Expression, Purification, and Activity Analysis of Chlorophyllide Oxidoreductase and Ni2+-Sirohydrochlorin a,c-Diamide Reductase.

Expression, Purification, and Activity Analysis of Chlorophyllide Oxidoreductase and Ni2+-Sirohydrochlorin a,c-Diamide Reductase.
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叶绿素氧化还原酶和Ni2-SiroHCla,c-二酰胺还原酶的表达、纯化和活性分析

DOI:
10.1007/978-1-4939-8864-8_8
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发表时间:
2019
影响因子:
--
通讯作者:
Jasper
Jasper
中科院分区:
--
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--
作者:
Jasper

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类氮酶在还原各种四吡咯分子的共轭环体系中起着至关重要的作用。所有细菌叶绿素的生物合成都涉及到绿色色素叶绿素内酯的C7-C8双键的双电子还原,这是由类氮酶的双组分金属酶叶绿素内酯氧化还原酶(COR)催化的;而在所有产甲烷微生物中,另一种类似氮酶的系统,CfbC/D,在辅酶f430生物合成过程中负责Ni2+-sirohydrochlorina,c-diamide的复杂六电子还原。本章第一部分介绍了COR组分(BchY/BchZ)2和BchX2的制备和纯化,COR活性的测定,以及三元COR配合物的捕获;第二部分描述了cfbc2和cfbd2均相和催化活性制剂的制备策略,以及提取反应产物Ni2+-六氢四氯胺- c-二胺的合适方法。
Nitrogenase-like enzymes play a vital role in the reduction of the conjugated ring systems of diverse tetrapyrrole molecules. The biosynthesis of all bacteriochlorophylls involves the two-electron reduction of the C7–C8 double bond of the green pigment chlorophyllide, which is catalyzed by the nitrogenase-like two-component metalloenzyme chlorophyllide oxidoreductase (COR); whereas in all methanogenic microbes, another nitrogenase-like system, CfbC/D, is responsible for the sophisticated six-electron reduction of Ni2+-sirohydrochlorina,c-diamide in the course of coenzyme F430biosynthesis. The first part of this chapter describes the production and purification of the COR components (BchY/BchZ)2and BchX2, the measurement of COR activity, and the trapping of the ternary COR complex; and the second part describes the strategy for obtaining homogenous and catalytically active preparations of CfbC2and CfbD2and a suitable method for extracting the reaction product Ni2+-hexahydrosirohydrochlorina,c-diamide.
DOI: 10.1093/oxfordjournals.jbchem.a022535
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