桔青霉 Uv-7 菌株固体发酵产葡萄糖氧化酶的分离与纯化  

Isolation and Purification of Glucose Oxidase Produced by Solid Fermentation of Penicillium Citrinum Uv-7 Strain

作　　者：赵芬芬 赵辉 白淑坤 杨洋[3] ZHAO Fenfen;ZHAO Hui;BAI Shukun;YANG Yang(Zhengzhou Yellow River Nursing Vocational College,Zhengzhou 450000,China;Zhengzhou Yuanchuang Gene Limited Company,Zhengzhou 450000,China;Guangxi University,Nanning 530004,China)

机构地区：[1]郑州黄河护理职业学院,河南郑州450000 [2]郑州源创基因科技有限公司,河南郑州450000 [3]广西大学,广西南宁530004

出　　处：《工业微生物》2024年第4期179-183,共5页Industrial Microbiology

基　　金：河南省科学技术厅项目(编号:232102310329);郑州黄河护理职业学院教育教学研究项目(编号:HHJY202401)。

摘　　要：葡萄糖氧化酶(Glucose oxidase,GOD)是一种需氧脱氢酶,能将葡萄糖氧化为葡萄糖酸和过氧化氢。本文采用紫外诱变筛选得到1株名为桔青霉Uv-7的葡萄糖氧化酶产生菌,以木薯叶为固体基质,采用优化后的最适培养基组分和发酵条件进行GOD的发酵生产。实验中,加入40 mL、pH 7.0的磷酸缓冲液抽提获取粗酶液,粗酶液经1∶0.5(V/V)的丙酮初步纯化,并经过Sephadex G75、Sephacryl S-200和DEAE-Sephadex A50离子交换层析,得到单组分的GOD,纯化倍数为23.46,比活力为158.3 U/mg,酶活回收率为21.06%。分子质量约为90 kDa,为单亚基酶。Glucose oxidase(GOD)is an aerobic dehydrogenation enzyme,which can catalyze the oxidation of茁-D-glucose to Dglucono-啄-lactone and hydrogen peroxide.A glucose oxidase producing strain named Penicillium citrinum UV-7 was obtained by Uv mutagenesis screening.Using cassava leaves as solid substrate,the optimal medium components and fermentation conditions were optimized for the fermentation production of GOD.After that,40 mL,pH 7.0 phosphate buffer was added to extract the crude enzyme solution.The crude enzyme solution was initially purified by 1∶0.5(V/V)of acetone.After ion exchange chromatography with Sephadex G-75,Superdex-200 and DEAE Sephadex A50,a single component GOD was obtained with a purification factor of 23.46,specific activity of 158.3 U/mg,and enzyme activity recovery rate of 21.06%.The purified GOD had a molecular weight of 90 kDa as a monomer.

关 键 词：葡萄糖氧化酶 桔青霉 木薯叶 分离 纯化 

分 类 号：TQ925.4[轻工技术与工程—发酵工程]