Solar-to-bioH2 production enhanced by homologous overexpression of hydrogenase in green alga Chlorella sp. DT
Solar-to-bioH2 production enhanced by homologous overexpression of hydrogenase in green alga Chlorella sp. DT
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DOI：
10.1016/j.ijhydene.2012.09.068
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发表时间：
2012-12-01
影响因子：
7.2
通讯作者：
Huang, Chieh-Chen
中科院分区：
文献类型：
--
作者：
Chien, Lee-Feng;Kuo, Ting-Ting;Huang, Chieh-Chen
绿色藻类能够通过叶绿体氢化酶（HydA）将太阳能从H+转化为生物氢（solar-to-H-2），叶绿体氢化酶（HydA）可以直接从铁氧还蛋白接受电子并产生H-2。HydA基因由核编码，只有在厌氧或缺硫条件下才能观察到它的转录和活性，本研究试图同源过表达HydA基因，以提高小球藻DT（Chlorella sp. DT，DT）在有氧和硫供应（+S）条件下的H-2产量。对DT hydA的部分基因组DNA和cDNA进行了核苷酸序列测定和克隆。将工程化的hydA基因编码区（hydAc）构建到质粒中，并由可在需氧条件下起作用的启动子驱动。用构建的质粒转化DT细胞，并验证所得转基因在其基因组DNA中含有同源hydAc片段。在有氧和+S条件下，可以观察到HyAc的RNA转录和HyA的蛋白表达。在相同条件下，与DT野生型相比，具有同源过表达HydA的DT转基因的H-2产量增加了7至10倍。版权所有（c）2012，氢能出版物有限责任公司。由爱思唯尔有限公司出版。保留所有权利。
Green algae are able to convert solar energy into biological hydrogen (solar-to-H-2) from H+ via chloroplast hydrogenase (HydA), which can accept electrons directly from ferredoxin and generates H-2. HydA is nuclear-encoded by the hydA gene, and its transcription and activity can only be observed under anaerobic or sulfur-deprived conditions.In this study we attempted to homologously overexpress hydA to enhance H-2 production in Chlorella sp. DT (DT) under aerobic and S-supplied (+S) conditions. The nucleotides of partial genomic DNA and cDNA of DT hydA were sequenced and cloned. The engineered coding region of the hydA gene (hydAc) was constructed into plasmids and driven by the promoters that can function under aerobic conditions. The DT cells were transformed with the constructed plasmids and the resultant transgenics were verified as containing homologous hydAc fragments in their genomic DNAs. Under aerobic and +S conditions, the RNA transcription of hyAc and the protein expression of HyA in the alga DT transgenics could be observed. The H-2 production of the DT transgenics with homologously overexpressed HydA increased by as much as 7 to 10-fold in comparison to the DT wild type under the same conditions. Copyright (c) 2012, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.