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Enhanced H2 Production and Redirected Metabolic Flux via Overexpression of fhlA and pncB in Klebsiella HQ-3 Strain

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Abstract

Genetic modifications are considered as one of the most important technologies for improving fermentative hydrogen yield. Herein, we overexpress fhlA and pncB genes from Klebsiella HQ-3 independently to enhance hydrogen molar yield. HQ-3-fhlA/pncB strain is developed by manipulation of pET28-Pkan/fhlA Kanr and pBBR1-MCS5/pncB Gmr as expression vectors to examine the synchronous effects of fhlA and pncB. Optimization of anaerobic batch fermentations is achieved and the maximum yield of biohydrogen (1.42 mol H2/mol of glucose) is produced in the range of pH 6.5–7.0 at 33–37 °C. Whole cell H2 yield is increased up to 40 % from HQ-3-fhlA/pncB, as compared with HQ-3-fhlA 20 % and HQ-3-pncB 12 % keeping HQ-3-C as a control. Mechanism of improved H2 yield is studied in combination with metabolic flux analysis by measuring glucose consumption and other metabolites including formate, succinate, 2,3 butanediol, lactate, acetate, ethanol, and hydrogen. The results suggest that under transient conditions, the increase in the total level of NAD by NAPRTase can enhance the rate of NADH-dependent pathways, and therefore, final distribution of metabolites is changed. Combined overexpression of fhlA and pncB eventually modifies the energy and carbon balance leading to enhanced H2 production from FHL as well as by NADH pathway.

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Acknowledgments

This work was supported by the Fundamental Research Funds for the Central Universities (HUST: 2010MS029) and the National Natural Science Foundation of P. R. China (NSFC) (Nos. 31070089 and 31170078). The authors are deeply indebted to the Analytical and Testing Center of Huazhong University of Science and Technology.

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Authors and Affiliations

  1. Key Laboratory of Molecular Biophysics, the Ministry of Education, Huazhong University of Science and Technology, Wuhan, 430074, People’s Republic of China

    Muhammad Jawed, Jian Pi, Li Xu, Houjin Zhang & Yunjun Yan

  2. College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, People’s Republic of China

    Li Xu, Houjin Zhang, Abdul Hakeem & Yunjun Yan

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  1. Muhammad Jawed
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  2. Jian Pi
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  3. Li Xu
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  5. Abdul Hakeem
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Correspondence to Yunjun Yan.

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Jawed, M., Pi, J., Xu, L. et al. Enhanced H2 Production and Redirected Metabolic Flux via Overexpression of fhlA and pncB in Klebsiella HQ-3 Strain. Appl Biochem Biotechnol 178, 1113–1128 (2016). https://doi.org/10.1007/s12010-015-1932-4

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