Identification, cloning and heterologous expression of active [NiFe]-hydrogenase 2 from Citrobacter sp. SG in Escherichia coli.

Identification, cloning and heterologous expression of active [NiFe]-hydrogenase 2 from Citrobacter sp. SG in Escherichia coli.
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柠檬酸杆菌 SG 活性 [NiFe]-氢化酶 2 在大肠杆菌中的鉴定、克隆和异源表达

DOI:
10.1016/j.jbiotec.2015.01.025
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发表时间:
2015
影响因子:
4.1
通讯作者:
Jeltsch A
Jeltsch A
中科院分区:
工程技术3区
文献类型:
--
作者:
Maier JAH;Ragozin S;Jeltsch A

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氢(H2)是一种潜在的替代能源载体,只在燃烧时产生水和热。今天,工业制氢主要使用以化石燃料为基础的热化学过程或电解水。因此,从生物质中生产氢气的生物技术方法是一个有趣的替代方案。在这里,我们介绍了一种新型的直接测氢系统,该系统使用了专门用于氢检测的半导体器件。利用该装置,分离到一株在好氧培养条件下大量产氢的细菌,并经16S核糖体DNA测序鉴定为柠檬酸杆菌。负责观察到的氢酶活性的酶通过3个层析纯化步骤进行了部分纯化,并通过肽质量指纹图谱鉴定为2型[NiFe]-氢酶。柠檬酸杆菌含操纵子的[NiFe]-氢酶2的表达在大肠杆菌允许的重组制氢中使用SG。在此确定的[NiFe]-氢酶2可能用于生物技术制氢。我们推测氢酶在柠檬酸细菌中的表达可能是对酸性条件下生长的一种适应。
Hydrogen (H2) is a potential alternative energy carrier which only produces water and heat upon combustion. Today, industrial hydrogen production mainly uses thermochemical processes based on fossil fuels or electrolysis of water. Therefore, biotechnological approaches to produce H2from biomass are an interesting alternative. We introduce here a novel direct hydrogen measurement system using a semiconducting device specific for hydrogen detection. Using this device, a bacterium producing considerable amounts of hydrogen under aerobic cultivation was isolated and identified by 16S ribosomal DNA sequencing asCitrobactersp. The enzyme responsible for the observed hydrogenase activity was partially purified by 3 chromatographic purification steps and could be identified by peptide mass fingerprinting to be a type 2 [NiFe]-hydrogenase. Expression of the [NiFe]-hydrogenase 2 containing operon fromCitrobactersp. SG inEscherichia coliallowed recombinant hydrogen production. The [NiFe]-hydrogenase 2 identified here may be useful for biotechnological hydrogen production. We speculate that the expression of the hydrogenase inCitrobactermay be an adaptation to growth in acidic conditions.
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