一株产中性蛋白酶菌株的筛选及其发酵产酶条件优化  

Screening of A Neutral Protease-producing Strain and Optimization of Fermentation Conditions

作　　者：李娜 附俊杰 刘军 温雪瓶 李丽 LI Na;FU Junjie;LIU Jun;WEN Xueping;LI Li(College of Bioengineering,Sichuan University of Science&Engineering,Yibin 644000,China;Key Laboratory of Solid-state Fermentation Resource Utilization in Sichuan Province,Yibin 644000,China)

机构地区：[1]四川轻化工大学生物工程学院,四川宜宾644000 [2]固态发酵资源利用四川省重点实验室,四川宜宾644000

出　　处：《食品工业科技》2023年第1期189-199,共11页Science and Technology of Food Industry

基　　金：固态发酵资源利用四川省重点实验室开放基金项目(2018GTY002)。

摘　　要：目前,低酶活及原料高成本严重制约了蛋白酶工业的发展,本研究采用透明圈法和摇瓶发酵法筛选出1株产中性蛋白酶的菌株,经形态学观察、生理生化实验以及16S rDNA序列分析鉴定为贝莱斯芽孢杆菌(Bacillus velezensis),并命名为ppr3,通过单因素实验和响应面试验对发酵培养基成分及发酵条件进行优化,结果表明:当发酵温度33℃,接种量8.5%(v/v),初始pH5.7,乳糖5.55 g/L、菜粕31.89 g/L、酵母浸粉7.09 g/L、吐温800.03%、MgSO_(4)0.04%、K_(2)HPO_(4)0.04%时,蛋白酶活为500 U/mL,相较于未优化前提高了132%。将菜粕作为培养基的发酵氮源,在一定程度上降低了成本,为农副产品的再利用提供了一个新途径,也为后续工业化生产奠定了研究基础。Currently,low enzyme activity and high raw material cost seriously restricted the development of the industrial production of protease.In this study,transparent circle and shaking flask fermentation methods were used to screen out a neutral protease-producing strain,which was identified as Bacillus velezensis by morphological observation,physiological and biochemical results and 16S rDNA analysis,and named ppr3.The medium composition and fermentation conditions for this strain to produce neutral protease were optimized by one-at-a-time strategy and response surface methodology.The results showed that:The fermentation temperature was 33℃,inoculation amount was 8.5%(v/v),initial pH was 5.7,lactose 5.55 g/L,rapeseed 31.89 g/L,yeast extract 7.09 g/L,tween-800.03%,MgSO_(4)0.04%,K_(2)HPO_(4)0.04%,the protease activity was 500 U/mL.Compared with before optimization,it increased by 132%.The addition of rapeseed meal as nitrogen source to the medium significantly reduced the fermentation cost and provided a new way for the reuse of agricultural and sideline products,which had major implications in protease industrial fermentation.

关 键 词：贝莱斯芽孢杆菌 筛选 蛋白酶活 产酶条件优化 

分 类 号：TQ925[轻工技术与工程—发酵工程]