自然发酵锦州小菜中乳酸菌的分离筛选
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摘要
通过探究自然发酵锦州小菜中乳酸菌的构成以及风味特征变化与微生物多样性之间的联系,为工业化生产提供依据。本研究对27份自然发酵锦州小菜的总酸含量、氨基酸态氮含量、亚硝酸盐含量、NaCl含量及菌落总数进行测定,根据采样地点的不同对样品进行差异性分析。结果发现,由于不同的发酵地点、条件、工艺而导致发酵液的成分含量存在显著差异(P<0.05),样品风味特征变化与小菜发酵过程中微生物变化存在直接关系。选择7份在样品成分含量中具有代表性特征的自然发酵锦州小菜进行乳酸菌分离筛选,分离出24株乳酸菌疑似菌株,初步鉴定10株为杆菌,14株为球菌。进一步采用16S rDNA序列分析对24株菌进行分子鉴定,通过序列分析进行属种鉴定。结果表明:24株菌均为乳酸菌,分别来自4个属10个种,10株乳杆菌属(Lactobacillus),8株肠球菌属(Enterococcus),3株链球菌属(Streptococcus),3株魏斯氏菌属(Weissella)。
This study was aimed at the composition of lactic acid bacteria and the relationship between changes in the flavor characteristics and the microbial diversity in naturally fermented Jinzhou pickle for the purpose of laying the basis for its industrial production. The contents of total acid, amino nitrogen, nitrite and NaCl and the total number of colonies were determined in 27 samples, and the differences among samples from different locations were analyzed. All investigated chemical components of pickle brines significantly varied with producing areas, fermentation conditions and methods (P < 0.05). We also found a direct association between changes in the flavor characteristics and changes in the bacterial community during the fermentation process. A total of 24 strains suspected of being lactic acid bacteria were isolated from 7 representative samples in terms of the chemical components. All these strains were identified as lactic acid bacteria by 16S rDNA sequence analysis, including 10 strains of Lactobacillus, 8 strains of Enterococcus, 3 strains of Streptococcus, and 3 strains of Weissella.
This study was aimed at the composition of lactic acid bacteria and the relationship between changes in the flavor characteristics and the microbial diversity in naturally fermented Jinzhou pickle for the purpose of laying the basis for its industrial production. The contents of total acid, amino nitrogen, nitrite and NaCl and the total number of colonies were determined in 27 samples, and the differences among samples from different locations were analyzed. All investigated chemical components of pickle brines significantly varied with producing areas, fermentation conditions and methods (P < 0.05). We also found a direct association between changes in the flavor characteristics and changes in the bacterial community during the fermentation process. A total of 24 strains suspected of being lactic acid bacteria were isolated from 7 representative samples in terms of the chemical components. All these strains were identified as lactic acid bacteria by 16S rDNA sequence analysis, including 10 strains of Lactobacillus, 8 strains of Enterococcus, 3 strains of Streptococcus, and 3 strains of Weissella.
引文
[1]王选志.锦州什锦小菜[J].中国酿造,1992,11(4):37-38.
[2]陈曾三.锦州虾油什锦小菜制作方法[J].中国调味品,2004(10):26-28.DOI:10.3969/j.issn.1000-9973.2004.10.008.
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[9]黄道梅.不同盐度的工业化盐渍萝卜多菌协同发酵过程中菌相变化与风味品质变化研究[D].成都:四川农业大学,2015.
[10]陈廷涛,张雁,陈于陇,等.发酵蔬菜风味形成机制及其分析技术的研究进展[J].中国食品学报,2014,14(2):217-224.
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[12]乌日娜.内蒙古传统酸马奶中乳杆菌的分离鉴定及16S rDNA序列同源性分析[D].呼和浩特:内蒙古农业大学,2005:14-15.
[13]孟令帅,张颖,邹婷婷,等.辣白菜中的乳酸菌分离鉴定[J].食品科学,2015,36(11):130-133.DOI:10.7506/spkx1002-6630-201511025.
[14]ISPIRLI H,DERTLI E.Isolation and characterisation of lactic acid bacteria from traditional koumiss and kurut[J].International Journal of Food Properties,2018,4:1-9.
[15]RIZZELLO C G,LORUSSO A,MONTEMURRO M,et al.Use of sourdough made with quinoa (Chenopodium quinoa) flour and autochthonous selected lactic acid bacteria for enhancing the nutritional,textural and sensory features of white bread[J].Food Microbiology,2016,56:1-13.DOI:10.1016/j.fm.2015.11.018.
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[18]卫生部.食品中氨基酸态氮的测定:GB 5009.235-2016[S].北京:中国标准出版社,2016.
[19]卫生部.食品中亚硝酸盐和硝酸盐的测定:GB 5009.33-2016[S].北京:中国标准出版社,2016.
