Sei sulla pagina 1di 14

Biotechnology Advances 24 (2006) 500 – 513

Research review paper

Industrial and biotechnological applications of laccases: A review
Susana Rodríguez Couto ⁎, José Luis Toca Herrera ⁎
Department of Chemical Engineering, Rovira i Virgili University, Av. Països Catalans 26, 43007 Tarragona, Spain
Received 20 January 2006; received in revised form 29 March 2006; accepted 1 April 2006
Available online 18 April 2006


Laccases have received much attention from researchers in last decades due to their ability to oxidise both phenolic and non-
phenolic lignin related compounds as well as highly recalcitrant environmental pollutants, which makes them very useful for their
application to several biotechnological processes. Such applications include the detoxification of industrial effluents, mostly from the
paper and pulp, textile and petrochemical industries, use as a tool for medical diagnostics and as a bioremediation agent to clean up
herbicides, pesticides and certain explosives in soil. Laccases are also used as cleaning agents for certain water purification systems, as
catalysts for the manufacture of anti-cancer drugs and even as ingredients in cosmetics. In addition, their capacity to remove
xenobiotic substances and produce polymeric products makes them a useful tool for bioremediation purposes. This paper reviews the
applications of laccases within different industrial fields as well as their potential extension to the nanobiotechnology area.
© 2006 Elsevier Inc. All rights reserved.

Keywords: Food industry; Industrial applications; Laccase; Nanobiotechnology; Pulp and paper industry; Textile industry


1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 501
2. Potential industrial and biotechnological applications of laccase enzyme . . . . . . . . . . . . . . . . . . . . . . . 501
2.1. Food industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 501
2.2. Pulp and paper industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 501
2.3. Textile industry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 505
2.4. Nanobiotechnology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 505
2.5. Other laccase applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 508
2.5.1. Soil bioremediation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 508
2.5.2. Synthetic chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 508
2.5.3. Cosmetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 508
3. Future outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 508
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 508
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 508

⁎ Corresponding authors. Tel.: +34 977 55 9617; fax: +34 977 55 9667.
E-mail addresses: (S. Rodríguez Couto), (J.L. Toca Herrera).

0734-9750/$ - see front matter © 2006 Elsevier Inc. All rights reserved.
S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513 501

