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Effect of anaerobic promoters on the microaerobic production of polyhydroxybutyrate (PHB) in recombinant Escherichia coli

Abstract

Nine anaerobic promoters were cloned and constructed upstream of PHB synthesis genes phbCAB from Ralstonia eutropha for the micro- or anaerobic PHB production in recombinant Escherichia coli. Among the promoters, the one for alcohol dehydrogenase (PadhE) was found most effective. Recombinant E. coli JM 109 (pWCY09) harboring PadhE and phbCAB achieved a 48% PHB accumulation in the cell dry weight after 48 h of static culture compared with only 30% PHB production under its native promoter. Sixty-seven percent PHB was produced in the dry weight (CDW) of an acetate pathway deleted (Δpta deletion) E. coli JW2294 harboring the vector pWCY09. In a batch process conducted in a 5.5-l NBS fermentor containing 3 l glucose LB medium, E. coli JW2294 (pWCY09) grew to 7.8 g/l CDW containing 64% PHB after 24 h of microaerobic incubation. In addition, molecular weight of PHB was observed to be much higher under microaerobic culture conditions. The high activity of PadhE appeared to be the reason for improved micro- or anaerobic cell growth and PHB production while high molecular weight contributed to the static culture condition.

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Acknowledgment

We are grateful to Professor Alexander Steinbüchel of the University of Münster in Germany for the generous donation of plasmid pBHR68 and to Keio collection, Japan, for kindly providing us with E. coli mutant strains. This research was supported by Natural Sciences Foundation of China Grant No. 30570024. Thanks must also be extended to National High Tech 863 Grant (Project Nos. 2006AA02Z242 and 2006AA020104), and National Basic Research Program of China (973 Program) No. 2007CB707804.

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Correspondence to Guo-Qiang Chen.

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Wei, X., Shi, Z., Yuan, M. et al. Effect of anaerobic promoters on the microaerobic production of polyhydroxybutyrate (PHB) in recombinant Escherichia coli . Appl Microbiol Biotechnol 82, 703–712 (2009). https://doi.org/10.1007/s00253-008-1816-4

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Keywords

  • PHB
  • Polyhydroxyalkanoates
  • Microaerobiosis
  • Anaerobic promoter
  • phbCAB
  • Alcohol dehydrogenase