Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 67, Issue 23, Pages 6532-6540Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.9b02646
Keywords
flavins; Escherichia coli; modular engineering; error-prone PCR
Funding
- National Natural Science Foundation of China [NSFC-21621004, NSFC-21776208, NSFC-21576200]
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In this work, modular engineering of Escherichia coli was peformed to improve flavin production and the conversion ratio of riboflavin (RE) to FMN/FAD. The RF operon and the bifunctional RF kinase/FAD synthetase were divided into two separate modules. The two modules were expressed at different levels to produce RF:ribF ratios ranging from 2:20 to 7:5. The best strain respectively produced 324.1 and 171.6 mg/L of FAD and FMN in shake flask fermentation, and the titers reached 1899.3 and 872.7 mg/L in a fed-batch process. Furthermore, error-prone PCR (epPCR) of the E. coli ribF gene was performed. The highest FMN production of the best mutant reached 586.1 mg/L in shake flask cultivation. Moreover, this mutant produced 1017.5 mg/L FMN with a greatly reduced proportion of FAD in fermenter culture. To the best of our knowledge, this is the highest production of FAD and FMN in a microbial fermentation process reported to date.
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