Structure of the Gene Encoding Cyclomaltodextrinase from Clostridium Thermohydrosulfuricum 39E and Characterization of the Enzyme Purified from Escherichia Coli

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1992 Aug 1

PMID 1644767

Citations 16

Authors

S M Podkovyrov

J G Zeikus

Affiliations

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Abstract

Clostridium thermohydrosulfuricum 39E, a gram-positive thermophilic anaerobic bacterium, produced a cyclodextrin (CD)-degrading enzyme, cyclodextrinase (CDase) (EC 3.2.1.54). The enzyme was purified to homogeneity from Escherichia coli cells carrying a recombinant multicopy plasmid that contained the gene encoding for thermophilic CDase. The purified enzyme was a monomer with an M(r) of 66,000 +/- 2,000. It showed the highest activity at pH 5.9 and 65 degrees C. The enzyme hydrolyzed alpha-, beta-, and gamma-CD and linear maltooligosaccharides to yield maltose and glucose. The Km values for alpha-, beta-, and gamma-CD were 2.5, 2.1, and 1.3 mM, respectively. The rates of hydrolysis for polysaccharides (starch, amylose, amylopectin, and pullulan) were less than 5% of the rate of hydrolysis for alpha-CD. The entire nucleotide sequence of the CDase gene was determined. The deduced amino acid sequence of CDase, consisting of 574 amino acids, showed some similarities with those of various amylolytic enzymes.

Citing Articles

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Mining for novel cyclomaltodextrin glucanotransferases unravels the carbohydrate metabolism pathway via cyclodextrins in Thermoanaerobacterales.

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Cyclodextrinase from Thermococcus sp expressed in Bacillus subtilis and its application in the preparation of maltoheptaose.

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Recombinant cyclodextrinase from Thermococcus kodakarensis KOD1: expression, purification, and enzymatic characterization.

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The characterisation of an alkali-stable maltogenic amylase from Bacillus lehensis G1 and improved malto-oligosaccharide production by hydrolysis suppression.

Abdul Manas N, Pachelles S, Mahadi N, Md Illias R PLoS One. 2014; 9(9):e106481.

PMID: 25221964 PMC: 4164441. DOI: 10.1371/journal.pone.0106481.

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