Study On The Biotransformation Of Chlor(o)-coumarins By Suspension Transgenic Hairy Root Of Polygonum Multiflorum

Objective:The purpose of this study is to study on the biotransformation of exogenous compound by the way of organic synthesis and bioconversion with transgenic plant,and to study the condition of biotransformation,the yield of products and to assess activity of the products,which is bid fair to provide infallible data for industrial produce pharmaceutical compound.Methods:The substrates of 4-phenyl-6-chloro-7-hydroxyl-coumarin(CBC01)and 4-methyl-6-chloro-7-hydroxyl-coumarin (CC01) were obtained by Pechmann reaction and were bioconvered by hairy root of transgenic Polygonum multiflorum Thunb.The products were detected by TLC and HPLC and isolated and purified by the column chromatography, recrystallization, etc. Their structures were elucidated by1H-NMR, 13C-NMR and MS. The optimal co-culture times of the products were determined by HPLC. The bioactive evaluation was done by the experiment of clearance of DPPH free radical in vitro.Results:The structures of products were elucidated identified as 4-phenyl-6-chloro-coumarin-7-O-β-D-glucopyranoside(CBCP 1),4-methyl-6-chloro-coumarin-7-O-β-D-glucopyranoside (CCP1).The DPPH free radical clearance IC50 of CCP1 and CBCP1 were 262.84 ug/ml,182.80 ug/ml which were much lower than the substrates of CBC01and CC01(The CC01 IC5Owas 390.12 ug/ml and CBC01 was higher than 400 ug/ml). In the hairy roots of P. multiflorum containing CBC01 and CC01 (10 mg/F), the yield of CBCP1 in the cultures was 65.4%in the 7th day for co-culture and the CCP1 was up to 90%in 5th day. However,the secretion rates of P. multiflorum for CBCP1 and CCP1 were extraordinary low.