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Efficient (R)-3-hydroxybutyrate production using acetyl CoA-regenerating pathway catalyzed by coenzyme A transferase

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Title: Efficient (R)-3-hydroxybutyrate production using acetyl CoA-regenerating pathway catalyzed by coenzyme A transferase
Authors: Matsumoto, Ken'ichiro Browse this author →KAKEN DB
Okei, Takehiro Browse this author
Honma, Inori Browse this author
Ooi, Toshihiko Browse this author →KAKEN DB
Aoki, Hirobumi Browse this author
Taguchi, Seiichi Browse this author →KAKEN DB
Keywords: Chiral synthesis
Production rate
Issue Date: Jan-2013
Publisher: Springer-Verlag
Journal Title: Applied Microbiology and Biotechnology
Volume: 97
Issue: 1
Start Page: 205
End Page: 210
Publisher DOI: 10.1007/s00253-012-4104-2
PMID: 22592551
Abstract: (R)-3-hydroxybutyrate [(R)-3HB] is a useful precursor in the synthesis of value-added chiral compounds such as antibiotics and vitamins. Typically, (R)-3HB has been microbially produced from sugars via modified (R)-3HB-polymer-synthesizing pathways in which acetyl-CoA is converted into (R)-3-hydroxybutyryl-coenzyme A [(R)-3HB-CoA] by β-ketothiolase (PhaA) and acetoacetyl-CoA reductase (PhaB). (R)-3HB-CoA is hydrolyzed into (R)-3HB by modifying enzymes or undergoes degradation of the polymerized product. In the present study, we constructed a new (R)-3HB-generating pathway from glucose by using propionyl-CoA transferase (PCT). This pathway was designed to excrete (R)-3HB by means of a PCT-catalyzed reaction coupled with regeneration of acetyl-CoA, the starting substance for synthesizing (R)-3HB-CoA. Considering the equilibrium reaction of PCT, the PCT-catalyzed (R)-3HB production would be expected to be facilitated by the addition of acetate since it acts as an acceptor of CoA. As expected, the engineered Escherichia coli harboring the phaAB and pct genes produced 1.0 g L^[-1] (R)-3HB from glucose, and with the addition of acetate into the medium, the concentration was increased up to 5.2 g L^[-1], with a productivity of 0.22 g L^[-1] h^[-1]. The effectiveness of the extracellularly added acetate was evaluated by monitoring the conversion of 13C carbonyl carbon labeled acetate into (R)-3HB using gas chromatography/mass spectrometry. The enantiopurity of (R)-3HB was determined to be 99.2% using chiral liquid chromatography. These results demonstrate that the PCT pathway achieved a rapid co-conversion of glucose and acetate into (R)-3HB.
Rights: The original publication is available at
Type: article (author version)
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 松本 謙一郎

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