Laccase multi-point covalent immobilization: characterization, kinetics, and its hydrophobicity applications

• Published in: Applied microbiology and biotechnology Vol. 107; no. 2-3; pp. 719 - 733
• Main Authors: Othman, Abdelmageed M., Sanromán, Ángeles, Moldes, Diego
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-02-2023; Springer; Springer Nature B.V
• Subjects: Biomedical and Life Sciences; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; Contact angle; Covalence; Enzymatic activity; Enzyme activity; Enzyme Stability; Enzymes; Enzymes, Immobilized - metabolism; Hydrogen-Ion Concentration; Hydrophobicity; Immobilization; Inactivation; Kinetics; Laccase; Laccase - metabolism; Life Sciences; Methods; Microbial Genetics and Genomics; Microbiology; Parameters; pH effects; Substrate preferences; Substrates; Thermodynamics; Water - chemistry; Wood; Spain; Thermodynamics; Wood hydrophobization; Immobead 150P; Covalent immobilization; Grafting; Laccase
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-022-12352-9

Laccase from
Myceliophthora thermophila
was immobilized using one-point and multi-point covalent attachment on both a native and a modified new commercial epoxy carrier (Immobead 150P). After 10 cycles of operation at pH 3.0 and temperature 70 °C, the multi-point covalently immobilized laccase on the modified Immobead 150P performed best in terms of immobilization characteristics, retaining 95% of its initial activity. Thermodynamic parameters of thermal inactivation emphasized the positive impact of the immobilization procedure. At 50 °C, the immobilized and free enzyme activity levels dropped by 27 and 73%, respectively, after 48 h of incubation. The immobilized enzyme enhanced its stability in alkaline conditions, resuming 95% of its original activity after 3 h at pH 9.0. Immobilization reduced substrate affinity because the free laccase’s
K
m
value was lower than that of the immobilized laccase. Finally, the application of immobilized laccase in an innovative wood treatment process was tested by grafting lauryl gallate (LG) in order to provide hydrophobic properties to the wood. The results showed a relative water contact angle of 85.7% for treated wood, whereas the control showed only 26.6%, after 4 min of contact between water and beechwood surface.
Key points
• Multi-point covalent immobilization of a commercial laccase on a commercial support.
• Enzymatic parameters generally improved by immobilization process.
• New application of immobilized laccase: enzymatic-assisted wood hydrophobization.