Partial Characterization of Fatty Acid Synthase of Propionibacterium shermanii

Abstract

Fractionated extracts of Propionibacterium shermanii were found to contain the readily dissociable type of fatty acid synthase complex. For optimal activity the partially purified synthase required the presence of acetyl-CoA, malonyl-CoA, NADH and NADPH as well as a cell-free fraction containing the acyl carrier protein (ACP). When ACP-containing fractions obtained from P. shermanii grown in the presence of [ l-'4C]pantothenate were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis, the radioactivity was located in a peak corresponding to a molecular weight of approx. 8500-9000. The biological activity of the ACP was relatively heat-stable and trypsin-resistant. When the labelled ACP-containing fraction was subjected to gel filtration on a calibrated Sephadex G-75 column, a radioactive peak corresponding to a molecular weight of 22 000 was obtained that could participate in the 14C0,-malonyl-CoA exchange reaction in the presence of the other requisite components.

A small, discrete acyl carrier protein is involved in de novo fatty acid biosynthesis in Streptomyces erythraeus

“…IsobutyrylCoA was synthesized from the corresponding anhydride. Propionibacterium shermanii was grown as in [5], and ACP [6] was purified from it by the method described in this paper for S. erythraeus ACP. [2-14C]Malonyl-CoA (55 Ci/mol) was from Amersham International.…”

Cloning, characterization, and high-level expression in Escherichia coli of the Saccharopolyspora erythraea gene encoding an acyl carrier protein potentially involved in fatty acid biosynthesis

“…This implies that the fatty acid synthase of S. erythraea may be patterned on the type II system found generally in prokaryotes, consisting of freely dissociable, monofunctional enzyme components. The best-studied system of this kind is that of Escherichia coli (see reference 52 for a recent review), but a discrete ACP is also apparently involved in fatty acid synthase from gram-positive bacteria more closely related to S. erythraea (1,27) and which, like S. erythraea, characteristically produce iso-and anteiso-terminally branched fatty acids. Alternatively, there may exist more than one type of fatty acid synthase activity, as in Euglena gracilis (13).…”

‘Phormidin’, a novel, better fatty acid synthase inhibitor from marine cyanobacteria, Phormidium Ambiguum

“…Crude FAS containing protein fraction was isolated according to the modified protocol of Ahmad et al (1981). Stock culture ofBacillus subtilis was inoculated in 100 mL LB broth pH 7.2 and cultured for 24 hrs at room temperature.…”