Construction of Recombinant Pichia pastoris Expressing Human SOD

VernacularTitle:表达人源SOD的毕赤酵母重组子的构建
Author: Xinming LI ¹ ; Ye SUN ; Chunling XIAO
Author Information

1. 沈阳医学院基础医学院病原生物学教研室
Keywords: Pichia pastoris; recombinant; human superoxide dismutase
From: Journal of China Medical University 2017;46(12):1117-1123
CountryChina
Language:Chinese
Abstract: Objective To optimize the expression of active human Cu,Zn-superoxide dismutase (SOD) in recombinant Pichia pastoris (P.pastoris).Methods A recombinant human SOD expression construct was generated and expressed in P.pastoris.The culture time,concentration of copper ions in the medium,and conditions for methanol induction were optimized,and SOD was preliminarily purified.Results Five P.pastoris recombinants that stably expressed human SOD were obtained.SOD could be purified from the culture supernarants of the recombinant strains by chromatography.The amount of SOD secreted into the supernatant was high with optimal activity,when the incubation time was 48 h,copper ion concentration in the medium was 0.5％,and methanol concentration was 1％.Conclusion An effective method for the expression of soluble human Cu,Zn-SOD in P.pastoris was established,and the enzyme produced using this method exhibited high activity.This method provides a foundation for further research on the production of human SOD by fermentation.