Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis

Area of Science:

• Biochemistry
• Molecular Biology
• Enzymology

Background:

• Cold-active lipases are crucial for industrial processes operating at low temperatures.
• Yarrowia lipolytica is a known source of industrially relevant enzymes.

Purpose of the Study:

• To clone and express the extracellular lipase gene LIPY8 from Yarrowia lipolytica.
• To characterize the biochemical properties of the recombinant lipase (LipY8p).

Main Methods:

• Gene cloning and expression using a baculovirus system.
• Purification of recombinant lipase via chromatographic techniques.
• Enzyme activity assays at various pH, temperatures, and substrate conditions.

Main Results:

• The recombinant lipase LipY8p was purified with a 25.7-fold increase in specific activity (1102.9 U/mg).
• The enzyme has a molecular mass of 40 kDa and optimal activity at pH 7.5 and 17°C.
• LipY8p shows maximum activity towards medium-chain (C10) esters and is influenced by metal ions, detergents, and organic solvents.

Conclusions:

• The characterized cold-active lipase LipY8p possesses properties suitable for biotechnological applications.
• Its stability and activity profile suggest utility in fine chemical synthesis, food processing, and detergent formulations.