Optimization of naringenin and p-coumaric acid hydroxylation using the native E. coli hydroxylase complex, HpaBC.
Flavonoids are a growing class of bioactive natural products with distinct and interesting bioactivity both in vitro and in vivo. The extraction of flavonoids from plant sources is limited by their low natural abundance and commonly results in a mixture of products that are difficult to separate. However, due to recent advances, the microbial production of plant natural products has developed as a promising alternative for flavonoid production. Through optimization of media, induction temperature, induction point, and substrate delay time, we demonstrate the highest conversion of naringenin to eriodictyol (62.7 ± 2.7 mg/L) to date, using the native E. coli hydroxylase complex, HpaBC. We also show the first evidence of in vivo HpaBC activity towards the monohydroxylated flavan-3-ol afzelechin with catechin product titers of 34.7 ± 1.5 mg/L. This work confirms the wide applicability of HpaBC towards realizing efficient de novo production of various orthohydroxylated flavonoids and flavonoid derived products in E. coli.
Dept. of Chemical and Biological Engineering, Rensselaer Polytechnic Inst., Troy, NY, 12180.
J Andrew
JA
Jones
Dept. of Chemical and Biological Engineering, Rensselaer Polytechnic Inst., Troy, NY, 12180.
Shannon M
SM
Collins
Dept. of Chemical and Biological Engineering, Rensselaer Polytechnic Inst., Troy, NY, 12180.
Victoria R
VR
Vernacchio
Dept. of Biological Sciences, Rensselaer Polytechnic Inst., Troy, NY, 12180.
Daniel M
DM
Lachance
Dept. of Chemical and Biological Engineering, Rensselaer Polytechnic Inst., Troy, NY, 12180.|Dept. of Biological Sciences, Rensselaer Polytechnic Inst., Troy, NY, 12180.
Mattheos A G
MA
Koffas
D002392:Catechin / Q000737:chemistry / Q000378:metabolism; D003373:Coumaric Acids / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D044950:Flavanones / Q000737:chemistry / Q000378:metabolism*; D005419:Flavonoids / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D006900:Hydroxylation; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000737:chemistry; D010636:Phenols / Q000737:chemistry / Q000378:metabolism; D011422:Propionates
D003373:Coumaric Acids; D044950:Flavanones; D005419:Flavonoids; D046912:Multiprotein Complexes; D010636:Phenols; D011422:Propionates; D002392:Catechin; D006899:Mixed Function Oxygenases; C026252:4-hydroxyphenylacetate 3-monooxygenase; C005273:naringenin; C495469:p-coumaric acid; C007619:eriodictyol; C510796:afzelechin
catechin; flavonoids; hydroxylation; non-P450 hydroxylase; pathway optimization