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Effects of NADH kinase on NADPH-dependent biotransformation processes in Escherichia coli

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Abstract

Sufficient supply of NADPH is one of the most important factors affecting the productivity of biotransformation processes. In this study, construction of an efficient NADPH-regenerating system was attempted using direct phosphorylation of NADH by NADH kinase (Pos5p) from Saccharomyces cerevisiae for producing guanosine diphosphate (GDP)-l-fucose and ε-caprolactone in recombinant Escherichia coli. Expression of Pos5p in a fed-batch culture of recombinant E. coli producing GDP-l-fucose resulted in a maximum GDP-l-fucose concentration of 291.5 mg/l, which corresponded to a 51 % enhancement compared with the control strain. In a fed-batch Baeyer–Villiger (BV) oxidation of cyclohexanone using recombinant E. coli expressing Pos5p, a maximum ε-caprolactone concentration of 21.6 g/l was obtained, which corresponded to a 96 % enhancement compared with the control strain. Such an increase might be due to the enhanced availability of NADPH in recombinant E. coli expressing Pos5p. These results suggested that efficient regeneration of NADPH was possible by functional expression of Pos5p in recombinant E. coli, which can be applied to other NADPH-dependent biotransformation processes in E. coli.

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Acknowledgments

This research was supported by Advanced Biomass R&D Center (ABC) (2011-0031359) and WCU (World Class University) program (R320110001018530) through the National Research Foundation of Korea (NRF) both funded by the Ministry of Education, Science and Technology. This work was also supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (2011-0003791). E.H. P. and M.D.K. were supported by the Ministry of Knowledge Economy (MKE) and the Korea Institute for Advancement of Technology (KIAT) through the Research and Development for Regional Industry.

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The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, 151-921, Korea

    Won-Heong Lee, Jin-Woo Kim & Jin-Ho Seo

  2. Department of Food Science and Biotechnology, Kangwon National University, Chuncheon, 200-701, Korea

    Eun-Hee Park & Myoung-Dong Kim

  3. Department of Food Science and Technology, Chungbuk National University, Cheongju, 361-763, Korea

    Nam Soo Han

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  1. Won-Heong Lee
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  2. Jin-Woo Kim
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  4. Nam Soo Han
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  5. Myoung-Dong Kim
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Correspondence to Myoung-Dong Kim or Jin-Ho Seo.

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Lee, WH., Kim, JW., Park, EH. et al. Effects of NADH kinase on NADPH-dependent biotransformation processes in Escherichia coli . Appl Microbiol Biotechnol 97, 1561–1569 (2013). https://doi.org/10.1007/s00253-012-4431-3

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  • DOI: https://doi.org/10.1007/s00253-012-4431-3

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