Molecular cloning and analysis of the ergopeptine assembly system in the ergot fungus Claviceps purpurea.
Claviceps purpurea produces the pharmacological important ergopeptines, a class of cyclol-structured alkaloid peptides containing D-lysergic acid. These compounds are assembled from D-lysergic acid and three different amino acids by the nonribosomal peptide synthetase enzymes LPS1 and LPS2. Cloning of alkaloid biosynthesis genes from C. purpurea has revealed a gene cluster including two NRPS genes, cpps 1 and cpps 2. Protein sequence data had assigned earlier cpps1 to encode the trimodular LPS1 assembling the tripeptide portion of ergopeptines. Here, we show by transcriptional analysis, targeted inactivation, analysis of disruption mutants, and heterologous expression that cpps 2 encodes the monomodular LPS2 responsible for D-lysergic acid activation and incorporation into the ergopeptine backbone. The presence of two distinct NRPS subunits catalyzing formation of ergot peptides is the first example of a fungal NRPS system consisting of different NRPS subunits.
Correia
Telmo
T
Institut für Botanik, Westfälische Wilhelms-Universität, Schlossgarten 3, D-48149 Münster, Germany.
Grammel
Nicolas
N
Ortel
Ingo
I
Keller
Ullrich
U
Tudzynski
Paul
P
D000595:Amino Acid Sequence; D002967:Claviceps / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D004876:Ergot Alkaloids / Q000096:biosynthesis* / Q000737:chemistry; D004926:Escherichia coli; D005800:Genes, Fungal / Q000235:genetics*; D008237:Lysergic Acid / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D010453:Peptide Synthases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D004876:Ergot Alkaloids; D021122:Protein Subunits; D010453:Peptide Synthases; C105940:lysergyl peptide synthetase; C104385:non-ribosomal peptide synthase; D008237:Lysergic Acid