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复杂金属酶异源生产的要求：膜结合[NiFe]氢化酶

Requirements for heterologous production of a complex metalloenzyme: the membrane-bound [NiFe] hydrogenase.

作者信息

Lenz Oliver, Gleiche Andrea, Strack Angelika, Friedrich Bärbel

机构信息

Institut für Biologie, Humboldt-Universität zu Berlin, Chausseestrasse 117, D-10115 Berlin, Germany.

出版信息

J Bacteriol. 2005 Sep;187(18):6590-5. doi: 10.1128/JB.187.18.6590-6595.2005.

DOI:10.1128/JB.187.18.6590-6595.2005
PMID:16159796
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1236620/
Abstract

By taking advantage of the tightly clustered genes for the membrane-bound [NiFe] hydrogenase of Ralstonia eutropha H16, broad-host-range recombinant plasmids were constructed carrying the entire membrane-bound hydrogenase (MBH) operon encompassing 21 genes. We demonstrate that the complex MBH biosynthetic apparatus is actively produced in hydrogenase-free hosts yielding fully assembled and functional MBH protein.

摘要

通过利用嗜麦芽窄食单胞菌H16膜结合[NiFe]氢化酶紧密聚集的基因，构建了携带包含21个基因的完整膜结合氢化酶（MBH）操纵子的广宿主范围重组质粒。我们证明，复杂的MBH生物合成装置在无氢化酶的宿主中被积极产生，产生完全组装且功能正常的MBH蛋白。

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本文引用的文献

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The complex between hydrogenase-maturation proteins HypC and HypD is an intermediate in the supply of cyanide to the active site iron of [NiFe]-hydrogenases.氢化酶成熟蛋白HypC和HypD之间的复合物是向[NiFe] -氢化酶活性位点铁供应氰化物过程中的一个中间体。
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The auxiliary protein HypX provides oxygen tolerance to the soluble [NiFe]-hydrogenase of ralstonia eutropha H16 by way of a cyanide ligand to nickel.辅助蛋白HypX通过镍的氰化物配体赋予嗜中性嗜麦芽窄食单胞菌H16的可溶性[NiFe]氢化酶耐氧性。
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Complete nucleotide sequence of pHG1: a Ralstonia eutropha H16 megaplasmid encoding key enzymes of H(2)-based ithoautotrophy and anaerobiosis.pHG1的完整核苷酸序列：一株真养产碱杆菌H16的巨型质粒，编码基于氢气的无机自养和厌氧生活的关键酶。
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Metal insertion into NiFe-hydrogenases.金属插入镍铁氢化酶中。
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