1. Introduction spite that LMS has been studied extensively there are still
unsolved problems concerned with mediator recycling,
Although oxidation reactions are essential in several cost and toxicity.
industries, most of the conventional oxidation technol- Laccases have been reviewed several times in recent
ogies have the following drawbacks: non-specific or years, generally with emphasis on narrow aspects. The
undesirable side-reactions and use of environmentally reviews by Messerschmidt (1993, 1997) and by
hazardous chemicals. This has impelled the search for Solomon et al. (1996) provide excellent summaries of
new oxidation technologies based on biological systems the enzymology and electron transfer mechanism of the
such as enzymatic oxidation. These systems show the laccases and a book edited by Messerschmidt (1997)
following advantages over chemical oxidation: enzymes contains a series of articles dealing with different aspects
are specific and biodegradable catalysts and enzyme of laccase kinetics and mechanism of action and the
reactions are carried out in mild conditions. possible roles of this enzyme. The aim of this review is
Enzymatic oxidation techniques have potential within to highlight the potential industrial and biotechnological
a great variety of industrial fields including the pulp and applications of laccase enzyme.
paper, textile and food industries. Enzymes recycling on
molecular oxygen as an electron acceptor are the most 2. Potential industrial and biotechnological applica-
interesting ones. Thus, laccase (benzenediol: oxygen tions of laccase enzyme
oxidoreductase; EC is a particularly promising
enzyme for the above-mentioned purposes. The laccase Table 1 shows different applications of laccases in
molecule is a dimeric or tetrameric glycoprotein, which the last two decades. Laccases find applications within
usually contains four copper atoms per monomer the following fields:
distributed in three redox sites (Gianfreda et al., 1999).
This enzyme catalyses the oxidation of ortho and 2.1. Food industry
paradiphenols, aminophenols, polyphenols, polyamines,
lignins and aryl diamines as well as some inorganic ions Laccases can be applied to certain processes that
coupled to the reduction of molecular dioxygen to water enhance or modify the colour appearance of food or
(Yaropolov et al., 1994; Solomon et al., 1996). beverage. In this way, an interesting application of
The reported redox potentials of laccases are lower laccases involves the elimination of undesirable phe-
than those of non-phenolic compounds, so these enzymes nolics, responsible for the browning, haze formation and
cannot oxidise such substances. However, it was shown turbidity development in clear fruit juice, beer and wine.
that in the presence of small molecules capable to act as Laccases are currently of interest in baking due to its
electron transfer mediators laccases were also able to ability to cross-link biopolymers. Thus, Selinheimo et
oxidise non-phenolic structures (Bourbonnais and Paice, al. (2006) showed that a laccase from the white-rot
1990; Call and Mücke, 1997), expanding, thus, the range fungus Trametes hirsuta increased the maximum
of compounds that can be oxidised by these enzymes. resistance of dough and decreased the dough extensi-
Laccase-mediated systems (LMS) have been applied to bility in both flour and gluten dough.
numerous processes such as pulp delignification (Bour- Recently, Minussi et al. (2002) have described the
bonnais et al., 1997; Bourbonnais et al., 1998; Crestini potential applications of laccase in different aspects of
and Argyropoulos, 1998; Li et al., 1999), oxidation of the food industry such as bioremediation, beverage
organic pollutants (Collins et al., 1996) and the processing, ascorbic acid determination, sugar beet
development of biosensors (Kulys et al., 1997; Trudeau pectin gelation, baking and as a biosensor. However,
et al., 1997; Kuznetsov et al., 2001) or biofuel cells they suggested that more studies of laccase production
(Palmore and Kim, 1999). Several organic and inorganic and immobilisation techniques at lower costs are needed
compounds have been reported as effective mediators for to improve the industrial application of this enzyme.
the above-mentioned purposes. These include thiol and
phenol aromatic derivatives, N-hydroxy compounds and 2.2. Pulp and paper industry
ferrocyanide, respectively. Claus et al. (2002) found that
the LMS enhanced dye decolourization and some dyes The industrial preparation of paper requires separa-
resistant to laccase degradation were decolourised. Lu and tion and degradation of lignin in wood pulp. Environ-
Xia (2004) have recently reviewed the applications of the mental concerns urge to replace conventional and
LMS, which comprise pulp bleaching, textile biofinishing polluting chlorine-based delignification/bleaching pro-
and environmental protection processes. However, de- cedures (Kuhad et al., 1997). Oxygen delignification
502 S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513

Table 1
Different laccase applications
Application Laccase source Reference
Decolourization of Aspergillus (genetically modified) Soares et al. (2001a)
dyes Aspergillus (genetically modified) Soares et al. (2001b)
Aspergillus niger Soares et al. (2002)
Cerrena unicolor Michniewicz et al. (2003)
Coriolopsis gallica Reyes et al. (1999)
Coriolopsis rigida Gómez et al. (2005)
Funalia trogii Ünyayar et al. (2005)
Irpex lacteus Kasinath et al. (2003)
Myceliophthora thermophila, Polyporus pinsitus, Trametes versicolor Claus et al. (2002)
Pleurotus eryngii, Pycnoporus cinnabarinus, T. versicolor Camarero et al. (2004)
Pleurotus ostreatus Hou et al. (2004)
P. ostreatus Palmieri et al. (2005)
P. cinnabarinus Mccarthy et al. (1999)
P. cinnabarinus Schliephake et al. (2000)
Sclerotium rolfsii, Trametes hirsuta Campos et al. (2001)
Streptomyces cyaneus Arias et al. (2003)
T. hirsuta Abadulla et al. (2000)
T. hirsuta Domínguez et al. (2005)
T. hirsuta Moldes et al. (2003)
T. hirsuta Rodríguez Couto et al.
T. hirsuta Rodríguez Couto et al.
T. hirsuta Rodríguez Couto et al. (2005)
T. hirsuta Rodríguez Couto et al. (2006)
T. hirsuta Rodríguez Couto and
Sanromán (2006)
T. hirsuta Rodríguez Couto and
Sanromán (2005)
T. hirsuta, T. versicolor Rodríguez Couto et al.
Trametes modesta Nyanhongo et al. (2002)
T. modesta Rehorek et al. (2004)
Trametes trogii Levin et al. (2005)
T. versicolor Maceiras et al. (2001)
T. versicolor Lorenzo et al. (2002)
T. versicolor Rodríguez Couto et al. (2002)
T. versicolor Peralta-Zamora et al. (2003)
T. versicolor Blánquez et al. (2004)
T. versicolor Tavares et al. (2004)
Trametes villosa Zille et al. (2003)
T. villosa Knutson and Ragauskas
Degradation of strain I-4 of the family Chaetomiaceae Saito et al. (2004)
xenobiotics Cladosporium sphaerospermum Potin et al. (2004)
Coprinus cinereus, Myceliophthora thermophila, P. pinsitus, Rhizoctonia solani Kulys et al. (2003)
C. gallica Pickard et al. (1999)
C. gallica Vandertol-Vanier et al. (2002)
Coriolus hirsutus Cho et al. (2002)
Coriolus versicolor Itoh et al. (2000)
C. versicolor Okazaki et al. (2002)
Myceliophtora thermophyla, Trametes pubescens Nicotra et al. (2004)
Panus tigrinus Zavarzina et al. (2004)
P. osteratus Eggen (1999)
P. ostreatus Hublik and Schinner (2000)
P. ostreatus, T. versicolor Keum and Li (2004)
P. cinnabarinus Mougin et al. (2002)
Pyricularia oryzae Lante et al. (2000)
S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513 503

