从浓香型大曲中分离1株扣囊复膜酵母菌,在混菌固态发酵条件下研究其对微生物结构及代谢物的影响。基于混菌固态发酵体系,在发酵温度/湿度分别为25 ℃/95%、28 ℃/90%、35 ℃/85%、38 ℃/80%、43 ℃/75%的条件下接种扣囊复膜酵母菌,分析其对混菌发酵体系微生物多样性、理化参数、酶活力、挥发性代谢物的影响。结果表明,在不同的发酵条件下扣囊复膜酵母菌的扰动会导致固态混菌培养体系中微生物物种丰富度降低,毕赤酵母属(Pichia)、横梗霉属(Lichtheimia)、肠杆菌属(Enterobacter)的相对丰度降低,芽孢杆菌属(Bacillus)、复膜酵母属(Saccharomycopsis)相对丰度增多。25 ℃/95%发酵条件下还原糖含量、糖化力、液化力、发酵力、酒化力有所提高,28 ℃/90%发酵条件下实验组的还原糖含量、糖化力、发酵力有明显增加,液化力在所有发酵条件下都有增强。35 ℃/85%、42 ℃/75% 发酵条件下吡嗪类化合物显著增多,25 ℃/95%、38 ℃/80% 发酵条件下醇类化合物增多,25 ℃/95%、35 ℃/85%发酵条件下酯类化合物增多。可见接种扣囊复膜酵母菌会对固态混菌体系中其他的微生物造成扰动进而影响体系中的微生物代谢。
A Saccharomycopsis fibuligera was isolated from strong flavor Baijiu Daqu. The effects of S. fibuligera on the microbial community structure and metabolism were studied in mixed solid-state fermentation. S. fibuligera was inoculated into mixed solid fermentation system. The effects of S. fibuligera on microbial diversity, physicochemical parameters, enzyme activity and volatile metabolites were analyzed, when the temperature and relative humidity of mixed solid fermentation system were controlled at 25 ℃/95%, 28 ℃/90%, 35 ℃/85%, 38 ℃/80% and 43 ℃/75%, respectively. The results showed that the bioturbation effect of S. fibuligera would lead to the decrease of microbial richness in the solid mixed fermentation system. The relative content of Pichia, Lichthemia and Enterobacter decreased, while that of Bacillus and Saccharomycosis increased. When the fermentation condition was 25 ℃/95%, the content of reducing sugar, saccharifying power, liquefying power, fermenting power and liquor-producing power increased. The content of reducing sugar, saccharifying power and fermentation power of all experimental groups increased significantly under 28 ℃/90%. The liquefaction power increased under all fermentation conditions. At 35 ℃/85% and 42 ℃/75%, the content of pyrazine compounds increased significantly. At 25 ℃/95% and 38 ℃/80%, alcohol compounds increased. Then, ester compounds increased at 25 ℃/95% and 35 ℃/85%. These results indicated that the bioturbation of S. fibuligera would have an influence on other microorganisms and the microbial metabolism in the mixed solid system.
[1] CHI Z M,CHI Z,LIU G L,et al.Saccharomycopsis fibuligera and its applications in biotechnology[J].Biotechnology Advances,2009,27(4):423-431.
[2] ZHENG X W,YAN Z,NOUT M J R,et al.Microbiota dynamics related to environmental conditions during the fermentative production of Fen-Daqu,a Chinese industrial fermentation starter[J].International Journal of Food Microbiology,2014,182-183:57-62.
[3] LYU X C,CHEN Z C,JIA R B,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.
[4] DU H,WANG X S,ZHANG Y H,et al.Exploring the impacts of raw materials and environments on the microbiota in Chinese Daqu starter[J].International Journal of Food Microbiology,2019,297:32-40.
[5] SU C,ZHANG K Z,CAO X Z,et al.Effects of Saccharomycopsis fibuligera and Saccharomyces cerevisiae inoculation on small fermentation starters in Sichuan-style Xiaoqu liquor[J].Food Research International,2020,137.DOI:10.1016/j.foodres.2020.109425.
