Heterologous expression and site-directed mutagenesis of endoglucanase CelA from Clostridium thermocellum.

doi:10.1007/s10529-007-9379-5

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Article

Heterologous expression and site-directed mutagenesis of endoglucanase CelA from Clostridium thermocellum.

Biotechnology letters - 1 Aug 2007

Yao Qiang, Sun Tingting, Chen Guanjun, Liu Weifeng

Abstract excerpt

The endoglucanase CelA from Clostridium thermocellum was strongly expressed in Bacillus subtilis. The enzyme was purified by Ni(2+)-affinity chromatography. Site-directed substitution of D278 with an asparagine or an alanine residue surprisingly did not decrease the apparent k(cat) value. Further substitutions of two other potentially critical residues, Y215 and D152, resulted in a 2-fold decrease in apparent...

Topics

Biotechnology
Catalytic Domain
Cellulase
Cellulose
Clostridium thermocellum
Escherichia coli
Gene Expression Regulation, Bacterial
Hydrogen-Ion Concentration
Kinetics
Models, Biological
Mutagenesis, Site-Directed
Mutation
Nickel

Source

Open in PubMed

Identifiers

PMID: 17520178
DOI: 10.1007/s10529-007-9379-5

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