#TITLE_ALTERNATIVE#

• Main Author: YOPA KRISTIA (NIM : 20513302), YOGI
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/24787
• Institution: Institut Teknologi Bandung

Lipase (triacylglycerol acylhydrolases, EC 3.1.3.3) is a lipolytic enzyme, catalyzing reaction of hydrolisys and or ester synthesize from triasilgliseride in interface phase between polar and nonpolar solvent. Lipase are commonly exist in nature produced by a variety of plants, animals and microorganisms. Lipases have many catalitycs profility so that enzyme be able to use in wide industrial field, especially in the field of biotechnology for many applications in the food industry, detergents, medical applications and enzymatic processes in the production of lipophilic chemicals. Most lipase, used in industries isolated from procaryotic microorganism, since lipase from bacteria shown an activity in the wide temperature and pH. Thermostability is desirable characteristic for commercial lipase, since reaction rate increases by increasing temperature, and the productivity improved at high temperatures. Therefore, exploration of lipase from other sources still conducted to find the new lipase that has other properties at high temperature. Some of termophilic bacteria from compost have been successfully cultivated in the laboratory, the gene encoding lipase were isolated. One of thermophilic bacteria was named by isolat AL17 and was identified by 16s RNA method and then the bacteria was Pseudoxanthomonas sp. This research was focus to further studies on the cloning and characterization of lipase gene from one of isolate bacteria of Pseudoxanthomonas sp. Thermostable lipase gen from Pseudoxanthomonas sp was cloned to pJET 1.2/blunt cloning vector. Alignment nucleotide sequence showed that lipase gen has 936 base paire with ATG as start codon and TAA as stop codon. This lipase gen has 98% similarity with lipase gen from Uncultured Pseudomonas sp. (GenBank No. AKA58891.1) in protein level and has similarity 99% with lipase from Uncultured Pseudomonas sp. clone LK_ITB4c (GenBank No. KP204885.1) in nucleotide level. Sub-cloning lipase gen from cloning recombinan plasmid pLipAL17-1.2 to expression vector pET-30a(+) has done. Lipase gene AL17 which is integrated to recombinan plasmid pET-30a(+) is placed after promotor T7 position. Insilico translation of lipase gen which present in expression vector pET-30a(+) (pLipAL17) showed that lipase recombinan protein (lipase AL17) has additional amino acid that is come from pET-30a(+) at N-terminal site which are 6x Histag, thrombin site, S-tag and enterokinase site. Recombinan plasmid was transformed to E.coli BL21(DE) for the expression of protein.