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Volume 143, Issue 10

Acetate kinase from : a highly specific enzyme that is actively transcribed during acidogenesis and solventogenesis Free

Abstract

Acetate kinase (ATP:phosphotransferase, EC 2.7.2.1) has been purified 294-fold from acid-producing cells of DSM 1731 to a specific activity of 1087 U mg (ADP-forming direction). The dimeric enzyme consisted of subunits with a molecular mass of 43 kDa. The molecular mass of the native acetate kinase was in the range 87-94 kDa as judged by gel filtration and native gel electrophoresis. The enzyme showed high specificity for the substrates acetate and ATP, and maximal activity was obtained with Mn as divalent cation. The presence of mercury compounds such as HgCl and -hydroxymercuribenzoate resulted in an essential loss of activity. The apparent values for acetate, Mg-ATP, acetyl phosphate, and Mg-ADP were 73, 0.37, 0.58 and 0.71 mM. An activity-staining procedure for detection of acetate kinase in crude cell extracts after separation on native polyacrylamide gels was developed. A DNA fragment encoding 246 bp of the acetate kinase gene of DSM 792 was cloned by a PCR-based approach. Northern blot analysis revealed transcription of the gene under acid- and solvent-producing conditions with no significant differences at the level of transcription.

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Keyword(s): acetate kinase , acidogenesis , ack gene , butyrate kinase and Clostridium acetobutylicum
© Society for General Microbiology 1997
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1997-10-01
2024-04-24
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Acetate kinase from Clostridium acetobutylicum: a highly specific enzyme that is actively transcribed during acidogenesis and solventogenesis
Microbiology 143, 3279 (1997); https://doi.org/10.1099/00221287-143-10-3279
/content/journal/micro/10.1099/00221287-143-10-3279
/content/journal/micro/10.1099/00221287-143-10-3279
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