Preliminary Research On Glycosylations Of Active Coumarins By Hairy Roots Of Polygonum Multiflorum Thunb

Objective:The paper studied on glycosylations of three active coumarins by hairy roots of Polygonum multiflorum Thunb for getting three water-soluble coumarin glycosides; then the enzymes of hairy root of P. Multiflorum Thunb were investigated by the way of biotransformation. Method:6-hydroxy-4-methylcoumarin (Al) daphnetin (A2),3-acetyl-5-hydroxymethyl-7-hydroxycoumarin (A3) 4-methyl-7-allyloxy coumarin (A4) were added into the media of hairy root of P. multiflorum Thunb then co-cultured for 7 d. The biotransfromed products were isolated by various chromatographic methods. The influences of co-cultured time on conversion ratio were investigated using HPLC. Results:Biotransformed product of A1,4-methylcoumarin-6-O-β-D-glucoside (A1P) was obtained; Biotransformed product of A2, daphnetin-8-O-β-D-glucoside (A2P) was obtained; Biotransformed product of A3,3-acetyl-5-hydroxymethyl-7-hydroxycoumarin-13-O-β-D-glucoside was obtained; A4 was Biotransformed successfully, two biotransformed products, 4-methyl-7-hydroxy coumarin (A4P1) and 4-allyl-coumarin-7-O-β-D-glucoside (A4P2) were obtained. The optimal co-cultured time of A1 in supension hairy root of P. Multiflorum thunb was 48 h with the highest biotransformation molar ratio of 76.3%, the sucrose medium (sucrose-only) can increase the biotransformation molar ratio of A2 to 72.44%, the optimal co-cultured time of A3 was 24 h with the highest biotransformation molar ratio of 58.4%. Conclusion:Three coumarins can be glycosylation by hairy root of P. Multiflorum Thunb; hairy roots of P. multiflorum Thunb contains hydrolytic enzymes.