高温大曲中高产四甲基吡嗪芽孢杆菌的筛选及其在酒醅堆积中的应用  

Screening of high yield tetramethylpyrazine Bacillus from high temperature Daqu and its application in the stacking of fermented grains

作　　者：史洪丽 余梦婷 王世昕 张鹏 刘慧敏 李春茂 舒南鑫 孙朋朋 龙代洲 郭松波 SHI Hongli;YU Mengting;WANG Shixin;ZHANG Peng;LIU Huimin;LI Chunmao;SHU Nanxin;SUN Pengpeng;LONG Daizhou;GUO Songbo(Kweichow Diaoyutai State Guest Distillery Co.,Ltd.,Zunyi 564501,China)

机构地区：[1]贵州钓鱼台国宾酒业有限公司,贵州遵义564501

出　　处：《中国酿造》2025年第3期59-65,共7页China Brewing

基　　金：贵州省科技支撑计划项目(黔科合成果[2023]一般151)。

摘　　要：该研究从高温大曲中筛选芽孢杆菌(Bacillus),通过耐高温试验、蛋白酶活力测定初筛,V-P试验及四甲基吡嗪含量分析复筛高产四甲基吡嗪芽孢杆菌,并经过形态学观察、生理生化试验及分子生物学技术鉴定筛选菌株,并采用气相色谱-质谱联用(GC-MS)仪分析筛选菌株发酵液中挥发性风味物质,并将其应用于酒醅堆积。结果表明,筛选出18株产蛋白酶菌株,高产四甲基吡嗪菌株D-13-11,其含量达339.99 mg/L,菌株D-13-11鉴定为贝莱斯芽孢杆菌(Bacillus velezensis)。GC-MS结果表明,共检出42种物质,其中,吡嗪类4种、酯类15种、醇类4种、醛酮类4种、酸类6种、酚类2种、烷烃类及硅氧烷类7种。吡嗪类化合物的相对含量最高,达40.14%,主要为四甲基吡嗪(24.84%)。将菌株D-13-11按接种量7%接种至堆积酒醅中,吡嗪类化合物的含量明显高于对照组,其中,四甲基吡嗪的含量达11.38μg/g,是对照组的71.13倍。In this study,Bacillus spp were screened from high temperature Daqu.The strains were preliminarily screened through high temperature resistance test and protease activity determination,rescreened by V-P test and tetramethylpyrazine content analysis.The screened strains identified through morphological observation,physiological and biochemical tests and molecular biology techniques.The volatile flavor substances in the fermentation liquid of the strains were analyzed by GC-MS,and the strains were applied to the stacking of fermented grains.The results showed that a total of 18 proteinase-producing strains and 1 high yield tetramethylpyrazine strain D-13-11 were screened,and the high yield tetramethylpyrazine strain D-13-11 was identified as Bacillus velezensis with its content as high as 339.99 mg/L.The GC-MS results showed that a total of 42 substances were detected,including pyrazines 4,esters 15,alcohols 4,aldehydes and ketones 4,acids 6,phenols 2,alkanes and siloxanes 7.The relative content of pyrazine compounds was the highest,reaching 40.14%,and the tetramethylpyrazine was the main compound(24.84%).The strain D-13-11 was inoculated into the stacking of fermented grains with inoculum 7%,and the relative content of pyrazine compounds was significantly higher than that of the control group.Among them,the content of tetramethylpyrazine was 11.38μg/g,which was 71.13 times that of the control group.

关 键 词：高温大曲 四甲基吡嗪 芽孢杆菌 筛选 鉴定 

分 类 号：TS262.3[轻工技术与工程—发酵工程]