ISOLATION, PARTIAL PURIFICATION, AND CHARACTERIZATION OF LIPASE FROM WHEAT GERM

• Main Author: Hashita, Putie
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/34584
• Institution: Institut Teknologi Bandung

Lipase is an enzyme which hydrolyzes triglycerides at the interface phases. As biocatalyst, lipases have been widely used in industry. Based on literature study wheat germ contains lipase enzyme. Wheat germ is a by-product of flour milling industry that has not been utilized yet. The goals of this study is to increase wheat germ value by using it as a raw material in the isolation of lipase and determining the character of the lipase. Isolation was carried out by grinding, acetone wash, and addition of HCl to obtain a crude extract. Partial purification was carried out by ammonium sulfate fractionation with three saturation levels of 0-35 %, 35-70 % and 70-100 %. Electrophotogram at Native-PAGE showed several protein bands. Protein concentration and lipase activity assays were performed to determine the fraction with the highest specific activity of lipase. The result show that 35-70 % fraction has the highest specific activity of 1,938 units/gram. This fraction showed optimum pH at 8 and 10. And Optimum temperature at 40 oC and 50 oC. Zymography of SDS-PAGE renaturation analysis showed that the active lipases have molecular weight at 17kDa and 34kDa.