产脂肪酶菌株的筛选、鉴定及产酶条件优化  

Screening and identification of lipase producing strains and optimization of lipase producing conditions

作　　者：吴子君 陈思沅 杜昭君 胡双 李海峰[1] WU Zijun;CHEN Siyuan;DU Zhaojun;HU Shuang;LI Haifeng(College of Biological Engineering,Henan University of Technology,Zhengzhou 450001,China)

机构地区：[1]河南工业大学生物工程学院,河南郑州450001

出　　处：《河南工业大学学报（自然科学版）》2021年第6期10-16,共7页Journal of Henan University of Technology：Natural Science Edition

基　　金：河南省自然科学基金项目(182300410034);河南省高等学校青年骨干教师培养计划(2019ggjs-88)。

摘　　要：为筛选出产高活性脂肪酶的菌株,应用中性红油脂培养基从发酵芝麻饼中初筛得到3株产脂肪酶菌株,采用对硝基苯酚法进行复筛,确定1株产酶活力较优的菌株K-2,经16S rRNA基因序列比对以及系统发育树的构建,确定该菌株为铜绿假单胞菌(Pseudomonas aeruginosa)。通过单因素试验考察其最适产酶发酵条件,结果表明:发酵培养基中最适碳源为淀粉,氮源为蛋白胨,初始pH值为7.0,最适接种量为3%,Cu^(2+)对菌株产脂肪酶有促进作用,最适培养时间为72 h;经单因素试验优化后,脂肪酶活力达到最大值,为262.8 U/mL。研究结果可以为脂肪酶的工业化生产提供菌种资源与条件参考。In this study, fermented sesame cake samples were selected to screen a bacterial strain producing lipase with a high activity using neutral red oil medium. Three lipase producing strains were isolated, and strain K-2 with a better lipase producing activity was screened by the p-nitrophenol quantitative method. After 16 S rRNA gene sequencing, alignment and phylogenetic tree construction, the strain was found to belong to the genus of Pseudomonas. Single factor experiment was used to investigate the effects of fermentation time, inoculum amount, carbon source, nitrogen source, metal ion, and initial pH value on the lipase production of the strain K-2. The results showed that when the fermentation time was 72 h, the lipase activity produced by strain K-2 was the highest, and the lipase enzyme activity was 5.7 U/mL. When the inoculation amount was 3%, the lipase activity produced by the strain K-2 was the highest, with a lipase enzyme activity of 5.3 U/mL. When Cu;was added in the medium, the production of lipase by this strain was promoted, and the lipase activity was 12.2 U/mL higher than that of the control group. Zn^(2+), Fe^(2+), K^(+) and Ca^(2+) all exerted inhibitory effects on the enzyme activity. The best carbon source in the fermentation medium was starch, and the lipase enzyme activity reached 13.5 U/mL. The most suitable nitrogen source was peptone, and when the amount of peptone was 10 g/L, the lipase enzyme activity of the strain was significantly increased and reached 243.7 U/mL. When the initial pH value was 7.0, the lipase activity was the highest(262.8 U/mL). After an optimization of fermentation culture conditions, the lipase enzyme-producing ability of strain K-2 significantly improved. This study can provide bacterial strain resources and cultural condition reference for the industrial production of lipase.

关 键 词：铜绿假单胞菌 脂肪酶 发酵条件 

分 类 号：TS201.3[轻工技术与工程—食品科学]