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ScienceDirect
Enzymatic conversion of lignin into renewable
chemicals
Timothy DH Bugg and Rahman Rahmanpour
The aromatic heteropolymer lignin is a major component of
plant cell walls, and is produced industrially from paper/pulp
manufacture and cellulosic bioethanol production. Conversion
of lignin into renewable chemicals is a major unsolved problem
in the development of a biomass-based biorefinery. The review
describes recent developments in the understanding of
bacterial enzymes for lignin breakdown, such as DyP
peroxidases, bacterial laccases, and beta-etherase enzymes.
The use of pathway engineering methods to construct
genetically modified microbes to convert lignin to renewable
chemicals (e.g. vanillin, adipic acid) via fermentation is
discussed, and the search for novel applications for lignin (e.g.
carbon fibre).
Address
Department of Chemistry, University of Warwick, Coventry CV4 7AL,
United Kingdom
Corresponding author: Bugg, Timothy DH (T.D.Bugg@warwick.ac.uk)

Current Opinion in Chemical Biology 2015, 29:1017

This review comes from a themed issue on Energy
Edited by Timothy DH Bugg and Michael Resch

http://dx.doi.org/10.1016/j.cbpa.2015.06.009
1367-5931/# 2015 Elsevier Ltd. All rights reserved.

The need to reduce global greenhouse gas emissions has

stimulated considerable interest in the generation of new
routes to fuels and chemicals from renewable sources,
especially from plant biomass [1]. For aromatic chemicals
used to make plastics, fine chemicals and materials, the
aromatic polymer lignin found in plant cell walls is
a readily available but challenging substrate [2,3]. This
article will review current research, challenges and prospects for bio-conversion of lignin into renewable chemicals.
Since the invention of the oil refinery in the late 19th
century, aromatic chemicals have been produced industrially from crude oil, mainly from catalytic reforming of
the naphtha fraction, and are used for a range of highvolume industrial applications [2], including plastics
from polystyrene (from styrene), polyethylene terephthalate (PET, from para-xylene), and phthalate resins
Current Opinion in Chemical Biology 2015, 29:1017

(from ortho-xylene). An aromatic component of 1520%

is needed in jet fuel, for which renewable sources are
needed [4]. As supplies of crude oil dwindle around the
world, alternative sustainable sources for these chemicals
must be found, for which the aromatic lignin polymer
found in plant biomass is an abundant raw material (see
Figure 1).

Lignin availability and structural types

Lignin is produced on a large scale from the pulp and
paper industry [5,6]. The Kraft process involves treatment of biomass with sodium hydroxide and sodium
sulphide, generating Kraft lignin (60100 ktons/yr) as a
by-product. The Sulfite process involves treatment with
aqueous sulphur dioxide, generating water-soluble lignosulfonates (1 Mtonne/yr). The soda lignin process
involves treatment with aqueous sodium hydroxide, generating a sulphur-free soda lignin (510 ktons/yr). The
Organosolv process uses an ethanolwater extraction
method to produce high purity cellulose, hemicellulose
and lignin streams from plant biomass, which generates
three ktons/yr Organosolv lignin (CIMV, Fr).
Potentially even larger amounts of lignin will be produced
from the industrial production of cellulosic bioethanol
production, which liberates lignin as a low-value byproduct. It has been estimated that the US bioethanol
industry alone will generate up to 60 Mtonne/yr lignin by
2022 [2], and large-scale bioethanol production sites at
Alagoas in Brazil (82 Ml/yr) and Crescentino in Italy
(20 Ml/yr) will also generate lignin-rich product streams.
Finding new product streams for lignin is therefore a
major unsolved challenge in creating a lignocellulosebased biorefinery.
However, it is also important to note that the chemical
properties of each type of lignin vary considerably.
Depending on the type of chemical pre-treatment used
in its production, industrial lignin may be highly
condensed, due to loss of benzylic hydroxyl groups in
the b-aryl ether structural unit, forming a benzylic cation
that can form new CC cross-linking bonds. Kraft lignin is
often highly condensed, and therefore contains greatly
reduced b-O-4 content, and also contains thiol groups
arising from the use of sodium sulphide (Na2S) used in
the Kraft process [3,5,7]. The Organosolv process uses a
milder biomass treatment method, hence Organosolv
lignin retains a higher proportion of b-O-4 linkages [8],
but to date the Organosolv process has only been commercialised at pilot scale [5]. There is also current interest
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Enzymatic conversion of lignin Bugg and Rahmanpour 11

