酿酒酵母胞质溶胶中多磷酸酶的纯化及性质

Purification and properties of polyphosphatase from Saccharomyces cerevisiae cytosol.

作者信息

Andreeva N, Kulakovskaya T, Sidorov I, Karpov A, Kulaev I

机构信息

Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow, Russia.

出版信息

Yeast. 1998 Mar 15;14(4):383-90. doi: 10.1002/(SICI)1097-0061(19980315)14:4<383::AID-YEA232>3.0.CO;2-S.

DOI:10.1002/(SICI)1097-0061(19980315)14:4<383::AID-YEA232>3.0.CO;2-S

PMID:9559546

Abstract

A homogenous polyphosphatase preparation was obtained from Saccharomyces cerevisiae cytosol with a 3.8% yield and 3540-fold purification. The enzyme hydrolysed polyphosphate (polyP) with various chain lengths, including polyP3, and split Pi off the end of the chain. It was inactive with respect to ATP, PPi, and p-nitrophenylphosphate. Its specific activity with polyP15 was 283 U/mg protein. The polyphosphatase was a monomeric protein with a molecular mass of 40 kDa. This enzyme was inactive without divalent cations and with Cu2+ and Ca2+. The ability of other divalent cations to activate the enzyme decreased in the following order; Co2+ > Mn2+ > Mg2+ > Zn2+. A kinetic model of the hydrolysis of polyP3 and action of Mg2+ is proposed.

摘要

从酿酒酵母胞质溶胶中获得了一种均一的多磷酸酶制剂，产率为3.8%，纯化倍数为3540倍。该酶能水解不同链长的多聚磷酸盐（polyP），包括多聚P3，并从链的末端裂解出无机磷酸（Pi）。它对ATP、焦磷酸（PPi）和对硝基苯磷酸酯无活性。其对多聚P15的比活性为283 U/mg蛋白质。该多磷酸酶是一种分子量为40 kDa的单体蛋白。该酶在没有二价阳离子以及有Cu2+和Ca2+存在时无活性。其他二价阳离子激活该酶的能力按以下顺序降低：Co2+ > Mn2+ > Mg2+ > Zn2+。提出了多聚P3水解和Mg2+作用的动力学模型。