Preliminary Study On Immobilization For Endo-Chitosanase And Its Properties

Immobilized enzyme technique is an important component of modern biotechnology.It has applied to broad fields,including iatrology,food,environment and analytical purposes,it is a research focus biological science.,The high active endo-type chitosanase is self-made in our laboratory,In this paper the condition and properties are be researched when the enzyme was immobilized.Objective: according to experiments,to choose the optimum carrier and method to made the immobilized endo-type chitosanase suitable for practical production.And to comparative study the properties of immobilized enzyme and free enzyme.Methods:1 The silica gel carrierprepare with high amino group content was prepared By modified method,to used in Immobilizing endo- chitosanase2 Through Analyzing the property of three carrier ---the aminated silica gel,aminated resin,and epoxided agarose sphere, we obtained the content of functional group in the unit mass carrier.3 We applyed the method of glutaric dialdehyde,method of cyanuric chloride,method of bromoacetic acid,method of carbodiimide and method of epoxidation to immobilization endo-chitosanase, and through comparing the recovery of protein \the recovery of enzyme activity\relative enzyme activity ,we obtained the best carrier and method to immobilize enzyme.4 We optimized the best of immobilization method above. optimization content was including the concentration \the best cross linking time\cross linking temperature \pH value and the concentration of cross linking agent.5 We study on the properties of immobilized endo- chitosanase,and comparison with free endo-type chitosanase.Results: Through checking the recovery of enzyme activity \ the recovery of protein\ stability , the endo-type chitosanase adsorbed on chitosan and cross-linked with EDC.HCl. The optimal parameters included: endo-type chitosanase dosage of 4.86IU/mL;0.06 mol/mL EDC.HCl solution ;under 8C, immobilization for 3h, The optimal pH was 6.5.The result of immobilized endo- chitosanase activity and ratio of recover erable activity were 0.87U/g(wet weight)and 58.43% respectively. Conclusion: The endo chitosanase adsorbed on chitosan and cross-linked with EDC.HCl, can be used to produce Low dispersity Chitosan oligosaccharide .but the immobilized enzyme and useful time remains to be further enhanced...