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Catalysts, Vol. 14, Pages 887: Enhanced Thermostability of Laccase from Myceliophthora thermophila Through Conjugation with mPEG-SC
Catalysts doi: 10.3390/catal14120887
Authors: Leonardo L. O. García Raissa H. S. Florindo Vivian Saez Robert Wojcieszak Jose Ramon Ivaldo Itabaiana Jr.
The search for more sustainable reaction conditions has been necessary to obtain more selective processes. In this context, laccases have gained great notoriety in recent years. However, these enzymes are unstable in organic solvents and have low thermal stability. Alternatively, conjugation with PEG (PEGylation) can be essential to overcome these problems. In this work, the commercial laccase from Myceliophthora thermophila (LacMT) was subjected to PEGylation with PEG functionalized as succinimidyl carbonate (mPEG-SC), followed by assessing its thermal stability and catalytic activity. Mono-PEGylated LacMT derivatives were obtained, with less than 50% of the enzyme remaining in its native form. In addition, 10% of the bi-PEGylated species was successfully obtained according to gel electrophoresis analysis. The PEGylated derivatives showed a significantly reduced ABTS oxidation activity (98 ± 3 U/mg) compared to the native LacMT (407 ± 9 U/mg) but higher than the control enzyme without PEGylation (51 ± 2 U/mg), demonstrating that the addition of activated PEG to the protein resulted in better protection against the harmful action of the pH change required in the process. PEGylated LacMT retained more than twice the initial activity of the native protein at 40 °C during 24 h. In addition, PEGylated LacMT exhibited kinetic changes, whereas the catalytic turnover rate (kcat) of the PEGylated enzyme was reduced by 27% compared to the control. These findings are being reported for the first time. This sets precedents for constructing efficient catalytic systems involving laccases since no immobilized biocatalyst or commercial conjugate contains these proteins.
