Abstract
By constructing the genomic library, a β-glucosidase gene, with a length of 2,382 bp, encoding 793 amino acids, designated bgla, is cloned from a marine bacterium Aeromonas sp. HC11e-3. The enzyme is expressed successfully in the recombinant host Escherichia coli BL21 (DE3) and purified using glutathione affinity purification system. It shows the optimal activity at pH 6, 55 °C and hydrolyzes aryl-glucoside specially. Ca(2+), Mn(2+), Zn(2+), Ba(2+), Pb(2+), Sr(2+) can activate the enzyme activity, whereas SDS, EDTA, DTT show slight inhibition to the enzyme activity. Homologous comparing shows that the enzyme belongs to glycosyl hydrolase family 3, exhibiting 46 % identity with a fully characterized glucosidase from Thermotoga neapolitana DSM 4359. Such results provide useful references for investigating other glucosidases in the glycosyl family 3 as well as developing glucosidases using in suitable industrial area.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aeromonas / enzymology*
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Aeromonas / genetics
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Aeromonas / isolation & purification
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Amino Acid Sequence
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Cations, Divalent / metabolism
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Chromatography, Affinity
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Cloning, Molecular
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Enzyme Activators / metabolism
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Enzyme Inhibitors / metabolism
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Enzyme Stability
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Escherichia coli / genetics
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Gene Expression
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Glucosidases / chemistry
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Glucosidases / genetics*
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Glucosidases / isolation & purification
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Glucosidases / metabolism*
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Hydrogen-Ion Concentration
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Metals / metabolism
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Models, Molecular
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Molecular Sequence Data
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Protein Conformation
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Seawater / microbiology
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Sequence Homology, Amino Acid
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Temperature
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Thermotoga neapolitana / enzymology
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Thermotoga neapolitana / genetics
Substances
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Cations, Divalent
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Enzyme Activators
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Enzyme Inhibitors
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Metals
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Recombinant Proteins
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Glucosidases