Survey of Intracellular Protein Extraction Methods from Pichia pastoris

Yuli Wang,Yangyong Lv,Le Wang,Yuansen Hu,Yang Lei,Hongwei Wei

doi:10.4236/wjet.2015.34c001

Abstract

The broken efficiency of cell wall and the release amount of Pichia pastoris intracellular protein under different cell breaking conditions were investigated in this paper. The results showed that broken efficiency using hot alkali combined with high-pressure homogenizing method was higher than that of enzyme hydrolysis, hot alkali treatment and high-pressure homogenation, respectively. Suspended medium had little effect on the broken efficiency of yeast cell, but had significant effect on the protein release yield. The results indicated that optimal condition for intracellular proteins extraction was 30% (wet weight, w/v) of yeast cells suspend in 50 mM phosphate buffer (pH 10.0), water bathed at 60?C for 2 hours, homogenized twice at 100 MPa pressure. The broken efficiency of Pichia pastoris cell could reach 87.6% and the protein yield was 35.48 g per 100 g cells.

Highlights

The methylotrophic yeast Pichia pastoris has become one of the most widely used expression systems for recombinant protein production (Jia-kun Dai et al, 2012) [1]
Yeast cell breaking methods have been reported in many literatures, such as ultrasonic method, hot acid treatment, biological enzymatic treatment (Yan Li et al, 2008; Ding-wei Cui et al, 2010; Su-qing Zhang and Gui-Lan Gao, How to cite this paper: Wang, Y.L., Hu, Y.S., Lei, Y., Lv, Y.Y., Wang, L. and Wei, H.W. (2015) Survey of Intracellular Protein Extraction Methods from Pichia pastoris
The results indicated that maximum efficiency of cell breaking was about 20%, and the released protein yield was lower

Summary

Introduction

The methylotrophic yeast Pichia pastoris has become one of the most widely used expression systems for recombinant protein production (Jia-kun Dai et al, 2012) [1]. Pichia pastoris contains rich proteins, ribonucleic acid (RNA), polysaccharide and other nutrients. Extraction of these intracellular materials from yeast cells is significant for the maximum utilization of yeast resource. Yeast cell breaking methods have been reported in many literatures, such as ultrasonic method, hot acid treatment, biological enzymatic treatment (2015) Survey of Intracellular Protein Extraction Methods from Pichia pastoris.

Methods

Results

Conclusion