一种传统发酵酵子细菌多样性及挥发性发酵产物分析
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摘要
通过分离纯化,结合16S rDNA克隆文库与扩增性rDNA限制酶切分析及Biolog碳源利用特性鉴定,得出一种传统发酵酵子中主要细菌群落及其构成比例依次为魏斯氏菌属(84.62%)、乳酸乳杆菌属(8.79%)、芽孢杆菌属(3.30%)、醋酸杆菌属(2.20%)和明串珠菌属(1.10%)。同时采用固相微萃取-气质联用法分析传统酵子制作馒头中的挥发性成分,共检测出59种挥发性风味物质,其中包括17种醇类物质、8种烃类物质、15种脂类物质、9种醛类物质、4种酮类物质、5种芳香类物质和1种其他类物质,较单纯酵母菌发酵的馒头风味物质种类丰富。结合定量和定性分析得出,醇类的含量和种类是区分两种馒头的主要标志物。
Microbial diversity of a traditional Jiaozi was studied by isolation and purification and 16 S r DNA Clone Library technique and Amplified r DNA Restriction Analysis(ARDRA) and Biolog carbon utilization analysis. The dominant bacterium were Weissella(84. 62%),Lactobacillus(8. 79%),Bacillus(3. 30%),Acetobacter(2. 20%)and Leuconostoc(1. 10%). The compositions of volatile components in Chinese steamed bread(CSB) fermented with the traditional Jiaozi were determined by solid phase microextraction(SPME) coupled with gas chromatography-mass spectrometry(GC-MS). 59 volatile compounds were detected in CBSs by Jiaozi including 17 kind of alcohols,8 kind of hydrocarbons,15 kind of esters,9 kind of aldehydes,4 kind of ketones,5 kind of aromatics and 1 kind of other compounds,which was more than in dry yeast starters. Combining quantitative and qualitative analysis,the content and variety of alcohols is the main marker to distinguish the two CSBs. This study revealed that the traditional Jiaozi had a complex composition of bacteria,and produced rich flavor compounds in CBS fermentation. This study supplied the basic data for industrialization of traditional staple food and supplied strain source for the further study of biological function research and supplied a new idea of the development of nutrition of traditional staple food.
Microbial diversity of a traditional Jiaozi was studied by isolation and purification and 16 S r DNA Clone Library technique and Amplified r DNA Restriction Analysis(ARDRA) and Biolog carbon utilization analysis. The dominant bacterium were Weissella(84. 62%),Lactobacillus(8. 79%),Bacillus(3. 30%),Acetobacter(2. 20%)and Leuconostoc(1. 10%). The compositions of volatile components in Chinese steamed bread(CSB) fermented with the traditional Jiaozi were determined by solid phase microextraction(SPME) coupled with gas chromatography-mass spectrometry(GC-MS). 59 volatile compounds were detected in CBSs by Jiaozi including 17 kind of alcohols,8 kind of hydrocarbons,15 kind of esters,9 kind of aldehydes,4 kind of ketones,5 kind of aromatics and 1 kind of other compounds,which was more than in dry yeast starters. Combining quantitative and qualitative analysis,the content and variety of alcohols is the main marker to distinguish the two CSBs. This study revealed that the traditional Jiaozi had a complex composition of bacteria,and produced rich flavor compounds in CBS fermentation. This study supplied the basic data for industrialization of traditional staple food and supplied strain source for the further study of biological function research and supplied a new idea of the development of nutrition of traditional staple food.
引文
[1]张国华,何国庆.我国传统馒头发酵剂的研究现状[J].中国食品学报,2012,12(11):115-120.
[2]杨敬雨,刘长虹.中国传统酵子的工业化[J].食品研究与开发,2007(2):164-166.
[3]ZHANG Jia-chao,LIU Wen-jun,SUN Zhi-hong,et al.Diversity of lactic acid bacteria and yeasts in traditional sourdoughs collected from western region in Inner Mongolia of China[J].Food Control,2011,22(5):767-774.
[4]马凯,华威,龚平,等.顶空-固相微萃取方法分析4种发酵剂制作馒头中挥发性风味物质[J].食品科学,2014,35(16):128-132.
[5]韩德权,孙庆申,李冰,等.复合发酵剂馒头和单一酵母馒头风味物质比较[J].食品科学,2012,33(2):240-242.
[6]邓璀,李志建,刘长虹,等.顶空固相微萃取-气质联用分析酵子发酵面团挥发性风味物质[J].食品科技,2015,40(11):124-130.
[7]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M].8版.北京:科学出版社,1984:729-805.
[8]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,1999:58-60.
[9]CLEENWERCK I,VANDEMEULEBROECKE K,JANSSENS D,et al.Re-examination of the genus Acetobacter,with descriptions of Acetobacter cerevisiae sp.nov.and Acetobacter malorum sp.nov.[J].International Journal of Systematic and Evolutionary Microbiology,2002,52:1 551-1 558.