Table 1 (continued )
Application Laccase source Reference
P. oryzae Carunchio et al. (2001)
Rhus vernicifera Moeder et al. (2004)
T. hirsuta Niku-Paavola and Viikari
T. hirsuta D10 Böhmer et al. (1988)
Trametes sp. Tanaka et al. (2001)
Trametes sp. Tanaka et al. (2003)
T. versicolor Collins et al. (1996)
T. versicolor Johannes et al. (1998)
T. versicolor Majcherczyk et al. (1998)
T. versicolor Johannes and Majcherczyk
T. versicolor Majcherczyk and Johannes
T. versicolor Castro et al. (2003)
T. versicolor Dodor et al. (2004)
T. villosa Fabbrini et al. (2001)
T. villosa Fukuda et al. (2001)
T. villosa Kang et al. (2002)
T. villosa Cantarella et al. (2003)
Trichophyton sp. LKY-7 Jung et al. (2003)
unspecified Zhang et al. (2002)
Biosensors Agaricus bisporus, A. niger, T. versicolor Timur et al. (2004)
Agaricus bisporus, R. vernicifera Rigidoporus lignosus, T. versicolor Vianello et al. (2004)
Aspergillus oryzae, Myceliophtora Thermophila, P. pinsitus Kulys and Vidziunaite (2003)
C. unicolor Jarosz-Wilkołazka et al.
C. unicolor Jarosz-Wilkołazka et al.
C. hirsutus Marko-Varga et al. (1995)
C. hirsutus Lisdat et al. (1997)
C. hirsutus Bauer et al. (1999)
C. hirsutus Kuznetsov et al. (2001)
C. hirsutus Freire et al. (2002)
C. hirsutus, R. vernicifera Gupta et al. (2003)
C. versicolor Gomes and Rebelo (2003)
P. ostreatus Leite et al. (2003)
P. oryzae Palmore and Kim (1999)
R. vernicifera Gardiol et al. (1996)
T. versicolor Leech and Daigle (1998)
T. versicolor Freire et al. (2001)
T. versicolor Gomes and Rebelo (2003)
T. versicolo Haghighi et al. (2003)
T.. versicolor Gomes et al. (2004)
T. versicolor Roy et al. (2005)
T. versicolor Ferry and Leech (2005)
Effluent treatment C. gallica Calvo et al. (1998)
Gliocladium virens Murugesan (2003)
Lentinula edodes D'Annibale et al. (1999)
L. edodes D'Annibale et al. (2000)
L. edodes Casa et al. (2003)
P. tigrinus D'Annibale et al. (2004)
P. ostreatus Aggelis et al. (2003)
Pleurotus spp. Tsioulpas et al. (2002)
Pycnoporus coccineus Jaouani et al. (2005)
R. vernicifera Durante et al. (2004)
Trametes sp. strain AH28-2 Xiao et al. (2003)
T. versicolor Jolivalt et al. (2000)
(continued on next page)
504 S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513

