BIOPROSPECTION OF MICROORGANISMS FROM BEKASAM FERMENTED FOOD THROUGH GENOMIC AND METABOLOMIC APPROACHES FOR IDENTIFICATION OF ANTI-INFLAMMATORY-RELATED COMPOUNDS

• Main Author: Azzahrah, Aisyah
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/79494
• Institution: Institut Teknologi Bandung

Bekasam is an Indonesian fermented product made from fish produced through a spontaneous fermentation process and has a sour taste, which comes from lactic acid bacteria (LAB). LAB are classified as probiotics that have many health benefits because they can produce anti-inflammatory compounds, antibiotics and other compounds. This study aims to characterize LAB isolate S44 from bekasam through metabolomic and genomic approaches for the exploration of potential compounds as anti-inflammatory agent. The research was conducted at the Research Center for Genetic Engineering, Research Organization for Life Sciences and Environment, National Research and Innovation Agency (BRIN). The research was carried out by characterization of the BAL S44 isolate, whole genome sequencing (WGS) analysis, characterization of metabolites through GC-MS, and analysis of biosynthetic pathways that produce potential metabolites. Microscopic observation of LAB isolate S44 has a morphological form of cocci, gram positive, the character of the growth curve starts from the logarithmic phase (log) at hour 5 to 15 and continued stationary phase. In the genomic approach, 16S rRNA sequencing and WGS results showed that LAB isolate S44 is Pediococcus acidilactici. The genome annotation of P. acidilactici S44 is known based on the assembly results with the P. acidilactici database (NCBI Taxonomy ID: 1254) with an Average Nucleotide Identity (ANI) value of 621 / 682 genes. Some of the anti-inflammatory substances discovered in isolate S44 include acetamide N-[4-[[2-methoxy-5-(1-methylethyl) phenyl] sulfonyl] amino] phenyl], cyclo (L-prolyl-L-valine), and 1-(2-Isopropoxybenzyl) piperazine found in the stationary phase. Molecular binding analysis showed hydrogen binding to the COX-2 active site and low binding affinity values for the three compounds using indomethacin as native ligand and diclofenac sodium as positive control. The results of absorption, distribution, metabolism, and excretion (ADME) analysis show that the 3 potential ligand compounds are safe to be used as drugs based on Lipinski's rule, while the results of molecular dynamics simulations show cyclo (L-prolyl-L-valine) is the best potential ligand in terms of radius of gyration (Rg), root mean square deviation (RMSD), root mean square fluctuation (RMSF), solvent accessible surface area (SASA), and hydrogen bonding determined for each receptor ligand complex. Furthermore, the annotation of Clusters of Orthologous Genes (COGs) showed the biosynthetic pathway of acetamide compound N - [ 4 - [ [ [ 2 – methoxy – 5 - (1 – methylethyl ) phenyl ] sulfonyl] amino] phenyl], cyclo(L-prolyl-L-valine).