Figure 1

Oil
Refinery

Fractions

Fuels

Chemicals

Petroleum C5 -C8

Petroleum
OH

Catalytic reforming

naphtha

Kerosene C6-C16

jet fuel

Diesel oil C8-C21

diesel

CO2H

CO2H

e
crude
oil
cellulose
O

OH

Bio-Refinery

O
HO

cellulosic
bioethanol

O
HO

OH

O
OH

OHC

CH2OH

OMe

Cellulosic
biorefinery

Biomass

CHO

OH

OH
CHO

O
HO
OH

OMe

vanillin
15 $/kg

OMe
OH

HO

OMe

Pulp/paper
manufacture

OMe
O

Biorefinery lignin
Kraft lignin
Lignosulfonate
Soda lignin

CO2H

ferulic acid
85 $/kg
OMe
OH
Current Opinion in Chemical Biology

Routes to aromatic chemicals from crude oil (a) and plant biomass (b).

in ionic liquid biomass treatment methods [9], which

generate an ionic liquid lignin preparation containing
reduced b-O-4 content, and some condensed structures
[10].

Challenges in lignin valorisation

There has been research into lignin valorisation since the
1980s [11], but there have been very few successful
examples of the conversion of lignin into higher value
aromatic products, due to a number of challenges, as
follows. First, Difficult bond cleavages. The linkages between aryl-C3 units in polymeric lignin are either CO
ether bonds, or CC bonds, which are not susceptible to
hydrolytic cleavage and hence chemically inert, so lignin
breakdown requires unusual chemistry or biochemistry.
Second, Physical properties. Native lignin from biomass,
and more native-like lignins such as Organosolv lignin,
are rather insoluble in water and organic solvents, though
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industrial Kraft lignins and lignosulfonates are more water-soluble. Access to the lignin polymer is therefore
challenging for both enzyme and chemical catalysts.
Third, Heterogeneous structure. Compared with cellulose,
the lignin polymer is inherently heterogeneous and irregular in structure, presenting a challenge for enzyme
biocatalysts, which are usually highly selective for a
particular substrate. Fourth, One lignin is not the same as
another lignin. The chemical structure of a lignin preparation is dependent on the method used to prepare the
lignin from biomass. In the case of industrial lignins, the
lignin structure is often condensed, making it more chemically inert, and the presence of sulfur in Kraft lignins
can poison chemical catalysts. Fifth, Repolymerisation vs.
depolymerisation. Often the catalysts for lignin depolymerisation will also catalyse the repolymerisation of radical
intermediates, resulting in competing repolymerisation (or
recondensation) of lignin fragments, generating higher
Current Opinion in Chemical Biology 2015, 29:1017

12 Energy

molecular weight products. Sixth, Generation of complex

mixtures of products. Since lignin is a complex, heterogeneous polymeric substrate, it is not surprising that chemical
or bio-catalytic processing of lignin would result in a rather
complex mixture of oligomeric fragments and low molecular weight products. Typically, complex mixtures of low
molecular weight aromatic products are produced by pyrolytic and chemocatalytic treatment of lignin, hence there is
a need for methods to simplify product mixtures by methods such as hydro-deoxygenation, to obtain a smaller
number of deoxygenated alkane [3].