Table 1 (continued )
Application Laccase source Reference
T. versicolor Edwards et al. (2002)
T. versicolor Lucas et al. (2003)
Biopulping Fomes fomentarius, Ganoderma collosum, Lentinus edades, Merulius tremellosus, Phlebia Bourbonnais et al. (1997)
radiata, P. ostreatus T. versicolor
C. versicolor Call and Mücke (1997)
Peniophora sp., Pycnoporus sanguineus, T. hirsuta, T. versicolor Kandioller and Christov
T. versicolor Archibald et al. (1997)
T. versicolor Crestini and Argyropoulos
unspecified Jacob et al. (1999)
unspecified Sealey et al. (1999)
unspecified Chakar and Ragauskas (2001)
unspecified Poppius-Levlin et al. (2001)
unspecified Tamminen et al. (2003)
Organic synthesis C. hirsuta Baker et al. (1996)
C. hirsutus Karamyshev et al. (2003)
P. cinnabarinus Mikolasch et al. (2002)
P. coccineus Uyama and Kobayashi (2002)
P. oryzae Setti et al. (1999)
T. versicolor Fritz-Langhals and Kunath
T. versicolor Akta et al. (2001)
T. versicolor Schäfer et al. (2001)
T. versicolor Akta and Tanyolaç (2003)
T. villosa Uchida et al. (2001)
Food industry Chinese rhus lacquer Huang et al. (1995)
Myceliophtora thermophili, P. pinsitius Micard and Thibault (1999)
P. cinnabarinus Georis et al. (2003)
T. hirsuta Kuuva et al. (2003)
T. versicolor Crecchio et al. (1995)
unspecified Mathiasen (1996)
unspecified Petersen and Mathiasen
unspecified Norsker et al. (2000)
Biobleaching C. versicolor Balakshin et al. (2001)
P. eryngii, P. cinnabarinus, T. versicolor Camarero et al. (2004)
P. cinnabarinus Georis et al. (2003)
T. versicolor Paice et al. (1995)
T. versicolor Archibald et al. (1997)
unspecified Balakshin et al. (2001)
unspecified Han et al. (2002)
Denim bleaching T. versicolor Pazarlıoglu et al. (2005)
unspecified Vinod (2001)

processes have been industrially introduced (Carter et development of LMS delignification technologies for
al., 1997), but pre-treatments of wood pulp with kraft pulps. In addition, laccase is more readily available
ligninolytic enzymes might provide milder and cleaner and easier to manipulate than both lignin peroxidase
strategies of delignification that are also respectful of the (LiP) and manganese-dependent peroxidase (MnP) and
integrity of cellulose (Kuhad et al., 1997). LMS has already found practical applications such as
Although extensive studies have been performed to the Lignozym®-process (Call and Mücke, 1997).
develop alternative bio-bleaching systems, few enzy- Several authors applied the LMS to pulp biobleach-
matic treatments exhibit the delignification/brightening ing (see Table 1). However, all these biobleaching
capabilities of modern chemical bleaching technologies. studies were focused on wood pulps and little is known
One of the few exceptions to this generalisation is the about the efficiency of the LMS on non-wood pulps
S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513 505

