Studies on Itaconate Metabolism

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Abstract
P. fluorescens cells were sonicated with [image]/15 phosphate buffer (pH 7.3), the extract passed through charcoal, dialyzed, treated with 0.2% protamine (pH 6.5), the supernatant fractionated by (NH4)2 SO4 (0.45-0.65 saturation) at pH 7, dialyzed mixed with Ca3 (PO4)2 gel, (pH 7.3), and refractlonated by (NH,4 SO4. The preparation possessed itaconyl and succinyl-CoA synthetase activity in the presence of CoA, adenosine triphosphate (ATP) and Mg2+. The enzymlc activity was adaptively increased when the cells were cultivated in the presence of itaconate but not D-glucose. The CoA-derivatlve synthesizing activity with succlnate and itaconate was about 4:1, which was approximately constant during purification steps. An identity of both CoA-derivative synthetases is suggested.