异化亚硝酸盐还原酶的分光光度测定法。

A spectrophotometric assay for dissimilatory nitrite reductases.

作者信息

Hulse C L, Tiedje J M, Averill B A

机构信息

Department of Chemistry, University of Virginia, Charlottesville 22901.

出版信息

Anal Biochem. 1988 Aug 1;172(2):420-6. doi: 10.1016/0003-2697(88)90464-2.

DOI:10.1016/0003-2697(88)90464-2

PMID:2847585

Abstract

A spectrophotometric assay for dissimilatory nitrite reductases has been developed utilizing mammalian cytochrome c (equine heart) as reductant and spectrophotometric agent. The copper-containing nitrite reductase from Achromobacter cycloclastes has been shown to have apparent Km's for reduced cytochrome c and nitrite of 86 +/- 5 and 5.63 +/- 0.03 microM, respectively. The heme cd-containing enzyme from Pseudomonas stutzeri was shown to have apparent Km's for reduced cytochrome c and nitrite of 260 +/- 60 and 1.8 +/- 0.8 microM, respectively. This assay represents a simple, general method for consistently evaluating the activity of the copper- and heme cd-containing nitrite reductases that are capable of utilizing readily available mammalian cytochrome c as electron donor and should be useful for mechanistic studies of these enzymes.

摘要

已开发出一种用于异化亚硝酸盐还原酶的分光光度测定法，该方法利用哺乳动物细胞色素c（马心）作为还原剂和分光光度试剂。已证明来自环裂无色杆菌的含铜亚硝酸盐还原酶对还原型细胞色素c和亚硝酸盐的表观Km值分别为86±5和5.63±0.03微摩尔。已证明来自施氏假单胞菌的含血红素cd的酶对还原型细胞色素c和亚硝酸盐的表观Km值分别为260±60和1.8±0.8微摩尔。该测定法是一种简单通用的方法，可用于持续评估能够利用易于获得的哺乳动物细胞色素c作为电子供体的含铜和含血红素cd的亚硝酸盐还原酶的活性，并且应该对这些酶的机理研究有用。