including those used for manufacturing specialty papers. processes based on laccases seems an attractive solution
In this sense, Camarero et al. (2004) explored the due to their potential in degrading dyes of diverse
potential of LMS to remove lignin-derived products chemical structure (Abadulla et al., 2000; Blánquez et
responsible for colour from a high-quality flax pulp. al., 2004; Hou et al., 2004), including synthetic dyes
They showed the feasibility of LMS to substitute currently employed in the industry (Rodríguez Couto et
chlorine-containing reagents in manufacturing of these al., 2004a, 2005).
high-price paper pulps. The use of laccase in the textile industry is growing
The capability of laccases to form reactive radicals in very fast, since besides to decolourise textile effluents as
lignin can also be used in targeted modification of wood commented above, laccase is used to bleach textiles and
fibers. For example, laccases can be used in the enzymatic even to synthetise dyes (Setti et al., 1999). Related to
adhesion of fibers in the manufacturing of lignocellulose- textile bleaching, in 1996 Novozyme (Novo Nordisk,
based composite materials such as fiberboards. Laccases Denmark) launched a new industrial application of
have been proposed to activate the fiberbound lignin laccase enzyme in denim finishing: DeniLite®, the first
during manufacturing of the composites, thus, resulting in industrial laccase and the first bleaching enzyme acting
boards with good mechanical properties without toxic with the help of a mediator molecule. Also, in 2001 the
synthetic adhesives (Felby et al., 1997; Hüttermann et al., company Zytex (Zytex Pvt. Ltd., Mumbai, India)
2001). Another possibility is to functionalise lignocellu- developed a formulation based on LMS capable of
losic fibers by laccases in order to improve the chemical or degrading indigo in a very specific way. The trade name
physical properties of the fiber products. Preliminary of the product is Zylite.
results have shown that laccases are able to graft various
phenolics acid derivatives onto kraft pulp fibers (Lund 2.4. Nanobiotechnology
and Ragauskas, 2001; Chandra and Ragauskas, 2002).
This ability could be used in the future to attach During the past two decades, bioelectrochemistry has
chemically versatile compounds to the fiber surfaces, received increased attention. Progress on bioelectrochem-
possibly resulting in fiber materials with completely novel istry has been integrated into analytical applications, e.g.
properties such as hydrophobicity or charge. in biosensors working as detectors in clinical and envi-
ronmental analysis (Haghighi et al., 2003). Since laccases
2.3. Textile industry are able to catalyse electron transfer reactions without
additional cofactors, their use has also been studied in
The textile industry accounts for two-thirds of the total biosensors to detect various phenolic compounds, oxygen
dyestuff market (Riu et al., 1998) and consumes large or azides (see Table 1). Moreover, biosensors for detection
volumes of water and chemicals for wet processing of of morphine and codeine (Bauer et al., 1999), catecho-
textiles. The chemical reagents used are very diverse in lamines (Lisdat et al., 1997; Leite et al., 2003; Ferry and
chemical composition, ranging from inorganic com- Leech, 2005), plant flavonoids (Jarosz-Wilkołazka et al.,
pounds to polymers and organic products (Mishra and 2004) and also for electroimmunoassay (Kuznetsov et al.,
Tripathy, 1993; Banat et al., 1996; Juang et al., 1996). 2001) have been developed. Nanotechnology contributes
There are more than 100,000 commercially available dyes to the development of smaller and more efficient
with over 7 × 105 t of dyestuff produced annually (Meyer, biosensors through controlled deposition and specific
1981; Zollinger, 2002). Due to their chemical structure adsorption of biomolecules on different types of surfaces,
dyes are resistant to fading on exposure to light, water and achieving micro and nanometer order. Hammond and
different chemicals (Poots and McKay, 1976; McKay, Whitesides (1995) have introduced a method to pattern
1979) and most of them are difficult to decolourise due to ultrathin ionic multilayer films with micron-sized features
their synthetic origin. onto surfaces building a patterned alkanethiol monolayer
Government legislation is becoming more and more with ionic functionality onto a gold surface. Typical mol-
stringent, especially in the more developed countries, ecules used in this process are shown in Fig. 1. Chen et al.
regarding the removal of dyes from industrial effluents (1998) showed a biotechnological application of such
(O'Neill et al., 1999). Concern arises, as several dyes are micropatterned surfaces: the production of islands of
made from known carcinogens such as benzidine and micrometer size of extracellular matrix, where the pattern
other aromatic compounds (Baughman and Perenich, of these islands could determine the position and
1988). Most currently existing processes to treat dye distribution of bovine and endothelial cells. The control
wastewater are ineffective and not economical (Cooper, of the nature and the density of the groups (e.g. alkys,
1995; Stephen, 1995). Therefore, the development of amides, alcohols) of a surface built with assembled
506 S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513

a) Polyanions Polycations

* *

NH 3+

SO 3 - PAH

* n*

* N+

COO - H3 C CH3


Phospholipids Alkanethiols

O +

O O–

Fig. 1. a) Polylectrolytes are currently used to build multilayers due to their different versatility. PSS: poly(styrene sulfonate); PAA: Poly(acrylic
acid), PAH: poly(allylamine hydrochlorid), PDADMAC: poly(diallyldimethylammonium chloride). b) Phospholipid and alkanethiols have the ability
to form bilayers and self-assembled monolayers.

