Purification and Biochemical Characterization of an endo-beta -1,3-Glucanase from Paenibacillus sp.

• Main Authors: Li-chi Huang, 黃欐淇
• Other Authors: none
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/41117803066718689654

碩士 === 大仁科技大學 === 生物科技研究所 === 96 === This research begins by screening the environment for native bacterial strains in the soil by testing for β-1,3-glucanase and chitinase activity. A colony with the clearest halo was picked up for further study. Identification of the strain was by way of the 16S rDNA. When this strain is cultured at 25℃contains insoluble beta-glucan pachyman the best thallus output and the enzyme specific activity was obtained. After collection and concentration of the enzyme was applied to hydrophobic interaction chromatography column for purification. The purified enzyme appeared homogeneously on gel of SDS-PAGE, and its apparent molecular mass was approximately 70 kDa. The enzyme exhibited an optimum activity at pH 8.0 and 50℃. The half-live of the enzyme was ~173 min at 50℃. Laminarin (a soluble beta-1,3-glucan) was the most favorable substrate followed by insoluble beta-1,3-glucans (curdlan, zymosan A, and pachyman) which had hydrolysis rates 8.5~29% of that of laminarin. The enzyme is an endo-beta-1,3-glucanase that was demonstrated by thin layer chromatography method. When this enzyme is applied at 2 μg/ml to Alternaria brassicicola could be reached 100% of the inhibitory effect. But the enzyme concentration in the 50 μg/ml inhibit the germination of spores of Botrytis cinerea and Pestalotiopsis eugeniae have the same effect.