earticle

논문검색

Poster 20

Metabolic Engineering of E. coli for Regiospecific Modifications of Naringenin to produce Astragalin

초록

영어

We successfully produced astragalin (AST) from regiospecific modifications of naringenin (NRN) in Escherichia coli BL21(DE3). The exogenously supplied NRN was converted into dihydrokaempferol (DHK) and then kaempferol (KMF) in the presence of flavanone-3-hydroxylase (f3h) and flavonone synthase (fls1) from Arabidopsis thaliana, respectively. KMF was further modified to produce AST by 3-O-glucosylation utilizing the endogeneous UDP-glucose in presence of UGT78K1 from Glycine max. To increase the intracellular UDP-glucose concentration by channeling the carbon flux toward UDP-glucose at the branch point of glucose-6-phosphate (G6P), the chromosomal glucose phosphate isomerase (pgi) and D-glucose-6-phosphate dehydrogenase (zwf) were knocked-out in E. coli BL21(DE3). The two enzymes directly involved in the synthesis of UDP-glucose from G6P, phosphoglucomutase (nfa44530) from Nocardia farcinia and glucose-1-phosphate uridylyltransferase (galU) from E. coli K12 were overexpressed, which successfully diverted the carbon flow from glycolysis to the synthesis of UDP-glucose. Furthermore, to prevent the dissociation of UDP-glucose into UDP and glucose, the UDP-glucose hydrolase (ushA) was deleted. The E. coliΔpgiΔzwfΔushA mutant harboring the UDP-glucose biosynthetic pathway and the aforementioned genes for the regiospecific glucosylation produced 109.3 mg/L (244μM) of AST representing 48.8% conversion from 500 μM of NRN in 60 h without any supplementation of extracellular UDP-glucose.

저자정보

  • Ramesh Prasad Pandey Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University, #100, Kalsan-ri, Tangjeon-myeon, Asan-si, Chungnam 336-708, South Korea
  • Sailesh Malla 2Laboratory of Molecular Biotechnology and Biomaterials, School of Chemical and Biological Engineering, Seoul National University, Seoul, South Korea, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark
  • Byung-Gee Kim Laboratory of Molecular Biotechnology and Biomaterials, School of Chemical and Biological Engineering, Seoul National University, Seoul, South Korea
  • Kyung Sohng Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University, #100, Kalsan-ri, Tangjeon-myeon, Asan-si, Chungnam 336-708, South Korea

참고문헌

자료제공 : 네이버학술정보

    함께 이용한 논문

      ※ 원문제공기관과의 협약기간이 종료되어 열람이 제한될 수 있습니다.

      0개의 논문이 장바구니에 담겼습니다.