生物资源2025，Vol.47Issue(4)：371-378,8.DOI:10.14188/j.ajsh.20250207001
蜡样芽胞杆菌产磷脂酶D分离提取及酶学性质研究
Isolation,extraction and enzymatic properties of phospholipase D produced by Bacillus cereus
摘要
Abstract
Phospholipase D(PLD)serves as a valuable biocatalytic resource with significant industrial potential for phospholipid modification and rare phospholipid production.In this study,PLD produced by the previously screened Ba-cillus cereus was isolated and extracted by salting-out and column chromatography.The enzymatic properties of PLD pro-duced by this bacterium were studied by changing conditions such as temperature and pH.Its molecular weight was deter-mined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Using phosphatidylcholine(PC)and L-serine as substrates,the characteristics of the transphosphatidylation reaction were studied.The results showed that after salting out,the precipitate achieved electrophoretic purity after two-step chromatography,with a specific enzyme activity of 40.06 U/mg.The optimum reaction pH for the hydrolase of PLD produced by this bacterium was 8.0,and the en-zyme activity was relatively stable when pH was from 7.0 to 10.0.The optimum reaction temperature was 50 ℃,and the enzyme activity was stable at 50 ℃ and below.Triton X-100 and sodium deoxycholate promoted the enzyme activi-ty,while ethylene diamine tetraacetic acid(EDTA)completely inhibited the hydrolase activity.Mg2+promoted the en-zyme activity,and Mn2+and Co2+inhibited the enzyme activity.The molecular weight was approximately 32 kDa.The study on the transphosphatidylation reaction indicates that this PLD has good transphosphatidylase activity with almost no production of the by-product phosphatidic acid(PA),demonstrating potential industrial application value.关键词
蜡样芽胞杆菌/磷脂酶D/分离提取/酶学性质Key words
Bacillus cereus/phospholipase D/isolation and extraction/enzymatic properties分类
生物科学引用本文复制引用
代书玲,刘影,范丽平,陈晶,徐莉..蜡样芽胞杆菌产磷脂酶D分离提取及酶学性质研究[J].生物资源,2025,47(4):371-378,8.基金项目
上海农林职业技术学院科研课题(KY2-0000-20-12) （KY2-0000-20-12）
