Control and reduction of n-butanol synthesis during Baijiu fermentation interfered with Clostridia in pit mud
GOU Wenjun1,2, FANG Fang1,2*
1(Science Center for Future Food,Jiangnan University,Wuxi 214122,China) 2(Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China)
Abstract: Higher alcohols (HAs) are flavor substances in Chinese Baijiu, but the high content of HAs makes the Chinese Baijiu spicy and may cause discomfort after drinking. This work aims to reduce n-butanol during fermentation by addition of strain with low n-butanol synthesis ability. Eight Clostridium strains were isolated from the pit mud of Luzhou-flavor Baijiu. Among them, Clostridium beijerinckii 6Y-1 produced the highest level of n-butanol, which was 1 013-8 956 times than other Clostridia. Clostridium tyrobutyricum ZY-4 produced the lowest level of n-butanol, and it could significantly reduce the n-butanol level of C. beijerinckii 6Y-1 by co-cultivation. In a simulated Chinese Baijiu underground fermentation system, C. tyrobutyricum ZY-4 reduced the amount of n-butanol by 30.05%, significantly increased the content of ethyl caproate and ethyl caprylate, and had no obvious impact on the amount of the main volatiles. It had been confirmed that C. tyrobutyricum ZY-4 produced acetic acid and butyric acid via branch metabolic pathways, thus reduced the n-butanol anabolism of C. beijerinckii 6Y-1. The results laid important theoretical fundamental and gave a good reference in working out strategies for reduction and control of n-butanol synthesis in mixed fermentation system.
勾文君,方芳. 窖泥梭菌扰动减控白酒发酵过程正丁醇生成[J]. 食品与发酵工业, 2021, 47(15): 43-49.
GOU Wenjun,FANG Fang. Control and reduction of n-butanol synthesis during Baijiu fermentation interfered with Clostridia in pit mud[J]. Food and Fermentation Industries, 2021, 47(15): 43-49.
孙金旭, 朱会霞,杨晓红,等.酵母添加量对酱香型白酒中杂油醇影响[J].食品研究与开发,2010,31(12):67-70.SUN J X,ZHU H X,YANG X H,et al.The increase the volume of different yeast inoculum concentration influence study on the Maotai-alcohol liquor in oil[J].Food Research and Development,2010,31(12):67-70.
[2]
ZHU J,SHI J L,LU Y,et al.Application of strains of Geotrichum spp.to decrease higher alcohols and to increase esters[J].Journal of the Institute of Brewing and Distilling,2016,122(1):147-155.
[3]
ZHOU Z L,NI W,JI Z W,et al.Development of a rapid method for determination of main higher alcohols in fermented alcoholic beverages based on dispersive liquid-liquid microextraction and gas chromatography-mass spectrometry[J].Food Analytical Methods,2020,13(3):591-600.
[4]
格绒泽仁, 皇甫洁,韩兴林,等.浓香型白酒饮后不适感关键高级醇类物质关联性判定新方法[J].食品与发酵工业,2019,45(14):191-195.GERONG Z R,HUANGFU J,HAN X L,et al.A method for determining associations between drinking discomforts and key higher alcohols in strong-aroma Baijiu[J].Food and Fermentation Industries,2019,45(14):191-195.
[5]
苟静瑜, 贾智勇,闫宗科,等.降低白酒中高级醇含量的研究进展[J].酿酒,2016,43(4):25-29.GOU J Y,JIA Z Y,YAN Z K,et al.Research progress in decreasing the contents of higher alcohols in Baijiu (liquor)[J].Liquor Making,2016,43(4):25-29.
[6]
游玲, 任羽,王涛,等.酵母对浓香型白酒糟醅中高级醇生成的影响[J].食品与发酵工业,2016,42(2):23-28.YOU L,REN Y,WANG T,et al.Effects of yeasts on higher alcohols forming in Chinese strong-flavored liquor producing[J].Food and Fermentation Industries,2016,42(2):23-28.
[7]
曾朝珍, 张永茂,康三江,等.发酵酒中高级醇的研究进展[J].中国酿造,2015,34(5):11-15.ZENG C Z,ZHANG Y M,KANG S J,et al.Research progress of higher alcohols in fermented wine[J].China Brewing,2015,34(5):11-15.
[8]
ZHANG W X,WU Z Y,ZHANG Q S,et al.Combination of newly developed high quality Fuqu with traditional Daqu for Luzhou-flavor liquor brewing[J].World Journal of Microbiology and Biotechnology,2009,25(10):1 721-1 726.
[9]
周玮婧, 江小明.气相色谱法测定不同香型白酒中醇类与醛类物质含量[J].中国酿造,2017,36(4):180-183.ZHOU W J,JIANG X M.Determination of the alcohols and the aldehydes contents in different flavor types of Baijiu (Chinese liquor) by gas chromatography[J].China Brewing,2017,36(4):180-183.
[10]
陈果忠, 李世锋,任列华,等.甘肃地产浓香型白酒香味成分分析与比较研究[J].酿酒科技,2017(4):46-50.CHEN G Z,LI S F,REN L H,et al.Comparative analysis of flavoring components of Nongxiang Baijiu produced in Gansu[J].Liquor-making Science and Technology,2017(4):46-50.
[11]
杜礼泉, 唐聪,刘玲,等.浓香型大曲酒生产中正丁醇生成量差异的原因[J].酿酒科技,2004(3):47-48.DU L Q,TANG C,LIU L,et al.Reasons of different content of normal butyl alcohol during the production of Luzhou-flavor Daqu liquor[J].Liquor-making Science and Technology,2004(3):47-48.
