[P006] Poster

1:00 PM - 1:50 PM JST(4:00 AM - 4:50 AM UTC)

[P03-316] Enhancing the thermostability of yeast lipase by semi-rational design and its application in hydrolysis of fats

○Carol Ping Han¹, Yan Ping Lim¹, Elvis Chua¹, Albert Xue¹, Wen Shan Yew¹ (1. National University of Singapore (Singapore))

Keywords:

Lipases,Thermostability,Triglycerides,Hydrolysis

Lipases are versatile biocatalysts with significant industrial applications in food processing, biodiesel production, and pharmaceutical synthesis. However, the limited thermostability of many native lipases restricts their use in high-temperature industrial processes. This study aimed to enhance the thermostability of a yeast lipase from Pseudozyma antarctica through semi-rational design while maintaining its catalytic regiospecificity. Potential thermostability-determining residues were identified using sequence alignment against orthologs with high thermotolerance and introduced into CL21 via site-directed mutagenesis. Of the six amino acid residues examined combinatorially, the double mutant S68G V167I sufficed to confer improved activity at 90°C compared to wild type. To explore the possibility for further enhancement in thermostability, site-saturated mutagenesis was performed at the two critical residues. The engineered lipase variants indicated a synergistic effect on the two sites and the optimal mutant retained 32% residual activity compared to 12% for S68G V167I. The thermostable lipase was successfully applied in the hydrolysis of solid triglycerides, demonstrating improved performance in processing substrates at elevated temperatures. This finding highlights the potential of engineered yeast lipases for industrial fat hydrolysis applications.