Abstract
In order to determine the potential of biochemical and structural features of Elaeis guineensis Jacq. oil palm mesocarp lipases, the LIP2 gene was isolated, expressed, purified and characterized through the Escherichia coli microbial recombinant system. Gene analysis of LIP2 revealed that it is composed of 1584 base pairs which are encoded in 528 amino acid residues with a molecular weight of around 57 kDa. LIP2 has distinctive lipolytic properties in terms of α/β fold and the catalytic triad for lipase. The LIP2 lipase was successfully expressed and purified from E. coli Rosetta (DE3) via affinity chromatography. The optimal temperature and pH for the lipase activity was 30 °C and a pH of 9, respectively. Stability was profoundly increased with the addition of metal ions (Ca2+, Mg2+, Mn+, and Ni+), along with organic solvents (ethanol and octanol). pNP myristate was the most suitable among all pNP esters. In biophysical characterization analysis, LIP2 has a thermal denaturing point at 66 °C, which mostly consists of random patterns (39.8%) followed by α-helix (30.3%), turns (23.8%) and β-sheet (6.2%). From the successful purification and characterization, the potential of oil palm mesocarp lipase was able to be further explored.
Highlights
Lipase is a versatile enzyme commonly used in industry, with applications as varied as yogurt and cheese fermentation, baking, laundry detergents, biocatalyst and alternative products [1]
The active site of a lipase contains a catalytic triad consisting of Serine—Histidine—Aspartic acid, but unlike most serine proteases, the active site is buried inside the structure covered with a lid
BLASTp Protein Data Bank (PDB)-based results revealed that the LIP2 lipase is homologous to most of the α/β hydrolase family
Summary
Lipase is a versatile enzyme commonly used in industry, with applications as varied as yogurt and cheese fermentation, baking, laundry detergents, biocatalyst and alternative products [1]. The protein structure of the lipases from plants has not been widely studied due to the low protein expression and laborious purification to produce, which tends to cause less purified protein compared to lipases expressed in microorganisms. Purification of plant lipases may cause damage and undesirable effects on the purified lipase protein through the downstream processing, since it is quite complex and tedious to extract plant lipases without getting other compounds such as high lipid and phenolic contents. These can cause conformation changes which eventually lead to different results in characterization and structural studies, especially in an analysis of their potential application in industry. Many plant lipases have been studied recently involving various methods of purification and characterization
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