期刊
CHEMBIOCHEM
卷 5, 期 4, 页码 500-507出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.200300783
关键词
biosynthesis; flavonoids; heterologous expression; natural products; polyketides
资金
- NIGMS NIH HHS [T32 GM08347] Funding Source: Medline
Flavonoids are important plant-specific secondary metabolites synthesized from 4-coumaroyl coenzyme A (CoA), derived from the general phenylpropanoid pathway, and three malonyl-CoAs. The synthesis involves a plant type III polyketide synthase, chalcone synthase. We report the cloning and coexpression in Escherichia coli of phenylalanine ammonia lyase, cinnamate-4-hydroxylose, 4-coumarate:CoA ligase, and chalcone synthase from the model plant Arabidopsis thaliana. Simultaneous expression of all four genes resulted in a blockage after the first enzymatic step caused by the presence of nonfunctional cinnamate-4-hydroxylose. To overcome this problem we fed exogenous 4-coumaric acid to induced cultures. We observed high-level production of the flavanone naringenin as a result. We were also able to produce phloretin by feeding cultures with 3-(4-hydroxyphenyl)propionic acid. Feeding with ferulic or caffeic acid did not yield the corresponding flavanones. We have also cloned and partially characterized a new tyrosine ammonia lyase from Rhodobacter sphaeroides. Tyrosine ammonia lyase was substituted for phenylalanine ammonia lyase and cinnamate-4-hydroxylose in our E. coli clones and three different growth media were tested. After 48 h induction, high-level production (20.8 mg L-1) of naringenin in metabolically engineered E. coli was observed for the first time.
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