GENOME EXPLORATION OF PRIESTIA FLEXA TO PRODUCE BIOPARTS OF VITAMIN B12

• Main Author: Rahmawati, Viana
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/71516
• Institution: Institut Teknologi Bandung

Vitamin B12, or cobalamin, is a cofactor in various metabolic processes not produced by the body. The production of vitamin B12 on an industrial scale has a process or process that is long, complex and expensive so that the production of vitamin B12 can be increased by obtaining bacteria that have the ability to produce more optimal vitamin B12.Recently, one of the methods that can be used for developing vitamin B12 products is by exploring the bacterial genome (genome mining) to produce bio parts. The obtained bio parts can produce vitamin B12 with a synthetic biology approach. Exploration of vitamin B12 from aquatic bacteria has not been widely carried out, and research has yet to be conducted on deep-sea water bacteria. Exploration was carried out using the Whole Genome Sequencing (WGS) method. The WGS method tends to be faster and more accurate and this method is important for identifying genes and bacteria produced from deep sea water and then continuing with the cloning process. This study aims to explore the genes that play a role in the biosynthesis of vitamin B12 in bacteria from Java Trench water samples from the 2019 TRIUMPH expedition as a source of bio parts for vitamin B12 production. Priestia flexa bacteria obtained from the WGS analysis showed six precursor genes for vitamin B12 biosynthesis, including the gltx, HemA, HemL, HemB, HemC and HemD genes. In this study, three genes, gltx, HemL and HemB, were successfully isolated, each measuring 396 bp, 507 bp and 981 bp from single isolate of Priestia flexa which was, confirmed by 16sRNA sequencing. The isolated genes were then cloned into the pGEMT- Easy vector, which was transformed into E. coli DH5-alpha. Analysis of the sequencing results of the gltx gene and the HemB gene showed a 100% similarity level with the Priestia flexa genome sequence, while the HemL gene only reached 99.4%. The genes that make up vitamin B12 can be used as bio parts for producing vitamin B12 using fermentation process with synthetic biological methods in future studies.