Hydrogen production by Escherichia coli containing a cloned hydrogenase gene from Citrobacter freundii |
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| Affiliation: | 1. Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan;1. Tokyo Research Laboratory, Japan Synthetic Rubber Co., Ltd., Kawasaki, Japan;1. Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;1. Microbial Biotechnology and Genomics, CSIR-Institute of Genomics and Integrative Biology (IGIB), Delhi University Campus, Mall Road, Delhi 110007, India;2. Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, Republic of Korea;1. Department of Horticulture Sciences, University of Mohaghegh Ardabili, Ardabil, Iran;2. Cellular and Molecular Research Center & Department of Medical Parasitology and Mycology, Urmia University of Medical Sciences, Urmia, Iran;3. Department of Plant Production and Genetics, Urmia University, Urmia, Iran;4. Centre for Climate and Environmental Protection, School of Water, Energy and Environment, Cranfield University, Cranfield MK43 0AL, UK;1. Biotechnology Division, CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram 695019, Kerala, India;2. Academy of Scientific and Innovative Research (AcSIR), CSIR-NIIST, Thiruvananthapuram 695019, Kerala, India;1. Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, Jülich, Germany;2. Institute of Complex Systems (ICS-4 (Cellular Biophysics)), Forschungszentrum Jülich, Jülich, Germany;3. Institute of Complex Systems (ICS-6 (Structural Biochemistry)), Forschungszentrum Jülich, Jülich, Germany |
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| Abstract: | Two hydrogenase genes of Citrobacter freundii complementing different Escherichia coli hyd mutations were cloned on the multicopy-plasmid pBR322. Recombinant plasmids pCBH2 and pCFH1 were obtained. Since hydrogenase activities of E. coli transformant HK-8 (pCBH2) and HK-7 (pCFH1) were much the same as E. coli C600 (wild type cells), the reduction in DNA size of recombinant plasmid pCBH2 (10.7 kb) was investigated. Reduced recombinant plasmids pCBH4 (6.2 kb) and pCBH6 (5.7 kb) were obtained, and a hydrogenase gene was found to be located on the 2.35 kb fragment between AvaI and EcoRI sites. Hydrogenase activity and hydrogen-evolving activity of E. coli HK-8 (pCBH4 or pCBH6) from sodium formate, sodium pyruvate or glucose were approximately 2-fold higher than those of E. coli C600 (wild type cells).On the other hand, a reduced recombinant plasmid pCBH10 (6.0 kb), which contained the adjacent DNA fragment (2.15 kb) to a hydrogenase gene, was obtained. Hydrogenase activity of E. coli C600 harboring pCBH10 was half that of E. coli C600. From these results we estimate that in plasmid pCBH2, the repressor gene suppressing the synthesis of hydrogenase might have been cloned together with a hydrogenase gene. |
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