This document summarizes a study investigating the enzymatic degradation of lignin model compounds by versatile peroxidase (VP). The key findings are: 1) VP can catalyze both the polymerization and depolymerization of lignin, with the reaction pathway depending on factors like pH, H2O2 concentration, and the functional groups on the lignin compounds. 2) Degradation of phenolic β-O-4 lignin dimers by VP proceeds through oxidation, followed by competing polymerization or depolymerization reactions depending on conditions. 3) The functional group at the 5 position of guaiacyl units influences whether polymerization or depolymerization occurs.

1. Understanding factors controlling depolymerization
and polymerization in catalytic degradation of ß-ether
linked model lignin compounds by versatile peroxidase
Outcomes
• In phenolic lignin dimers the VP first produces a neutral
radical via oxidation of the 4-OH position, followed by
polymerization and depolymerization reactions.
• Selection between polymerization and depolymerization
reaction pathways was found to be dependent on the
functional group at the 5 position of the guaiacyl group
(G5).
• The degradation pathway of phenolic β-O-4 was identified
as Cα-aryl cleavage rather than Cα-Cβ.
The effect of H2O2 concentration on the conversion of the
phenolic G-O-4 dimer (red) to monomer (blue) and insoluble
polymer (green) at pH 4.5 (left) and at pH 3.0 (right).
Zeng et al. (2017) ”Understanding factors controlling depolymerization and polymerization in catalytic degradation of
β-ether linked model lignin compounds by versatile peroxidase.” Green Chemistry, DOI: 10.1039/C6GC03379B
Background
• Enzyme catalyzed breakdown of lignin is hindered by the competition
between polymerization and depolymerization reaction pathways.
• Understanding the factors that drive these reactions toward
depolymerization is critical to developing processes for lignin
valorization.
Significance
• New insights into the reaction conditions and structural
features of lignin that facilitate its depolymerization to
smaller fragments
Versatile peroxidase catalyzes both
polymerization and depolymerization of lignin.
Approach
• Analyzed the effects of reaction conditions (pH, addition of H2O2 and
mediators) on the enzymatically catalyzed cleavage of several lignin
β-ether compounds using versatile peroxidase (VP) from B. adusta.
• Performed quantum chemistry calculations of free energy changes of
relevant chemical reactions and of electron spin density distributions
of radical species.
Depolymerization
Repolymerization
G
G S
OCH3
O
HO
OH
O
OCH3
OH
OH
OCH3
O
OCH3

2. Strategy for extending the stability of bio-oil
derived phenolic oligomer via mild hydrotreatment
with ionic liquid stabilized nanoparticles
Kim et al. (2017) “Strategy for extending the stability of bio-oil derived phenolic oligomer via mild
hydrotreatment with ionic liquid stabilized nanoparticles.” Chemsuschem, DOI: 10.1002/cssc.201601515
Background
• Development of catalytic transformations and
processes is essential to utilize bio-oil and lignin
derivatives
• Metal nanoparticles (NPs) stabilized in ionic liquid (IL)
are promising for catalytic hydrotreating of bio-oil and
phenolics
Approach
• Ruthenium NPs were synthesized with copolymers in
1-ethyl-3-methylimidazolium acetate
• Mild hydrotreating of phenolic oligomer was performed
in the presence of synthesized NP catalyst at 100 °C
for 6 hrs with the goal of producing a stable phenolics
Outcomes
• Hydrotreating of phenolic oligomer over NPs in IL
significantly increased aliphatic carbons, resulting in
alkylphenol units with improved thermal stability
• The catalyst system employed in this work was highly
effective in stabilizing reactive phenolic oligomer
Significance
• The findings of this work provide insight into
hydrotreating mechanisms of phenolic oligomer and
whole bio-oil, which will be useful for development of
sustainable processes in the future
OH
H3CO
OH
O
OCH3
O
OH
OCH3
O
N N
N
O
x y
OH
H3CO
OH
O
OCH3
OH
OH
OCH3
O
Mild hydrotrea ng
The formation of aliphatic carbon compounds during the hydrotreating of
phenolic oligomers in the presence of ILs and NPs

3. Expression of Aspergillus niger CAZymes is
determined by compositional changes in wheat
straw generated by hydrothermal or ionic liquid
pretreatments
Daly et al. (2017) “Expression of Aspergillus niger CAZymes is determined by compositional changes in wheat straw
generated by hydrothermal or ionic liquid pretreatments.” Biotechnology for Biofuels, DOI: 10.1186/s13068-017-0700-9
Background
• Fungi are major degraders of lignocellulose in nature and are
the main source of the costly enzymes used to saccharify pre-
treated lignocellulose in the production of second-generation
biofuels.
• There is limited understanding of the responses of fungi to
substrates that are pretreated as well as the temporal aspects
of the response.
Approach
• We investigate the response of Aspergillus niger to untreated
and pretreated substrates in a temporally extensive manner,
and are the first to report the fungal response to ionic liquid
pretreated substrates and the response of Aspergilli to
Miscanthus.
Outcomes
• The datasets demonstrate that pretreatment, substrate and
time each have major influence on the fungal responses to
lignocellulose.
• the transcript levels in A. niger correlated with the changes in
substrate composition brought about by the pretreatments
Significance
• Understanding these complex responses of fungi to
pretreated substrates facilitates identification of better
saccharifying enzymes and reduced enzyme production
costs.
a) The number of CAZy genes that encode plant-polysaccharide active CAZymes
and is significantly induced in cultures with untreated and pretreated straw
compared to the Glu 48 h b) The proportion of transcripts from CAZy genes that
encode plant-polysaccharide active CAZymes is expressed as FPKM value. c)
Subset of the MFuzz clusters from the clustering of all genes from the KMS
time-course with number of genes belonging to the indicated categories.