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甘薯叶片蔗糖酶的分离纯化及其部分性质

唐云明, 黎南, 陈定福

西南师范大学生命科学系.四川,重庆 630715

摘要

纯化酶经聚丙烯酰胺凝胶电泳显示单一蛋白带,SDS-PAGE显示一条蛋白带,其亚基分子量为39.8kD。用Sephacryl S-200凝胶过滤测得全酶的分子量为79.4kD,该酶由两个相同亚基组成。其表现K_(m)为12mmol/L,v_(max)为99.5mg还原糖mg~(-1)protein h~(-1)。

关键词： 甘薯叶片; 蔗糖酶; 纯化; 性质

Isolation,purification and some properties of invertase from sweet potato

Tang Yunming, Li Nan, Chen Dingfu

Department of Life Sciences,Southwest China Normal University.Chongqing 630715,Sichuan

Abstract

Invertase was isolated and purified from the extract of sweet potato (Ipomeoa batatas Lam.) leaves by ammonium sulphate precipitation, DEAE-Sepharose column chromatography, and Sephacry1 S-200 gel filtration. It was purified 282.1-fold with an activity recovery of 22.7%. The purified invertase preparation was homogeneous as judged by polyacrylamide gel electrophoresis and SDS-PAGE. The molecular weight of the native invertase was determined by gel filtration to be 79.4 kD. The invertase consists of two identical subnuits with an apparent molecular weight of 39.8 kD as determined by SDS-PAGE(Fig.3B).The invertase followed typical Michaelis-Menten Kinetics with an apparent K_(m) of 12 mmol/L for sucrose. The V_(max) of the invertase was 99.5 mg reducing sugar mg~(-1) protein h~(-1). The enzyme was shown to be stable between pH 4. 0 and 6. 0 and to have an optimum pH of 5.0 for its activity. The temperature optimum was 60℃.

Key words: Sweet potato leaves;Invertase;Purification;Properties

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引用本文：

唐云明, 黎南, 陈定福. 甘薯叶片蔗糖酶的分离纯化及其部分性质[J]. 生理学报 1996; 48 (1): .

Tang Yunming, Li Nan, Chen Dingfu. Isolation,purification and some properties of invertase from sweet potato. Acta Physiol Sin 1996; 48 (1): (in Chinese with English abstract).