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- Title
Heterologous expression and site-directed mutagenesis of endoglucanase CelA from Clostridium thermocellum.
- Authors
Qiang Yao; Tingting Sun; Guanjun Chen; Weifeng Liu
- Abstract
The endoglucanase CelA from Clostridium thermocellum was strongly expressed in Bacillus subtilis. The enzyme was purified by Ni2+-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 k cat value for Y215P and complete loss of activity for D152N.
- Subjects
CLOSTRIDIUM; MUTAGENESIS; BACILLUS subtilis; SUBSTITUTION reactions; CHROMATOGRAPHIC analysis
- Publication
Biotechnology Letters, 2007, Vol 29, Issue 8, p1243
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-007-9379-5