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Microbial production of R-3-hydroxybutyric acid by recombinant E. coli harboring genes of phbA, phbB, and tesB

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Abstract

Production of R-3-hydroxybutyric acid (3HB) was observed when genes of β-ketothiolase (PhbA), acetoacetyl CoA reductase (PhbB), and thioesterase II (TesB) were jointly expressed in Escherichia coli. TesB, generally regarded as a medium chain length acyl CoA thioesterase, was found, for the first time, to play an important role for transforming short chain length 3-hydroxybutyrate-CoA to its free fatty acid, namely, 3HB. E. coli BW25113 (pSPB01) harboring phbA, phbB, and tesB genes produced approximately 4 g/l 3HB in shake flask culture within 24 h with glucose used as a carbon source. Under anaerobic growth conditions, 3HB production was found to be more effective, achieving 0.47 g 3HB/g glucose compared with only 0.32 g 3HB/g glucose obtained from aerobic process. When growth was conducted on sodium gluconate, 6 g/l 3HB was obtained. In a 24-h fed-batch growth process conducted in a 6-l fermentor containing 3 l glucose mineral medium, 12 g/l 3HB was produced from 17 g/l cell dry weight (CDW). This was the highest 3HB productivity achieved by a one-stage fermentation process for 3HB production.

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Acknowledgements

The strain E. coli BW25113 was kindly donated by Dr. BL Wanner of Purdue University, USA. Plasmid pBHR69 was donated by Professor A. Steinbüchel of the University of Muenster, Germany. This research was supported by the Natural Sciences Foundation of China Grant Nos. 20334020 and 30570024. Thanks must also be extended to the Hi-Tech Research and Development Program of China (863 Program) Grant No. 2006AA02Z242 and the National Basic Research Program of China (973 Program) No. 2007CB707804.

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Correspondence to Guo-Qiang Chen.

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Liu and Ouyang contributed equally to the paper.

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Liu, Q., Ouyang, SP., Chung, A. et al. Microbial production of R-3-hydroxybutyric acid by recombinant E. coli harboring genes of phbA, phbB, and tesB . Appl Microbiol Biotechnol 76, 811–818 (2007). https://doi.org/10.1007/s00253-007-1063-0

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