In Vitro Investigation of Crosstalk between Fatty Acid and Polyketide Synthases in the Andrimid Biosynthetic Assembly Line.
Andrimid (Adm) synthase, which belongs to the type II system of enzymes, produces Adm in Pantoea agglomerans. The adm biosynthetic gene cluster lacks canonical acyltransferases (ATs) to load the malonyl group to acyl carrier proteins (ACPs), thus suggesting that a malonyl-CoA ACP transacylase (MCAT) from the fatty acid synthase (FAS) complex provides the essential AT activity in Adm biosynthesis. Here we report that an MCAT is essential for catalysis of the transacylation of malonate from malonyl-CoA to AdmA polyketide synthase (PKS) ACP in vitro. Catalytic self-malonylation of AdmA (PKS ACP) was not observed in reactions without MCAT, although many type II PKS ACPs are capable of catalyzing self-acylation. This lack of self-malonylation was explained by amino acid sequence analysis of the AdmA PKS ACP and the type II PKS ACPs. The results show that MCAT from the organism's FAS complex can provide the missing AT activity in trans, thus suggesting a protein-protein interaction between the fatty acid and polyketide synthases in the Adm assembly line.
Department of System Chemotherapy and Molecular Sciences, Division of Bioinformatics and Chemical Genomics, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo, Kyoto, 606-8501, Japan.
Fumihiro
F
Ishikawa
Department of System Chemotherapy and Molecular Sciences, Division of Bioinformatics and Chemical Genomics, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo, Kyoto, 606-8501, Japan.
Hiroyasu
H
Sugimoto
Department of System Chemotherapy and Molecular Sciences, Division of Bioinformatics and Chemical Genomics, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo, Kyoto, 606-8501, Japan.
Hideaki
H
Kakeya
D051080:Acyl-Carrier Protein S-Malonyltransferase / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D020636:Pantoea / Q000201:enzymology / Q000235:genetics; D011090:Polyenes / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*; D054730:Protein Interaction Domains and Motifs; D011758:Pyrroles / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D011090:Polyenes; D011758:Pyrroles; D011994:Recombinant Proteins; C088069:andrimid; D048630:Polyketide Synthases; D051080:Acyl-Carrier Protein S-Malonyltransferase; D064429:Fatty Acid Synthases
antibiotics; biosynthesis; fatty acid synthases; polyketide synthases; protein-protein interactions