The mmgA gene from Bacillus subtilis encodes a degradative acetoacetyl-CoA thiolase |
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Authors: | Jason J. Reddick Jayme K. Williams |
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Affiliation: | (1) Department of Chemistry and Biochemistry, The University of North Carolina at Greensboro, P.O. Box 26170, Greensboro, NC 27402-6170, USA |
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Abstract: | Early in sporulation, the mother cell compartment of Bacillus subtilis transcribes the mother cell metabolic gene (mmg) operon. The gene mmgA was assigned by other workers using sequence homology as an acetyl-CoA acetyltransferase [E.C. 2.3.1.9]. The gene was overexpressed in Escherichia coli, and the protein was purified by Ni2+-affinity chromatography. However, the expected MmgA-catalyzed biosynthesis of acetoacetyl-CoA from acetyl-CoA was undetectable by a standard UV assay, HPLC, and mass spectrometry. These methods indicated a preference for the reverse degradative thiolytic reaction, with a k cat of 80 s−1, and a K m of 70 and 50 μM for CoA and acetoacetyl-CoA, respectively. |
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Keywords: | Bacillus subtilis Sporulation metabolism Thiolase |
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