Cloning, Expression and Property Analysis of Arabinosidase in Pichia pastoris
- VernacularTitle:重组阿拉伯糖苷酶在毕赤酵母中的分泌表达及其活性分析
- Author:
Dong-Sheng YAO
;
Hui-Mei TAN
;
Hui HUANG
;
Da-Ling LIU
;
Chun-Fang XIE
;
- Publication Type:Journal Article
- Keywords:
Armillariella tabescens Arabinosidase Pichia pastoris Secrete expression
- From:
China Biotechnology
2006;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.
