CHARACTERIZATION AND ISOLATION OF VITAMIN B12 BIOSYNTHETIC GENES IN PRIESTIA FLEXA JT4

• Main Author: K. A. Tambunan, Thresia
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/74829
• Institution: Institut Teknologi Bandung

Vitamin B12 is an important vitamin needed by the body, a deficiency of vitamin B12 can cause megaloblastic anemia to nerve damage. The production of vitamin B12 is not environmentally friendly and the steps are too complex, while the production of vitamin B12 through bacterial fermentation takes a long time and also expensive. An alternative method that can be used to develop production of vitamin B12 is by exploring the bacterial genome to produce bioparts. Bioparts then can be used for the production of vitamin B12 on an industrial scale using a gene heterologous expression approach. Studies about vitamin B12 from aquatic bacteria has not much done yet. This research aims to explore aquatic bacteria from the 2019 TRIUMPH expedition, namely Priestia flexa JT4. The results of Whole Genome Sequencing (WGS) of Priestia flexa JT4 from previous studies reported that there are 6 precursor genes and 11 biosynthetic genes of vitamin B12. Of the 11 biosynthetic vitamin B12 genes, 8 of them (cobF, cobG, cobI, cobJ, cobK, cobL, cobM, cobNST) were not detected in the WGS results. In this study, 7 of the 8 genes were successfully characterized and isolated from Priestia flexa JT4 with each size cobF 780 bp, cobG 1104 bp, Recognize702 bp, cobJ 1623 bp, cobK 768 bp, cobL 1197 bp, and cobM 777 bp. The isolated genes encoding for vitamin B12 can be used as bioparts for the production of vitamin B12 supplements. This study also succeeded in confirming the presence of vitamin B12 in Priestia flexa JT4 qualitatively using HPLC. Production of vitamin B12 by Priestia flexa JT4 is carried out aerobically in contrast to other bacteria which are carried out anaerobically.