Efficient production of (2S)-flavanones by Escherichia coli containing an artificial biosynthetic gene cluster.
For the fermentative production of plant-specific flavanones (naringenin, pinocembrin) by Escherichia coli, a plasmid was constructed which carried an artificial biosynthetic gene cluster, including PAL encoding a phenylalanine ammonia-lyase from a yeast, ScCCL encoding a cinnamate/coumarate:CoA ligase from the actinomycete Streptomyces coelicolor A3(2), CHS encoding a chalcone synthase from a licorice plant and CHI encoding a chalcone isomerase from the Pueraria plant. The recombinant E. coli cells produced (2S)-naringenin from tyrosine and (2S)-pinocembrin from phenylalanine. When the two subunit genes of acetyl-CoA carboxylase from Corynebacterium glutamicum were expressed under the control of the T7 promoter and the ribosome-binding sequence in the recombinant E. coli cells, the flavanone yields were greatly increased, probably because enhanced expression of acetyl-CoA carboxylase increased a pool of malonyl-CoA that was available for flavanone synthesis. Under cultural conditions where E. coli at a cell density of 50 g/l was incubated in the presence of 3 mM tyrosine or phenylalanine, the yields of naringenin and pinocembrin reached about 60 mg/l. The fermentative production of flavanones in E. coli is the first step in the construction of a library of 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. aa36248@mail.ecc.u-tokyo.ac.jp
Kaneko
Masafumi
M
Funa
Nobutaka
N
Kawasaki
Hisashi
H
Kojima
Hiroyuki
H
Ohnishi
Yasuo
Y
Horinouchi
Sueharu
S
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D044950:Flavanones / Q000096:biosynthesis* / Q000235:genetics; D017343:Genes, Plant / Q000502:physiology*; D005813:Genes, Synthetic; D005818:Genetic Engineering; D008316:Malonyl Coenzyme A / Q000378:metabolism
D044950:Flavanones; D008316:Malonyl Coenzyme A; C005273:naringenin