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[11]马凯,安云鹏,武会娟,等.一株优质传统馒头发酵酵素中乳酸菌CGMCC10264的分离与鉴定[J].食品安全质量检测学报,2016,7(9):3 720-3 725.
[12]李树高.影响馒头品质因素的探讨[J].粮食加工,2010,35(4):72-73.
[13]刘同杰,李云,吴诗榕,等.传统酸面团中细菌与酵母菌的分离与鉴定[J].现代食品科技,2014,30(9):114-120,148.
[14]姚国强,李慧,高鹏飞,等.乳酸菌在发酵酸面团中的研究与应用[J].中国食品学报,2013,13(3):163-170.
[15]姚笛,孙大庆,李洪飞.东北粘豆包酸面团菌群多样性与自制酸面团中菌群变化规律的研究[J].现代食品科技,2015,31(5):90-95.
[16]ZHONG Guo-hua,HE Guo-qing.Predominant bacteria diversity in Chinese traditional sourdough[J].Journal ofFood Science,2013,78(8):1 218-1 223.
[17]LIU Tong-jie,LI Yun,CHEN Jian-cai,et al.Prevalence and diversity of lactic acid bacteria in Chinese traditional sourdough revealed by culture dependent and pyrosequencing approaches[J].LWT-Food Science and Technology,2016,68:91-97.
[18]COLLINS M D,SAMELIS J,METAXOPOULOS J,et al.Taxonomic studies on some leuconostoc-like organisms from fermented sausages:description of a new genus Weissella for the Leuconostoc paramesenteroides group of species[J].Journal of Applied Bacteriology,1993,75(6):595-603.
[19]王海娟,戴雨珂,潘渠.魏斯氏菌的研究现状[J].成都医学院学报,2014,9(6):747-750.
[20]张红,胡青平.探究馒头用酸面团中自然发酵种的最初来源[J].食品工业科技,2013,34(17):185-188.
[21]ZHENG Xiao-wei,Tabrizi M R,NOUT M,et al.Daqu—a traditional Chinese liquor fermentation starter[J].Journal of the Institute of Brewing,2011,117(1):82-90.
[22]尹艳丽,王金水,杨森,等.酸面团中的微生物群落及发酵过程中的代谢反应[J].食品工业科技,2014,35(19):368-372.
[23]苏东海,李自红,苏东民,等.固相微萃取分析传统老酵头馒头挥发性物质[J].食品研究与开发,2011,32(6):94-97.
[24]LE V D,ZHENG Xiao-wei,CHEN jing-yu,et al.Characterization of volatile compounds in Fen-Daqu a traditional Chinese liquor fermentation starter[J].Journal of the Institute of Brewing,2012,118(1):107-113.
[25]HEIMAN M L,GREENWAY F L.A healthy gastrointestinal microbiome is dependent on dietary diversity[J].Molecular Metabolism,2016,5(5):317-320.
[26]乜兰春,孙建设,陈华君,等.苹果不同品种果实香气物质研究[J].中国农业科学,2006,39(3):641-646.
[27]SALMINEN S,von Wright A.Lactic acid bacteria microbiology and functional aspects[M].New York:Marcel Dekker,1999:1-72.
[28]丁必权,李秋鹏,季秀玲,等.高温短小芽孢杆菌及其产高温淀粉酶的初步研究[J].食品工业科技,2010,31(9):185-187:190.
[29]刘青娥,钟仙龙.柑桔果醋酿造用菌种分离及其生产性能测定[J].广东农业科学,2015,42(20):80-85.
[30]THIELE C,GANZLE M G,VOGEL R F.Contribution of sourdough lactobacilli,yeast,and cereal enzymes to the generation of amino acids indough relevant for bread flavor[J].Cereal Chemistry,2002,79(1):45-51.
[31]LIU S Q.Practical implications of lactose and pyruvate metabolism bylactic acid bacteria in food and beverage fermentations[J].International Journal of Food Microbiology,2003,83(2):115-131.
[2]杨敬雨,刘长虹.中国传统酵子的工业化[J].食品研究与开发,2007(2):164-166.
[3]ZHANG Jia-chao,LIU Wen-jun,SUN Zhi-hong,et al.Diversity of lactic acid bacteria and yeasts in traditional sourdoughs collected from western region in Inner Mongolia of China[J].Food Control,2011,22(5):767-774.
[4]马凯,华威,龚平,等.顶空-固相微萃取方法分析4种发酵剂制作馒头中挥发性风味物质[J].食品科学,2014,35(16):128-132.
[5]韩德权,孙庆申,李冰,等.复合发酵剂馒头和单一酵母馒头风味物质比较[J].食品科学,2012,33(2):240-242.
