Stepwise Synthesis of Quercetin Bisglycosides Using Engineered Escherichia coli
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
Synthesis of flavonoid glycoside is difficult due to diverse hydroxy groups in flavonoids and sugars. As such, enzymatic synthesis or biotransformation is an approach to solve this problem. In this report, we used stepwise biotransformation to synthesize two quercetin bisglycosides (quercetin 3-O-glucuronic acid 7-O-rhamnoside [Q-GR] and quercetin 3-Oarabinose 7-O-rhamnoside [Q-AR]) because quercetin O-rhamnosides contain antiviral activity. Two sequential enzymatic reactions were required to synthesize these flavonoid glycosides. We first synthesized quercetin 3-O-glucuronic acid [Q-G], and quercetin 3-Oarabinose [Q-A] from quercetin using E. coli harboring specific uridine diphopsphate glycosyltransferase (UGT) and genes for UDP-glucuronic acid and UDP-arabinose, respectively. With each quercetin 3-O-glycoside, rhamnosylation using E. coli harboring UGT and the gene for UDP-rhamnose was conducted. This approach resulted in the production of 44.8 mg/l Q-GR and 45.1 mg/l Q-AR. This stepwise synthesis could be applicable to synthesize various natural product derivatives in case that the final yield of product was low due to the multistep reaction in one cell or when sequential synthesis is necessary in order to reduce the synthesis of byproducts.
Abstract
Synthesis of flavonoid glycoside is difficult due to diverse hydroxy groups in flavonoids and sugars. As such, enzymatic synthesis or biotransformation is an approach to solve this problem. In this report, we used stepwise biotransformation to synthesize two quercetin bisglycosides (quercetin 3-O-glucuronic acid 7-O-rhamnoside [Q-GR] and quercetin 3-Oarabinose 7-O-rhamnoside [Q-AR]) because quercetin O-rhamnosides contain antiviral activity. Two sequential enzymatic reactions were required to synthesize these flavonoid glycosides. We first synthesized quercetin 3-O-glucuronic acid [Q-G], and quercetin 3-Oarabinose [Q-A] from quercetin using E. coli harboring specific uridine diphopsphate glycosyltransferase (UGT) and genes for UDP-glucuronic acid and UDP-arabinose, respectively. With each quercetin 3-O-glycoside, rhamnosylation using E. coli harboring UGT and the gene for UDP-rhamnose was conducted. This approach resulted in the production of 44.8 mg/l Q-GR and 45.1 mg/l Q-AR. This stepwise synthesis could be applicable to synthesize various natural product derivatives in case that the final yield of product was low due to the multistep reaction in one cell or when sequential synthesis is necessary in order to reduce the synthesis of byproducts.
참고문헌 (20)
- Jones P, "Glycosyltransferases in secondary plant metabolism: tranquilizers and stimulant controllers", Planta, 213, pp.164~174 (2001)
- Osmani SA, "Substrate specif icity of plant UDP-dependent glycosyltransferase predicted from crystal structures and homology modeling", Phytochemitsry, 70, pp.325~347 (2009)
- Panche AN, "Flavonoids: an overview", J. Nutr. Sci, 5, pp.e47~ (2016)
- Veitch NC, "Flavonoids and their glycosides, including anthocyanins", Nat. Prod. Rep, 28, pp.1626~1695 (2011)
- Kim BG, "Biosynthesis and production of glycosylated flavonoids in Escherichia coli: current state and perspectives", Appl. Microbiol. Biotechnol, 99, pp.2979~2988 (2015)
- 한송이, "Molecular cloning and characterization of a flavonoid glucosyltransferase from Byungkyool (Citrus platymamma hort. ex Tanaka)", Applied Biological Chemistry, 60(1), pp.49~55 (2017), 한국응용생명화학회
- Bowles D, "Glycosyltransferases: managers of small molecules", Curr. Opin. Plant Biol, 8, pp.254~263 (2005)
- 조아라, "Biotransformation of quercetin to quercetin 3-O-gentiobioside using engineered Escherichia coli", Applied Biological Chemistry, 59(5), pp.689~693 (2016), 한국응용생명화학회
- Pabst M, "Nucleotide and nucleotide sugar analysis by liquid chromatography-electrospray ionization-mass spectrometry on surface-conditioned porous graphitic carbon", Anal. Chem, 82, pp.9782~9788 (2010)
- Han SH, "Synthesis of flavonoid O-pentosides by Escherichia coli through engineering nucleotide sugar synthesis pathway and glycosyltransferase", Appl. Env. Microbiol, 80, pp.2754~2762 (2014)
- Kim BG, "Production of bioactive flavonol rhamnosides by expression of plant genes in Escherichia coli", J. Agr. Food Chem, 60, pp.11143~11148 (2012)
- Choi HJ, "Antiviral activity of quercetin 7-rhamnoside against porcine epidemic diarrhea virus", Antiviral Res, 81, pp.77~81 (2009)
- Song JH, "Quercetin 7-rhamnoside reduces porcine epidemic diarrhea virus replication via independent pathway of viral induced reactive oxygen species", Virol. J, 8, pp.460~ (2011)
- Kim SY, "Metabolic engineering of Escherichia coli for the biosynthesis of flavonoid O-glucuronides and flavonoid O-galactoside", Appl. Microbiol. Biotechnol, 99, pp.2233~2242 (2015)
- Kim HJ, "Regioselective synthesis of flavonoid bisglycosides using Escherichia coli harboring two glycosyltransferases", Appl. Microbiol. Biotechnol, 97, pp.5275~5282 (2013)
- Sim GY, "Bacterial synthesis of N-hydroxycinnamoyl phenethylamines and tyramines", Microb. Cell Fact, 14, pp.162~ (2015)
- Urgaonkar S, "Synthesis of kaempferitrin", J. Org. Chem, 72, pp.4582~4585 (2007)
- Yang Y, "Efficient synthesis of kaempferol 3,7-O-bisglycosides via successive glycosylation with glycosyl ortho-alkynylbenzoates and trifluoroacetimidates", Tetrahedron Lett, 53, pp.2773~2776 (2012)
- Jones JA, "Complete biosynthesis of anthocyanins using E. coli polycultures", MBio, 8, pp.e00621-17~ (2017)
- Kim B-G, "Biological synthesis of quercetin 3-O-N-acetylglucosamine conjugate using engineered Escherichia coli expressing UGT78D2", Appl. Microbiol. Biotechnol, 93, pp.2447~2453 (2012)