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Application of acetyl-CoA acetyltransferase (AtoAD) in Escherichia coli to increase 3-hydroxyvalerate fraction in poly(3-hydroxybutyrate-co-3-hydroxyvalerate)

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Abstract

Polyhydroxyalkanoate (PHA) is a family of biodegradable polymers, and incorporation of different monomers can alter its physical properties. To produce the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB-co-3HV)) containing a high level of 3-hydroxyvalerate (3HV) by altering acetyl-CoA pool levels, we overexpressed an acetyl-CoA acetyltransferase (atoAD) in an engineered E. coli strain, YH090, carrying PHA synthetic genes bktB, phaB, and phaC. It was found that, with introduction of atoAD and with propionate as a co-substrate, 3HV fraction in PHA was increased up to 7.3-fold higher than a strain without atoAD expressed in trans (67.9 mol%). By the analysis of CoA pool concentrations in vivo and in vitro using HPLC and LC-MS, overexpression of AtoAD was shown to decrease the amount of acetyl-CoA and increase the propionyl-CoA/acetyl-CoA ratio, ultimately resulting in an increased 3HV fraction in PHA. Finally, synthesis of P(3HB-co-3HV) containing 57.9 mol% of 3HV was achieved by fed-batch fermentation of YJ101 with propionate.

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Acknowledgements

This study was supported by the National Research Foundation of Korea (NRF-2016R1D1A1B03932301 and NRF-2015M1A5A1037196). This paper was written as part of Konkuk University’s research support program for its faculty on sabbatical leave in 2016. Consulting service from the Microbial Carbohydrate Resource Bank (MCRB, Seoul, Korea) was kindly appreciated.

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Authors and Affiliations

  1. Department of Bioengineering, College of Engineering, Konkuk University, Seoul, South Korea

    Jong-Min Jeon, Hyun-Joong Kim, Shashi Kant Bhatia, Hyung-Min Seo, Jung-Ho Kim, Hyung-Yeon Park, Dahye Lee & Yung-Hun Yang

  2. School of Chemical and Biological Engineering, Seoul National University, Kwanak-Gu, 151-742, Seoul, South Korea

    Changmin Sung

  3. Department of Bioengineering, University of Massachusetts Dartmouth, 285 Old Westport Road, North Dartmouth, MA, 02747-2300, USA

    Christopher J. Brigham

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  1. Jong-Min Jeon
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Correspondence to Yung-Hun Yang.

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Jeon, JM., Kim, HJ., Bhatia, S. et al. Application of acetyl-CoA acetyltransferase (AtoAD) in Escherichia coli to increase 3-hydroxyvalerate fraction in poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Bioprocess Biosyst Eng 40, 781–789 (2017). https://doi.org/10.1007/s00449-017-1743-9

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