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The Research of Xylanase Hydrolysing Chitosan

Author: GaoWei
Tutor: DingWenPing
School: Wuhan Polytechnic University
Course: Food, oil and vegetable protein
Keywords: Chitosan Xylanase Enzymatic Properties Separation and purification
CLC: TQ28
Type: Master's thesis
Year: 2008
Downloads: 86
Quote: 1
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Abstract


Many domestic and foreign research reports indicate that it has been found that there are 37 kinds of enzymes capable of non-specificity of hydrolyzed chitosan, including cellulase, papain, hemicellulase, lipase, lysozyme, etc.. The xylanase is a hemicellulase, few studies of the hydrolysis chitosan. This thesis, to further clarify the role of the xylanase enzyme hydrolysis of chitosan mechanism has important academic significance, and has important applications for the enzymatic production of low molecular weight chitosan. Firstly xylanase enzymes and other four enzymatic hydrolysis of chitosan activity were compared, results show that: the highest activity of xylanase hydrolysis of chitosan. The best conditions of the enzyme hydrolysis of chitosan: pH4.75, temperature 60 ℃, 15% of the amount of the enzyme. The substrate concentration does not exceed 20g / L, the hydrolysis process of the xylanase of chitosan is consistent with Michaelis-Menten equation, where, Vmax = 15.3μmol/min, · L and Km = 3.5g / L,. This also shows that in the case of a sufficiently low concentration of the substrate, xylanase hydrolysis process of the chitosan is an enzymatic reaction of a single substrate, each enzyme molecule is only one binding site, the reaction to form a network only compounds. Determination of the molecular weight distribution of the hydrolyzate on the basis of understanding of the enzymatic kinetics using the Sepharose CL-6B gel chromatography, and other several enzymatic hydrolysis product of chitosan and its molecular weight distribution for comparison, the results show that: The molecular weight of the chitosan after the xylanase hydrolysis 6h after concentrated in about 8.6kDa; inactivated xylanase hydrolysis 6h chitosan molecular weight concentrated in the 18.4kDa;, Aspergillus niger cellulase liquid source hydrolysis after 0.5h chitosan molecular weight of 11.9kDa; Penicillium sources of cellulose hydrolysis of the enzyme powder after 0.5h chitosan molecular weight of 13.3kDa; the Aspergillus niger sources cellulose hydrolysis of the enzyme powder 0.5h post Chitosan The molecular weight of 12.6kDa. Finally, the commercial sources of xylanase solution purification. Purification process in the enzyme activity loss is small, the loss of xylanase activity 5200U, the chitosan activity loss 3000U. Using SDS-PAGE and HPLC analysis of the enzyme component. From the results, xylanase mainly by the following molecular weight component: 55 kDa, 34kDa. Specifically which one component of the hydrolysis of chitosan effect is unclear, but also the need for further research and discussion.

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