[6]邓璀,李志建,刘长虹,等.顶空固相微萃取-气质联用分析酵子发酵面团挥发性风味物质[J].食品科技,2015,40(11):124-130.
[7]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M].8版.北京:科学出版社,1984:729-805.
[8]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,1999:58-60.
[9]CLEENWERCK I,VANDEMEULEBROECKE K,JANSSENS D,et al.Re-examination of the genus Acetobacter,with descriptions of Acetobacter cerevisiae sp.nov.and Acetobacter malorum sp.nov.[J].International Journal of Systematic and Evolutionary Microbiology,2002,52:1 551-1 558.
[10]BJORKROTH K J,SCHILLINGER U,GEISEN R,et al.Taxonomic study of Weissella confusa and description of Weissella cibaria sp.nov.,detected in food and clinical samples[J].International Journal of Systematic and Evolutionary Microbiology,2002,52(Pt1):141-148.
[11]马凯,安云鹏,武会娟,等.一株优质传统馒头发酵酵素中乳酸菌CGMCC10264的分离与鉴定[J].食品安全质量检测学报,2016,7(9):3 720-3 725.
[12]李树高.影响馒头品质因素的探讨[J].粮食加工,2010,35(4):72-73.
[13]刘同杰,李云,吴诗榕,等.传统酸面团中细菌与酵母菌的分离与鉴定[J].现代食品科技,2014,30(9):114-120,148.
[14]姚国强,李慧,高鹏飞,等.乳酸菌在发酵酸面团中的研究与应用[J].中国食品学报,2013,13(3):163-170.
[15]姚笛,孙大庆,李洪飞.东北粘豆包酸面团菌群多样性与自制酸面团中菌群变化规律的研究[J].现代食品科技,2015,31(5):90-95.
[16]ZHONG Guo-hua,HE Guo-qing.Predominant bacteria diversity in Chinese traditional sourdough[J].Journal ofFood Science,2013,78(8):1 218-1 223.
[17]LIU Tong-jie,LI Yun,CHEN Jian-cai,et al.Prevalence and diversity of lactic acid bacteria in Chinese traditional sourdough revealed by culture dependent and pyrosequencing approaches[J].LWT-Food Science and Technology,2016,68:91-97.
[18]COLLINS M D,SAMELIS J,METAXOPOULOS J,et al.Taxonomic studies on some leuconostoc-like organisms from fermented sausages:description of a new genus Weissella for the Leuconostoc paramesenteroides group of species[J].Journal of Applied Bacteriology,1993,75(6):595-603.
[19]王海娟,戴雨珂,潘渠.魏斯氏菌的研究现状[J].成都医学院学报,2014,9(6):747-750.
[20]张红,胡青平.探究馒头用酸面团中自然发酵种的最初来源[J].食品工业科技,2013,34(17):185-188.
[21]ZHENG Xiao-wei,Tabrizi M R,NOUT M,et al.Daqu—a traditional Chinese liquor fermentation starter[J].Journal of the Institute of Brewing,2011,117(1):82-90.
[22]尹艳丽,王金水,杨森,等.酸面团中的微生物群落及发酵过程中的代谢反应[J].食品工业科技,2014,35(19):368-372.
[23]苏东海,李自红,苏东民,等.固相微萃取分析传统老酵头馒头挥发性物质[J].食品研究与开发,2011,32(6):94-97.
[24]LE V D,ZHENG Xiao-wei,CHEN jing-yu,et al.Characterization of volatile compounds in Fen-Daqu a traditional Chinese liquor fermentation starter[J].Journal of the Institute of Brewing,2012,118(1):107-113.
[25]HEIMAN M L,GREENWAY F L.A healthy gastrointestinal microbiome is dependent on dietary diversity[J].Molecular Metabolism,2016,5(5):317-320.
[26]乜兰春,孙建设,陈华君,等.苹果不同品种果实香气物质研究[J].中国农业科学,2006,39(3):641-646.
[27]SALMINEN S,von Wright A.Lactic acid bacteria microbiology and functional aspects[M].New York:Marcel Dekker,1999:1-72.
[28]丁必权,李秋鹏,季秀玲,等.高温短小芽孢杆菌及其产高温淀粉酶的初步研究[J].食品工业科技,2010,31(9):185-187:190.
[29]刘青娥,钟仙龙.柑桔果醋酿造用菌种分离及其生产性能测定[J].广东农业科学,2015,42(20):80-85.
[30]THIELE C,GANZLE M G,VOGEL R F.Contribution of sourdough lactobacilli,yeast,and cereal enzymes to the generation of amino acids indough relevant for bread flavor[J].Cereal Chemistry,2002,79(1):45-51.
[31]LIU S Q.Practical implications of lactose and pyruvate metabolism bylactic acid bacteria in food and beverage fermentations[J].International Journal of Food Microbiology,2003,83(2):115-131.