ANALYSIS AND ISOLATION OF LPAAT ENCODING GENE FORM MARINE GREEN MICROALGAE

• Main Author: Ulfa, Syafiqa
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/49221
• Institution: Institut Teknologi Bandung

Microalgae have potential as world's main energy source in addition to fossil-based fuel because of its ability to store abundant triacylglycerols (TAG). One of the factors that support this ability is the high activity of the enzyme lysopophospatidic acid acyltransferase (LPAAT) in TAG biosynthetic pathway in microalgae cells. This study aims to isolate LPAAT encoding gene from local Indonesian marine microalgae (GSI) strain. For this reason, LPAAT analysis from literature or GenBank has been carried out to design the primers for isolating LPAAT encoding gene by PCR. In silico analysis towards LPAAT protein structure has been conducted to understand the molecular structure of LPAAT enzyme. The results show that there are domain similarities between LPAAT with lysophospholipid acyltransferase (LPLAT) and C-terminus acyltransferase protein families. Catalytic sides of LPAAT enzyme are H-88, P-167, and P-198. Isolation of total DNA from microalgae GSI was carried out using CTAB (cetyl trimethyl ammonium bromide) method. DNA extract with concentration of 196.15 ng/µL have absorbance at wavelength of 260/280 and 260/230 of 1.828 and 1.995, respectively. The 18S rDNA-V4 marker gene confirms the microalgae GSI as green microalgae (chlorophyta), which phylogenetically has the highest association with genus Parachlorella. The LPAAT encoding gene with length of ± 2,000 base pairs was isolated by PCR from the microalgae GSI. The pure extract of LPAAT encoding gene has a concentration of 31.250 ng/µL.