title: An iterative, bimodular nonribosomal peptide synthetase that converts anthranilate and tryptophan into tetracyclic asperlicins.
abstract: The bimodular 276 kDa nonribosomal peptide synthetase AspA from Aspergillus alliaceus, heterologously expressed in Saccharomyces cerevisiae, converts tryptophan and two molecules of the aromatic β-amino acid anthranilate (Ant) into a pair of tetracyclic peptidyl alkaloids asperlicin C and D in a ratio of 10:1. The first module of AspA activates and processes two molecules of Ant iteratively to generate a tethered Ant-Ant-Trp-S-enzyme intermediate on module two. Release is postulated to involve tandem cyclizations, in which the first step is the macrocyclization of the linear tripeptidyl-S-enzyme, by the terminal condensation (CT) domain to generate the regioisomeric tetracyclic asperlicin scaffolds. Computational analysis of the transannular cyclization of the 11-membered macrocyclic intermediate shows that asperlicin C is the kinetically favored product due to the high stability of a conformation resembling the transition state for cyclization, while asperlicin D is thermodynamically more stable.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, 420 Westwood Plaza, Los Angeles, CA 90095, USA.
authors.forename: Xue
authors.initials: X
authors.lastname: Gao
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Jiménez-Osés
authors.forename: Moon Seok
authors.initials: MS
authors.lastname: Choi
authors.forename: Kendall N
authors.initials: KN
authors.lastname: Houk
authors.forename: Yi
authors.initials: Y
authors.lastname: Tang
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Walsh
mesh_terms: D001230:Aspergillus / Q000201:enzymology; D001570:Benzodiazepinones / Q000737:chemistry / Q000378:metabolism*; D055162:Biocatalysis; D003500:Cyclization; D020035:Holoenzymes / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D009842:Oligopeptides / Q000378:metabolism; D010453:Peptide Synthases / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012995:Solubility; D013237:Stereoisomerism; D014364:Tryptophan / Q000378:metabolism*; D062367:ortho-Aminobenzoates / Q000378:metabolism*
chemical_list: D001570:Benzodiazepinones; D020035:Holoenzymes; D009842:Oligopeptides; D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D014364:Tryptophan; C057101:asperlicin C; C057102:asperlicin D; D010453:Peptide Synthases
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