醇化烟叶附生产蛋白酶细菌分离筛选及活性评估  

Isolation,Screening and Activity Evaluation of Protease Producing Bacteria from Alcoholized Tobacco Leaves

作　　者：梁伟[1] 刘鸿[1] 邹克兴[1] 陈义昌[1] 王小东[2] 孙会忠[2] 蔡联合[1] 苏赞[1] 胡逸超 LIANG Wei;LIU Hong;ZOU Kexing;CHEN Yichang;WANG Xiaodong;SUN Huizhong;CAI Lianhe;SU Zan;HU Yichao(Technical Center of Guangxi China Tobacco Industry Co.,Ltd.,Nanning Guangxi 530000;College of Agriculture,Henan University of Science and Technology,Luoyang Henan 471023)

机构地区：[1]广西中烟工业有限责任公司技术中心,广西南宁530000 [2]河南科技大学农学院,河南洛阳471023

出　　处：《安徽农学通报》2023年第3期21-25,共5页Anhui Agricultural Science Bulletin

基　　金：广西中烟工业有限责任公司科技计划项目(功能微生物提升真龙卷烟原料醇化质量技术研究,GXZYCX2021B002)。

摘　　要：本文以醇化烟叶为材料,采用平板分离技术分离培养出26株细菌活体纯培养物,依次编号为GXZY1~GXZY26,纯化菌株通过牛奶琼脂培养基的鉴别培养,初筛出GXZY3、GXZY7和GXZY93株具有明显的水解圈,初步定性为产蛋白酶功能细菌。对GXZY3、GXZY7和GXZY9菌株连续60 h的连续发酵实验,发现3株菌发酵液中的蛋白酶活性均呈现先高后低的变化规律,酶活性的最大值均出现在发酵后18 h;GXZY9菌株发酵液中的酶活性持续保持高位,最大值达到18.2 U/g,且酶活性下降幅度最小,而GXZY3和GXZY7则在发酵36 h后持续下降到较低水平。对菌株GXZY13的多相鉴定结果表明,其隶属于芽孢杆菌属(Bacillus),命名为Bacillus subtilis GXZY9。菌株GXZY9对烤烟蛋白质含量具有较强的调控功能,可作为烟叶醇化微生物制剂开发的良好备用菌株。Twenty-six strains of pure bacteria in vivo were isolated and cultured from alcoholized tobacco leaf by plate separation technology,which were numbered GXZY1-GXZY26.The purified strains were identified by milk AGAR medium and GXZY3,GXZY7 and GXZY9 had obvious hydrolytic circles,and were preliminarily identified as protease-producing functional bacteria.The protease activity of GXZY3,GXZY7 and GXZY9 strains in the fermentation broth of GXZY3,GXZY7 and GXZY9 strains increased at first and then decreased,and the maximum value of enzyme activity occurred at 18 h after fermentation.The enzyme activity in the fermentation broth of GXZY9 strain remained high,reaching the maximum value of 18.2 U/g,and the enzyme activity decreased the least,while the enzyme activity of GXZY3 and GXZY7 continued to decrease to a low level after 36 h of fermentation.Polyphase identification of strain GXZY13 showed that it belonged to Bacillus subtilis GXZY9.Strain GXZY9 had strong regulation function on protein content of flue-cured tobacco and could be used as a good standby strain for the development of tobacco mellow microbial preparation.

关 键 词：烟叶醇化 附生菌 蛋白酶 筛选 

分 类 号：S47[农业科学—农业昆虫与害虫防治]