高产酯化酶细菌的复合诱变选育及固态发酵条件优化  

Combinatorial mutagenesis of high-yielding esterase bacteria and optimization of solid-state fermentation

作　　者：赵志军[1] 张艳珠 刘延波[1] 周平平 葛少华[2] 孙西玉[1] ZHAO Zhijun;ZHANG Yanzhu;LIU Yanbo;ZHOU Pingping;GE Shaohua;SUN Xiyu(Liquor College,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China;Baofeng Liquor Industry Co.Ltd,Pingdingshan 467500,China;College of Food and Pharmaceutical Science,Ningbo University,Ningbo 315211,China)

机构地区：[1]河南牧业经济学院酒业学院,河南郑州450046 [2]宝丰酒业有限公司,河南平顶山467500 [3]宁波大学食品与药学学院,浙江宁波315211

出　　处：《食品与发酵工业》2021年第2期174-181,共8页Food and Fermentation Industries

基　　金：河南省重大科技专项(181100211400);河南牧业经济学院重点学科项目;河南省科技攻关项目(202102110130)。

摘　　要：为了获得高产酯化酶的细菌,该研究以先期分离获得的贝莱斯芽胞杆菌(Bacillus velezensis)Nz 1.201为出发菌株,利用紫外线(ultraviolet,UV)及硫酸二乙酯(diethyl sulfate,DES)复合诱变技术进行诱变育种。采用透明圈法初筛、酯化酶酶活力测定法复筛,选育出高产突变菌株D27。经过5次连续传代培养验证其遗传性能较稳定,测定其菌株酶活力可达62.45 U/100g,比初始菌株提高了66%。对D27菌株进行固态发酵,在单因素试验的基础上,以酯化酶酶活力为响应值,采用Box-Behnken试验设计法进行响应面优化。结果表明,该突变菌株D27的最佳培养条件为:培养温度33℃、培养时间6 d、菌种接种量3%、原料含水量73%,此时,酯化酶酶活力最高为89.52 U/100g,较初始菌株提高了137%。研究表明,突变菌株D27在固态培养条件下具有较高的产酯化酶能力,在白酒生产中有着一定的应用潜力。In order to obtain bacteria with high esterase production,combinatorial mutagenesis using ultraviolet(UV)and diethyl sulfate(DES)technology was applied to previous isolated strain Bacillus velezensis Nz 1.201.The high-yielding mutant strain D27 was selected by the transparent circle method for primary screening and the esterase activity measurement for rescreening.After five consecutive subcultures,its genetic performance was relatively stable.The enzyme activity reached 62.45 U/100 g,which was 66%higher than that of the original strain.Mutant strain D27 was further used in solid-state fermentation.Based on single factor experiment,Box-Behnken design-response surface methodology was used to optimize culture conditions for mutant D27 with esterase activity as the response value.The results showed that the optimal culture conditions were:33℃,6 days,strain inoculation volume of 3%,and raw material moisture content of 73%.Under the optimal condition,the highest esterase activity was 89.52 U/100 g,an increase of 137%compared with the initial strain.Studies have shown that the mutant strain D27 has a higher ability to produce esterase under solid-state fermentation,and has certain application potential in liquor production.

关 键 词：产酯化酶细菌 复合诱变 固态发酵 响应面优化 酯化酶 

分 类 号：TS262.3[轻工技术与工程—发酵工程]