Mining and Engineering a Thermostable Lipase from Baijiu Daqu

Area of Science:

• Enzymology
• Protein Engineering
• Biotechnology

Background:

• Thermostable lipases are crucial for industrial applications, especially in the food sector.
• Identifying and engineering lipases with enhanced thermal stability is a key area of research.
• Baijiu Daqu is a potential source for novel enzymes with desirable properties.

Purpose of the Study:

• To identify and characterize a thermotolerant lipase (rTaLip) from Baijiu Daqu.
• To engineer thermostable mutants of rTaLip with improved enzymatic properties.
• To elucidate the structural basis for enhanced stability and activity.

Main Methods:

• Lipase identification and expression in Komagataella phaffii.
• Rational protein engineering to create thermostable mutants.
• Enzymatic property determination and molecular dynamics simulations.
• Application testing in palm oil hydrolysis.

Main Results:

• Two thermostable lipase mutants (V219C/D222C/S73F/V141T and V219C/D222C/S73F/V141D) were successfully engineered.
• Mutants displayed 2.2-fold and 4.1-fold longer half-lives at 65 °C compared to wild-type.
• Catalytic efficiency increased by 455% and 119% in the mutants.
• One mutant enhanced palm oil hydrolysis efficiency from 63.01% to 88.77%.

Conclusions:

• Rational engineering significantly enhances lipase thermostability and catalytic activity.
• Combining enzyme discovery from traditional sources with protein engineering is effective for industrial enzyme development.
• Engineered lipases show promise for applications in harsh industrial environments, such as food processing.