Purification of Pseudomonas sp. Lipase by Continuous Elution Electrophoresis Based on Pb2+ Precipitation Method
Introduction Lipases are enzymes that play a crucial role in various biochemical processes, particularly in the hydrolysis of lipids. Pseudomonas sp. lipase is a well-known enzyme that has been widely studied for its industrial applications, including in the food, pharmaceutical, and biofuel industries. Purification of lipases is essential to enhance their catalytic efficiency and stability for industrial applications. In this study, we aim to purify Pseudomonas sp. lipase using continuous elution electrophoresis based on the Pb2+ precipitation method.
Methods The purification process begins with the culture of Pseudomonas sp. to obtain the crude enzyme extract. The crude extract is then subjected to centrifugation to remove cell debris, followed by ammonium sulfate precipitation to concentrate the enzyme. Next, the enzyme is dialyzed against a buffer solution to remove excess salts. The Pb2+ precipitation method is then employed to selectively precipitate the lipase. The precipitate is dissolved in a suitable buffer and subjected to continuous elution electrophoresis using an electrophoretic apparatus. The purified lipase fractions are collected and analyzed for purity and enzyme activity.
Results The continuous elution electrophoresis based on the Pb2+ precipitation method resulted in the purification of Pseudomonas sp. lipase. The purity of the purified lipase was confirmed by SDS-PAGE analysis, which showed a single band corresponding to the molecular weight of the lipase. The specific activity of the purified lipase was also determined, indicating a significant increase compared to the crude extract.
Conclusion In conclusion, continuous elution electrophoresis based on the Pb2+ precipitation method is an effective technique for the purification of Pseudomonas sp. lipase. This method offers several advantages, including high purification efficiency, simplicity, and scalability. The purified lipase can be used for various industrial applications, including in the food, pharmaceutical, and biofuel industries.
