Characterization, Comparative Genomics and Genome Mining for Antibiotics and Secondary Metabolite of two Actinomycetales isolates

Abstract

Actinomycetes are ubiquitous Gram (+) bacteria commonly found to have high G+C content and best known for their metabolic by-products and novel enzymes [1]. Isolates CCMMD2014 & MRMD2014 were co-cultured from soil impacted by a rusty fire hydrant in Woods Hole, MA. The Streptomyces sp. and Curtobacterium sp. isolates were identified by marker genes for 16S rRNA, rpoB, xylose isomerase, tryptophan synthase beta chain and Cytochrome P450 monooxygenase. Both isolates showed lactic acid fermentation and urease activity. The co-isolates were separated by selective culturing with antibiotics. In addition, whole genome sequencing revealed distinct inherent metabolic pathways in each culture that allowed for mutually exclusive selective culture conditions. Assembly was done using HGAP3 with Celera8 assembler using SMRT portal [2,3]. Annotation was done using the RAST server [4], with 7540 and 3969 CDS for Streptomyces sp. and Curtobacterium sp. respectively being revealed by AMIGene and BASys [5,6]. Subsequently, antiSMASH [7], was used to predict 52 and 26 secondary metabolite biosynthetic clusters that included genes for lantipeptides, terpenes, siderophores, polyketide synthases type I and II, bacteriocin and nonribosomal peptide synthase genes for Streptomyces sp. and Curtobacterium sp. respectively. The isolates have genes of potentially beneficial traits that could help study, among others, the role of fimbrial adhesins and iron in biofilm formation and investigation on natural products.