Early catalytic steps of Euglena gracilis chloroplast type II fatty acid synthase.
Euglena gracilis is a very ancient eukaryote whose chloroplast acquisition and evolution has been independent of higher plants. The organism in unique in possessing two de novo fatty acid synthases, a true multienzyme complex of great size in the cytosol and a plastid-localized type II fatty acid synthase composed of discrete enzymes and acyl carrier protein (ACP). The enzymology of the early steps of fatty acid biosynthesis differed in the Euglena type II fatty acid synthase compared to those of Escherichia coli and plants. The enzymes of Euglena participating in both priming and elongation reactions to form a new carbon-carbon bond were acetyl-CoA-ACP transacylase, malonyl-CoA-ACP transacylase, and beta-ketoacyl-ACP synthase I. The effects of inhibitors on the three different enzymes were noted. All carbon-carbon bond formation was inhibited by cerulenin. Although neither fatty acid biosynthesis nor any of the isolated enzymes were sensitive to diisopropylphosphofluoridate, the three Euglena enzymes studied were sensitive to different sulfhydryl-alkylating agents. Acetyl-ACP supported fatty acid biosynthesis as effectively as did comparable amounts of ACPSH and acetyl-CoA. There was no evidence for a beta-ketoacyl-ACP synthase III for priming such as has been reported in type II fatty acid synthase of higher plants and bacteria. The roles of the acetyl-CoA-ACP transacylase and beta-ketoacyl-ACP synthase I appear to be unique in the type II fatty acid synthase of Euglena. Acetyl-CoA-ACP transacylase, malonyl-CoA-ACP transacylase, and beta-ketoacyl-ACP synthase I were separated from one another and shown to have different molecular weights.
Worsham
L M
LM
Department of Biochemistry, James H. Quillen College of Medicine, East Tennessee State University, Johnson City 37614-0581.
Williams
S G
SG
Ernst-Fonberg
M L
ML
D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / Q000378:metabolism; D000123:Acetyltransferases / Q000378:metabolism; D051051:Acyl-Carrier Protein S-Acetyltransferase; D051080:Acyl-Carrier Protein S-Malonyltransferase; D000217:Acyltransferases / Q000378:metabolism; D000818:Animals; D005056:Euglena gracilis / Q000201:enzymology*; D064429:Fatty Acid Synthases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005227:Fatty Acids / Q000096:biosynthesis; D007527:Isoenzymes / Q000378:metabolism
D005227:Fatty Acids; D007527:Isoenzymes; D000217:Acyltransferases; D000123:Acetyltransferases; C078182:beta-ketoacyl-acyl carrier protein synthase I; D051051:Acyl-Carrier Protein S-Acetyltransferase; D051080:Acyl-Carrier Protein S-Malonyltransferase; D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase; D064429:Fatty Acid Synthases