[20]国家质量监督检验检疫总局,国家标准化管理委员会.食品中氯化钠的测定:GB/T 12457-2008[S].北京:中国标准出版社,2008.
[21]国家卫生和计划生育委员会,国家食品药品监督管理总局.食品微生物学检验?菌落总数测定:GB 4789.2-2016[S].北京:中国标准出版社,2016.
[22]尹利端.几种泡菜的安全性研究与优良乳酸菌的分离与鉴定[D].北京:中国农业大学,2005.
[23]ELINA S,ELISA L,JOHANNA B.Lactic acid bacteria in dried vegetables and spices[J].Food Microbiology,2016,53:110-114.
[24]李巧玉,方芳,堵国成,等.魏斯氏菌在发酵食品中的应用[J].食品发酵与工业,2017(10):241-247.DOI:10.13995/j.cnki.11-1802/ts.014934.
[25]刘晓辉,陈顺,高晓梅,等.酸菜中乳酸菌的分离筛选与鉴定研究[J].中国酿造,2009,28(2):62-64.DOI:10.3969/j.issn.0254-5071.2009.02.019.
[26]张香美,赵玉星,闫晓晶,等.1株具抑菌和抗氧化活性乳酸菌的筛选及鉴定[J].食品科学,2018,39(2):93-98.DOI:10.7506/spkx1002-6630-201802015.
[27]李欣,武俊瑞,田甜,等.大庆自然发酵酸菜中乳酸菌的分离鉴定及耐酸菌株初步筛选[J].食品科学,2014,35(1):150-154.DOI:10.7506/spkx1002-6630-201401029.
[28]王磊,张洁,赵国忠.东北酸菜优良乳酸菌筛选及抑菌特性研究[J].中国酿造,2017,36(7):48-52.
[29]ADESULU-DAHUNSI A T,SANNI A I,JEYARAM K.Rapid differentiation among Lactobacillus,Pediococcus and Weissella species from some Nigerian indigenous fermented foods[J].Food Science and Technology,2017,77:39-44.DOI:10.1016/j.lwt.2016.11.007.
[30]LIU A,LIU G,HUANG C,et al.The bacterial diversity of ripened Guang’yuan Suancai and in vitro,evaluation of potential probiotic lactic acid bacteria isolated from Suancai[J].LWT-Food Science and Technology,2017,85:175-180.
[31]卫生部.酱腌菜卫生标准:SB/T 10439-2007[S].北京:中国标准出版社,2007.
[32]农业部.绿色食品酱腌菜:NY/T 437-2000[S].北京:中国标准出版社,2000.
[33]张庆芳,迟乃玉,郑燕,等.乳酸菌降解亚硝酸盐机理的研究[J].食品与发酵工业,2002,28(8):27-31.DOI:10.3321/j.issn:0253-990X.2002.08.007.
[34]黄丽慧,张雁,陈于陇,等.发酵蔬菜中亚硝酸盐消长规律及调控技术的研究进展[J].食品科学,2013,34(5):303-307.
[2]陈曾三.锦州虾油什锦小菜制作方法[J].中国调味品,2004(10):26-28.DOI:10.3969/j.issn.1000-9973.2004.10.008.
[3]曲玲童,牛文静,孙征,等.锦州盐渍小菜张耐盐乳酸菌的筛选与产酸性能研究[J].中国酿造,2012,31(6):83-87.DOI:10.3969/j.issn.0254-5071.2012.06.022.
[4]孙慧君,张颖,王洪玉,等.传统发酵锦州小菜中主要微生物多样性分析[J].食品科学,2017,38(2):15-19.DOI:10.7506/spkx1002-6630-201702003.
[5]丛敏,李欣蔚,武俊瑞,等.PCR-DGGE分析东北传统发酵酸菜中乳酸菌多样性[J].食品科学,2016,37(7):78-82.DOI:10.7506/spkx1002-6630-201607015.
[6]曹碧璇,胡滨,刘爱平.利用16S rRNA基因克隆文库分析东北自然发酵酸菜中细菌多样性[J].食品与发酵工业,2015,41(11):76-80.DOI:10.13995/j.cnki.11-1802/ts.201511014.
[7]曹振辉,刘永仕,潘洪彬,等.乳酸菌的益生功能作用机制研究进展[J].食品工业科技,2015,36(8):366-370;377.DOI:10.13386/j.issn1002-0306.2015.24.072.
[8]杨荣玲,肖更生,吴晓玉,等.我国蔬菜发酵加工研究进展[J].保鲜与加工,2006(2):15-18.DOI:10.3969/j.issn.1009-6221.2006.02.008.
[9]黄道梅.不同盐度的工业化盐渍萝卜多菌协同发酵过程中菌相变化与风味品质变化研究[D].成都:四川农业大学,2015.
[10]陈廷涛,张雁,陈于陇,等.发酵蔬菜风味形成机制及其分析技术的研究进展[J].中国食品学报,2014,14(2):217-224.
