食品工业科技2024，Vol.45Issue(20)：157-166,10.DOI:10.13386/j.issn1002-0306.2023110321
高产壳聚糖酶菌株筛选鉴定、条件优化及酶学性质分析
Screening,Identification and Conditions Optimization of High Chitosanase Producing Strains,and Analysis of Its Enzymatic Properties
摘要
Abstract
To screen and identify high chitosanase producing strains and to optimize the fermentation condition and analyze the enzymatic properties of the target strain.In this study,high chitosanase-producing strain was screened from the sediment of shrimp ponds in Beihai,Guangxi through primary and secondary screening.The strain underwent identification,the enzyme production process of the strain was optimized through single-factor,Plackett-Burman and orthogonal optimization methods.The enzymatic properties and products of chitosanase produced by the strain were studied.The results showed that the strain of Gxun-5.3.3 with high chitosanase production was obtained by screening and was identified as Bacillus cereus.The optimal components for the enzyme-producing medium of B.cereus Gxun-5.3.3 was maltose 5 g/L,yeast powder 3 g/L,chitosan 15 g/L,and the initial pH and fermentation time were 6.0 and 84 h,respectively.Under the optimized conditions,the chitosanase activity reached 62.03±1.18 U/mL,which was 55.58％higher before optimization(39.87±1.24 U/mL).The protease produced by strain Gxun-5.3.3 had maximum activity at 60 ℃and optimum pH at 5.0,respectively.Metal ions Fe2+and Mn2+had a significant(P＜0.05)activation effect on chitosanase,but Al3+and Cu2+had a strong inhibition.The enzymatic chitosan products were determined by TLC(thin layer chromatography)as chito-oligosaccharides with polymerization degree 2～4,indicating that the chitosanase produced by the strain was acting on endo-oligosaccharides.This study would provide a theoretical basis for the chitosanase production of B.cereus Gxun-5.3.3 and the enzymatic preparation of chito-oligosaccharides.关键词
壳聚糖酶/蜡样芽孢杆菌/条件优化/酶学性质/壳寡糖Key words
chitosanase/Bacillus cereus/condition optimization/enzymatic properties/chito-oligosaccharides分类
轻工纺织引用本文复制引用
梁颖,刘睿,宋超东,谢俊杰,殷豆豆,许霞,杨登峰,张红岩,申乃坤..高产壳聚糖酶菌株筛选鉴定、条件优化及酶学性质分析[J].食品工业科技,2024,45(20):157-166,10.基金项目
广西科技重点研发计划项目(桂科AB21196019,桂科AB21220020) （桂科AB21196019,桂科AB21220020）
国家自然科学基金项目(31660022,32060020) （31660022,32060020）
大学生创新创业训练计划项目(202110608014,S202210608164,S202210608156),广西民族大学实验技术创新与实验室管理研究项目(2023MDSY002) （202110608014,S202210608164,S202210608156）
广西海洋天然产物与组合生物合成化学重点实验室开放基金资助. （）
