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Inactivation of dhaD and dhaK abolishes by-product accumulation during 1,3-propanediol production in Klebsiella pneumoniae

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Journal of Industrial Microbiology & Biotechnology

Abstract

1,3-Propanediol (1,3-PD) can be used for the industrial synthesis of a variety of compounds, including polyesters, polyethers, and polyurethanes. 1,3-PD is generated from petrochemical and microbial sources. 1,3-Propanediol is a typical product of glycerol fermentation, while acetate, lactate, 2,3-butanediol, and ethanol also accumulate during the process. Substrate and product inhibition limit the final concentration of 1,3-propanediol in the fermentation broth. It is impossible to increase the yield of 1,3-propanediol by using the traditional whole-cell fermentation process. In this study, dhaD and dhaK, the genes for glycerol dehydrogenase and dihydroxyacetone kinase, respectively, were inactivated by homologous recombination in Klebsiella pneumoniae. The dhaD/dhaK double mutant (designated TC100), selected from 5,000 single or double cross homologous recombination mutants, was confirmed as a double cross by using polymerase chain reaction. Analysis of the cell-free supernatant with high-performance liquid chromatography revealed elimination of lactate and 2,3-butanediol, as well as ethanol accumulation in TC100, compared with the wild-type strain. Furthermore, 1,3-propanediol productivity was increased in the TC100 strain expressing glycerol dehydratase and 1,3-PDO dehydrogenase regulated by the arabinose PBAD promoter. The genetic engineering and medium formulation approaches used here should aid in the separation of 1,3-propanediol from lactate, 2,3-butanediol, and ethanol and lead to increased production of 1,3-propanediol in Klebsiella pneumoniae.

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Acknowledgments

This work was supported by grants from the National Science Council (NSC-97-2320-B-320-012-MY3) which were really appreciated.

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Authors and Affiliations

  1. Department of Laboratory, Medicine and Biotechnology, Tzu Chi University, College of Medicine, 701 Section 3, Zhongyang Road, Hualien, 970, Taiwan, ROC

    Yu-Tze Horng, Kai-Chih Chang & Po-Chi Soo

  2. Department of Medical Biotechnology and Laboratory Science, Chang Gung University, 259 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan, 333, Taiwan, ROC

    Yu-Tze Horng

  3. Graduate School of Biotechnology and Bioengineering, Yuan Ze University, 135 Yuan-Tung Road, Chung-Li, 320, Taiwan, ROC

    Ta-Chung Chou, Chung-Jen Yu, Chih-Ching Chien & Yu-Hong Wei

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  1. Yu-Tze Horng
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  2. Kai-Chih Chang
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Correspondence to Po-Chi Soo.

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Yu-Tze Horng and Kai-Chih Chang contributed equally to this work

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Horng, YT., Chang, KC., Chou, TC. et al. Inactivation of dhaD and dhaK abolishes by-product accumulation during 1,3-propanediol production in Klebsiella pneumoniae . J Ind Microbiol Biotechnol 37, 707–716 (2010). https://doi.org/10.1007/s10295-010-0714-9

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  • DOI: https://doi.org/10.1007/s10295-010-0714-9

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