao G F,et al.Role of lactic acid bacteria on the yogurt flavour:A review[J].International Journal of Food Properties,2017,20(1):S316-S330.
[6]Azam M,Mohsin M,Ijaz H,et al.Lactic acid bacteria in traditional fermented Asian foods[J].Pakistan Journal of Pharmaceutical Sciences,2017,30(5):1803-1814.
[7]Wu Q L,Shan N P.High gamma-aminobutyric acid production from lactic acid bacteria:Emphasis on Lactobacillus brevis as a functional dairy starter[J].LWT-Food Science and Technology,2017,57(17):3661-3672.
[8]邸维,张兰威,易华西,等.乳酸菌胞外多糖结构及其功能多样性的研究进展[J].中国乳品工业,2017,45(5):32-37.
[9]López-Cuellar M D R,Rodríguez-Hernández A I,Chavarría-Hernández N.LAB bacteriocin applications in the last decade[J].Biotechnology&Biotechnological Equipment,2016,30(6):1039-1050.
[10]曹佳璐,张列兵.韩国泡菜乳酸菌研究进展[J].中国食品学报,2017,17(10):184-192.
[11]王玉荣,孙永坤,代凯文,等.基于单分子实时测序技术的3个当阳广椒样品细菌多样性研究[J].食品工业科技,2018(2):108-112.
[12]苗乘源,郑琳,程雅韵,等.朝鲜族传统米酒中的乳酸菌多样性分析[J].延边大学农学学报,2016,38(3):248-250.
[13]Lv X C,Chen Z C,Jia R,et al.Microbial community structure and dynamics during the traditional brewing of Fuzhou Hong Qu glutinous rice wine as determined by culture-dependent and culture-independent techniques[J].Food Control,2015,57:216-224.
[14]靳志敏,段艳,通力嘎,等.降胆固醇乳酸菌的筛选及其在内蒙古发酵香肠中的应用[J].食品科学,2015,36(5):99-103.
[15]周书楠,席修璞,董蕴,等.琚湾酸浆面浆水细菌多样性评价[J].中国酿造,2018,37(1):49-53.
[16]Zhang D,Li W,Zhang S,et al.Evaluation of the impact of DNA extraction methods on BAC bacterial community composition measured by denaturing gradient gel electrophoresis[J].Letters in Applied Microbiology,2011,53(1):44-49.
[17]Ponce-Soto G Y,Aguirre-Von-Wobeser E,Eguiarte L E,et al.Enrichment experiment changes microbial interactions in an ultra-oligotrophic environment[J].Frontiers in Microbiology,2015,6:246.
[18]Ke C R,Li Z Y,Tao W Q,et al.Impacts of chloride deicing salt on bulk soils,fungi,and bacterial populations surrounding the plant rhizosphere[J].Applied Soil Ecology,2013,72(5):69-78.
[19]张振东,赵慧君,沈馨,等.米酒曲细菌多样性研究[J].中国微生态杂志,2018,30(6):640-646.
[20]王丹丹,倪慧,赵慧君,等.凤窝酒曲中乳酸菌的分离及其作用下的米酒品质评价[J].2018,37(6):80-84.
[21]陈芝兰,杨吉霞,李梦寒,等.西藏地区传统发酵乳中乳酸菌多样性及微生物数量分析[J].食品科学,2013,34(17):140-145.
[22]Mveobiang A,Mestdagh M,Portaels F.DNA isolation from chloroform/methanol-treated mycobacterial cells without lysozyme and proteinase K[J].Biotechniques,2001,30(2):272-274.
[23]张晓辉,杨靖鹏,王少军,等.浆水中细菌多样性分析及乳酸菌的分离鉴定[J].食品科学,2017,38(4):70-76.
[24]孟宪刚,李雪萍,李建宏,等.浆水中乳酸菌分离鉴定及其代谢特性的初步研究[J].食品工业科技,2015,36(1):181-186.
[25]Qiu Y L,Hanada S,Ohashi A,et al.Syntrophorhabdus aromaticivorans gen.nov.,sp.nov.,the first cultured anaerobe capable of degrading phenol to acetate in obligate syntrophic assciations with a hydrogenotrophic methanogen[J].Applied and Environmental Microbiology.2008,74(7):2051-2058.
[26]Tohno M,Kitahara M,Irisawa T,et al.Lactobacillus silagei sp.nov.,isolated from orchardgrass silage[J].International Journal of Systematic and Evolutionary Microbiology,2013,63(12):4613-4618.
[27]Roos S,Engstrand L,Jonsson H,et al.Lactobacillus gastricus sp.nov.,Lactobacillus antri sp.nov.,Lactobacillus kalixensis sp.nov.and Lactobacillus ultunensis sp.nov.,isolated from human stomach mucosa[J].International Journal of Systematic and Evolutionary Microbiology,2005,55(1):77-82.
[28]Tohno M,Kitahara M,Inoue H,et al.Weissella oryzae sp.nov.,isolated from fermented rice grains[J].International Journal of Systematic and Evolutionary Microbiology,2013,63(4):1417-1420.
[29]Baele M,Vancannevt M,Devriese L A,et al.Lactobacillus ingluviei sp.nov.,isolated from the intestinal tract of pigeons[J].International Journal of Systematic and Evolutionary Microbiology,2003,53(1):133-136.
[30]Volkhov D V,Amselle M,Beck B J,et al.Lactobacillus brantae sp.nov.,isolated from faeces of Canada geese(Branta canadensis)[J].International Journal of Systematic and Evolutionary Microbiology,2011,62(9):2068-2076.
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