Enzymatic Hydrolysis Of Polysaccharide From Ophiopogon Japonicus And Preparation Of Its Products

In this paper, polysaccharide OPF-1 was purified from Ophiopogon japonicus, and the primary structure of OPF-1 was studied, based on monosaccharide composition analysis of OPF-1. Finally, the hydrolysis of OPF-1 by bio-enzyme and its products were analyzed.Crud polysaccharide extracted from Ophiopogon japonicus with water was precipitated by ethanol. The precipitated polysaccharide was deproteinized by method of Sevage. Then a single component of OPF-1 with a homogenous molecular weight ascertained by TLC, gelcolumn chromatography and ultraviolet spectroscopy, was further separated and purified from polysaccharide using Sephadex G-100.To study the Monosaccharide composition of OPF-1, the complete acid hydrolysates of OPF-1 was analyzed by HPLC. The results showed that OPF-1 was mainly composed of glucose and fructose. Respectively, the concentration of fructose and glucose in the 20 uL sample injected into HPLC were 1.671 mg/mL and 0.116 mg/mL by standard curve method. Hence the molar ratio of fructose and glucose in OPF-1 was 16:1.Based on analysis of the products from Malaprade reaction and Smith degradation of OPF-1 by GC and IR, the structure of OPF-1 was studied. OPF-1, whose relative molecular mass was about 48kDa, was mainly polymerized by furan fructose through glycosides of 2?1 or 2?6. The small amount of glucose in OPF-1 may be linked to the non-reducing terminate.Using Sephadex G-100 and Sephadex G-50, OP-1, OP-2 and OP-3 were obtained from hydrolyastes of OPF-1 by the enzyme from Absidia sp.O8s, and their relative molecular weights were approximately determined as 32 kDa?9 kDa?1 kDa, respectively. From analysis of HPLC, OP-1?OP-2?OP-3 consisted of fructose?fructose?glucose and fructose respectively. The results showed that the macromolecules OPF-1 could be transformed into smaller ones by the enzyme from Absidia sp.O8s, whose relative molecular weights was below 10000.