为生产具有西藏地区特色的青稞小曲产品,确保青稞酒产品风味的稳定性,采用传统培养法对收集的21个青稞小曲样品中的微生物进行分离纯化和分子生物学鉴定,利用透明圈法初筛和制曲复筛选择优良菌株。结果表明,西藏青稞小曲中共分离出278株菌,主要包括蜡样芽孢杆菌、沙福芽孢杆菌、短小芽孢杆菌、苏云金芽孢杆菌、解淀粉芽孢杆菌和Kocuria marina 6种细菌,扣囊复膜孢酵母、酿酒酵母、Saccharomycopsis malanga、米根霉、卷枝毛霉、总状毛霉、白囊耙齿菌、伞状毛霉菌、米曲霉、黄曲霉、黑曲霉、Coriolopsis trogii和毛栓菌13种真菌。复筛得到糖化力为(1 382±77.38)mg/(g·h)的米根霉M12,中性蛋白酶活力为(1 035.56±40.09)μg/(g·min)的解淀粉芽孢杆菌Q4。该研究结果为优化制曲工艺和提升青稞酒品质提供了研究基础。
This study aimed to produce highland barley Xiaoqu products with Tibetan characteristics while ensuring flavor stability of highland barley wine products. The traditional culture method was used to separate, purify and identify microorganisms collected from 21 highland barley Xiaoqu samples, followed by transparent circle and starter-making screening. The results showed that a total of 278 strains were isolated and purified from Tibetan highland barley Xiaoqu, including six species of bacteria: Bacillus cereus, Bacillus safensis, Bacillus pumilus, Bacillus thuringiensis, Bacillus amyloliquefaciens, and Kocuria marina. Besides, there were 13 fungi identified, including Saccharomycopsis fibuligera, Saccharomyces cerevisiae, Saccharomycopsis malanga, Rhizopus oryzae, Coriolopsis trogii, Mucor circinelloides, Mucor racemosus, Irpex lacteus, Lichtheimia corymbifera, Aspergillus oryzae, Aspergillus flavus, Aspergillus niger, and Trametes hirsute. R. oryzae M12 with a saccharifying power of (1 382±77.38) mg/(g·h) and B. amyloliquefaciens Q4 with a protease activity of (1 035.56±40.09) μg/(g·min) were obtained by secondary screening. This study provides a research basis for optimizing starter-making process and improving the quality of highland barley wine.
[1] 杜木英.西藏青稞酒发酵微生物及酿造技术研究[D].重庆:西南大学,2008.
[2] 王丽华.西藏传统青稞酒的生产菌株选育及生产技术研究[D].重庆:西南大学,2008.
[3] LIU Z B,WANG Z Y,LYU X C,et al. Comparison study of the volatile profiles and microbial communities of Wuyi Qu and Gutian Qu, two major types of traditional fermentation starters of Hong Qu glutinous rice wine[J].Food Microbiology,2018,69:105-115.
[4] GOU M,WANG H Z,YUAN H W,et al.Characterization of the microbial community in three types of fermentation starters used for Chinese liquor production[J].Journal of the Institute of Brewing,2015,121(4):620-627.
[5] 郭小芳,王燕鸽,刘帅,等.西藏青稞酒曲中真菌的分离与鉴定[J].西藏大学学报(自然科学版),2014, 29(2):37-43.
[6] 李玉英,赵文娟,陈占秀,等.青稞酒生产工艺优化研究进展[J].酿酒,2018,45(5):50-52.
[7] ZHENG X W,YAN Z,HAN B Z,et al. Complex microbiota of a Chinese “Fen” liquor fermentation starter (Fen-Daqu), revealed by culture-dependent and culture-independent methods[J].Food Microbiology,2012,31(2):293-300.
[8] 刘杰雄,陈号,陆雯,等.淀粉酶高产菌株的筛选及其酶活的测定[J].食品工程,2010(1):45-47.
[9] 余培斌.改善绍兴黄酒麦曲品质的初步研究[D].无锡:江南大学,2013.
[10] 任清,侯昌.北宗黄酒麦曲微生物的分离鉴定[J].食品科学,2017,38(4):83-88.
[11] 翟磊,信春晖,许玲,等.芝麻香型白酒高温大曲产脂肪酶菌株的筛选与鉴定[J].食品与发酵工业,2015,41(12):36-39.
[12] 苏畅,窦晓,叶新,等.基于ITS4/5 rRNA区序列对不同时期大曲中霉菌的分离与鉴定[J].现代食品科技,2018,34(3):54-58.
[13] 毛祥,黄丹,沈才萍,等.酱香型大曲中产淀粉酶菌的分离鉴定及发酵特性研究[J].中国酿造,2015,34(12):24-27.
[14] 杜娟,艾小委,张乐怡,等.不同酒曲生料液态酿酒比较及优势真菌分离鉴定[J].蚌埠学院学报,2018,38(2):7-10.
[15] 王舸楠,马文瑞,全莉,等.酿酒葡萄DNA的提取方法[J].食品与发酵工业,2018,44(12):118-122.
[16] 刘茗铭,周阳子,袁乐梅,等.酒曲中高产糖化酶霉菌的筛选及其固态发酵产酶条件优化[J].食品与发酵工业,2018,44(10):118-123.
[17] 叶磊.小曲中优良根霉的分离筛选及产酶条件研究[D].重庆:西南大学,2009.
[18] 武顺,王德良,闫寅卓.产吡嗪类细菌的麸曲制作工艺优化[J].中国酿造,2016,35(2):13-17.
[19] 罗惠波,谢军,黄治国,等.纯种根霉麸曲制曲工艺优化研究[J].四川理工学院学报(自然科学版),2015,28(5):7-11.
[20] 沈怡方.白酒生产技术全书[M].北京:中国轻工业出版社,1998.
[21] 满都拉,刘昊,郑逸飞,等.不同地区小曲中可培养细菌的筛选及鉴定[J].中国酿造,2018,37(10):61-65.
[22] 梁香,吴天祥,敖锐,等.黄酒优良糖化菌株的筛选及其酶活性能的研究[J].酿酒科技,2015,249(3):12-15.
[23] ZHENG Xiaowei,YAN Zheng,NOUT M J,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.
[24] WANG C L,SHI D J,GONG G L.Microorganisms in Daqu: a starter culture of Chinese Maotai-flavor liquor[J].World Journal of Microbiology and Biotechnology,2008,24(10):2 183-2 190.
[25] CAI H Y,ZHANG T,ZHANG Q,et al.Microbial diversity and chemical analysis of the starters used in traditional Chinese sweet rice wine[J].Food Microbiology,2018,73:319-326.
[26] 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.
[27] 王旭亮,王异静,王德良,等.白酒发酵高糖化性能霉菌的筛选及鉴定[J].酿酒科技,2012,219(9):22-28.
[28] 黄家驷,刘盾,陈玄阳,等.浓香型高温酒曲中蛋白酶产生菌的筛选与鉴定[J].安徽农学通报,2017,23(21):29-31.
[29] 张彩莹,肖连冬.生物化学实验[M].北京:化学工业出版社,2009:10-67.
[30] 张中义,畅晓霞,王琦.大曲酒糖化阶段一株优势细菌分离鉴定[J].中国酿造,2009,28(7):47-49.