Enzymes for lignin breakdown

The bio-degradation of lignin by lignin-degrading
microbes has been described as enzymatic combustion,
where the oxidising potential of hydrogen peroxide or
molecular oxygen by ligninolytic peroxidase enzymes or
laccase enzymes respectively is exploited to oxidise aromatic units [11]. White-rot fungi such as Phanerochaete
chrysosporium produce an arsenal of extracellular lignin
peroxidases, Mn peroxidases and laccases for lignin
breakdown, which have been extensively reviewed
[12,13]. The development of fungal biocatalysts for
large-scale lignin depolymerisation has been hampered
by the challenges of fungal protein expression, therefore,
in recent years there has been renewed interest in the
identification of bacterial biocatalysts for lignin depolymerisation [14]. Bacterial lignin degradation activity has
been best characterised in actinobacteria, a- and g-proteobacteria, and in some cases related bacteria have been
reported in the gut of wood-infesting termites and beetles
[14]. A recent review of lignin-degrading phenotypes and
genotypes lists 22 actinobacteria, 10 a-proteobacteria and
11 g-proteobacterial strains with lignin degradation phenotypes, but also seven firmicutes, four b-proteobacteria,
one d-proteobacterium, one bacteroides and one archaeal
strain [15].
Until recently, the enzymology of bacterial lignin degradation was not well understood [14]. Using a spectroscopic assay method, Ahmad et al. have shown lignin
degradation activity in the extracellular fraction of two
known aromatic degraders, Rhodococcus jostii RHA1 and
Pseudomonas putida [16]. Bioinformatic analysis of unannotated peroxidase genes in the R. jostii RHA1 genome
sequence led to the identification of two dye-decolorizing
peroxidase (DyP) genes dypA and dypB, with homologues
in other lignin-oxidising bacteria [17]. Gene deletion
studies revealed that a DdypB mutant shows significant
decreased lignin degradation activity, and recombinant
DypB catalyses oxidative CaCb cleavage of a b-aryl
ether lignin model compound, and also oxidises Mn2+
[17]. The structure of DypB from R. jostii RHA1 shows a
ferredoxin-like fold, similar to other DyP structures [18].
Replacement of active site Asn-246 in R. jostii DypB by
Ala has been found to increase the kcat for Mn2+ oxidation
80-fold, and a Mn2+ binding site has been identified by
Current Opinion in Chemical Biology 2015, 29:1017

X-ray crystallography, as shown in Figure 2a [19]. Remarkably, the R. jostii RHA1 DypB protein is packaged
into a nanocompartment formed by 60 protein subunits
encoded by an adjacent encapsulin gene, and encapsulation increases its lignin oxidation activity [20].
The DyPs, discovered nearly a decade ago, form a distinct
superfamily of peroxidases, due to their specific primary
and tertiary structures and unique reaction characteristics
[21]. Phylogenetic analyses have led to the classification
of DyPs into four subfamilies (AD), DyPs from bacteria
belong to the A, B and C subfamilies (see Figure 2b),
whereas fungal enzymes are found in the D subfamily
[22]. A DyP1B enzyme from Gram-negative Pseudomonas
fluorescens Pf-5 has been reported to show activity for
oxidation of Kraft lignin and Mn2+, and in the presence
of Mn2+ releases an oxidised lignin dimer from wheat
straw lignocellulose [23]. Heme-dependent lignin-oxidising enzymes have also been identified in soil bacterium
Amycolatopsis sp. 75iv2 ATCC 39116 [24]. A Dyp type C
peroxidase enzyme has been identified from the same
strain which shows Mn2+ oxidation activity with much
higher catalytic efficiency than R. jostii DypB, approaching the activity of fungal Mn peroxidase enzymes [25].
Laccase enzymes from fungi have been utilized in different areas such as wood and paper industries, environmental applications and biosensors, and have activity for lignin
oxidation [12,13], but few prokaryotic laccases have been
discovered. Bioinformatic analysis of >2200 complete
and draft bacterial genomes and four metagenomic datasets has identified >1200 putative bacterial genes for
laccase-like enzymes, of which 76% showed the existence
of putative signal sequences, implying that the encoded
enzymes might be exported from the cytoplasm [26].
Bacterial laccases are especially prevalent in actinobacteria [27], but are also found in a-, b- and g-proteobacteria
[15]. Laccase enzymes from Streptomyces coelicolor A3(2),
Streptomyces lividans TK24, Streptomyces viridosporus T7A,
and Amycolatopsis sp. 75iv2 have been characterised kinetically, and were found to catalyse Ca oxidation of
lignin model compounds [28]. Deletion of the laccase
gene from S. coelicolor A3(2) was found to significantly
diminish the ability to form acid-precipitable lignin
(APPL), supporting a role in lignin oxidation in vivo,
but treatment of ethanosolv lignin in vitro led to a higher
molecular weight product, due to competing repolymerisation of lignin fragments [28]. A multicopper oxidase
similar to Pseudomonas stutzeri CopA has also been identified from a novel screen of metagenomic DNA libraries
using an ermR reporter gene that responds to aromatic
lignin degradation products [29]. This multicopper oxidase was found to oxidise a polymeric lignin substrate to
generate 2,6-dimethoxybenzene-1,4-diol as a major product [29]. Fungal laccase enzymes have been shown to be
effective biocatalysts for delignification of wood and
biomass feedstocks, using a suitable laccase mediator
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Enzymatic conversion of lignin Bugg and Rahmanpour 13

