GENOME EXPLORATION OF VIRGIBACILLUS SALARIUS TO PRODUCE BIOPART OF SQUALENE SYNTHASE (SQS)

• Main Author: Mufliyati, Lily
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/84502
• Institution: Institut Teknologi Bandung

The biodiversity of Indonesia has become the habitat of a variety of unique microorganisms with many benefits for human life, one of which is Virgibacillus salarius. The moderate halofilic bacterium V. salarius, isolated from marine corals, has the sqs gene, which encodes the squalene-producing enzyme (SQS) in its genome and potentially produces squalene. Squalene is useful for a variety of industries, such as additional vaccinations in the medical field, cosmetic products in the beauty field, and nutritional supplements in the nutraceutical field. Therefore, the study aims to isolate and clone the sqs gene from the V. salarius genome data produced in previous studies, as well as characterize the gene in silico. The genome sequencing of Whole Genome Sequencing (WGS) V. salarius is annotated using KEGG Onthology. The characterization of the sqs gene in the WGS, including sequence length and transcription factor, is done using FGENESB. A comparative analysis was conducted by looking for the percentage similarity of the protein sequence of the SQS V. salarius gene with six squalene-producing bacteria of the Bacillaceae family and one of the Planococcaceae family as outgroups based on the sequence obtained from NCBI using ClustalX, the identification of motifs and domains on each suspected protein by MEME-Suites and CD-Search, as well as the construction of the phylogenetic tree of the suspected SQS protein sequencing with MEGA-X. The result of the sqs gene isolation in V. salarius is cloned into the pGEMT plasmid and transformed into Escherichia coli DH? bacteria. The resulting sqs gene is sequenced, and the sequence is confirmed through Pairwise Sequence Alignment with a sequence predicted by the WGS. The sqs gene that is isolated from V. salarius measures 822 bp and is forecast to have the ERG9 superfamily domain on the protein of SQS.