scholarly journals Site-Directed Mutagenesis and Expression of the Soluble Form of the Family IIIa Cellulose Binding Domain from the Cellulosomal Scaffolding Protein of Clostridium cellulovorans

2005 ◽  
Vol 187 (20) ◽  
pp. 7146-7149 ◽  
Author(s):  
Koichiro Murashima ◽  
Akihiko Kosugi ◽  
Roy H. Doi
Keyword(s):  
Binding Domain ◽  
Site Directed Mutagenesis ◽  
Scaffolding Protein ◽  
Soluble Form ◽  
Directed Mutagenesis ◽  
Cellulose Binding Domain ◽  
Binding Studies ◽  
Clostridium Cellulovorans ◽  
The Family ◽  
Cellulose Binding

ABSTRACT The planar and anchoring residues of the family IIIa cellulose binding domain (CBD) from the cellulosomal scaffolding protein of Clostridium cellulovorans were investigated by site-directed mutagenesis and cellulose binding studies. By fusion with maltose binding protein, the family IIIa recombinant wild-type and mutant CBDs from C. cellulovorans were expressed as soluble forms. Cellulose binding tests of the mutant CBDs indicated that the planar strip residues played a major role in cellulose binding and that the anchoring residues played only a minor role.

scholarly journals Characterization of EngF from Clostridium cellulovorans and Identification of a Novel Cellulose Binding Domain

1998 ◽  
Vol 64 (3) ◽  
pp. 1086-1090 ◽  
Author(s):  
Akihiko Ichi-ishi ◽  
Salah Sheweita ◽  
Roy H. Doi
Keyword(s):  
Amino Acids ◽  
Cellulose Degradation ◽  
Binding Activity ◽  
Binding Domain ◽  
Cellulose Binding Domain ◽  
Binding Studies ◽  
Endoglucanase Activity ◽  
Clostridium Cellulovorans ◽  
C Terminus ◽  
Cellulose Binding

ABSTRACT The physical and enzymatic properties of noncellulosomal endoglucanase F (EngF) from Clostridium cellulovorans were studied. Binding studies revealed that the Kd and the maximum amount of protein bound for acid-swollen cellulose were 1.8 μM and 7.1 μmol/g of cellulose, respectively. The presence of cellobiose but not glucose or maltose could dissociate EngF from cellulose. N- and C-terminally truncated enzymes showed that binding activity was located at some site between amino acid residues 356 and 557 and that enzyme activity was still present when 20 amino acids but not 45 amino acids were removed from the N terminus and when 32 amino acids were removed from the C terminus; when 57 amino acids were removed from the C terminus, all activity was lost. EngF showed low endoglucanase activity and could hydrolyze cellotetraose and cellopentaose but not cellotriose. Activity studies suggested that EngF plays a role as an endoglucanase during cellulose degradation. Comparative sequence analyses indicated strongly that the cellulose binding domain (CBD) is different from previously reported CBDs.

Binding Site Analysis of Cellulose Binding Domain CBDN1from Endoglucanse C ofCellulomonas fimiby Site-Directed Mutagenesis†

Biochemistry ◽  
2000 ◽  
Vol 39 (30) ◽  
pp. 8844-8852 ◽  
Author(s):  
Jeff Kormos ◽  
Philip E. Johnson ◽  
Emmanual Brun ◽  
Peter Tomme ◽  
Lawrence P. McIntosh ◽  
...  
Keyword(s):  
Binding Site ◽  
Binding Domain ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Cellulose Binding Domain ◽  
Site Analysis ◽  
Cellulose Binding

scholarly journals Characterization of the cellulose-binding domain of the Clostridium cellulovorans cellulose-binding protein A.

1993 ◽  
Vol 175 (18) ◽  
pp. 5762-5768 ◽  
Author(s):  
M A Goldstein ◽  
M Takagi ◽  
S Hashida ◽  
O Shoseyov ◽  
R H Doi ◽  
...  
Keyword(s):  
Binding Protein ◽  
Protein A ◽  
Binding Domain ◽  
Cellulose Binding Domain ◽  
Clostridium Cellulovorans ◽  
Cellulose Binding
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