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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

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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(ARTP-UV)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/mL，比原始菌株S5-4的产酯酶能力提高了30。45％。
ARTP-UV compound mutation breeding and fermentation conditions optimization of high-yield esterase Lactococcus garvieae
Using Lactococcus garvieae S5-4 preserved from the pre-screening of the laboratory as the starting strain,the original strain S5-4 was mutat-ed 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.

esterification enzymeLactococcus garvieaecompound mutagenesisresponse surface methodfermentation conditions optimization

蔡岭肸、陈晓松、邹伟、汤秀娟

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四川轻化工大学生物工程学院,四川 宜宾 644000

酿酒生物技术及应用四川省重点实验室,四川 宜宾 644000

乐山职业技术学院,四川乐山 614000

酯酶 格氏乳球菌 复合诱变 响应面法 发酵条件优化

泸州老窖研究生创新基金四川轻化工大学研究生创新基金资助项目四川轻化工大学研究生创新基金资助项目四川轻化工大学校级大学生创新创业训练计划项目

LJCX2022-2Y2023222Y2023234cx2022126

2024

10.11882/j.issn.0254-5071.2024.02.019

中国酿造
中国调味品协会 北京食品科学研究院

中国酿造

CSTPCD北大核心
影响因子：0.759
ISSN：0254-5071
年,卷(期)：2024.43(2)
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