ISOLATION AND CHARACTERIZATION LIPASE ENZYME OF THERMOPHILIC BACTERIA ISOLATE AL89

• Main Author: NUR AFIFAH (NIM:90516005), DEVIYANTHI
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/26544
• Institution: Institut Teknologi Bandung

Lipase (Triacylglicerol acylhydrolase, E.C. 3.1.1.3) is an important enzyme in the <br /> <br /> biotechnological industries such as food, dairy, detergents, and health industries. Lipase might <br /> <br /> be isolated from almost all living things, either higher organisms however microbes are the <br /> <br /> potential organisms as enzyme sources like of animals and plants or microorganisms such as <br /> <br /> fungi and bacteria. Lipase produced by bacteria is usually extracellular enzyme. In this study, <br /> <br /> the lipase was isolated from compost isolated microbes. The purposed at the study to obtain <br /> <br /> lipase from isolate AL89. Isolate AL89 has known closed to Pseudoxanthomonas taiwanensis. <br /> <br /> The crude extract of lipase was produced by incubating of the bacterial culture 55 °C for 19 <br /> <br /> hours with speed of 150 rpm. The crude extract was partially purified using acetone. <br /> <br /> Fractionation was carried out at concentrations of acetone 0-20 %, 20-40 % and 40-60 %. The <br /> <br /> specific activity was determined by the activity test of lipase hydrolysis with the substrate of <br /> <br /> para-nitrophenil palmitat (pNPP). Determination of protein content was used by Bradford <br /> <br /> method. The result showed that the higest specific activity is 0.0971 U/mg protein from the <br /> <br /> fraction of 0-20 % with incubation of 19 hours. Lipase from isolate AL89 showed pH optimum at <br /> <br /> 9 and 55 °C. In addition the enzyme substrate preference of para-nitrophenil laurat (pNPL). <br /> <br /> Using zymografi analysis showed that the active protein at the size of 70 kDa. Based on the <br /> <br /> results show that lipase from isolate AL89 is alkalotolerant and thermotolerant