monolayers has been used succesfully to investigate the on the cathode of biofuel cells that could provide power,
non-specific adsorption of proteins (Sigal et al., 1998). for example, for small transmitter systems (Chen et al.,
Regarding laccases, the immobilisation has an important 2001; Calabrese et al., 2002). Biofuel cells are extremely
influence on the biosensor sensitivity (Freire et al., 2001). attractive from an environmental point of view because
Martele et al. (2003) have shown that micropatterning is electrical energy is generated without combusting fuel,
an efficient method for the immobilisation of laccases on a thus, providing a cleaner source of energy. Fig. 2a shows
solid surface in order to develop a multi-functional bio- different functionalised flat surfaces built with polymers
sensor. Also, Roy et al. (2005) found that cross-linked and self-assembled monolayers (SAMs) that can be used
enzyme crystals (CLEC) of laccase from Trametes ver- to adsorb and immobilise proteins or other biomolecules.
sicolor could be used in biosensor applications with great The layer-by-layer technique (LbL) (Decher, 1997)
advantage over the soluble enzyme. More recently, can be used to build macromolecular structures down to
Cabrita et al. (2005) have immobilised laccase from nanometer control leading to surfaces of well-defined
Coriolus versicolor on N-Hydroxysuccinimide-terminat- thickness (see Fig. 2a). Recently, flat polyelectrolyte
ed self-assembled monolayers on gold. This procedure multilayers built by alternating adsorption of oppositely
could be useful for the further development of biosensors. charged polyelectrolytes have been used to recrystallise
In addition, an enzyme electrode based on the co-immo- bacterial proteins making the building of artificial cell
bilisation of an osmium redox polymer and a laccase from walls possible (Toca-Herrera et al., 2005). The LbL
T. versicolor on glassy carbon electrodes has been applied technique has also been used to build hollow polyelec-
to ultrasensitive amperometric detection of the catechol- trolyte capsules after core removal (Donath et al., 1998).
amine neurotransmitters dopamine, epinephrine and Further application of the sequential adsorption of
norepinephrine, attaining nanomolar detection limits oppositely charged polyelectrolytes onto enzyme crystal
(Ferry and Leech, 2005). Laccase can also be immobilised templates would permit their encapsulation. Caruso et
S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513 507

Enzymes, Proteins


Si, Au, mica, glass Substrate

Enzyme, protein
Enzyme, protein


Si, Au, mica, glass Au Substrate

Molecules to degrade


Polyelectrolyte wall

Protein layer

Enzyme, biomolecules

Polyelectrolyte wall
Colloidal particle

Fig. 2. a) 2D supramacromolecular structures that can be used to immobilise biomolecules. Several structures are suitable: polylectrolyte multilayer,
micropatterning and self-assembled monolayers (SAMs). b) 3D supramacromolecular structures that can be used to build microreactors and
immobilise biomolecules. In the first case, hollow polelectrolyte shells can host proteins inside, permitting the diffusion of molecules through the
shell wall. A colloidal particle covered by polyelectrolytes (and phospholipids) can host proteins and/or other types of functional molecules.

al. (2000) showed that the encapsulated enzyme could lower than 6 the macromolecules permeate into the
retain 100% of its activity after incubation for 100 min capsule interior. In this way, the authors showed how to
with protease. The permeability properties of the wall open and close the capsule wall in a reversible way. This
capsule are important for the proper function of the mechanism together with the LbL encapsulation
encapsulated enzyme. Antipov et al. (2002) investigated technique permits the development of microreactors
the permeability properties of hollow polyelectrolyte Also, colloidal particles covered with polyelectrolytes
multilayer capsules as a function of pH and salt and phospholipids have been used to host and activate
concentration. It was shown that the capsule wall was rubella virus (Fischlechner et al., 2005). This type of
closed to a pH value of 8 and higher, but at pH values system is shown in Fig. 2b.
508 S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513

2.5. Other laccase applications tion of this biocatalyst in heterologous hosts and also
their modification by chemical means or protein engi-
2.5.1. Soil bioremediation neering to obtain more robust and active enzymes.
Polycyclic aromatic hydrocarbons (PAHs) together On the other hand, the development of an effective
with other xenobiotics are a major source of contami- system for laccase immobilisation also deserves great at-
nation in soil, therefore, their degradation is of great tention. Immobilisation could be achieved by chemical
importance for the environment. The catalytic properties modification of the substrates. Hence, micropatterning,
of laccases can be used to degrade such compounds. SAMs and LbL techniques can be used to functionalise flat
Thus, laccases were able to mediate the coupling of and curved surfaces in order to have specific adsorption.
reduced 2,4,6-trinitrotoluene (TNT) metabolites to an Laccase encapsulation with polyelectrolytes will be used
organic soil matrix, which resulted in detoxification of as a microreactor for catalytic reactions by changing the
the munition residue (Durán and Esposito, 2000). permeability properties of the capsule wall. Since the
Moreover, PAHs, which arise from natural oil deposits general goal is to obtain stable catalysts with long life times
and utilisation of fossil fuels, were also found to be and low cost, we think that the combination of these
degraded by laccases (Pointing, 2001). Recently, techniques will enhance: i) the adsorption of laccase on a
Nyanhongo et al. (in press) showed that a laccase suitable substrate, ii) the lifetime of the laccase activity and
from Trametes modesta was involved in immobilisation iii) reutilisation of the substrate/laccase product. Our
of TNT degradation products. research group is currently working in this direction.

