高通量测序技术分析酱香型白酒下造沙轮次的微生物多样性
|
摘要
下沙、造沙是酱香型白酒的基础轮次,为探究酱香型白酒下沙、造沙轮次发酵过程中微生物群落结构的变化、微生物多样性与样品之间的关系,本研究采用了高通量测序对酒醅中细菌、真菌多样性进行了全面解析。结果表明,下沙、造沙分别检测到细菌203个属和184个属,真菌160个属和109个属。下沙、造沙期间细菌优势菌属均为乳酸菌,堆积发酵结束,Weissella paramesenteroides、Pediococcus acidilactici、Lactobacillus panis占比可达80%以上,窖内7 d时的菌群结构发生较大变化,至窖内发酵结束,Lactobacillus panis、Lactobacillus sp.、Lactobacillus acetotolerans占比可达90%以上;下沙、造沙期间的真菌优势菌属均为酵母菌,下沙堆积到入窖,Pichia kudriavzevii均占绝对优势,占比在90%左右,造沙窖内发酵时,Saccharomyce scerevisiae大量繁殖,至窖内发酵结束达到22.32%,Pichia kudriavzevii占比仍达48.78%。该研究结果揭示,酱香型白酒下沙、造沙发酵过程中微生物的多样性,确定了下沙、造沙的主要微生物种类,反映了不同阶段微生物群落特点,为深入研究酿造机理提供参考依据。
The grains feeding at the first/second time is the basic fermentation cycle of Jiangxiang Baijiu. In order to explore the change in microbial community structure, and the relations between microbial diversity and samples in the 1 st and the 2 nd fermentation cycle, high-throughput sequencing technology was used to analyze the diversity of bacteria and fungi in fermented grains. The results showed that, 203 genera of bacteria and 160 genera of fungi were detected in the first fermentation cycle, and 184 genera of bacteria and 109 genera of fungi were detected in the second fermentation cycle. There were significant differences in microbial community structure between different fermentation stages. The main bacteria groups were lactic acid bacteria in both fermentation cycles.After accumulation fermentation ended, the ratio of Weissella paramesenteroides, Pediococcus acidilactici and Lactobacillus panis was over 80 %. Great change occurred in bacteria structure on the 7 th day of the fermentation; after the ending of in-pit fermentation,the ratio of Lactobacillus panis, Lactobacillus sp., and Lactobacillus acetotolerans was over 90 %. The main fungi groups were yeasts in both fermentation cycles. The ratio of Pichia kudriavzevii was about 90 % in the first fermentation cycle. Saccharomyce scerevisiae began to multiply during the 2 nd fermentation cycle and its ratio reached up to 22.32 %, and the ratio of Pichia kudriavzevii was still 48.78 % at the end of the 2 nd fermentation cycle. The research revealed the diversity of microbes in the 1 st and the 2 nd fermentation cycles of Jiangxiang Baijiu, and reflected the characteristics of microbial community structure in different fermentation stages, which could provide references for further study of the fermenting mechanism of Jiangxiang Baijiu.
The grains feeding at the first/second time is the basic fermentation cycle of Jiangxiang Baijiu. In order to explore the change in microbial community structure, and the relations between microbial diversity and samples in the 1 st and the 2 nd fermentation cycle, high-throughput sequencing technology was used to analyze the diversity of bacteria and fungi in fermented grains. The results showed that, 203 genera of bacteria and 160 genera of fungi were detected in the first fermentation cycle, and 184 genera of bacteria and 109 genera of fungi were detected in the second fermentation cycle. There were significant differences in microbial community structure between different fermentation stages. The main bacteria groups were lactic acid bacteria in both fermentation cycles.After accumulation fermentation ended, the ratio of Weissella paramesenteroides, Pediococcus acidilactici and Lactobacillus panis was over 80 %. Great change occurred in bacteria structure on the 7 th day of the fermentation; after the ending of in-pit fermentation,the ratio of Lactobacillus panis, Lactobacillus sp., and Lactobacillus acetotolerans was over 90 %. The main fungi groups were yeasts in both fermentation cycles. The ratio of Pichia kudriavzevii was about 90 % in the first fermentation cycle. Saccharomyce scerevisiae began to multiply during the 2 nd fermentation cycle and its ratio reached up to 22.32 %, and the ratio of Pichia kudriavzevii was still 48.78 % at the end of the 2 nd fermentation cycle. The research revealed the diversity of microbes in the 1 st and the 2 nd fermentation cycles of Jiangxiang Baijiu, and reflected the characteristics of microbial community structure in different fermentation stages, which could provide references for further study of the fermenting mechanism of Jiangxiang Baijiu.
