Abstract
The gene coding for D-lactate dehydrogenase (D-LDH) from Pediococcus acidilactici DSM 20284 was cloned and expressed in E. coli. The recombinant enzyme was purified by nickel-affinity chromatography. It converted phenylpyruvic acid (PPA) to 3-phenyllactic acid maximally at 30°C and pH 5.5 with a specific activity of 140 and 422 U/mg for PPA and pyruvate, respectively. The K(m), turnover number (k(cat)), and catalytic efficiency (k(cat)/K(m)) for PPA were 2.9 mM, 305 s(-1), and 105 mM(-1) s(-1), respectively.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Biotransformation
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Cats
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Chromatography, Affinity
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Cloning, Molecular
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Enzyme Stability
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Escherichia coli / enzymology
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Escherichia coli / genetics
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Gene Expression
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Hydrogen-Ion Concentration
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Kinetics
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Lactate Dehydrogenases / chemistry
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Lactate Dehydrogenases / genetics*
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Lactate Dehydrogenases / metabolism*
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Lactates / metabolism*
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Pediococcus / enzymology*
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Pediococcus / genetics*
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Phenylpyruvic Acids / metabolism*
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Temperature
Substances
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Lactates
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Phenylpyruvic Acids
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Recombinant Proteins
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3-phenyllactic acid
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Lactate Dehydrogenases
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D-lactate dehydrogenase
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phenylpyruvic acid