Cloning, nucleotide sequence, and expression of the Clostridium thermocellum cellodextrin phosphorylase gene and its application to synthesis of cellulase inhibitors

The cellodextrin phosphorylase (CDP) gene of Clostridium thermocellum was cloned and sequenced. The nucleotide sequence of the insert of a positive clone contained an open reading frame of 2940 bp encoding a polypeptide of 980 amino acid residues with a calculated molecular mass of 111,182 daltons....

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Bibliographic Details
Published in:Journal of fermentation and bioengineering 1998-01, Vol.85 (2), p.144-149
Main Authors: Kawaguchi, Takashi, Ikeuchi, Yasuo, Tsutsumi, Noriko, Kan, Akihiko, Sumitani, Jun-Ichi, Arai, Motoo
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biology of microorganisms of confirmed or potential industrial interest
Biotechnology
cellodextrin phosphorylase
cellulase inhibitor
CLONACION MOLECULAR
CLONAGE MOLECULAIRE
cloning
CLOSTRIDIUM THERMOCELLUM
enzymatic synthesis
Escherichia coli
EXPRESION GENICA
expression
EXPRESSION DES GENES
FOSFOPIROLASA
Fundamental and applied biological sciences. Psychology
GENE EXPRESSION
Genetics
Mission oriented research
MOLECULAR CLONING
NUCLEOTIDE SEQUENCE
PHOSPHORYLASE
SECUENCIA NUCLEOTIDICA
SEQUENCE NUCLEOTIDIQUE
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Summary:The cellodextrin phosphorylase (CDP) gene of Clostridium thermocellum was cloned and sequenced. The nucleotide sequence of the insert of a positive clone contained an open reading frame of 2940 bp encoding a polypeptide of 980 amino acid residues with a calculated molecular mass of 111,182 daltons. Escherichia coli cells harboring the plasmid for expression produced CDP protein accounting for 30% of the total cellular proteins with an activity of 59.9 units/ml culture, which corresponded to 0.93 mg/ml culture. The expressed CDP could be used to synthesize cellulase inhibitors as celloligosaccharide analogues using glucose-1-phosphate as a glucose donor and 4- O-β- d-glucopyranosyl-1-deoxynojirimycin or 6- O-β-cellobiosyl-1-deoxynojirimycin as an acceptor.
ISSN:0922-338X
DOI:10.1016/S0922-338X(97)86758-X