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Expression of 1,3-propanediol oxidoreductase and its isoenzyme in Klebsiella pneumoniae for bioconversion of glycerol into 1,3-propanediol

  • Applied Genetics and Molecular Biotechnology
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Abstract

In the Klebsiella pneumoniae reduction pathway for 1,3-propanediol (1,3-PD) synthesis, glycerol is first dehydrated to 3-hydroxypropionaldehyde (3-HPA) and then reduced to 1,3-PD with NADH consumption. Rapid conversion of 3-HPA to 1,3-PD is one of the ways to improve the yield of 1,3-PD from glycerol and to avoid 3-HPA accumulation, which depends on enzyme activity of the reaction and the amount of reducing equivalents available from the oxidative pathway of glycerol. In the present study, the yqhD gene, encoding 3-propanediol oxidoreductase isoenzyme from Escherichia coli and the dhaT gene, encoding 3-propanediol oxidoreductase from K. pneumoniae were expressed individually and co-expressed in K. pneumoniae using the double tac promoter expression plasmid pEtac-dhaT-tac-yqhD. The three resultant recombinant strains (K. pneumoniae/pEtac-yqhD, K. pneumoniae/pEtac-dhaT, and K. pneumoniae/pEtac-dhaT-tac-yqhD) were used for fermentation studies. Experimental results showed that the peak values for 3-HPA production in broth of the three recombinant strains were less than 25% of that of the parent strain. Expression of dhaT reduced formation of by-products (ethanol and lactic acid) and increased molar yield of 1,3-PD slightly, while expression of yqhD did not enhance molar yield of 1,3-PD, but increased ethanol concentration in broth as NADPH participation in transforming 3-HPA to 1,3-PD allowed more cellular NADH to be used to produce ethanol. Co-expression of both genes therefore decreased by-products and increased the molar yield of 1,3-PD by 11.8%, by catalyzing 3-HPA conversion to 1,3-propanediol using two cofactors (NADH and NADPH). These results have important implications for further studies involving use of YqhD and DhaT for bioconversion of glycerol into 1,3-PD.

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Acknowledgements

This work was supported by National Programs for High Technology Research and Development of China (2006AA020103 and 2009AA02Z210).

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Authors and Affiliations

  1. Key Laboratory of Industrial Biotechnology of Ministry of Education, Research Center of Industrial Microbiology, School of Biotechnology and State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province, 214122, People’s Republic of China

    Bin Zhuge, Cheng Zhang, Huiying Fang & Jian Zhuge

  2. Department of Biotechnology and Food Technology, Durban University of Technology, P.O. Box 1334, Durban, 4000, South Africa

    Kugen Permaul

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  1. Bin Zhuge
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  2. Cheng Zhang
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  3. Huiying Fang
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  4. Jian Zhuge
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  5. Kugen Permaul
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Correspondence to Bin Zhuge.

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Zhuge, B., Zhang, C., Fang, H. et al. Expression of 1,3-propanediol oxidoreductase and its isoenzyme in Klebsiella pneumoniae for bioconversion of glycerol into 1,3-propanediol. Appl Microbiol Biotechnol 87, 2177–2184 (2010). https://doi.org/10.1007/s00253-010-2678-0

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  • DOI: https://doi.org/10.1007/s00253-010-2678-0

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