Combinatorial biosynthesis of flavones and flavonols in Escherichia coli.
(2S)-Flavanones (naringenin and pinocembrin) are key intermediates in the flavonoid biosynthetic pathway in plants. Recombinant Escherichia coli cells containing four genes for a phenylalanine ammonia-lyase, cinnamate/coumarate:CoA ligase, chalcone synthase, and chalcone isomerase, in addition to the acetyl-CoA carboxylase, have been established for efficient production of (2S)-naringenin from tyrosine and (2S)-pinocembrin from phenylalanine. Further introduction of the flavone synthase I gene from Petroselinum crispum under the control of the T7 promoter and the synthetic ribosome-binding sequence in pACYCDuet-1 caused the E. coli cells to produce flavones: apigenin (13 mg/l) from tyrosine and chrysin (9.4 mg/l) from phenylalanine. Introduction into the E. coli cells of the flavanone 3beta-hydroxylase and flavonol synthase genes from the plant Citrus species led to production of flavonols: kaempferol (15.1 mg/l) from tyrosine and galangin (1.1 mg/l) from phenylalanine. The combinatorial biosynthesis of the flavones and flavonols in E. coli is promising for the construction of a library of various flavonoid compounds and un-natural flavonoids in bacteria.
Miyahisa
Ikuo
I
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan.
Funa
Nobutaka
N
Ohnishi
Yasuo
Y
Martens
Stefan
S
Moriguchi
Takaya
T
Horinouchi
Sueharu
S
D002957:Citrus / Q000201:enzymology / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D047309:Flavones / Q000096:biosynthesis* / Q000737:chemistry; D044948:Flavonols / Q000096:biosynthesis* / Q000737:chemistry; D006899:Mixed Function Oxygenases / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics; D028528:Petroselinum / Q000201:enzymology / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D015202:Protein Engineering
D047309:Flavones; D044948:Flavonols; D010940:Plant Proteins; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; C065998:flavone synthase I; C048797:flavanone 3-dioxygenase; C085313:flavonol synthase