2.5.2. Synthetic chemistry Acknowledgments

In the future laccases may also be of great interest in
synthetic chemistry, where they have been proposed to SRC and JLTH are Ramón y Cajal Senior Research
be applicable for oxidative deprotection (Semenov et al., Fellows. Therefore, the authors thank the Spanish
1993) and production of complex polymers and medical Ministry of Education and Science for promoting the
agents (Xu, 1999 and Refs. therein, Mai et al., 2000; Ramón y Cajal Programme.
Uyama and Kobayashi, 2002; Kurisawa et al., 2003;
Nicotra et al., 2004). Recently, Mustafa et al. (2005) References
synthetised phenolic colourants by using an industrial
laccase named Suberase® (Novo Nordisk A/S, Bags- Aaslyng D, Rorbaek K, Sorensen NH, (29.11.1996). An ezyme for
dying keratinous fibres. Int Pat Apl WO9719998.
vaerdt, Denmark). Abadulla E, Tzanov T, Costa S, Robra KH, Cavaco-Paulo A, Gübitz
G. Decolorization and detoxification of textile dyes with a laccase
2.5.3. Cosmetics from Trametes hirsuta. Appl Environ Microbiol 2000;66:
The cosmetic world has not been indifferent to the 3357–62.
application of laccase: for example, laccase-based hair Aggelis G, Iconomou D, Christouc M, Bokas D, Kotzailias S, Christou G,
et al. Phenolic removal in a model olive oil mill wastewater using
dyes are less irritant and easier to handle than current hair Pleurotus ostreatus in bioreactor cultures and biological evaluation of
dyes, since laccases replace H2O2 as an oxidising agent in the process. Water Res 2003;37:3897–904.
the dye formulation (Roure et al., 1992; Aaslyng et al., Akta N, Tanyolaç A. Reaction conditions for laccase catalyzed
1996; Lang and Cotteret, 1999). More recently, cosmetic polymerization of catechol. Bioresour Technol 2003;87:209–14.
Akta N, Çiçek H, Tapınar ÜA, Kibarer G, Kolankaya N, Tanyolaç A.
and dermatological preparations containing proteins for
Reaction kinetics for laccase-catalyzed polymerization of 1-
skin lightening have also been developed (Golz-Berner et naphthol. Bioresour Technol 2001;80:29–36.
al., 2004). Antipov A, Sukhorukov GB, Leporatti S, Radtchenko IL, Donath E,
Möhwald H. Polyelectrolyte nultilayer capsule permeability control.
3. Future outlook Colloids Surf A Physicochem Eng Asp 2002;198-200:535–41.
Archibald FS, Bourbonnais R, Jurasek L, Paice MG, Reid ID. Kraft
pulp bleaching and delignification by Trametes versicolor.
The most important obstacles to commercial appli- J Biotechnol 1997;53:215–36.
cation of laccases are the lack of sufficient enzyme stocks Arias ME, Arenas M, Rodríguez J, Soliveri J, Ball AS, Hernández M.
and the cost of redox mediators. Marked progress has Kraft pulp biobleaching and mediated oxidation of a nonphenolic
been made over the last years to solve these problems and substrate by laccase from Streptomyces cyaneus CECT 3335. Appl
it is expected that laccases will be able to compete with Environ Microbiol 2003;69:1953–8.
Baker WL, Sabapathy K, Vibat M, Lonergan G. Lactase catalyzes
other processes such as elemental chlorine-free (ECF) formation of an indamine dye between 3-methyl-2-benzothiazoli-
and totally chlorine-free (TCF) bleaching. Thus, efforts none hydrazone and 3-dimethylaminobenzoic acid. Enzyme
have to be made in order to achieve cheap overproduc- Microb Technol 1996;18: 90–4.
S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513 509

