Eukaryotic Expression and Purification Technology of Glucosidase in Pichia Pastoris

Abstract

Glucosidase is one of the most important hydrolases in the carbohydrate metabolism pathway in human body. It is noteworthy that glucosidase was raising high attention in numerous researches, attributed to its functional values and biological activities. The expression, secretion, extraction, purification, and quantitative analysis of glucosidase occupies an important position in the research of glucosidase. The Pichia pastoris expression system has been developed for 50 years. With its unique advantages, the expression system has successfully expressed hundreds of heterologous proteins. This article briefly describes the process of high-efficiency expression, extraction and purification of glucosidase in the P. pastoris expression system, focusing on the expression, separation and purification of glucosidase in P. pastoris, as well as the quantitative and activity analysis methods of glucosidase. Affinity chromatography is used for the separation and purification of crude enzyme solution, and UV–visible spectroscopy can be used for enzyme quantification and activity analysis.

Keywords

• Pichia pastoris
• glucosidase
• eukaryotic expression
• protein purification
• p-NPG