Heard, S. C. | bioRxiv Science

bioRxiv2022

Biosynthesis of the Fungal Nonribosomal Peptide Penilumamide A and Biochemical Characterization of a Pterin-Specific Adenylation Domain

We report the characterization of the penilumamide A biosynthetic gene cluster from the marine-derived fungus Aspergillus flavipes CNL-338. In vitro reconstitution studies demonstrated that three Plm nonribosomal peptide synthetases encoding four modules are required for constructing the lumazine-containing tripeptide. Further investigations using dissected adenylation domains determined substrate specificity for methionine and anthranilic acid and led to the first biochemical characterization of an adenylation domain with selectivity for a pterin-derived building block. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=106 SRC="FIGDIR/small/505926v1_ufig1.gif" ALT="Figure 1"> View larger version (17K): org.highwire.dtl.DTLVardef@dfc4aeorg.highwire.dtl.DTLVardef@e9a4fdorg.highwire.dtl.DTLVardef@14ee7beorg.highwire.dtl.DTLVardef@1508021_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

bioRxiv2022

Genome Mining and Biochemical Characterization of a Bifunctional Type I Diterpene Synthase from a Marine-Derived Fungus

We report the identification of the tnd biosynthetic cluster from the marine-derived fungal strain Aspergillus flavipes CNL-338 and in vivo characterization of a cryptic type I diterpene synthase. Heterologous expression of the bifunctional terpene synthase TndC in Saccharomyces cerevisiae led to the discovery of a new diterpene backbone, talarodiene, harboring a benzo[a]cyclopenta[d]cyclooctane tricyclic fused ring system. The cyclization mechanism that converts geranylgeranyl diphosphate to the tricyclic hydrocarbon skeleton was investigated using 13C-labeling studies and stable isotope tracer experiments showed the biotransformation of talarodiene into talaronoid C.

bioengineering↗