Production and Characterization of Laccase from Botrytis cinerea 61-34

Area of Science:

• Biochemistry
• Enzymology
• Mycology

Background:

• Botrytis cinerea produces extracellular enzymes.
• Laccases are copper-containing oxidase enzymes with diverse applications.
• Constitutive enzyme expression can be advantageous for production.

Purpose of the Study:

• To purify and characterize extracellular laccase from Botrytis cinerea strain 61-34.
• To assess the enzyme's properties for potential biotechnological use.

Main Methods:

• Cultivation of Botrytis cinerea on a defined medium.
• Enzyme purification using hydrophobic interaction chromatography.
• Biochemical characterization including molecular weight, pI, optimal pH and temperature, and kinetic parameters.

Main Results:

• High yield (80 mg/L) of purified, homogeneous extracellular laccase obtained.
• The enzyme is a monomeric glycoprotein (M(infr) 74,000, pI 4.0) with 49% carbohydrate content.
• Optimal activity at pH 3.5 and 60°C, with a K(infm) of 100 µM for 2,6-dimethoxyphenol and specific activity of 9.1 mkat/mg.
• Demonstrated stability in SDS and urea solutions, and a broad substrate range.

Conclusions:

• A facile purification protocol yields substantial amounts of active Botrytis cinerea laccase.
• The enzyme's stability and catalytic properties support its potential in various biotechnology applications.
• Further research into specific industrial applications is warranted.