IDENTIFICATION OF A PUTATIVE ACTIVE-SITE RESIDUE IN THE EXO-BETA-(1,3)-GLUCANASE OF CANDIDA-ALBICANS

被引:24
作者
CHAMBERS, RS [1 ]
WALDEN, AR [1 ]
BROOKE, GS [1 ]
CUTFIELD, JF [1 ]
SULLIVAN, PA [1 ]
机构
[1] UNIV OTAGO,DEPT BIOCHEM,POB 56,DUNEDIN,NEW ZEALAND
关键词
SITE-DIRECTED MUTAGENESIS; EXO-BETA-(1,3)-GLUCANASE; CANDIDA-ALBICANS;
D O I
10.1016/0014-5793(93)81022-R
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Recombinant exo-beta-(1-3)-glucanase from Candida albicans was expressed in Saccharomyces cerevisiae and purified. The enzyme contains a number of short blocks of sequence homology with several genes for cellulases of the family A glycanases including the conserved sequence motif NEP which has previously been shown to be important in the catalytic function of several cellulases. Site directed mutagenesis of this glutamic acid residue in the 1,3 glucanase (E230D, E230Q) decreased the enzymatic activity 15,000- and 400-fold, respectively. This suggests that the E of the NEP participates in catalysis of the exoglucanase and other related glycanases.
引用
收藏
页码:366 / 369
页数:4
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