Construction of heterologous gene expression cassettes for the development of recombinant <Emphasis Type="Italic">Clostridium beijerinckii</Emphasis> |
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Authors: | Young Hoon Oh Gyeong Tae Eom Kyoung Hee Kang Jeong Chan Joo Young-Ah Jang Jae Woo Choi Bong Keun Song Seung Hwan Lee Si Jae Park |
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Institution: | 1.Center for Bio-based Chemistry, Division of Convergence Chemistry,Korea Research Institute of Chemical Technology,Taejon,Republic of Korea;2.Center for Industrial Chemical Biotechnology, Ulsan Research and Business Development Division,Korea Research Institute of Chemical Technology,Ulsan,Republic of Korea;3.Department of Biological and Chemical Engineering,Hongik University,Chungnam,Republic of Korea;4.Department of Biotechnology and Bioengineering, College of Engineering,Chonnam National University,Kwangju,Republic of Korea;5.Department of Environmental Engineering and Energy,Myongji University,Yongin,Republic of Korea |
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Abstract: | Gene-expression cassettes for the construction of recombinant Clostridium beijerinckii were developed as potential tools for metabolic engineering of C. beijerinckii. Gene expression cassettes containing ColE1 origin and pAMB origin along with the erythromycin resistance gene were constructed, in which promoters from Escherichia coli, Lactococcus lactis, Ralstonia eutropha, C. acetobutylicum, and C. beijerinckii are examined as potential promoters in C. beijerinckii. Zymogram analysis of the cell extracts and comparison of lipase activities of the recombinant C. beijerinckii strains expressing Pseudomonas fluorescens tliA gene suggested that the tliA gene was functionally expressed by all the examined promoters with different expression level. Also, recombinant C. beijerinckii expressing C. beijerinckii secondary alcohol dehydrogenase by the constructed expression cassettes successfully produced 2-propanol from glucose. The best promoter for TliA expression was the R. eutropha phaP promoter while that for 2-propanol production was the putative C. beijerinckii pta promoter. Gene expression cassettes developed in this study may be useful tools for the construction of recombinant C. beijerinckii strains as host strains for the valuable chemicals and fuels from renewable resources. |
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