Propionyl-CoA dependent biosynthesis of 2-hydroxybutyrate containing polyhydroxyalkanoates in metabolically engineered Escherichia coli

Si Jae Park, Kyoung Hee Kang, Hyuk Lee, A. Reum Park, Jung Eun Yang, Young Hoon Oh, Bong Keun Song, Jonggeon Jegal, Seung Hwan Lee, Sang Yup Lee

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

We have previously reported in vivo biosynthesis of 2-hydroxyacid containing polyesters including polylactic acid (PLA), poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)], and poly(3-hydroxybutyrate-co-2-hydroxybutyrate-co-lactate) [P(3HB-co-2HB-co-LA)] employing metabolically engineered Escherichia coli strains by the introduction of evolved Clostridium propionicum propionyl-CoA transferase (PctCp) and Pseudomonas sp. MBEL 6-19 polyhydroxyalkanoate (PHA) synthase 1 (PhaC1Ps6-19). In this study, we further engineered in vivo PLA biosynthesis system in E. coli to synthesize 2HB-containing PHA, in which propionyl-CoA was used as precursor for 2-ketobutyrate that was converted into 2HB-CoA by the sequential actions of Lactococcus lactis (d)-2-hydroxybutyrate dehydrogenase (PanE) and PctCp and then 2HB-CoA was polymerized by PhaC1Ps6-19. The recombinant E. coli XL1-blue expressing the phaC1437 gene, the pct540 gene, and the Ralstonia eutropha prpE gene together with the panE gene could be grown to 0.66g/L and successfully produced P(70mol%3HB-co-18mol%2HB-co-12mol%LA) up to the PHA content of 66wt% from 20g/L of glucose, 2g/L of 3HB and 1g/L of sodium propionate. Removal of the prpC gene in the chromosome of E. coli XL1-blue could increase the mole fraction of 2HB in copolymer, but the PHA content was decreased.The metabolic engineering strategy reported here suggests that propionyl-CoA can be successfully used as the precursor to provide PHA synthase with 2HB-CoA for the production of PHAs containing 2HB monomer.

Original languageEnglish
Pages (from-to)93-98
Number of pages6
JournalJournal of Biotechnology
Volume165
Issue number2
DOIs
StatePublished - May 2013

Keywords

  • 2-Ketobutyrate
  • 2HB containing PHA
  • Metabolic engineering
  • Propionyl-CoA

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