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Production of (2S)-sakuranetin from (2S)-naringenin in Escherichia coli by strengthening methylation process and cell resistance.
- Source :
-
Synthetic & Systems Biotechnology . Dec2022, Vol. 7 Issue 4, p1117-1125. 9p. - Publication Year :
- 2022
-
Abstract
- (2S)-Sakuranetin is a 7-O-methylflavonoid that has anticancer, antiviral, and antimicrobial activities. Methylation process is involved in biosynthesizing (2S)-sakuranetin from (2S)-naringenin, in which S-adenosylmethionine (SAM) serves as the methyl donor. In this study, after methyl donor and substrate inhibition were identified as limiting factors for (2S)-sakuranetin biosynthesis, an efficient (2S)-sakuranetin-producing strain was constructed by enhancing methyl donor supply and cell tolerance to (2S)-naringenin. Firstly, PfOMT3 from Perilla frutescens was selected as the optimal flavonoid 7-O-methyltransferase (F7-OMT) for the conversion of (2S)-naringenin to (2S)-sakuranetin. Then, the methylation process was upregulated by regulating pyridoxal 5′ - phosphate (PLP) content, key enzymes in methionine synthesis pathway, and the availability of ATP. Furthermore, genes that can enhance cell resistance to (2S)-naringenin were identified from molecular chaperones and sRNAs. Finally, by optimizing the fermentation process, 681.44 mg/L of (2S)-sakuranetin was obtained in 250- mL shake flasks. The titer of (2S)-sakuranetin reached 2642.38 mg/L in a 5-L bioreactor, which is the highest titer ever reported. This work demonstrates the importance of cofactor PLP in methylation process, and provides insights to biosynthesize other O-methylated flavonoids efficiently in E. coli. [ABSTRACT FROM AUTHOR]
- Subjects :
- *METHYLATION
*ANTINEOPLASTIC agents
*BIOSYNTHESIS
*BIOCHEMISTRY
*ESCHERICHIA coli
Subjects
Details
- Language :
- English
- ISSN :
- 20971206
- Volume :
- 7
- Issue :
- 4
- Database :
- Academic Search Index
- Journal :
- Synthetic & Systems Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 161091259
- Full Text :
- https://doi.org/10.1016/j.synbio.2022.07.004