[11]武俊瑞,李欣,李晓忱,等.自然发酵酸菜汁中乳杆菌的分离鉴定[J].食品科学,2012,33(15):191-194.
[12]乌日娜.内蒙古传统酸马奶中乳杆菌的分离鉴定及16S rDNA序列同源性分析[D].呼和浩特:内蒙古农业大学,2005:14-15.
[13]孟令帅,张颖,邹婷婷,等.辣白菜中的乳酸菌分离鉴定[J].食品科学,2015,36(11):130-133.DOI:10.7506/spkx1002-6630-201511025.
[14]ISPIRLI H,DERTLI E.Isolation and characterisation of lactic acid bacteria from traditional koumiss and kurut[J].International Journal of Food Properties,2018,4:1-9.
[15]RIZZELLO C G,LORUSSO A,MONTEMURRO M,et al.Use of sourdough made with quinoa (Chenopodium quinoa) flour and autochthonous selected lactic acid bacteria for enhancing the nutritional,textural and sensory features of white bread[J].Food Microbiology,2016,56:1-13.DOI:10.1016/j.fm.2015.11.018.
[16]WITTMANN C,LIAO J C,RUIZ-RODRíGUEZ L,et al.Lactic acid bacteria[M]//Industrial biotechnology.Wiley-VCH Verlag GmbH&Co.KGaA,2017:395-451.
[17]国家质量监督检验检疫总局,国家标准化管理委员会.食品中总酸的测定:GB/T 12456-2008[S].北京:中国标准出版社,2008.
[18]卫生部.食品中氨基酸态氮的测定:GB 5009.235-2016[S].北京:中国标准出版社,2016.
[19]卫生部.食品中亚硝酸盐和硝酸盐的测定:GB 5009.33-2016[S].北京:中国标准出版社,2016.
[20]国家质量监督检验检疫总局,国家标准化管理委员会.食品中氯化钠的测定:GB/T 12457-2008[S].北京:中国标准出版社,2008.
[21]国家卫生和计划生育委员会,国家食品药品监督管理总局.食品微生物学检验?菌落总数测定:GB 4789.2-2016[S].北京:中国标准出版社,2016.
[22]尹利端.几种泡菜的安全性研究与优良乳酸菌的分离与鉴定[D].北京:中国农业大学,2005.
[23]ELINA S,ELISA L,JOHANNA B.Lactic acid bacteria in dried vegetables and spices[J].Food Microbiology,2016,53:110-114.
[24]李巧玉,方芳,堵国成,等.魏斯氏菌在发酵食品中的应用[J].食品发酵与工业,2017(10):241-247.DOI:10.13995/j.cnki.11-1802/ts.014934.
[25]刘晓辉,陈顺,高晓梅,等.酸菜中乳酸菌的分离筛选与鉴定研究[J].中国酿造,2009,28(2):62-64.DOI:10.3969/j.issn.0254-5071.2009.02.019.
[26]张香美,赵玉星,闫晓晶,等.1株具抑菌和抗氧化活性乳酸菌的筛选及鉴定[J].食品科学,2018,39(2):93-98.DOI:10.7506/spkx1002-6630-201802015.
[27]李欣,武俊瑞,田甜,等.大庆自然发酵酸菜中乳酸菌的分离鉴定及耐酸菌株初步筛选[J].食品科学,2014,35(1):150-154.DOI:10.7506/spkx1002-6630-201401029.
[28]王磊,张洁,赵国忠.东北酸菜优良乳酸菌筛选及抑菌特性研究[J].中国酿造,2017,36(7):48-52.
[29]ADESULU-DAHUNSI A T,SANNI A I,JEYARAM K.Rapid differentiation among Lactobacillus,Pediococcus and Weissella species from some Nigerian indigenous fermented foods[J].Food Science and Technology,2017,77:39-44.DOI:10.1016/j.lwt.2016.11.007.
[30]LIU A,LIU G,HUANG C,et al.The bacterial diversity of ripened Guang’yuan Suancai and in vitro,evaluation of potential probiotic lactic acid bacteria isolated from Suancai[J].LWT-Food Science and Technology,2017,85:175-180.
[31]卫生部.酱腌菜卫生标准:SB/T 10439-2007[S].北京:中国标准出版社,2007.
[32]农业部.绿色食品酱腌菜:NY/T 437-2000[S].北京:中国标准出版社,2000.
[33]张庆芳,迟乃玉,郑燕,等.乳酸菌降解亚硝酸盐机理的研究[J].食品与发酵工业,2002,28(8):27-31.DOI:10.3321/j.issn:0253-990X.2002.08.007.
[34]黄丽慧,张雁,陈于陇,等.发酵蔬菜中亚硝酸盐消长规律及调控技术的研究进展[J].食品科学,2013,34(5):303-307.