Figure 2

(a)

(b)

0.115

0.128
0.136
0.190

0.022
0.019
0.306

A
0.000

Burkholderia terrae
Rhodococcus josti RHA1 DyPB
0 022 Rhodococcus wratislaviensis
0.044 Rhodococcus opacus
0.059
Pseudomonas fluorescens DyP2B
0.057
Pseudomonas chlororaphis O6
0.079
Pseudomonas brassicacearum
0.348
Pseudomonas fluorescens DyPA
0.233
Thermobifida fusca DyPA
0.244
Nocardiopsis dassonvillei
0.163
Rhodococcus RHA1 DyPA
0.183
Mycobacterium smegmatis
0.165
Amycolatopsis DyP2C
0.038
Amycolatopsis vancoresmycina
0.136
0.037
Amycolatopsis mediterranei
0.216
Rhizobium leguminosarum

0.025

0.069

0.078
0.032
0.161

0.025

0.239

Pseudomonas fluorescens DyP1B

0.125

0.024

0.220
Current Opinion in Chemical Biology

Bacterial DyP peroxidases. (a) Structure of Rhodococcus jostii RHA1 DypB, showing the location of the Mn2+ binding site (Glu-156, Glu-215,
Thr-231, Glu-239, in pink) close to the heme cofactor (in red) and catalytic residues Asp-153 and Arg-244 (in blue). (b). Phylogenetic cladogram for
bacterial DyP peroxidases, showing DyP groups A, B, and C; enzymes mentioned in the text are starred.

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Current Opinion in Chemical Biology 2015, 29:1017

14 Energy

[30], hence bacterial laccases might have interesting

applications for delignification, as well as lignin valorisation.
Bacterial b-etherase enzymes have also been recently
discovered that catalyse a glutathione-dependent cleavage reaction on lignin model compounds. This class of
b-etherase enzymes were previously identified in Sphingobium SYK-6, a strain capable of breakdown of a number
of lignin model compounds, and were shown to be
dependent on glutathione, whose nucleophilic thiol sidechain was used to cleave the ether linkage of b-aryl ether
model compounds [31]. Picart et al. have recently identified four b-etherase enzymes in Novosphingobium strains,
which catalyse b-ether cleavage on lignin model compounds, and also show activity towards a fluorescently
labelled polymeric lignin substrate [32]. Gall et al. have
also characterised b-etherase enzymes from Sphingobium
SYK-6 [33] and from two Novosphingobium strains [34],
demonstrating that different sub-classes show opposite
stereospecificity for the b-aryl ether b-carbon centre:
LigE-type enzymes are selective for the R-enantiomer,
while LigF-type enzymes are selective for the S-enantiomer
[33,34].
Table 1 summarises the properties of the currently known
bacterial enzymes for lignin breakdown, only some of
which are reported to be active with polymeric lignin,
rather than lignin model compounds. It seems likely that
a group of oxidative enzymes (and accessory proteins) are
needed to break down such a complex substrate, so new
genome sequence data and metagenomic data are likely
to identify further enzymes. The genome of an Enterobacter lignolyticus strain which shows activity for lignin
degradation under anaerobic conditions contains many
glutathione S-transferase enzymes which might be related to the LigEF b-etherase enzymes, as well as two

putative laccases [35], while the genome of a Cupriavidus

basilensis B-8 strain which oxidises Kraft lignin contains
one putative laccase, and a range of aromatic degradation
gene clusters [36].

