杂色曲霉β-葡萄糖苷酶分离纯化及酶学性质研究  

Isolation,purification and enzymatic properties of β-glucosidase from Aspergillus versicolor

作　　者：付跃 李书安 石娟娟 罗奉奉 FU Yue;LI Shuan;SHI Juanjuan;LUO Fengfeng(College of Chemical and Biological Engineering,Hechi University,Hechi,Guangxi 546300,China;Guangxi Colleges Universities Key Laboratory of Exploitation and Utilization of Microbial and Botanical Resources,Hechi,Guangxi 546300,China;Agricultural Biotechnology Application Research Center,Hechi University,Hechi,Guangxi 546300,China)

机构地区：[1]河池学院化学与生物工程学院,广西河池546300 [2]微生物及植物资源开发利用广西高校重点实验室,广西河池546300 [3]河池学院农业生物技术应用研究中心,广西河池546300

出　　处：《中国饲料》2024年第13期48-54,共7页China Feed

基　　金：广西高校中青年教师科研基础能力提升项目(2020KY15018);河池学院重点科研项目(2021XJZD006);广西自然科学基金项目(2020GXNSFAA297218)。

摘　　要：试验旨在从杂色曲霉发酵液中分离纯化β-葡萄糖苷酶,并对其进行酶学性质研究。利用ENrich Q,10×100离子交换色谱法对杂色曲霉产β-葡萄糖苷酶分离纯化,采用SDS-PAGE测定其分子质量。结果表明:该β-葡萄糖苷酶的分子质量约为88.5 kDa,纯化倍数为53.44,回收率为70.78%,比酶活为1438.11 U/mg。该β-葡萄糖苷酶最适反应温度为50℃,最适反应pH为5.5;在温度20~50℃,pH 5.0~7.0时有较好的稳定性。Mn^(2+)对该酶具有明显的促进作用;Co^(2+)、Cu^(2+)、Cd^(2+)对该酶表现出抑制作用;而Na^(+)、Mg^(2+)、K^(+)、Ca^(2+)、Ba^(2+)对该酶无明显影响。该酶以水杨苷为底物时,米氏常数K_(m)为0.48 mg/mL,最大反应速率V_(m)为0.04 mg/(min·mL)。试验从杂色曲霉发酵液中成功分离出一种β-葡萄糖苷酶,为纤维素类饲料的应用提供了数据支撑。The experiment aimed to isolate and purify β-glucosidase from Aspergillus versicolor fermentation broth and study the enzymatic properties of β-glucosidase.β-glucosidase was isolated from Aspergillus versicolor and purified by ENrich Q,10×100 anion exchange chromatography.The molecular weight was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the properties of the enzyme were studied.The results showed that the β-glucosidase with molecular mass 88.5kDa was obtained after purification.The purification fold,recovery and specific enzyme activity were53.44,70.78%and 1438.11 U/mg,respectively.The optimum reaction temperature and pH of the β-glucosidase were 50℃and 5.5,respectively,and the enzyme had good stability at 20~50℃and pH 5.0~7.0.The metal ions Mn^(2+)could promote the β-glucosidase activity,while metal ions Co^(2+),Cu^(2+),Cd^(2+)could inhibit the β-glucosidase activity,whereas Na~+,Mg^(2+),K~+,Ca^(2+),Ba^(2+)had no effect.With salicylin as substrate,the K_(mwas) 0.48 mg/mL and the V_(mwas) 0.04 mg/(min·mL).The study isolated a strain from the fermentation broth of Aspergillus versicolor successfully and its enzymatic properties were studied,providing data support for the application of cellulose based feed.

关 键 词：杂色曲霉 Β-葡萄糖苷酶 分离纯化 酶学性质 

分 类 号：S816.3[农业科学—饲料科学]