Research On Isolation Of Ophiopogon Polysaccharide With Enzymic Hydrolysis

This paper adopt a method of extraction of water and alcohol precipitation to abstract Ophiopogon polysaccharide,in. order to studying on how to improve the capability of biolog-ical enzymatic method to hydrolyze Ophiopogon polysaccharide,we conduct a study on the fermentation and enzyme reaction conditions as well as enzyme properties.The optimal conditions for the enzyme cultivation and the enzyme reaction were explored in:beginning with Absidia sp. O84s, inducing by20%soybean in the medium, and cultivating for6days to reaching the higher activity, fermentation temperature is30℃; the concentration substrate of the enzyme reaction is1%(10mg/mL),pH5.0,reaction temperature is40℃,reaction time is lOhours.The method of viscosity determines the different materials to the important results.It is that the average molecular weight of Ophiopogon polysaccharide is66836.5, its enzymic hydrolysates is46783.9after the enzyme fermentation,and its enzymic hydrolysates is39474.5after the enzyme reaction,the empirical constant is K=8.88×10-4.This paper also concerns the purification and reaction characters of this glycosidases from microorganism Absidia sp.084s, and M.W. of hydrolytic enzyme of Ophiopogon polysaccharide was73.1kDa. The optimum reaction characters of enzymes shows that optimum condition:pH5.0; the range of stabilization is pH4.0～6.0;the optimum temperature is40℃; the range of stabilization is20～40℃; five metalion of Ca2+、Na+、K+、Mg2+、Zn2+can’t effecte the enzyme activity, Cu2+、Fe can inhibit the enzyme activity;the Vmax is1.276mmol/L·h, the Km is0.1459mmol/L, theMichaelis-Mentonequation is1/V=0.11439/[S]+0.7837;the enzyme reaction characteristics of the hydrolytic enzyme of Ophiopogon polysaccharide show that it can hydrolysis the starch glucosidic bond in some extent and can’t hydrolysis the cellobiose’s and CMC’s β-glucosyl group.