酒曲中高产蛋白酶菌株的筛选及其发酵培养基优化  

Screening and fermentation medium optimization of high-yield protease-producing strain from Jiuqu

作　　者：赵华[1] 任青霞 张敏 王艺会 杨贞耐[1] ZHAO Hua;REN Qingxia;ZHANG Min;WANG Yihui;YANG Zhennai(Beijing Engineering and Technology Research Center of Food Additives,Beijing Advanced Innovation Center for Food Nutrition and Human Health,Beijing Technology and Business University,Beijing 100048,China;Key Laboratory of Agro-Products Primary Processing,Academy of Agricultural Planning and Engineering,MARA,Beijing 100125,China)

机构地区：[1]北京工商大学北京食品营养与健康高精尖创新中心北京市食品添加剂工程技术研究中心,北京100048 [2]农业农村部规划设计研究院农业农村部产地初加工重点实验室,北京100125

出　　处：《中国酿造》2023年第5期157-164,共8页China Brewing

基　　金：国家重点研发计划项目(2018YFC1604302-2)。

摘　　要：从传统酒曲中分离鉴定高产胞外蛋白酶的菌株,探究其生长特性,通过酪蛋白平板法、福林酚法筛选高产蛋白酶菌株,并通过形态特征、生理生化试验、16S rDNA序列分析和种特异性基因分析鉴定菌株。采用单因素试验、Plackett-Burman试验、最陡爬坡试验及Box-Behnken试验优化其发酵培养基。结果表明,分离筛选出一株高产蛋白酶菌株(编号为JQ-2)被鉴定为枯草芽孢杆菌(Bacillus subtilis)。菌株JQ-2最佳发酵培养基为:马铃薯浸出粉5.4 g/L、麦芽浸膏1.8 g/L、牛肉膏10.0 g/L。在此优化条件下,菌株JQ-2的胞外蛋白酶活性为154.75 U/mL,是未优化培养基的14.77倍。High-yield extracellular protease-producing strains were isolated and identified from traditional Jiuqu and their growth characteristics were explored.High-yield protease-producing strains were screened by casein plate method and Folin-phenol method,and the strains were identified by morphological characteristics,physiological and biochemical tests,16S rDNA sequence analysis and species specific gene analysis.The fermentation medium was optimized by single factor tests,Plackett-Burman tests,the steepest climbing tests and Box-Behnken tests.The results showed that a high-yield protease-producing strain was isolated and screened(numbered as JQ-2),and identified as Bacillus subtilis.The optimal fermentation medium of the strain JQ-2 was as follows:potato extract powder 5.4 g/L,malt extract 1.8 g/L and beef extract 10.0 g/L.Under these optimal conditions,the extracellular protease activity of the strain JQ-2 was 157.75 U/ml,which was 14.77 times that of unoptimized medium.

关 键 词：芽孢杆菌 蛋白酶 分离鉴定 发酵培养基 响应面优化 

分 类 号：TQ920[轻工技术与工程—发酵工程]