Understanding pulp delignification by Laccase−Mediator Systems through isolation and characterization of lignin−carbohydrate complexes

The effects and mechanism of pulp delignification by laccases in the presence of redox mediators have been investigated on unbleached eucalyptus kraft pulp treated with laccases from Pycnoporus cinnabarinus (PcL) and Myceliophthora thermophila (MtL) and 1-hydroxybenzotriazole (HBT) and methyl syringate (MeS) as mediators, respectively. Determination of the corrected κ number in eucalyptus pulps after the enzymatic treatments revealed that the PcL−HBT system exhibited a more remarkable delignification effect than the MtL−MeS system. To obtain further insight, lignin−carbohydrate complexes were fractionated and subsequently characterized by nuclear magnetic resonance, thioacidolysis (followed by gas chromatography and size exclusion chromatography), and pyrolysis−gas chromatography−mass spectrometry (pyrolysis−GC−MS) analyses before and after the enzymatic treatments and their controls. We can conclude that the laccase−mediator treatments altered the lignin structures in such a way that more lignin was recovered in the xylan−lignin fractions, as shown by Klason lignin estimation, with smaller amounts of both syringyl (S) and guaiacyl (G) uncondensed units, as shown by thioacidolysis and gas chromatography, especially after the PcL−HBT treatment. The laccase−mediator treatment produced oxidation at Cα and cleavage of Cα and Cβ bonds in pulp lignin, as shown by pyrolysis−GC−MS. The general mechanism of residual lignin degradation in the pulp by laccase−mediator treatments is discussed in light of the results obtained.This study was financially supported by the EU BIORENEW Project (NMP2-CT-2006-026456). Novozymes (Bagsvaerd, Denmark) and Beldem (Andenne, Belgium) are acknowledged for the M. thermophila and P. cinnabarinus laccases, respectively. J. Romero (ENCE) is acknowledged for providing the eucalyptus pulp samples and E. D. Babot for performing the enzymatic treatments. J.R. acknowledges the CSIC JAE-Doc contract of the program “Junta para la Ampliación de Estudios” cofinanced by the European Social Fund (ESF).Peer reviewe