[6] WANG X D,QIU S Y,LI P,et al.Analysis of microbial community structure in traditional and automated Moutai-flavor Daqu[J].Journal of the American Society of Brewing Chemists,2019,77(2):140-146.
[7] JUNG M J,NAM Y D,ROH S W,et al.Unexpected convergence of fungal and bacterial communities during fermentation of traditional Korean alcoholic beverages inoculated with various natural starters[J].Food Microbiology,2012,30(1):112-123.
[8] 周森, 韩培杰,胡佳音,等.牛栏山二锅头酿造过程中的真菌多样性分析[J].食品工业科技,2018,19(1):127-130;136.
ZHOU S,HAN P J,HU J Y,et al.Analysis of fungal diversity in Niulanshan Erguotou brewing process[J].Science and Technology of Food Industry,2018,39(1):127-130;136.
[9] 周森,胡佳音,崔洋,等.应用高通量测序技术解析清香型大曲微生物多样性[J].中国食品学报,2019,19(6):244-250.
ZHOU S,HU J Y,CUI Y,et al.Microbial diversity analysis of flight-flavor Daqu using high-throughput sequencing[J].Journal of Chinese Institute of Food Science and Technology,2019,19(6):244-250.
[10] 马凯,崔哲男,郑晓卫,等.汾酒大曲可培养真菌多样性的初步分析[J].中国酿造,2011,30(8):19-21.
MA K,CUI Z N,ZHENG X W,et al.Diversity of culturable fungi in Fen-Daqu[J].China Brewing,2011,30(8):19-21.
[11] ZHENG X W,TABRIZI M R,NOUT M J R,et al.Daqu-a traditional Chinese liquor fermentation starter[J].Journal of the Institute of Brewing,2012,117(1):82-90.
[12] HE G,HUANG J,WU C,et al.Bioturbation effect of fortified Daqu on microbial community and flavor metabolite in Chinese strong-flavor liquor brewing microecosystem[J].Food Research International,2019,129:108 851.
[13] WANG H Y,GAO Y B,FAN Q W,et al.Characterization and comparison of microbial community of different typical Chinese liquor Daqus by PCR-DGGE[J].Letters in Applied Microbiology,2011,53(2):134-140.
[14] YANG J G,DOU X,MA Y Y,et al.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.
[15] JULIA M N,LEONTION G,SANTIAGO A,et al.Structural and functional analysis of hybrid enzymes generated by domain shuffling between Saccharomyces cerevisiae (var.diastaticus) Sta1 glucoamylase and Saccharomycopsis fibuligera Bgl1 β-glucosidase[J].Applied Microbiology and Biotechnology,2011,89(1):121-130.
[16] 黄晓宁, 刘晶晶,韩北忠,等.基于酶学特性筛选大曲来源芽孢杆菌用于强化酿酒[J/OL].食品科学:2021.http://kns.cnki.net/kcms/detail/11.2206.TS.20200722.1708.234.html.
HUANG X N,LIU J J,HAN B Z,et al.Screening of bacillus from Daqu based on enzymatic characteristics for enhanced brewing [J/OL].Food Science:2021.http://kns.cnki.net/kcms/detail/11.2206.TS.20200722.1708.234.html.
[17] HE G,HUANG J,ZHOU R,et al.Effect of fortified Daqu on the microbial community and flavor in Chinese strong-flavor liquor brewing process[J].Frontiers in Microbiology,2019,10.DOI:10.3389/fmicb.2019.00056.
[18] 张丽杰, 张怀志,徐岩.枯草芽孢杆菌Nr.5和底物添加促进酱油中吡嗪类物质合成[J].食品与发酵工业,2020,46(21):1-8.
ZHANG L J,ZHANG H Z,XU Y.Pyrazines synthesis in soy sauce promoted by Bacillus subtilis Nr.5 and substrate addition[J].Food and Fermentation Industry,2020,46 (21):1-8.
