Abstract
A hyperthermostable glycoside hydrolase family 51 (GH51) α-L-arabinofuranosidase from Thermotoga petrophila RKU-1 (TpAraF) was cloned, overexpressed, purified and characterized. The recombinant enzyme had optimum activity at pH 6.0 and 70°C with linear α-1,5-linked arabinoheptaose as substrate. The substrate cleavage pattern monitored by capillary zone electrophoresis showed that TpAraF is a classical exo-acting enzyme producing arabinose as its end-product. Far-UV circular dichroism analysis displayed a typical spectrum of α/β barrel proteins analogously observed for other GH51 α-L-arabinofuranosidases. Moreover, TpAraF was crystallized in two crystalline forms, which can be used to determine its crystallographic structure.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Arabinose / metabolism
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Bacteria / enzymology*
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Circular Dichroism
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Cloning, Molecular
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Crystallization
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DNA, Bacterial / chemistry
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DNA, Bacterial / genetics
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Enzyme Stability
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Gene Expression
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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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Hot Temperature
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Hydrogen-Ion Concentration
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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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Sequence Analysis, DNA
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Substrate Specificity
Substances
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DNA, Bacterial
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Recombinant Proteins
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Arabinose
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Glycoside Hydrolases
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alpha-N-arabinofuranosidase