[12]
ZHANG C,LI T G,HE J Z.Characterization and genome analysis of a butanol-isopropanol-producing Clostridium beijerinckii strain BGS1[J].Biotechnology for Biofuels,2018,11(1):280-291.
[13]
LAN E I,LIAO J C.Microbial synthesis of n-butanol,isobutanol,and other higher alcohols from diverse resources[J].Bioresource Technology,2013,135:339-349.
[14]
LI S B,HUANG L,KE C Z,et al.Pathway dissection,regulation,engineering and application:Lessons learned from biobutanol production by Solventogenic clostridia[J].Biotechnology for Biofuels,2020,13:39-64.
[15]
陈继良, 蔡林洋.生物发酵法生产丁醇的优化策略[J].中国酿造,2020,39(4):13-18.CHEN J L,CAI L Y.Optimization strategy for butanol production by biological fermentation[J].China Brewing,2020,39(4):13-18.
[16]
林俊涵, 邱东凤,林晨.丁醇产生菌育种研究进展[J].中国生物工程杂志,2014,34(12):118-128.LIN J H,QIU D F,LIN C.Development in breeding of butanol producing strain[J].China Biotechnology,2014,34(12):118-128.
[17]
张兴亚, 林玲,蒋予箭.黄酒中高级醇含量控制的研究进展[J].中国酿造,2011,30(10):13-17.ZHANG X Y,LIN L,JIANG Y J.Research progress of controlling higher alcohols in rice wine[J].China Brewing,2011,30(10):13-17.
[18]
罗惠波, 苟云淩,叶光斌,等.影响白酒中高级醇生成的工艺条件研究[J].中国酿造,2011,30(8):87-90.LUO H B,GOU Y L,YE G B,et al.Process conditions influencing the formation of higher alcohols in liquor spirit[J].China Brewing,2011,30(8):87-90.
[19]
JIANG J,LIU Y C,LI H H,et al.Modeling and regulation of higher alcohol production through the combined effects of the C/N ratio and microbial interaction[J].Journal of Agricultural and Food Chemistry,2019,67(38):10 694-10 701.
[20]
ZHANG J R,DU G C,CHEN J,et al.Characterization of a Bacillus amyloliquefaciens strain for reduction of citrulline accumulation during soy sauce fermentation[J].Biotechnology Letters,2016,38(10):1 723-1 730.
[21]
田源, 孔小勇,方芳.浓香型白酒发酵过程微生物合成正丙醇途径解析[J].微生物学报,2020,60(7):1 421-1 432.TIAN Y,KONG X Y,FANG F.Microbial n-propanol synthesis during Luzhou-flavor liquor fermentation[J].Acta Microbiologica Sinica,2020,60(7):1 421-1 432.
[22]
HU X L,DU H,XU Y.Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production[J].International Journal of Food Microbiology,2015,214:116-122.
[23]
周金虎, 李良,方尚玲,等.窖泥中高产愈创木酚类功能菌的筛选与鉴定[J].食品与发酵工业,2019,45(3):64-70.ZHOU J H,LI L,FANG S L,et al.Screening and identification of functional boateria with high yield guaiacol in the mud[J].Food and Fermentation Industries,2019,45(3):64-70.
[24]
芦丽, 宫旭.静态顶空气相色谱法测定保健酒中的高级醇类和酯类[J].中国药师,2019,22(12):2 354-2 358.LU L,GONG X.Determination of purified alcohols and esters in health wine by static headspace-gas chromatography[J].China Pharmacist,2019,22(12):2 354-2 358.
[25]
马瑞,欧阳嘉,李鑫,等.高效液相色谱法同时测定生物质乳酸发酵液中有机酸及糖类化合物[J].色谱,2012,30(1):62-66.MA R,OUYANG J,LI X,et al.Simultaneous determination of organic acids and saccharides in lactic acid fermentation broth from biomass using high performance liquid chromatography[J].Chinese Journal of Chromatography,2012,30(1):62-66.
[26]
周智友,许张柯,张庆华,等.丙酮丁醇产生菌的筛选、鉴定及其产丁醇性能优化[J].食品与发酵工业,2019,45(4):31-35;41.ZHOU Z Y,XU Z K,ZHANG Q H,et al.Screening and identification of acetone butanol-producing strains and process optimization for butanol production[J].Food and Fermentation Industries,2019,45(4):31-35;41.
[27]
刘佳乐,周朝晖,李铁桥,等.酱油发酵过程强化嗜盐四联球菌对酱油品质的影响[J].食品与发酵工业,2020,46(17):96-106.LIU J L,ZHOU Z H,LI T Q,et al.Effect of Tetragenococcus halophilus on soy sauce quality prepared by fermentation[J].Food and Fermentation Industries,2020,46(17):96-106.
[28]
LI L,AI H X,ZHANG S X,et al.Enhanced butanol production by coculture of Clostridium beijerinckii and Clostridium tyrobutyricum[J].Bioresource Technology,2013,143:397-404.
[29]
ZHENG Y N,LI L Z,XIAN M,et al.Problems with the microbial production of butanol[J].Journal of Industrial Microbiology and Biotechnology,2009,36(9):1 127-1 138.
[30]
胡晓龙. 浓香型白酒窖泥中梭菌群落多样性与窖泥质量关联性研究[D].无锡:江南大学,2015.HU X L.Illuminating the correlation between anaerobic clostridial community diversity and quality of pit mud used for the production of Chinese strong-flavor liquor[D].Wuxi:Jiangnan University,2015.
2021, Vol. 47 
