Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 80, Issue 10, Pages 3072-3080Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.03986-13
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Funding
- Major State Basic Research Development Program of China [2012CB720806]
- National Natural Science Foundation of China [31370130]
- Fundamental Research Funds for the Central Universities [JUSRP51307A]
- Foundation for the Author of National Excellent Doctoral Dissertation of People's Republic of China (FANEDD) [2011046]
- New Century Excellent Talents in University [NCET-12-0876]
- 111 Project [111-2-06]
- Natural Science Foundation of Jiangsu Province [BK2011004, BK2010150]
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The health benefits of flavonoids for humans are increasingly attracting attention. Because the extraction of high-purity flavonoids from plants presents a major obstacle, interest has emerged in biosynthesizing them using microbial hosts. Eriodictyol is a flavonoid with anti-inflammatory and antioxidant activities. Its efficient synthesis has been hampered by two factors: the poor expression of cytochrome P450 and the low intracellular malonyl coenzyme A (malonyl-CoA) concentration in Escherichia coli. To address these issues, a truncated plant P450 flavonoid, flavonoid 3 '-hydroxylase (tF3 ' H), was functionally expressed as a fusion protein with a truncated P450 reductase (tCPR) in E. coli. This allowed the engineered E. coli to produce eriodictyol from L-tyrosine by simultaneously coexpressing the fusion protein with tyrosine ammonia lyase (TAL), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI). In addition, metabolic engineering was employed to enhance the availability of malonyl-CoA so as to achieve a new metabolic balance and rebalance the relative expression of genes to enhance eriodictyol accumulation. This approach made the production of eriodictyol 203% higher than that in the control strain. By using these strategies, the production of eriodictyol from L-tyrosine reached 107 mg/liter. The present work offers an approach to the efficient synthesis of other hydroxylated flavonoids from L-tyrosine or even glucose in E. coli.
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