高产酯酶格氏乳球菌的ARTP-UV复合诱变选育及发酵条件优化  

ARTP-UV compound mutation breeding and fermentation conditions optimization of high-yield esterase Lactococcus garvieae

作　　者：蔡岭肸 陈晓松 邹伟 汤秀娟 CAI Lingxi;CHEN Xiaosong;ZOU Wei;TANG Xiujuan(College of Bioengineering,Sichuan University of Science&Engineering,Yibin 644000,China;Liquor Brewing Biotechnology and Application Key Laboratory of Sichuan Province,Yibin 644000,China;Leshan Vocational&Technical College,Leshan 614000,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]酿酒生物技术及应用四川省重点实验室,四川宜宾644000 [3]乐山职业技术学院,四川乐山614000

出　　处：《中国酿造》2024年第2期125-133,共9页China Brewing

基　　金：泸州老窖研究生创新基金项目(LJCX2022-2);四川轻化工大学研究生创新基金资助项目(Y2023222,Y2023234);四川轻化工大学校级大学生创新创业训练计划项目(cx2022126)。

摘　　要：以实验室前期筛选保藏的格氏乳球菌(Lactococcus garvieae)S5-4为出发菌株,采用多轮次常压室温等离子体-紫外(ARTPUV)复合诱变对出发菌株S5-4进行诱变选育,以产酯酶酶活为评价指标,得到一株高产酯酶突变株ARUV3-26,其产酯酶活力达到(25.88±0.02)U/mL,较原始菌株S5-4提高了64.42%。经15代传代测试后,突变菌株ARUV3-26产酯酶活力稳定在(23.51±0.02)~(23.56±0.02)U/mL,与原始菌株S5-4相比提高了49.36%~49.68%。生理耐受性试验表明,突变株ARUV3-26与原始菌株S5-4相比,温度耐受性、pH耐受性、葡萄糖耐受性、乙醇耐受性、丁酸和己酸耐受性均有提升。通过单因素试验优化最佳培养基组分为:葡萄糖20 g/L,蛋白胨15 g/L,NaCl 5 g/L。通过单因素和响应面试验优化突变株ARUV3-26发酵条件为发酵时间3.0 d、发酵温度为30℃、初始pH值为7.0。此优化条件下,突变菌株ARUV3-26的产酯酶活力为(30.67±0.17)U/m L,比原始菌株S5-4的产酯酶能力提高了30.45%。Using Lactococcus garvieae S5-4 preserved from the pre-screening of the laboratory as the starting strain,the original strain S5-4 was mutated by multiple rounds of atmospheric and room temperature plasma-ultraviolet(ARTP-UV)compound mutagenesis.Esterase activity was used as evaluation index,and a high-yield esterase mutant strain ARUV3-26 was obtained,with an esterase activity of(25.88±0.02)U/ml,which was 64.42%higher than the original strain S5-4.After 15 generations of passage testing,the esterase activities of the mutant strain ARUV3-26 remained stable at(23.51±0.02)to(23.56±0.02)U/ml,which was 49.36%to 49.68%higher than that of the original strain S5-4.Physiological tolerance tests showed that the temperature tolerance,pH tolerance,glucose tolerance,ethanol tolerance,butyric acid and caproic acid tolerance of the mutant strain ARUV3-26 were improved compared with the original strain S5-4.Through single-factor tests optimization,the optimal medium compositions were obtained as follows:glucose 20 g/L,peptone 15 g/L,and NaCl 5 g/L.Through single-factor and response surface experiments,the optimal fermentation conditions were determined as follows:fermentation time 3.0 d,fermentation temperature 30℃,and initial pH value 7.0.Under these optimal conditions,the esterase activity of the mutant strain ARUV3-26 was(30.67±0.17)U/ml,which was 30.45%higher than the esterase production capacity of the original strain S5-4.

关 键 词：酯酶 格氏乳球菌 复合诱变 响应面法 发酵条件优化 

分 类 号：TS261.1[轻工技术与工程—发酵工程] TS262.3[轻工技术与工程—食品科学与工程]