Manipulating lignin degradation pathways to

produce fuels and chemicals
As an alternative to using enzymes in vitro to convert
polymeric lignin into chemicals, pathway engineering
could potentially be used to engineer the lignin degradation pathways of a microbe that possesses the metabolic
capability to break down lignin, in order to produce useful
chemicals. Some recent examples suggest that this may
be a practical way forward. While our knowledge of
microbial lignin degradation pathways is still incomplete
[37], it seems likely that the initial lignin oxidation
products are processed via downstream aromatic degradation pathways that are well understood, such as the
b-ketoadipate pathway involving intradiol cleavage of
protocatechuic acid to 3-carboxymuconic acid, as shown
in Figure 3 [38]. Protocatechuic acid and catechol are
also metabolised via extradiol cleavage in some bacteria
[37,38], hence these metabolites are key branchpoints for
aromatic degradation.
There are indications that vanillic acid is an intermediate
in G unit breakdown by lignin-degrading bacteria, since it
has been detected as a metabolite from lignin breakdown
[14,37]. Sainsbury et al. have shown deletion of the
vanillin dehydrogenase gene on the vanillin degradation
pathway in R. jostii RHA1 gave a mutant strain which
accumulated vanillin, a high value chemical used in the
food/flavour industry, at up to 96 mg/L after six days when
grown on minimal media containing 2.5% wheat straw
lignocellulose, together with 4-hydroxybenzaldehyde and
ferulic acid [39], as shown in Figure 3. Primary metabolism can also be used to generate useful bioproducts:

Table 1
Summary of bacterial enzymes for lignin breakdown. NR, not reported
Enzyme

Bacterium

Cofactor

Co-substrate

Low MW
substrates

Lignin model compounds

Rhodococcus
jostii RHA1
Pseudomonas
fluorescens
Amycolatopsis
sp 75iv2
Streptomyces
coelicolor
Pseudomonas
stutzeri
Sphingobium
SYK6
Novosphingobium
sp.

Heme Fe

H2O2

ABTS, Mn(II)

b-O-4

Heme Fe

H2O2

ABTS, Mn(II)

NR

Heme Fe

H2O2

ABTS, Mn(II)

b-O-4

NR

NR

Cu

O2

ABTS, DMP

b-O-4

Ca oxidation

Ethanosolv

Higher MW

[28]

Cu

O2

ABTS, DMP

b-O-4

NR

HP lignin

Aromatic
monomer

[29]

Glutathione

b-O-4

NR

[33]

Glutathione

b-O-4

b-Ether
cleavage
b-Ether
cleavage

Fluorescent lignin

[32]

Substrate
DypB

Dyp2
Laccase
CopA
Etherase

Current Opinion in Chemical Biology 2015, 29:1017

Polymeric lignin

Reaction
CaCb
cleavage

Substrate

Products
[17]

Kraft lignin
Lignocellulose

Ref

Lignin dimer

[23]
[25]

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Enzymatic conversion of lignin Bugg and Rahmanpour 15

Figure 3

CO2H

Hemicellulose

OCH3
OH

OH
HO

OAr

G type
lignin

CHO

CO2H

OCH3
OH

OH

OAr

H type
lignin

CO2H

OCH3

protocatechuate
3,4-dioxygenase

OH

CO2H

OH

CO2H
CO2H

OH

OH

OH
catechol

H2/cat
HO2C

CO2H

O
triacylglycerol
lipids

CO2H

cis,cismuconic acid

X
OH

CO2H

CO2H
CO2H
CO2H

OH

OH

vanillin

CHO

HO

-ketoadipic
acid

protocatechuic
acid

OCH3
OH

-oxidation

TCA cycle

O
O

O
O

polyhydroxy
butyrate

SCoA

C8H17 O
O

malonyl CoA

O
O
n
Current Opinion in Chemical Biology

Production of bioproducts from microbial degradation pathways, illustrating schematically the metabolic pathways responsible for accumulation of
small molecule products vanillin or cis,cis-muconic acid via gene deletion (in blue), and the accumulation of triacylglycerol lipids or
polyhydroxybutyrate via primary metabolism (in green).

