Analysis of the catalytic center of Cyclomaltodextrinase from Thermoanaerobacter ethanolicus 39E

The amino acid sequences of cyclomaltodextrinase (CDase) from Thermoanaerobacter ethanolicus 39E (formerly Clostridium thermohydrosulfuricum 39E) and other amylolytic enzymes were compared by using linear alignment and hydrophobic cluster analysis. Two Asp and one Glu residue, which were considered...

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Bibliographic Details
Published in:FEBS letters 1993-02, Vol.317 (3), p.259-262
Main Authors: Podkovyrov, Sergey M., Burdette, Douglas, Zeikus, J.Gregory
Format: Article
Language:English
Subjects:
Active center
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Bacteria, Anaerobic - enzymology
Base Sequence
Binding Sites
Biological and medical sciences
CDase
CGTase
Conserved Sequence
cyclodextrin
cyclodextrin glycosyltransferase
Cyclomaltodextrinase
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Glycoside Hydrolases - chemistry
Glycoside Hydrolases - genetics
Glycoside Hydrolases - metabolism
Gram-Positive Asporogenous Rods, Irregular - enzymology
HCA
Hydrolases
Hydrophobic cluster analysis
Molecular Sequence Data
Mutagenesis, Site-Directed
Sequence Homology, Amino Acid
Site-directed mutagenesis
Thermoanaerobacter ethanolicus
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Summary:The amino acid sequences of cyclomaltodextrinase (CDase) from Thermoanaerobacter ethanolicus 39E (formerly Clostridium thermohydrosulfuricum 39E) and other amylolytic enzymes were compared by using linear alignment and hydrophobic cluster analysis. Two Asp and one Glu residue, which were considered to be the catalytic residues of the compared enzymes according to crystallographic or protein engineering experiments, were also conserved in CDase. Asp 325, Asp 421 and Glu 354 of the CDase were individually replaced by means of site-directed mutagenesis. The mutant enzymes completely lost activity, suggesting that these residues play an important role in catalysis.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(93)81288-B