引文
[1] WANG X D, BAN S D, HU B D, et al. Bacterial diversity of Moutai-flavour Daqu based on high-throughput sequencing method[J]. Journal of the Institute of Brewing,2017,123(6):138-143.
[2]杨旭剑,吕云怀.浅谈茅台酒生产工艺的系统性[J].酿酒科技, 2007(6):54-58.
[3]李长江,沈才洪,张宿义,等.武陵酱香型白酒工艺创新——特殊润粮工艺的研究(第一报)[J].酿酒科技, 2009(7):40-42.
[4]杨飞,沈才洪,张洪远,等.酱香型白酒造沙轮次堆积新工艺研究[J].酿酒科技,2012(4):35-38.
[5] WANG H Y, ZHANG X J, ZHAO L P, et al. Analysis and comparison of the bacterial community in fermented grains during the fermentation for two different styles of Chinese liquor[J]. Journal of industrial microbiology&biotechnology, 2008,35(6):603-609.
[6]赵亮,王莉,汪地强,等.白酒微生物群落研究技术现状与二代测序数据分析方略[J].酿酒科技,2016(7):88-96.
[7]关统伟,向慧平,王鹏昊,等.基于高通量测序的郫县豆瓣不同发酵期细菌群落结构及其动态演替[J].食品科学,2018,39(4):106-111.
[8]邢敏钰,杜海,徐岩.芝麻香型白酒发酵过程中乳酸菌多样性及其演替规律[J].微生物学通报,2018,45(1):19-28.
[9]吴莉莉,王海燕,徐岩,等.酱香型与清香型白酒发酵过程中乳酸菌菌群的差异性分析[J].微生物学通报,2013(12):2182-2188.
[10] SUN W N, XIAO H Z, PENG Q, et al. Analysis of bacterial diversity of Chinese Luzhou-flavor liquor brewed in different seasons by Illumina Miseq sequencing[J]. Annals of microbiology,2016,66(3):1293-1301.
[11] YANG J G, DOU X, MA Y Y. Diversity and dynamic succession of microorganisms during Daqu preparation for Luzhou-flavour liquor using second-generation sequencing technology[J]. Journal of The Institute of Brewing, 2018,124(4):498-507.
[12]栗连会.泸型酒酒醅中乳酸菌和乳酸降解菌的多样性和代谢特性[D].无锡:江南大学, 2016:10-20.
[13]刘雄,甘兴,罗立新.基于16S/18S rRNA基因文库技术研究酿醋大曲固态发酵过程中微生物群落结构演变[J].酿酒科技, 2017(1):42-47.
[14]陈良强,杨帆,王和玉,等.基于细菌群落构成对高温大曲类别的判别分析[J].酿酒科技,2016(9):48-50.
[15]李申奥.兼香型白酒高温大曲微生物群落演替规律的研究[D].武汉:华中农业大学,2016:20-30.
[16]许焰,吴群,徐岩.酱香型白酒酿造主要功能菌株对拜耳接合酵母的作用[J].微生物学通报,2018,45(1):42-53.
[17] LIU H L, SUN B G. Effect of fermentation processing on the flavor of Baijiu[J].Journal of agricultural and food chemistry, 2018,66(22):5425-5432.
[18]林建春,吴群,徐岩.清香型白酒发酵过程中酵母群落结构及其与尿素代谢的关系[J].微生物学通报,2017,44(11):2522-2529.
[19] WANG X S, DU H, ZHANG Y, et al. Environmental microbiota drives microbial succession and metabolic profiles during Chinese liquor fermentation[J]. Appl environ microbiol, 2018,84(4):9-17.
[20] GUO J H, SUN L H, GUO H W, et al. Effect of cellulase-producing bacteria on fungi community structure and ester generation in Chinese liquor fermenting grains[J]. Journal of the American Society of Brewing Chemists, 2018,76(2):130-140.
[21] LIU P L, XIONG X M, WANG S, et al. Population dynamics and metabolite analysis of yeasts involved in a Chinese miscellaneous-flavor liquor fermentation[J].Annals of microbiology, 2017,67(8):553-565.
[22]胡佳音,周森,赵卫鹏,等.清、浓、酱三种大曲真菌多样性初步分析[J].酿酒科技, 2016(8):87-90.
[23]刘婷婷.白云边酒酿造微生物分析及东方伊萨酵母发酵特性研究[D].武汉:武汉工业学院,2011.
[24]杨建刚,苏畅,窦晓,等.泸型酒发酵过程中酵母菌演替规律及其对部分风味分子形成的影响[J].食品科学,2018,39(18):166-172.
[25]唐洁,王海燕,徐岩.酿酒酵母和异常毕赤酵母混菌发酵对白酒液态发酵效率和风味物质的影响[J].微生物学通报,2012,39(7):921-930.
