ISSN:
1432-072X
Keywords:
Clostridium kluyveri
;
Acetyl-CoA content of C. kluyveri
;
CoASH content of C. kluyveri
;
Regulatory function of the acetyl-CoA/CoASH ratio
;
Isotope assay of acetyl-CoA
;
Quantitative determination of CoASH with 14C-N-ethyl-maleimide
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract CoASH and some of its acyl derivatives, especially acetyl-SCoA, occupy a central position in the energy metabolism of the anaerobic Clostridium kluyveri, both as intermediates and as regulatory effectors. The steady state concentrations of these compounds were determined in growing cultures of this organism using an anaerobic and fast deproteinization technique and radio isotope assays. Acetyl-SCoA was determined as [1-14C]citrate formed in the presence of [4-14C]oxaloacetate and citrate synthase; 0.49 μmol/g cell wet wt. were found CoASH, CoAS-SCoA after borohydride reduction, and total acyl derivatives of coenzyme A after hydrolysis of the thiol esters were converted to thioethers with [2,3-14C]N-ethylmaleimide and brought to radiochemical purity by chromatographic methods. While disulfides of coenzyme A were undetectable, 0.13 μmol CoASH and 1.17 μmol of total acyl-SCoA per g wet wt. were found. These data are consistent with the regulatory scheme of the energy metabolism of C. kluyveri previously proposed.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00425942
