Abstract
An alginate lyase gene, algA, encoding a new poly β-D-mannuronate (polyM)-specific alginate lyase AlgA, was cloned from Pseudomonas sp. E03. The recombinant AlgA with (His)6-tag, consisting of 364 amino acids (40.4 kDa),was purified using Ni-NTA Sepharose. The purified lyase had maximal activity (222 EU/mg) at pH 8 and 30 °C and also maintained activity between pH 7-9 and below 45 °C. It exclusively and endolytically depolymerized polyM by β-elimination into oligosaccharides with degrees of polymerization (DP) of 2-5. Due to its high substrate specificity, AlgA could be a valuable tool for production of polyM oligosaccharides with low DP and for determining the fine structure of alginate.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacterial Proteins / chemistry*
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Bacterial Proteins / genetics
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism
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Enzyme Stability
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Escherichia coli / genetics
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Hydrogen-Ion Concentration
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Polysaccharide-Lyases / chemistry*
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Polysaccharide-Lyases / genetics
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Polysaccharide-Lyases / isolation & purification
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Polysaccharide-Lyases / metabolism
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Pseudomonas / enzymology*
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Pseudomonas / genetics
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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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Substrate Specificity
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Temperature
Substances
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Bacterial Proteins
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Recombinant Proteins
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Polysaccharide-Lyases
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poly(beta-D-mannuronate) lyase