TITLE:
Survey of Intracellular Protein Extraction Methods from Pichia pastoris
AUTHORS:
Yuli Wang, Yuansen Hu, Yang Lei, Yangyong Lv, Le Wang, Hongwei Wei
KEYWORDS:
Pichia pastoris, High Temperature and Alkali Treatment, High Pressure Homogeniza-Tion, Cell Wall Breaking, Protein Extraction
JOURNAL NAME:
World Journal of Engineering and Technology,
Vol.3 No.4C,
December
17,
2015
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.