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Construction of a Novel Expression System in Klebsiella pneumoniae and its Application for 1,3-Propanediol Production

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Abstract

A novel expression system of Klebsiella pneumoniae was developed in order to improve 1,3-propanediol (1,3-PD) production using a K. pneumoniae–Escherichia coli shuttle vector pET28a consisting of the kanamycin-resistance gene promoter Pkan. The recombinant plasmid pETPkan-cat carrying the chloramphenicol acetyltransferase gene cat as selectable marker was constructed to test the availability of the promoter Pkan in K. pneumoniae. The results showed that the chloramphenicol acetyltransferase was apparently expressed in K. pneumoniae, and the recombinant strain had a high-level resistance to chloramphenicol, suggesting that the promoter Pkan was efficient in K. pneumoniae. Then, the expression system was applied to the expression of 1,3-PD oxidoreductase in K. pneumoniae. The enzyme was over-expressed, and the recombinant K. pneumoniae showed a nearly 3.0-fold decrease in peak level of the intermediary metabolite 3-hydroxypropionaldehyde and an increase of 16.5% in yield of 1,3-PD with respect to the wild-type strain. From these results, the first reported expression system has paved the way for improvement of 1,3-PD production and will be available and efficient for other heterologous gene expression in K. pneumoniae.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (20676053), National Programs for High Technology Research and Development of China (2006AA020103 and 2007AA02Z207), Program for New Century Excellent Talents in University (No. NCET-07-0380), and National Basic Research Program (973 Program; No. 2007CB707800).

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Authors and Affiliations

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

    Zheng Ma, Zhiming Rao, Bin Zhuge, Huiying Fang, Xiangru Liao & Jian Zhuge

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  1. Zheng Ma
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  2. Zhiming Rao
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  3. Bin Zhuge
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  4. Huiying Fang
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  5. Xiangru Liao
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  6. Jian Zhuge
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Correspondence to Zhiming Rao.

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Ma, Z., Rao, Z., Zhuge, B. et al. Construction of a Novel Expression System in Klebsiella pneumoniae and its Application for 1,3-Propanediol Production. Appl Biochem Biotechnol 162, 399–407 (2010). https://doi.org/10.1007/s12010-009-8743-4

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