The cellulose-binding domain of cellobiohydrolase Cel7A from Trichoderma reesei is also a thermostabilizing domain

被引：31

作者：

Hall, Melanie ^{[1
]}

Rubin, Jonathan ^{[1
]}

Behrens, Sven H. ^{[1
]}

Bommarius, Andreas S. ^{[1
,2
]}

机构：

[1] Georgia Inst Technol, Sch Chem & Biomol Engn, Parker H Petit Inst Bioengn & Biosci, Atlanta, GA 30332 USA

[2] Georgia Inst Technol, Sch Chem & Biochem, Atlanta, GA 30332 USA

来源：

JOURNAL OF BIOTECHNOLOGY | 2011年 / 155卷 / 04期

关键词：

Cel7A; Cellulose-binding domain; Thermostability; Thermostabilizing domain; Dynamic light scattering; Melting point; DYNAMIC LIGHT-SCATTERING; FUNGAL CELLULASES; MUTAGENESIS; EXPRESSION; STABILITY; RESONANCE; IMPROVE; ENZYMES; PH;

D O I：

10.1016/j.jbiotec.2011.07.016

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The thermostability of cellobiohydrolase I Cel7A from Trichoderma reesei was investigated using dynamic light scattering. While the whole enzyme displayed a melting point of 59 degrees C, the catalytic domain obtained via papain-catalyzed proteolysis was shown to denature at 51 degrees C and the cellulose-binding domain (with linker attached) melted at 65-66 degrees C. This variation in individual melting temperatures is proposed to account for the full retention of binding capacity of Cel7A at 50 degrees C, along with a loss of catalytic activity observed for the catalytic domain alone. Thus, the cellulose-binding domain of Cel7A acts as a thermostabilizing domain for the enzyme. The effect of reducing agents on the protein melting behavior was also investigated. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：370 / 376

页数：7

共 40 条

[1] Thermostability Enhancement of Clostridium thermocellum Cellulosomal Endoglucanase Cel8A by a Single Glycine Substitution [J].

Anbar, Michael ;

Lamed, Raphael ;

Bayer, Edward A. .

CHEMCATCHEM, 2010, 2 (08) :997-1003

[2]

[Anonymous], 1991, Laser light scattering

[3] Modeling cellulase kinetics on lignocellulosic substrates [J].

Bansal, Prabuddha ;

Hall, Melanie ;

Realff, Matthew J. ;

Lee, Jay H. ;

Bommarius, Andreas S. .

BIOTECHNOLOGY ADVANCES, 2009, 27 (06) :833-848

[4]

Berne B.J., 1976, DYNAMIC LIGHT SCATTE

[5] The relationship between thermal stability and pH optimum studied with wild-type and mutant Trichoderma reesei cellobiohydrolase Cel7A [J].

Boer, H ;

Koivula, A .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2003, 270 (05) :841-848

[6] Improved biocatalysts by directed evolution and rational protein design [J].

Bornscheuer, UT ;

Pohl, M .

CURRENT OPINION IN CHEMICAL BIOLOGY, 2001, 5 (02) :137-143

[7] The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics [J].

Cantarel, Brandi L. ;

Coutinho, Pedro M. ;

Rancurel, Corinne ;

Bernard, Thomas ;

Lombard, Vincent ;

Henrissat, Bernard .

NUCLEIC ACIDS RESEARCH, 2009, 37 :D233-D238

[8]

Chodankar S., 2007, Journal of Physics: Condensed Matter, V19, P1

[9] STRUCTURES AND MECHANISMS OF GLYCOSYL HYDROLASES [J].

DAVIES, G ;

HENRISSAT, B .

STRUCTURE, 1995, 3 (09) :853-859

[10] THE 3-DIMENSIONAL CRYSTAL-STRUCTURE OF THE CATALYTIC CORE OF CELLOBIOHYDROLASE-I FROM TRICHODERMA-REESEI [J].

DIVNE, C ;

STAHLBERG, J ;

REINIKAINEN, T ;

RUOHONEN, L ;

PETTERSSON, G ;

KNOWLES, JKC ;

TEERI, TT ;

JONES, TA .

SCIENCE, 1994, 265 (5171) :524-528

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