Anthranilate-activating modules from fungal nonribosomal peptide assembly lines.
Fungal natural products containing benzodiazepinone- and quinazolinone-fused ring systems can be assembled by nonribosomal peptide synthetases (NRPS) using the conformationally restricted beta-amino acid anthranilate as one of the key building blocks. We validated that the first module of the acetylaszonalenin synthetase of Neosartorya fischeri NRRL 181 activates anthranilate to anthranilyl-AMP. With this as a starting point, we then used bioinformatic predictions about fungal adenylation domain selectivities to identify and confirm an anthranilate-activating module in the fumiquinazoline A producer Aspergillus fumigatus Af293 as well as a second anthranilate-activating NRPS in N. fischeri. This establishes an anthranilate adenylation domain code for fungal NRPS and should facilitate detection and cloning of gene clusters for benzodiazepine- and quinazoline-containing polycyclic alkaloids with a wide range of biological activities.
Ames
Brian D
BD
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Walsh
Christopher T
CT
D000595:Amino Acid Sequence; D001232:Aspergillus fumigatus / Q000235:genetics / Q000378:metabolism; D002250:Carbon Radioisotopes; D003001:Cloning, Molecular; D017931:DNA Primers; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D005658:Fungi / Q000235:genetics / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000737:chemistry; D062367:ortho-Aminobenzoates / Q000378:metabolism*
D002250:Carbon Radioisotopes; D017931:DNA Primers; D005656:Fungal Proteins; D011994:Recombinant Proteins; D062367:ortho-Aminobenzoates; C031385:anthranilic acid