华侨大学学报（自然科学版）2016，Vol.37Issue(2)：224-229,6.DOI:10.11830/ISSN.1000-5013.2016.02.0224
阿魏酸酯酶 O42807在毕赤酵母 GS115中的表达
Expression of Feruloyl Esterase O42807 in Pichia pastoris GS115
摘要
Abstract
This paper studied the expression of the feruloyl estrase O42807 in Pichia pastoris GS115 and the enzymatic properties of the recombinant feruloyl estrase.Based on the amino acids of feruloyl estrase O42807,the gene (fae)of fer-uloyl estrase was synthesized chemically.The secreted expression vector pPIC9K-fae was constructed by the ligation of the fae gene into the shuttle vector pPIC9K.The plasmid pPIC9K-fae was linearized and then electrotransformed into Pichia pastoris GS115.Afterwards,the recombinant strain with high level of feruloyl estrase activity was obtained through activity screening.The SDS-PAGE result showed a single band in the fermentation supernatant.The molecular weight of the recombinant feruloyl estrase was about 42 ku and the enzyme activity was 78.49 nkat·mL-1 .The specific activity of the recombinant feruloyl estrase was 524.38 nkat·mg-1 .The results also showed that the optimal reaction temperature was 50 ℃,and stable from 40 ℃ to 45 ℃.And the optimal pH was from 5.0 to 5.5,stable at pH 6.0.Fur-thermore,the enzymatic activity was slightly enhanced by K+ ,Ca2 + ,Na+ ,whereas it was slightly inhibited by Fe2 + , Zn2 + ,and strongly inhibited by Mn2 + ,Cu2 + .关键词
阿魏酸酯酶/表达/毕赤酵母/酶学性质Key words
feruloyl esterase/expression/Pichia pastoris/enzymatic properties分类
生物科学引用本文复制引用
陈云华,李慧,张光亚,葛慧华,李夏兰..阿魏酸酯酶 O42807在毕赤酵母 GS115中的表达[J].华侨大学学报（自然科学版）,2016,37(2):224-229,6.基金项目
福建省厦门市科技计划项目（2014S006）；福建省发改委投资基金资助项目 （）
