Abstract
The gene coding for the beta-glycosidase from the archaeon Sulfolobus solfataricus has been overexpressed in Escherichia coli. The enzyme was purified to homogeneity with a rapid purification procedure employing a thermal precipitation as a crucial step. The final yield was 64% and the purification from the thermal precipitation was 5.4-fold. The expressed enzyme shows the same molecular mass, thermophilicity, thermal stability, and broad substrate specificity, with noticeable exocellobiase (glucan 1,4-beta-D-glucosidase) activity, of the enzyme purified from S. Solfataricus. We provide evidence that the beta-glycosidase can assume its functional state in E. coli without the contribution of N-epsilon-methylated lysine residues.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Amino Acids / analysis
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Base Sequence
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Enzyme Stability
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Gene Expression Regulation, Bacterial / genetics
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Glycoside Hydrolases / chemistry*
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Glycoside Hydrolases / genetics
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Glycoside Hydrolases / isolation & purification
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Glycoside Hydrolases / metabolism
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Hydrolysis
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Kinetics
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Lysine / analogs & derivatives
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Lysine / analysis
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Molecular Sequence Data
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Protein Processing, Post-Translational
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Sequence Analysis
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Substrate Specificity
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Sulfolobus / enzymology*
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Temperature
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Thermodynamics
Substances
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Amino Acids
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Recombinant Proteins
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epsilon-N-methyllysine
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Glycoside Hydrolases
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Lysine