Production of biohydrogen by heterologous expression of oxygen-tolerant Hydrogenovibrio marinus [NiFe]-hydrogenase in Escherichia coli
Production of biohydrogen by heterologous expression of oxygen-tolerant Hydrogenovibrio marinus [NiFe]-hydrogenase in Escherichia coli
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DOI：
10.1016/j.jbiotec.2011.07.007
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发表时间：
2011-09-20
影响因子：
4.1
通讯作者：
Cha, Hyung Joon
中科院分区：
文献类型：
--
作者：
Kim, Jaoon Y. H.;Jo, Byung Hoon;Cha, Hyung Joon
氢酶对氧的敏感性是高效生物制氢的关键问题。本研究首次在大肠杆菌中异源表达了海洋细菌海洋氢基因弧菌的耐氧[NiFe]-氢酶。表达海洋嗜血杆菌[NiFe]-氢酶的重组大肠杆菌BL21具有产氢活性，而亲本菌株不产氢。重组海洋梭菌氢酶的生物活性需要镍和铁两种元素。与我们先前报告中描述的重组大肠杆菌[NiFe]-氢酶1相比，重组H.marinus[NiFe]-氢酶在体外显示出1.6-1.7倍的产氢活性。重要的是，H.marinus[NiFe]氢酶在涉及表面曝气量和混合气体中氧比例变化的分析中表现出相对较好的耐氧性。具体地说，在含氧量为5-10%(v/v)的气体环境中，重组H.marinus[NiFe]-氢酶产生的氢气类似于大肠杆菌[NiFe]-氢酶1的7-9倍。此外，在正常的好氧纯化条件下，纯化的海参[NiFe]-氢酶的放氢活性接近28.8nmolH-2/(min mg蛋白)。基于这些结果，我们认为耐氧性海洋梭菌[NiFe]-氢酶可以用于体内和体外生物制氢，而不需要严格的厌氧设施。(C)2011爱思唯尔B.V.保留所有权利。
Oxygen sensitivity of hydrogenase is a critical issue in efficient biological hydrogen production. In the present study, oxygen-tolerant [NiFe]-hydrogenase from the marine bacterium, Hydrogenovibrio marinus, was heterologously expressed in Escherichia coli, for the first time. Recombinant E. coli BL21 expressing H. marinus [NiFe]-hydrogenase actively produced hydrogen, but the parent strain did not. Recombinant H. marinus hydrogenase required both nickel and iron for biological activity. Compared to the recombinant E. coli [NiFe]-hydrogenase 1 described in our previous report, recombinant H. marinus [NiFe]-hydrogenase displayed 1.6- to 1.7-fold higher hydrogen production activity in vitro. Importantly, H. marinus [NiFe]hydrogenase exhibited relatively good oxygen tolerance in analyses involving changes of surface aeration and oxygen proportion within a gas mixture. Specifically, recombinant H. marinus [NiFe]-hydrogenase produced similar to 7-to 9-fold more hydrogen than did E. coli [NiFe]-hydrogenase 1 in a gaseous environment containing 5-10% (v/v) oxygen. In addition, purified H. marinus [NiFe]-hydrogenase displayed a hydrogen evolution activity of similar to 28.8 nmol H-2/(min mg protein) under normal aerobic purification conditions. Based on these results, we suggest that oxygen-tolerant H. marinus [NiFe]-hydrogenase can be employed for in vivo and in vitro biohydrogen production without requirement for strictly anaerobic facilities. (C) 2011 Elsevier B.V. All rights reserved.