CHARACTERIZATION AND BIOPROSPECTING PRIESTIA FLEXA USING IN SILICO METHODS BASED ON WHOLE GENOME SEQUENCING (WGS) DATA

• Main Author: Natanael, Yosua
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/68490
• Institution: Institut Teknologi Bandung

Deep sea is an ecosystem for many living bacteria. Bacteria tend to produce secondary metabolites to adapt to their environment. Currently, secondary metabolites from marine bacteria are widely used in various industrial fields because they have biological activities that are beneficial for industry. In Indonesia, no studies have been conducted to explored Indonesia deep sea microbes. Therefore, the aim of this study was to analyze the genomic characteristics of Indonesia deep sea microbes, Priestia flexa and analyze biosynthetic gene clusters (BGCs) from the Priestia flexa genome. The method used in this study included the morphological characterization of P. flexa with Gram staining, taxonomic determination using the overall genome relatedness indices (OGRI), namely FastANI (ANI value) and GGDC (dDDH), constructing a phylogenetic tree using OrthoFinder and visualization of tree construct using iTOL. After that, the quality of the sequencing data was determined using the CheckM tool and the general characterization of the genome using Biophyton. Pangenomic analysis was performed on the genus Priestia using GET_HOMOLOGUES software and genomic island analysis was performed using IslandViewer. Analysis of secondary metabolite gene clusters was carried out using antiSMASH V6.0 tools and visualization of reaction pathways using KEGG Reconstructor. Priestia flexa is rod-shaped and Gram-negative bacteria. From the results of ANI and dDDH calculations, as well as the results of phylogenetic analysis, the bacterial isolates were Priestia flexa bacteria. The quality of the sequenced genome obtained is in the "high-quality draft" category. The size of P. flexa genome is 4.02Mb, with 5,135 genes and 5,185 CDS. Genes in the genome were successfully mapped into COG (Cluster of Orthologous Groups). The pangenome of genus Priestia is "open" pangenome with a value of ? -2.66. P. flexa has 21 genomic islands with 413 genes in it. Secondary metabolite gene cluster analysis showed that P. flexa has 8 secondary metabolite-producing gene clusters divided into Cyclic-lactone-autoinducer/RiPP-like, NRPS/NRPS-like, siderophore, lassopeptide, terpene, NRPS, and PKS (type III). The terpene gene cluster that produces carotenoids in the form of lycopene has complete genes and has high economic value, so that this gene cluster can be utilized for industrial production processes.