TY - JOUR
T1 - Investigating E. coli Coculture for Resveratrol Production with 13C Metabolic Flux Analysis
AU - Hong, Jaeseung
AU - Im, Dae Kyun
AU - Oh, Min Kyu
N1 - Funding Information:
This study was supported by the National Research Foundation of Korea funded by the Korean Government (2012M1A2A2026560 and 2017R1A2B4008758).
Publisher Copyright:
Copyright © 2020 American Chemical Society.
PY - 2020/3/18
Y1 - 2020/3/18
N2 - Resveratrol, a phytoalexin produced by plants, has several beneficial effects in humans. It can be produced using Escherichia coli by introducing only three heterologous genes: TAL, 4CL, and STS. However, the resveratrol synthesis pathway requires two precursors, tyrosine and acetyl-CoA, which are produced by two branched central metabolic pathways. Therefore, overexpression of these genes in E. coli results in the production of only trace amounts of resveratrol. In this study, we attempted to produce resveratrol via coculture of two engineered strains in which the two metabolic pathways are activated. The first strain was engineered to produce p-coumaric acid using tyrosine as a precursor, which can be synthesized by the pentose phosphate pathway. The second strain produced resveratrol by combining p-coumaric acid from the first strain and malonyl-CoA synthesized from acetyl-CoA, which is produced by the glycolytic pathway. In total, 55.7 mg/L of resveratrol was produced from 20 g/L of glucose via coculture of these two strains in glucose minimal medium without any supplements. The metabolic fluxes in each of the strains producing resveratrol were successfully investigated by 13C metabolic flux analysis. The results showed that the balance between the citric acid cycle and the malonyl-CoA supply node was important for resveratrol production.
AB - Resveratrol, a phytoalexin produced by plants, has several beneficial effects in humans. It can be produced using Escherichia coli by introducing only three heterologous genes: TAL, 4CL, and STS. However, the resveratrol synthesis pathway requires two precursors, tyrosine and acetyl-CoA, which are produced by two branched central metabolic pathways. Therefore, overexpression of these genes in E. coli results in the production of only trace amounts of resveratrol. In this study, we attempted to produce resveratrol via coculture of two engineered strains in which the two metabolic pathways are activated. The first strain was engineered to produce p-coumaric acid using tyrosine as a precursor, which can be synthesized by the pentose phosphate pathway. The second strain produced resveratrol by combining p-coumaric acid from the first strain and malonyl-CoA synthesized from acetyl-CoA, which is produced by the glycolytic pathway. In total, 55.7 mg/L of resveratrol was produced from 20 g/L of glucose via coculture of these two strains in glucose minimal medium without any supplements. The metabolic fluxes in each of the strains producing resveratrol were successfully investigated by 13C metabolic flux analysis. The results showed that the balance between the citric acid cycle and the malonyl-CoA supply node was important for resveratrol production.
KW - C-MFA
KW - E. coli
KW - coculture
KW - metabolic engineering
KW - resveratrol
UR - http://www.scopus.com/inward/record.url?scp=85082095420&partnerID=8YFLogxK
U2 - 10.1021/acs.jafc.9b07628
DO - 10.1021/acs.jafc.9b07628
M3 - Article
C2 - 32079399
AN - SCOPUS:85082095420
SN - 0021-8561
VL - 68
SP - 3466
EP - 3473
JO - Journal of agricultural and food chemistry
JF - Journal of agricultural and food chemistry
IS - 11
ER -