Determination of Intracellular Concentration of Acyl-Coenzyme A Esters for Metabolic Profiling Clostridium acetobutylicum

A method for determination of intracellular acyl‐coenzyme A esters in Clostridium acetobutylicum (CA) by high performance liquid chromatography (HPLC) was developed and validated. In our experiment, two important intermediates acyl‐coenzyme A esters including acetyl‐CoA, butyryl‐CoA could be baseline separated on a Zobax‐C18 column with mobile phase composed of the acetonitrile and phosphate buffer (pH 5.0). Samples treated with freeze‐thaw and protein precipitated by addition of trichloroacetic acid could be directly injected for determination of acetyl‐CoA, butyryl‐CoA with a recovery higher than 73%. With this method, the metabolite profiling of the acyl‐coenzyme A esters of CA was obtained. The comparison of the metabolite profiling between the model strain ATCC 824 and a mutant strain EA 2018, which produces higher butanol than the former was performed. Some useful information was concluded for understanding the mechanism of CA for selectively producing the solvent.