Transglycosylation activity of α-D-galactosidase from Trichoderma reesei -: An investigation of the active site

被引:40
作者
Eneyskaya, EV
Golubev, AM
Kachurin, AM
Savel'ev, AN
Neustroev, KN [1 ]
机构
[1] Petersburg Nucl Phys Inst, Mol & Radiat Biophys Div, St Petersburg 188350, Russia
[2] St Petersburg Tech Univ, Dept Biophys, St Petersburg 195251, Russia
基金
俄罗斯基础研究基金会;
关键词
alpha-D-galactosidase; Trichoderma reesei; transglycosylation products; alkyl galactosides; p-nitrophenyl alpha-D-galactopyranoside;
D O I
10.1016/S0008-6215(97)00229-2
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
The transglycosylation reaction catalyzed by alpha-D-galactosidase from the mycelial fungus Trichoderma reesei was studied using p-nitrophenyl alpha-D-galactopyranoside (PNPG). An aliphatic alcohol or the substrate itself can be an acceptor of the galactose residue in this reaction. The transglycosylation products were identified as alkyl galactosides in the case of alcohols or as galactobioside and galactotrioside in the case of PNPG. The transglycosylation rates follow a first-order equation with respect to the alcohol concentrations except for methanol. Affinities of some substrates were estimated from their K-i values in the reaction of the enzyme with PNPG. Transglycosylation of the substrate suggests a model for the enzyme active center. It is proposed that the active center includes two galactose-binding sites and a hydrophobic site. (C) 1998 Elsevier Science Ltd.
引用
收藏
页码:83 / 91
页数:9
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