Rhodococcus opacus DSM1069 and PD630 are known to

accumulate triglyceride lipids from primary metabolism
under nitrogen-limiting conditions, and have been shown
to grow on minimal media containing ultrasonicated
ethanol organosolv lignin as carbon source, generating
triglyceride lipids at 4 mg/g us-EOL (or 20 mg/L) after a
nine day fermentation [40].
P. putida is a well-known aromatic degrader that also
possesses activity for lignin breakdown [16]. Linger
et al. have shown that the ability of P. putida to accumulate
polyhydroxyalkanoate (PHA) biopolyesters under nitrogen-limiting conditions can be harnessed to convert lignin
from alkaline pre-treated liquor (APL) into PHAs in a
48 h fermentation [41]. Pathway engineering has also
been used in P. putida to accumulate cis,cis-muconic acid
from aromatic degradation, via blockage of the protocatechuate cleavage pathway, and re-routing via catechol
cleavage [42]. Using p-coumaric acid as a carbon source, a
yield of 13.5 g/L was obtained in a fed-batch bioreactor
after 78 h, whereas using alkali-APL a yield of 0.7 g/L was
obtained after 24 h [42]. Cis,cis-muconic acid can then be
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converted via catalytic hydrogenation into adipic acid,

thereby providing a renewable route to adipic acid from
lignin [42]. As we improve our understanding of lignin
degradation, it should be possible to use pathway engineering in suitable microbial hosts to generate other
useful products from microbial lignin breakdown, hence
it is interesting that replacement of the genes encoding
the catechol intradiol pathway in P. putida by genes
encoding a corresponding extradiol pathway leads to an
increase in yield of pyruvate from aromatic substrates
[43]. A selective hydroxamic acid inhibitor of protocatechuate 3,4-dioxygenase has also been developed, as an
alternative chemical approach to manipulating bacterial
lignin degradation [44].

Industrial applications for lignin

There is interest in finding new applications for polymeric lignins, as well as depolymerised lignin breakdown
products. Jin et al. have used lignin derived from corn
bioethanol production to make phenol-formaldehyde adhesive [45], while Ramires et al. have used Organosolv
lignin from sugarcane bagasse to prepare phenolic resins
Current Opinion in Chemical Biology 2015, 29:1017

16 Energy

[46]. In principle polymeric lignin could be a template for

synthesis of new biomaterials. It has been shown that
Kraft lignin can be modified by atom transfer radical
polymerisation (ATRP), changing the thermostability
and water solubility of the polymer [47]. Polyurethane
foams can also be made from either Organosolv or Kraft
lignin, with stronger foams being formed from Organosolv
lignin [48]. There is interest in the conversion of lignin
into carbon fibre, however, there are challenges in generating high quality carbon fibre, notably that highly purified lignin fractions are needed, due to the heterogeneity
of most lignin preparations [49]. In a related application,
lignin has also been converted into porous carbon, suitable for supercapacitor electrode materials [50]. The high
carbon content of lignin makes it potentially interesting
for conversion into nanomaterials, and Yiamsawas et al.
have recently shown that lignin nanocontainers can be
formed from water-soluble lignosulfonates in inverse
micro-emulsions [51].

Acknowledgement
The authors thank the University of Warwick for funding a Chancellors
Scholarship to R.R.

References and recommended reading

Papers of particular interest, published within the period of review,
have been highlighted as:
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8.

Bauer S, Sorek H, Mitchell VD, Ibanez AB, Wemmer DE:

Characterization of Miscanthus giganteus lignin isolated
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Mora-Pale M, Meli L, Doherty TV, Linhardt RJ, Dordick JS: Room

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Current Opinion in Chemical Biology 2015, 29:1017

24. Brown ME, Walker MC, Nakashige TG, Iavarone AT, Chang MCY:
Discovery and characterization of heme enzymes from
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25. Brown ME, Barros T, Chang MCY: Identification and

characterization of a multifunctional dye peroxidase from a
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The Amycolatopsis sp 75iv2 Dyp2 shows much higher activity for Mn(II)
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Gerlt JA: Roles of small laccases from Streptomyces in lignin
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This paper characterises in detail the activity of some bacterial laccase
enzymes, and demonstrates that they have some role in lignin degradation.
www.sciencedirect.com

Enzymatic conversion of lignin Bugg and Rahmanpour 17

29. Strachan CR, Singh R, VanInsberghe D, Ievdokymenko K,

Budwill K, Mohn WW, Eltis LD, Hallam SJ: Metagenomic
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32. Picart P, Muller C, Mottweiler J, Wiermans L, Bolm C, Dominguez

de Maria P, Schallmey A: From gene towards selective biomass
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lignin-like polymers. ChemSusChem 2014, 7:3164-3171.
This paper (and the following reference) demonstrates that b-etherase
enzymes could be useful biocatalysts for lignin breakdown.
33. Gall DL, Kim H, Lu F, Donohoe TJ, Noguera DR, Ralph J:

