BIOCHEMICAL PROPERTIES OF LIPASE/ESTRASE GDSL FROM BACILLUS AQUIMARIS MKSC 6.2

• Main Author: AL MUQADASI NIM: 10513023 , MUTAWAKIL
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/29415
• Institution: Institut Teknologi Bandung

Bacteria can produce class lipolytic enzymes that play a role in breaking fat into glycerol and fatty acids. Lipolytic enzyme groups are carboxylesterase (EC 3.1.1.1) and true Lipase (EC 3.1.1.3). Carboxyesterase can hydrolyze short-chain ester that water-soluble. True Lipase can hydrolyze long-chain triglycerides and some types of phospholipases that are not water-soluble. Bacillus aquimaris MKSC 6.2 from the sea water of Merak Kecil Island, Banten, West Java has been identified to have the coding gene of GDSL enzyme. This study aims to express the coding gene of GDSL from Bacillus aquimaris MKSC 6.2 in E. coli BL21 (DE3) and analyze the product of this gene expression in pET30a. The GDSL protein was successfully expressed with a 0.2 mM IPTG (isopropyl β-D-1-thiogalactopiranoside) inducer for 2 hours. The activity of GDSL protein was tested with several derived fatty acid molecules. The GDSL protein is active to nitrophenols (p-NP) butyrate, p-NP valerate, pNP caprylate, and p-NP decanoate. As many as 100% of p-NP decanoate can be catalyzed by recombinant GDSL protein from Bacillus aquimaris MKSC 6.2. However, the activity of this protein on p-NP butyrate, p-NP valerate and p-NP caprylate is less than 40%. Based on these results, the recombinant GDSL protein from Bacillus aquimaris MKSC 6.2 belongs to the group of lipase (EC 3.1.1.3).