Expression of Flammeovirga agarivorans Alginate Lyase in Pichia pastoris
Putative alginate lyase encoding gene of Flammeovirga agarivorans was optimized,synthesized and secretory expressed in Pichia pastoris.The protein concentration of the expressed product reached 0.23 mg/mL.The optimum pH and temperature of the expressed alginate lyase was 9.0 and 40 ℃,respectively,and enzymatic activity reached 400.00 U/mL under these reaction conditions.The enzyme was thermostable at no more than 40 ℃.Substrate specific analysis showed that the enzyme could hydrolyze both poly-mannuronic acid(polyM)and poly-guluronic acid(polyG),but the hydrolysis ability of polyG was stronger.Sodium alginate was hydrolyzed by this enzyme and the products were analyzed by high performance liquid chromatography and mass spectrometry.The results showed that the main products were alginate oligosaccharides with polymerization degree from 2 to 4.The alginate lyase obtained in this study can be used for the enzymatic scale preparation of alginate oligosaccharides.
万方数据
维普
