Abstract
The function of the mlr6787 gene from Mesorhizobium loti MAFF303099 has been identified. This gene encodes 2-(acetamidomethylene)succinate hydrolase, an enzyme involved in the catabolism of pyridoxal 5'-phosphate (vitamin B 6). This enzyme was overexpressed in Escherichia coli, purified to homogeneity, and characterized. 2-(Acetamidomethylene)succinate hydrolase catalyzes the hydrolysis of 2-(acetamidomethylene)succinate to yield succinic semialdehyde, acetic acid, carbon dioxide, and ammonia. The k cat and K M for this reaction were 0.6 s (-1) and 143 microM, respectively. The enzyme was shown to utilize the E isomer of 2-(acetamidomethylene)succinate.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Alphaproteobacteria / enzymology*
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Alphaproteobacteria / genetics*
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism*
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Cloning, Molecular
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DNA Primers
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Escherichia coli / enzymology
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Hydrolases / metabolism*
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Kinetics
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Pseudomonas / enzymology
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Pyridoxal Phosphate / metabolism*
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Recombinant Proteins / metabolism
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Succinate-Semialdehyde Dehydrogenase / isolation & purification
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Succinate-Semialdehyde Dehydrogenase / metabolism
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gamma-Aminobutyric Acid / analogs & derivatives
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gamma-Aminobutyric Acid / metabolism
Substances
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Bacterial Proteins
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DNA Primers
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Recombinant Proteins
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gamma-Aminobutyric Acid
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Pyridoxal Phosphate
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Succinate-Semialdehyde Dehydrogenase
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2-(acetamidomethylene)succinate hydrolase, Mesorhizobium loti
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Hydrolases
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succinic semialdehyde