Abstract
The report is the first of purification, overproduction, and characterization of a unique γ-butyrobetainyl CoA synthetase from soil-isolated Agrobacterium sp. 525a. The primary structure of the enzyme shares 70-95% identity with those of ATP-dependent microbial acyl-CoA synthetases of the Rhizobiaceae family. As distinctive characteristics of the enzyme of this study, ADP was released in the catalytic reaction process, whereas many acyl CoA synthetases are annotated as an AMP-forming enzyme. The apparent Km values for γ-butyrobetaine, CoA, and ATP were, respectively, 0.69, 0.02, and 0.24 mM.
Keywords:
4-trimethylaminobutyrate-CoAligase; ADP-forming acyl CoA synthetase; l-carnitine; γ-butyrobetaine; γ-butyrobetaine-CoA ligase.
MeSH terms
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Acyl Coenzyme A / chemistry
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Acyl Coenzyme A / metabolism*
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Adenosine Diphosphate / chemistry
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Adenosine Diphosphate / metabolism*
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Agrobacterium / enzymology*
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Agrobacterium / genetics
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Amino Acid Sequence
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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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Betaine / analogs & derivatives*
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Betaine / chemistry
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Betaine / metabolism
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Carnitine / chemistry
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Carnitine / metabolism*
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Cloning, Molecular
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Coenzyme A Ligases / genetics
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Coenzyme A Ligases / isolation & purification
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Coenzyme A Ligases / metabolism*
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gene Expression
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Hydrogen-Ion Concentration
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Kinetics
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Sequence Alignment
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Sequence Homology, Amino Acid
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Soil Microbiology*
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Substrate Specificity
Substances
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Acyl Coenzyme A
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Bacterial Proteins
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gamma-butyrobetainyl-CoA
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Betaine
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gamma-butyrobetaine
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Adenosine Diphosphate
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Coenzyme A Ligases
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Carnitine