title: Discovery of indolotryptoline antiproliferative agents by homology-guided metagenomic screening.
abstract: Natural product discovery by random screening of broth extracts derived from cultured bacteria often suffers from high rates of redundant isolation, making it ever more challenging to identify novel biologically interesting natural products. Here we show that homology-based screening of soil metagenomes can be used to specifically target the discovery of new members of traditionally rare, biomedically relevant natural product families. Phylogenetic analysis of oxy-tryptophan dimerization gene homologs found within a large soil DNA library enabled the identification and recovery of a unique tryptophan dimerization biosynthetic gene cluster, which we have termed the bor cluster. When heterologously expressed in Streptomyces albus, this cluster produced an indolotryptoline antiproliferative agent with CaMKIIδ kinase inhibitory activity (borregomycin A), along with several dihydroxyindolocarbazole anticancer/antibiotics (borregomycins B-D). Similar homology-based screening of large environmental DNA libraries is likely to permit the directed discovery of new members within other previously rare families of bioactive natural products.
authors.affiliation: Laboratory of Genetically Encoded Small Molecules, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Fang-Yuan
authors.initials: FY
authors.lastname: Chang
authors.forename: Sean F
authors.initials: SF
authors.lastname: Brady
mesh_terms: D006916:5-Hydroxytryptophan / Q000235:genetics*; D001483:Base Sequence; D001688:Biological Products / Q000737:chemistry*; D053898:Biosynthetic Pathways / Q000235:genetics*; D002243:Carbolines / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D019281:Dimerization; D055808:Drug Discovery / Q000379:methods; D015723:Gene Library; D005820:Genetic Testing / Q000379:methods; D013058:Mass Spectrometry; D054892:Metagenome / Q000235:genetics*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D005810:Multigene Family / Q000235:genetics; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D012987:Soil / Q000032:analysis*; D013302:Streptomyces
chemical_list: D001688:Biological Products; D002243:Carbolines; D017931:DNA Primers; D012987:Soil; C009804:tryptoline; D006916:5-Hydroxytryptophan; C110531:calmodulin dependent protein kinase II phosphatase; D010749:Phosphoprotein Phosphatases