Cloning, sequencing and overexpression of a Sinorhizobium meliloti M5N1CS carboxymethyl-cellulase gene.
The EndS encoding sequence was isolated from Sinorhizobium meliloti M5N1CS DNA. Comparisons between the deduced amino acid sequence of the mature EndS (337 amino acids, molecular mass 36,418 Da, isoelectric point 4.92) and those of published beta-glycanases showed that this enzyme belongs to family 5 of the glycoside hydrolases. The protein was overproduced in Escherichia coli using a T7 expression system. When the purified overexpressed EndS protein was tested on cellulose-type components, the best substrate was CM-cellulose.
Michaud
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Laboratoire des Polysaccharides Microbiens et Végétaux, Université de Picardie Jules Verne, IUT d'Amiens, Génie Biologique, Avenue des Facultés, Le Bailly, 80 025 Amiens cedex, France. Philippe.Michaud@iut.u-picardie.fr
Belaich
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A
Courtois
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B
Courtois
J
J
D001483:Base Sequence; D002480:Cellulase; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000235:genetics*; D016962:Sinorhizobium meliloti / Q000201:enzymology / Q000235:genetics*
D017931:DNA Primers; D006026:Glycoside Hydrolases; D002480:Cellulase; C019826:carboxymethylcellulase