川西彝族传统酸菜汁中乳酸菌的分离鉴定与特性分析
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摘要
川西彝族传统酸菜因其制作工艺独特,在没有盐胁迫的环境中乳酸菌成为优势菌,说明其菌群具有较强的生长繁殖能力和抑菌能力。为将川西彝族传统酸菜汁中的功能微生物运用于低盐发酵蔬菜,运用溶钙圈法从川西彝族传统酸菜汁中分离出41株乳酸菌。采用牛津杯法,以大肠杆菌和金黄色葡萄球菌为指示菌,筛选出6株抑菌能力较好的菌株。通过生长情况、产酸能力分析及温度、pH、盐浓度耐受性的研究,最终筛选出1株综合性能较好的乳酸菌B1。该菌表现出较强的繁殖能力和抑菌性,大肠杆菌和金黄色葡萄球菌的抑菌圈大小分别为22.63±0.23、21.83±0.18,且温度、pH耐受性强,适合作为低盐发酵蔬菜的发酵剂。经分子生物学鉴定,结果表明该菌为植物乳杆菌(Lactobacillus plantarum)。
Traditional Yi sauerkraut in western Sichuan has unique processing technology,in which Lactobacillus is still the dominant bacterium without the salt stress.It indicates that Lactobacillus in Yi sauerkraut juice has strong growth and propagation ability and antibacterial efficiency.In order to apply the functional microorganism in the traditional Yi sauerkraut juice in western Sichuan to the low-salt fermented vegetables,41 strains of Lactobacillusare isolated from traditional Yi sauerkraut juice by the method of dissolving calcium circle.By using the Oxford cup method,6 strains of bacteria with good antibacterial efficiency are screened out with Escherichia coli and Staphylococcus aureus as the indicator bacteria.Through determining the growth curve,analyzing the acid production capacity and studying the tolerance of temperature,pH and salt,a strain of Lactobacillus B1 with better comprehensive performance is finally screened out.The bacterium show strong reproduction ability and antibacterial efficiency,and the inhibition zone diameters of Escherichia coli and Staphylococcus aureus are 22.63±0.23 and 21.83±0.18 respectively,and the temperature and pH tolerance are strong,it shows that this bacterium is suitable to be used as fermentation agent for low-salt fermented vegetables.The molecular biological identification results show that the bacterium is Lactobacillus plantarum.
Traditional Yi sauerkraut in western Sichuan has unique processing technology,in which Lactobacillus is still the dominant bacterium without the salt stress.It indicates that Lactobacillus in Yi sauerkraut juice has strong growth and propagation ability and antibacterial efficiency.In order to apply the functional microorganism in the traditional Yi sauerkraut juice in western Sichuan to the low-salt fermented vegetables,41 strains of Lactobacillusare isolated from traditional Yi sauerkraut juice by the method of dissolving calcium circle.By using the Oxford cup method,6 strains of bacteria with good antibacterial efficiency are screened out with Escherichia coli and Staphylococcus aureus as the indicator bacteria.Through determining the growth curve,analyzing the acid production capacity and studying the tolerance of temperature,pH and salt,a strain of Lactobacillus B1 with better comprehensive performance is finally screened out.The bacterium show strong reproduction ability and antibacterial efficiency,and the inhibition zone diameters of Escherichia coli and Staphylococcus aureus are 22.63±0.23 and 21.83±0.18 respectively,and the temperature and pH tolerance are strong,it shows that this bacterium is suitable to be used as fermentation agent for low-salt fermented vegetables.The molecular biological identification results show that the bacterium is Lactobacillus plantarum.
引文
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[14]林松洋,郝利民,刘鑫,等.乳酸菌耐盐分子机制研究进展[J].食品科学,2018,39(3):295-301.
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[19]敖晓琳,张小平,史令,等.四川泡菜中两株优良乳酸菌的鉴定及不同发酵条件对其发酵泡菜品质的影响[J].食品科学,2011,32(11):152-156.
[20]张锐,吴祖芳,沈锡权,等.榨菜低盐腌制过程的微生物群落结构与动态分析[J].中国食品学报,2011,11(3):175-180.
[2]丛敏.酸菜自然发酵过程中化学成分及乳酸菌变化规律的研究[D].沈阳:沈阳农业大学,2016.
[3]邱亚利.凉山彝族酸菜的制作工艺及旅游开发价值研究[J].食品研究与开发,2014,35(19):68-71.
[4]陈晓平,刘华英,魏小川,等.自然发酵酸菜汁中乳酸菌的分离筛选与鉴定研究[J].食品科学,2006,27(2):91-94.
[5]凌代文.乳酸细菌分类鉴定及实验方法[M].北京:中国轻工业出版社,1999.
[6]王颉,李标,李彦涛,等.泡菜中亚硝酸盐降解乳酸菌的筛选与鉴定[J].食品与药品,2014,16(2):112-116.
[7]Ao X,Zhang X,Shi L,et al.Identification of lactic acid bacteria in traditional fermented yak milk and evaluation of their application in fermented milk products[J].Journal of Dairy Science,2012,95(3):133-139.
[8]Mufandaedza J,Viljoen B C,Feresu S B,et al.Antimicrobial properties of lactic acid bacteria and yeast-LAB cultures isolated from traditional fermented milk against pathogenic Escherichia coli and Salmonella enteritidis strains[J].International Journal of Food Microbiology,2006,108(1):147-152.
[9]谢芳,杨承剑,唐艳,等.水牛乳中乳酸菌的分离及其抑菌特性研究[J].食品工业,2016,37(12):172-175.
[10]张晓娟,夏珊,陈洁,等.四川泡菜降解亚硝酸盐优良功能菌种的筛选、鉴定及其性能初探[J].中国调味品,2016,41(7):73-77.
[11]胡博涵,吴晖,赖富饶,等.耐盐乳酸菌的筛选及其在剁辣椒发酵中的应用[J].食品与机械,2014,30(1):51-54.
[12]李豪,章霞,张静,等.草莓果酒酵母菌的筛选、鉴定及耐受性研究[J].中国酿造,2017,36(2):85-88.
[13]范志平,王修俊,程艳波,等.泡菜中优良乳酸菌的筛选鉴定及生长特性的研究[J].中国调味品,2014,39(3):10-14.
[14]林松洋,郝利民,刘鑫,等.乳酸菌耐盐分子机制研究进展[J].食品科学,2018,39(3):295-301.
[15]Lehner A,Tasara T,Stephan R.16SrRNA gene based analysis of Enterobacter sakazakii strains from different sources and development of a PCR assay for identification[J].Bmc Microbiology,2004,4(1):1-7.
[16]李凤梅,王晓,张双灵,等.自然发酵酸菜汁中乳酸菌分离鉴定[J].中国酿造,2008(5):33-35.
[17]辛博.浆水接种发酵中亚硝酸盐的控制研究[D].西安:陕西科技大学,2014.
[18]张兰威.乳酸菌优良菌株的选育及直投式酸奶发酵剂的研制[D].哈尔滨:东北农业大学,2002.
[19]敖晓琳,张小平,史令,等.四川泡菜中两株优良乳酸菌的鉴定及不同发酵条件对其发酵泡菜品质的影响[J].食品科学,2011,32(11):152-156.
[20]张锐,吴祖芳,沈锡权,等.榨菜低盐腌制过程的微生物群落结构与动态分析[J].中国食品学报,2011,11(3):175-180.