Journal
BIORESOURCE TECHNOLOGY
Volume 287, Issue -, Pages -Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2019.121484
Keywords
Yarrowia lipolytica; Crotonic acid; Butanol-forming route; Pyruvate dehydrogenase; White biotechnology
Funding
- Hubei Provincial Natural Science Foundation of China [2018CFB358]
- Research Fund for the Doctoral Program of Hubei University of Technology [BSQD2017033]
- Beijing Laboratory for Food Quality and Safety, Beijing Technology and Business University
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In this study, Y. lipolytica was engineered to produce crotonic acid via the butanol-forming route. Firstly, the crotonase and 3-hydroxybutyryl-CoA dehydrogenase genes from Clostridium beijerinckii, and the thioesterase gene from Bacteroides thetaiotaomicron were heterologously expressed in Y. lipolytica, the engineered strain LZJ001 accumulated 62.3 +/- 4.2 mg/L of crotonic acid. Secondly, the acetyl-CoA acetyltransferase from Saccharomyces cerevisiae was overexpressed, the derived recombinant strain LZJ002 produced 123.5 +/- 6.8 mg/L of crotonic acid. Finally, the pyruvate dehydrogenase from Escherichia coll. was additionally expressed, giving the fully engineered strain LZJ004 that produced 220.0 +/- 8.2 mg/L of crotonic acid in shaking-flask culture, which represents a 3.5-fold increase over LZJ001 strain. The approach described here paves the way for environmentally friendly and large-scale industrial production of crotonic acid.
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