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Construction of heterologous gene expression cassettes for the development of recombinant Clostridium beijerinckii

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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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Acknowledgments

This work was supported by the R&D Program of MOTIE/KEIT (10049674) and 2014 Research Fund of Myongji University.

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Authors and Affiliations

  1. Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, 141 Gajeong-ro, Yuseong-gu, P.O. Box 107, Taejon, 305-600, Republic of Korea

    Young Hoon Oh, Kyoung Hee Kang, Jeong Chan Joo, Young-Ah Jang, Jae Woo Choi & Bong Keun Song

  2. Center for Industrial Chemical Biotechnology, Ulsan Research and Business Development Division, Korea Research Institute of Chemical Technology, 45 Jongga-ro, Jung-gu, Ulsan, 681-802, Republic of Korea

    Gyeong Tae Eom

  3. Department of Biological and Chemical Engineering, Hongik University, Chungnam, 339-701, Republic of Korea

    Jae Woo Choi

  4. Department of Biotechnology and Bioengineering, College of Engineering, Chonnam National University, 77 Yongbong-ro, Buk-gu, Kwangju, 500-757, Republic of Korea

    Seung Hwan Lee

  5. Department of Environmental Engineering and Energy, Myongji University, 116 Myongji-ro, Cheoin-gu, Yongin, Gyeonggi-do, 449-728, Republic of Korea

    Si Jae Park

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  1. Young Hoon Oh
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  2. Gyeong Tae Eom
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Correspondence to Seung Hwan Lee or Si Jae Park.

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Oh, Y.H., Eom, G.T., Kang, K.H. et al. Construction of heterologous gene expression cassettes for the development of recombinant Clostridium beijerinckii . Bioprocess Biosyst Eng 39, 555–563 (2016). https://doi.org/10.1007/s00449-016-1537-5

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  • DOI: https://doi.org/10.1007/s00449-016-1537-5

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