Rhodobacter Sphaeroides Biotransformation Of The Mistletoe Fingerprinting And Chemical Composition Of The Study

Objective To study the activities in vitro and HPLC fingerprints of different original places of mistletoe biotransformated by Rhodobacter sphaeroides, photosynthetic bacteria (PSB). To separate and identify the chemical components from the n-butanol ether extraction part of phytosynthetic bacteria transforming Viscum coloratum (Kom.) Nakai (PSBT). To study bioactivities of the chemical constitiuents.Methods HPLC was used for establishing fingerprints of PSBT from different original places which were compared with those not transformated. PSBT's anti-tumor activity was determined using MTT method.The efficiency of antioxidance of PSBT was determined by the DPPH·assay. The chemical components of PSBT were separated and purified through sillcal gel, Sephadex LH-20 and HPLC, which were characterized by chemical reaction, UV,1H-NMR and 13C-NMR, along with comparing the data with those reported in literatures. Antioxidant acivities of the separated compounds were tested. The interactions of homoeriodictyol-7-O-β-D-apiosyl-(1â†'2)-β-D-glucopyranoside (HAG) and serum albumin were studied under the simulative human physiological condition by ultraviolet spectroscopy, fluorescence spectrophotometry and synchronous fluorescence spectroscopy. The binding parameters of HAG and SA were obtained.Results HPLC fingerprints were established, three original places of the Mistletoe transformated by PSB slightly increased the anti-tumor activity compared with Misletoe, The chemical constituents of PSBT had a strong scavenging effect on DPPH free radical. Chemical constituents changed, mistletoe from Dongbei changed most evidently. Eight compounds were isolated and identified as follows:rhamnazin-3-O-β-D-apiosyl-(1â†'2)-β-D-[6"-(3- hydroxy -3-methyglutaric methyl ester)-]-glucoside(â… ), rhamnazin-3-O-β-D-apiosyl-(1â†'2)-β-D-[6"-(3-hydroxy-3-methyglutarate)-]-glucoside(â…¡), isorhamnetin-3-O-β-D-glucoside(â…¢), quercetin-3-O-β-D-glucoside(â…£), homoeriodictyol(â…¤), homoeriodictyol-7-O-β-D-glucoside(â…¥), homoerio-dictyol-7-O-β-D-apiosyl-(1â†'2)-β-D-glucoside(â…¦) and homoeriodictyol-7-O-β-D-apiosyl-(1â†'5)-β-D-apiosyl-(1â†'2)-β-D-glucoside(â…§). Compoundâ… was a new compound, compoundâ…£hasn't been obtained from mistletoe yet,compoundâ… andâ…£were speculated as different components transformated by PSB. Compoundâ… -â…§have strong antioxidant activities. The results of ultraviolet and fluorescence spectrometry demonstrated that the endogenous fluorescence of BSA and HSA were significantly quenched by HAG and the main mechanism of fluorescence quenching was static quenching. The binding parameters of HAG and SA were as follows:for BSA and HSA, the binding constants K were 3.03×104 L·mol-1 and 6.22×104 L·moL-1, the binding sites n were 0.858 and 0.911, the binding distances r were 2.88 nm and 3.09 nm, respectively, and the interaction was mainly driven by H-bond and Van der Waals. It was showed by synchronal fluorescence technique that HAG had effect on the conformation of BSA and HSA.Conclusion:HPLC fingerprints of PSBT and mistletoe were established. The study provided a theoretical basis for the development of modern biological agents PSBT, offered reference data for its quality control and material basis research of pharmacy efficacy. The chemical constituents and bioacitivities of PSBT were studied. New compound contributes to the study of PSBT's transformation mechanism.