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 ofβ-glucosidase from Aspergillus versicolor
付跃 1李书安 2石娟娟 2罗奉奉1
作者信息
- 1. 河池学院化学与生物工程学院,广西河池 546300;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;河池学院农业生物技术应用研究中心,广西河池 546300
- 2. 河池学院化学与生物工程学院,广西河池 546300
- 折叠
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.Mn2+对该酶具有明显的促进作用;Co2+、Cu2+、Cd2+对该酶表现出抑制作用;而Na+、Mg2+、K+、Ca2+、Ba2+对该酶无明显影响.该酶以水杨苷为底物时,米氏常数Km为0.48 mg/mL,最大反应速率Vm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.
Abstract
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 88.5 kDa was obtained after purification.The purification fold,recovery and specific enzyme activity were 53.44,70.78％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 Mn2+could promote the β-glucosidase activity,while metal ions Co2+,Cu2+,Cd2+could inhibit the β-glucosidase activity,whereas Na+,Mg2+,K+,Ca2+,Ba2+had no effect.With salicylin as substrate,the Km was 0.48 mg/mL and the Vm was 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Key words
Aspergillus versicolor/β-glucosidase/isolation and purification/enzymatic property引用本文复制引用
基金项目
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)
出版年
2024
国家科技期刊平台
NSTL
万方数据