Functions of malonate decarboxylase subunits from Pseudomonas putida.
Malonate decarboxylase from Pseudomonasputida is composed of five subunits, alpha, beta, gamma, delta, and epsilon. Two subunits, delta and epsilon, have been identified as an acyl-carrier protein (ACP) and malonyl-CoA:ACP transacylase, respectively. Functions of the other three subunits have not been identified, because recombinant subunits expressed in Escherichia coi formed inclusion bodies. To resolve this problem, we used a coexpression system with GroEL/ES from E. coli, and obtained active recombinant subunits. Enzymatic analysis of the purified recombinant subunits showed that the alpha subunit was an acetyl-S-ACP:malonate ACP transferase and that the betagamma-subunit complex was a malonyl-S-ACP decarboxylase.
Chohnan
Shigeru
S
Department of Bioresource Science, College of Agriculture, Ibaraki University, 3-21-1 Chuo, Ami, Ibaraki 300-0393, Japan. chohnan@mx.ibaraki.ac.jp
Akagi
Kayo
K
Takamura
Yoshichika
Y
D000213:Acyl Carrier Protein / Q000378:metabolism; D051080:Acyl-Carrier Protein S-Malonyltransferase; D000217:Acyltransferases / Q000737:chemistry / Q000378:metabolism; D002262:Carboxy-Lyases / Q000378:metabolism*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D029968:Escherichia coli Proteins; D054889:Fatty Acid Synthase, Type II; D010957:Plasmids / Q000235:genetics; D016958:Pseudomonas putida / Q000201:enzymology*; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000213:Acyl Carrier Protein; D017931:DNA Primers; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D000217:Acyltransferases; D051080:Acyl-Carrier Protein S-Malonyltransferase; C494968:fabD protein, E coli; D002262:Carboxy-Lyases; C071852:malonate decarboxylase; D054889:Fatty Acid Synthase, Type II