Stereochemical features of glutathione-dependent enzymes
in the Sphingobium sp. strain SYK-6 b-aryl etherase pathway.
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This paper highlights the stereospecificity of the b-etherase enzyme family.
34. Gall DL, Ralph J, Donohoe TJ, Noguera DR: A group of sequencerelated sphingomonad enzymes catalyzes cleavage of b-aryl
ether linkages in lignin b-guaiacyl and b-syringyl ether dimers.
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35. De Angelis KM, DHaeseleer P, Chivian D, Fortney JL,
Khudyakov J, Simmons B, Woo H, Arkin AP, Davenport KW,
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36. Shi Y, Chai L, Tang C, Yang Z, Zhang H, Chen R, Chen Y, Zheng Y:
Characterization and genomic analysis of kraft lignin
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37. Bugg TDH, Ahmad M, Hardiman EM, Rahmanpour R: Pathways
for degradation of lignin in bacteria and fungi. Nat Prod Rep
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38. Bugg TDH, Winfield CJ: Enzymatic cleavage of aromatic rings:
mechanistic aspects of the catechol dioxygenases and
later enzymes of bacterial aromatic degradation pathways.
Nat Prod Rep 1998, 15:513-530.
39. Sainsbury PD, Hardiman EM, Ahmad M, Otani H, Seghezzi N,
 Eltis LD, Bugg TDH: Breaking down lignin to high-value
chemicals: the conversion of lignocellulose to vanillin in
a gene deletion mutant of Rhodococcus jostii RHA1.
ACS Chem Biol 2013, 8:2151-2156.
This was the first demonstration that microbial lignin degradation pathways could be manipulated to produce aromatic chemicals from lignin via
fermentation.

www.sciencedirect.com

40. Kosa M, Ragauskas AJ: Lignin to lipid bioconversion by

oleaginous Rhodococci. Green Chem 2013, 15:2070-2074.
41. Linger JG, Vardon DR, Guarneri MT, Karp EM, Hunsinger GB,

Franden MA, Johnson CW, Chupka G, Strathmann TJ, Pienkos PT,
Beckham GT: Lignin valorization through integrated biological
funnelling and chemical catalysis. Proc Natl Acad Sci U S A
2014, 111:12013-12018.
This was a nice demonstration of how bacterial degradation of lignin
could be harnessed, via primary metabolism, to produce valuable biosynthetic products.
42. Vardon DR, Franden MA, Johnson CW, Karp EM, Guarneri MT,

Linger JG, Salm MJ, Strathman TJ, Beckham GT: Adipic
acid production from lignin. Energy Environ Sci 2015,
8:617-628.
A nice example of pathway engineering in Pseudomonas putida to
produce a metabolite that can be converted chemically into adipic acid.
43. Johnson CW, Beckham GT: Aromatic catabolic pathway
selection for optimal production of pyruvate and lactate from
lignin. Metab Eng 2015, 28:240-247.
44. Sainsbury PD, Mineyeva Y, Mycroft Z, Bugg TDH: Chemical
intervention in bacterial lignin degradation pathways:
development of selective inhibitors for intradiol and extradiol
catechol dioxygenases. Bioorg Chem 2015, 60:102-109.
45. Jin Y, Cheng X, Zheng Z: Preparation and characterization of
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hydrolysis lignin. Biores Technol 2010, 101:2046-2048.
46. Ramires EC, Megiatto JD Jr, Gardrat C, Castellan A, Frollini E:
Valorization of an industrial organosolv-sugarcane bagasse
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composites reinforced with sisal fibers. Biotech Bioeng 2010,
107:612-621.
47. Kim YS, Kadla JF: Preparation of a thermoresponsive ligninbased biomaterial through atom transfer radical
polymerization. Biomacromolecules 2010, 11:981-988.
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50. Jeon J-W, Zhang L, Lutkenhaus JL, Laskar DD, Lemmon JP,
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et al.: Controlling porosity in lignin-derived nanoporous
carbon for supercapacitor applications. ChemSusChem 2015,
8:428-432.
51. Yiamsawas D, Baier G, Thines E, Landfester K, Wurm FR:
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4:11661-11663.

Current Opinion in Chemical Biology 2015, 29:1017