[2]杨旭剑,吕云怀.浅谈茅台酒生产工艺的系统性[J].酿酒科技, 2007(6):54-58.
[3]李长江,沈才洪,张宿义,等.武陵酱香型白酒工艺创新——特殊润粮工艺的研究(第一报)[J].酿酒科技, 2009(7):40-42.
[4]杨飞,沈才洪,张洪远,等.酱香型白酒造沙轮次堆积新工艺研究[J].酿酒科技,2012(4):35-38.
[5] WANG H Y, ZHANG X J, ZHAO L P, et al. Analysis and comparison of the bacterial community in fermented grains during the fermentation for two different styles of Chinese liquor[J]. Journal of industrial microbiology&biotechnology, 2008,35(6):603-609.
[6]赵亮,王莉,汪地强,等.白酒微生物群落研究技术现状与二代测序数据分析方略[J].酿酒科技,2016(7):88-96.
[7]关统伟,向慧平,王鹏昊,等.基于高通量测序的郫县豆瓣不同发酵期细菌群落结构及其动态演替[J].食品科学,2018,39(4):106-111.
[8]邢敏钰,杜海,徐岩.芝麻香型白酒发酵过程中乳酸菌多样性及其演替规律[J].微生物学通报,2018,45(1):19-28.
[9]吴莉莉,王海燕,徐岩,等.酱香型与清香型白酒发酵过程中乳酸菌菌群的差异性分析[J].微生物学通报,2013(12):2182-2188.
[10] SUN W N, XIAO H Z, PENG Q, et al. Analysis of bacterial diversity of Chinese Luzhou-flavor liquor brewed in different seasons by Illumina Miseq sequencing[J]. Annals of microbiology,2016,66(3):1293-1301.
[11] YANG J G, DOU X, MA Y Y. Diversity and dynamic succession of microorganisms during Daqu preparation for Luzhou-flavour liquor using second-generation sequencing technology[J]. Journal of The Institute of Brewing, 2018,124(4):498-507.
[12]栗连会.泸型酒酒醅中乳酸菌和乳酸降解菌的多样性和代谢特性[D].无锡:江南大学, 2016:10-20.
[13]刘雄,甘兴,罗立新.基于16S/18S rRNA基因文库技术研究酿醋大曲固态发酵过程中微生物群落结构演变[J].酿酒科技, 2017(1):42-47.
[14]陈良强,杨帆,王和玉,等.基于细菌群落构成对高温大曲类别的判别分析[J].酿酒科技,2016(9):48-50.
[15]李申奥.兼香型白酒高温大曲微生物群落演替规律的研究[D].武汉:华中农业大学,2016:20-30.
[16]许焰,吴群,徐岩.酱香型白酒酿造主要功能菌株对拜耳接合酵母的作用[J].微生物学通报,2018,45(1):42-53.
[17] LIU H L, SUN B G. Effect of fermentation processing on the flavor of Baijiu[J].Journal of agricultural and food chemistry, 2018,66(22):5425-5432.
[18]林建春,吴群,徐岩.清香型白酒发酵过程中酵母群落结构及其与尿素代谢的关系[J].微生物学通报,2017,44(11):2522-2529.
[19] WANG X S, DU H, ZHANG Y, et al. Environmental microbiota drives microbial succession and metabolic profiles during Chinese liquor fermentation[J]. Appl environ microbiol, 2018,84(4):9-17.
[20] GUO J H, SUN L H, GUO H W, et al. Effect of cellulase-producing bacteria on fungi community structure and ester generation in Chinese liquor fermenting grains[J]. Journal of the American Society of Brewing Chemists, 2018,76(2):130-140.
[21] LIU P L, XIONG X M, WANG S, et al. Population dynamics and metabolite analysis of yeasts involved in a Chinese miscellaneous-flavor liquor fermentation[J].Annals of microbiology, 2017,67(8):553-565.
[22]胡佳音,周森,赵卫鹏,等.清、浓、酱三种大曲真菌多样性初步分析[J].酿酒科技, 2016(8):87-90.
[23]刘婷婷.白云边酒酿造微生物分析及东方伊萨酵母发酵特性研究[D].武汉:武汉工业学院,2011.
[24]杨建刚,苏畅,窦晓,等.泸型酒发酵过程中酵母菌演替规律及其对部分风味分子形成的影响[J].食品科学,2018,39(18):166-172.
[25]唐洁,王海燕,徐岩.酿酒酵母和异常毕赤酵母混菌发酵对白酒液态发酵效率和风味物质的影响[J].微生物学通报,2012,39(7):921-930.