ISOLATION, PURIFICATION AND CHARACTERIZATION OF LIPASE FROM Staphylococcus aureus AND Staphylococcus

• Main Author: Telussa, Ivonne
• Format: Theses
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/35455
• Institution: Institut Teknologi Bandung

Lipase is an important enzyme in the field of biotechnology. This enzyme is used widely in industries, such as in food, milk, detergent, paper pulp, and pharmaceuticals. Several types of lipase-producing microorganisms are potential to be used as a source of lipase. This study conducted the isolation of lipase-producing bacteria from coco butter subtitute and biochemical characterization of the lipase. The isolate of lipace-producing bacteria used in this study was Staphylococcus epidermisis, while the Staphylococcus aureus used as a comparator that produced an lipase and was active to lipid at high temperature. Staphylococcus epidermisis and Staphylococcus aureus grew very rapidly at a temperature of 37 º C and lipase produced shown maximum activity at the end of the exponential phase. Lipase from Staphylococcus aureus and Staphylococcus epidermisis was an extracellular enzyme excreted into the medium for bacterial growth. This enzyme was partially purified using ammonium sulfate fractionation to produce five fractions, namely the fraction of 0-20%, 20-40%, 40-60%, 60-80% and 80-100%. The activity tests using the substrate para-nitrophenyl palmitate revealed that the fraction with highest activity for Staphylococcus epidermisis was a fraction of 20-40%, and for Staphylococcus aureus was in the fraction of 0-20%. The results of biochemical characterization shown that the lipase of Staphylococcus epidermisis and Staphylococcus aureus have optimum activity at a temperature of 70 º C and pH 7. Further, the analysis of molecular masses using Sodium dedosil sulfate polyacrylamide gel electrophoresis (SDS PAGE) on proteins precipitated with 0-20% ammonium sulphate shown two protein bands of Staphylococcus aureus with a molecular mass of 41 and 49 kDa, and one protein band with molecular mass of 56 kDa identified for proteins precipitated with 20-40% ammonium sulfate from Staphylococcus epidermisis. Based on this result, it can be concluded that lipase produced by Staphylococcus epidermisis and Staphylococcus aureus bacterias was different.