Research On The Separation And Purification Of Snail's Xylanase

The xylanase is a kind of complex compound of enzyme system, which was hydrolyzed by xylan into the wooden two sugars, the wooden widowed sugar, a few pectin sugars and xylose. It was widely existed in the nature of bacterium, fungus and mollusk, moreover, it had the widespread application prospect in the papermaking, feed, food, manufacture of drugs and so on industries. As the huge latent application value of xylanase, it has become the hot spot of domestic and foreign research, and has made much important progress.This paper took the xylanase in snail's liver as the object of study, and has studied the extraction, the separation, the purification, the appraisal and the partial physics and chemistry of the snail liver xylanase systematically,which provided the research foundation for analyzing and determining the protein sequence of snail's xylanase, moreover the regulation and expression of gene list.The liver of snail after getting rid of membrane, grinding, rupturing cell and degreasing, then carried on the extraction and centrifugation with the buffer solution of sodium hydrogen phosphate - potassium dihydrogen phosphate of 1/15 mol/L(pH = 5.29) under the condition of 4℃, and obtained the coarse enzyme fluid of xylanase in snail's liver, which was salted out with the saturated ammonium sulfate solution by Stage separation at 0℃. Sephadex G-50 Desalinization; DEAE- Sephadex G-150 ionic exchange chromatographic analysis separation; Sephadex G-150 molecular sieve chromatographic analysis purification; Polyacrylamide gelatin preparation electrophoresis. Prepares the snail liver xylanase by electrophoresis . After partial physics and chemistry nature determination, this enzyme relative molecular weight approximately 34.4Kda; Most suitable reaction temperature for 55℃; Most suitable pH is 5.29; The most suitable substrate is the xylan, and has partly enzymolysis activeness toβ- mannan, does not have the enzymolysis function to carboxymethylcellulose nearly; When determination snail xylanase enzymolysis activeness, the enzyme reaction time not suitable to be surpasses for 1.5 hours, substrate concentration not suitable oversized or too small, and take 0.65% xylan as suitable.This research has designed separation-purification technology route and the plan, can Can satisfy the request well for the separation and the purification. By the electrophoresis preparation's snail xylanase, achieves the request to determine sequence and to prepare the monoclonal antibody.