Balakshin M, Chen C-L, Gratzl JS, Kirkman AG, Jakob H. Bio- wastewater by an enzymatic treatment and its impact on durum wheat
bleaching of pulp with dioxygen in laccase-mediator system—effect (Triticum durum Desf.) germinability. Chemosphere 2003;50:959–66.
of variables on the reaction kinetics. J Mol Catal B Enzym Castro AIRP, Evtuguin DV, Xavier AMB. Degradation of biphenyl
2001;16:205–15. lignin model compounds by laccase of Trametes versicolor in the
Banat IM, Nigam P, Singh D, Marchant R. Microbial decolorization of presence of 1 hydroxybenzotriazole and heteropolyanion
textile-dye-containing effluents: a review. Bioresour Technol [SiW11VO40]5−. J Mol Catal B Enzym 2003;22:13–20.
1996;58:217–27. Chakar FS, Ragauskas AJ. Formation of quinonoid structures in
Bauer CG, Kuhn A, Gajovic N, Skorobogatko O, Holt PJ, Bruce NC, laccase-mediator reactions. In: Argyropoulos DS, editor. Oxidative
et al. New enzyme sensors for morphine and codeine based on delignification chemistry fundamentals and catalysis. ACS sym-
morphine dehydrogenase and laccase. Fresenius' J Anal Chem posium series. USA: Oxford University Press; 2001. p. 444–55.
1999;364:179–83. Chandra RP, Ragauskas AJ. Evaluating laccase-facilitated coupling of
Baughman GL, Perenich TA. Fate of dyes in aquatic systems: I phenolic acids to high-yield kraft pulps. Enzyme Microb Technol
solubility and partitioning of some hydrophobic dyes and related 2002;30:855–61.
compounds. Environ Toxicol Chem 1988;7:183–99. Chen CS, Mrkisch M, Huang S, Whistesides GM, Ingber DE.
Blánquez P, Casas N, Font X, Gabarrell M, Sarrá M, Caminal G, et al. Micropatterned surfaces for control of cell shape, position, and
Mechanism of textile metal dye biotransformation by Trametes function. Biotechnol Prog 1998;14:356–63.
versicolor. Water Res 2004;38:2166–72. Chen T, Barton SC, Binyamin G, Gao Z, Zhang Y, Kim H-H, et al. A
Böhmer S, Messner K, Srebotnik E. Oxidation of phenanthrene by a miniature biofuel cell. J Am Chem Soc 2001;123:8630–1.
fungal laccase in the presence of 1-hydroxybenzotriazole and Cho S-J, Park SJ, Lim JS, Rhee YH, Shin KS. Oxidation of polycyclic
unsaturated lipids. Biochem Biophys Res Commun 1988;244: aromatic hydrocarbons by laccase of Coriolus hirsutus. Biotechnol
233–8. Lett 2002;24:1337–40.
Bourbonnais R, Paice MG. Oxidation of non-phenolic substrates: an Claus H, Faber G, König H. Redox-mediated decolorization of syn-
expanded role of laccase in lignin biodegradation. FEBS Lett thetic dyes by fungal laccases. Appl Microbiol Biotechnol
1990;267:99­102. 2002;59: 672–8.
Bourbonnais R, Paice MG, Freiermuth B, Bodie E, Borneman S. Collins PJ, Kotterman MJJ, Field JA, Dobson ADW. Oxidation of
Reactivities of various mediators and laccases with kraft pulp and anthracene and benzo[a]pyrene by laccases from Trametes
lignin model compounds. Appl Environ Microbiol 1997;63:4627–32. versicolor. Appl Environ Microbiol 1996;62:4563–7.
Bourbonnais R, Leech D, Paice MG. Electrochemical analysis of the Cooper P. Removing colour from dye house wastewater. Asian Textile
interactions of laccase mediators with lignin model compounds. J 1995;3:52–6.
Biochim Biophys Acta 1998;1379:381–90. Crecchio C, Ruggiero P, Pizzigallo MDR. Polyphenoloxidases immo-
Cabrita JF, Abrantes LM, Viana AS. N-Hydroxysuccinimide-termi- bilized in organic gels: properties and applications in the detoxifi-
nated self-assembled monolayers on gold for biomolecules cation of aromatic compounds. Biotechnol Bioeng 1995;48:585–91.
immobilisation. Electrochim Acta 2005;50:2117–24. Crestini C, Argyropoulos DS. The early oxidative biodegradation steps
Calabrese BS, Pickard M, Vazquez-Duhalt R, Heller A. Electroreduc- of residual kraft lignin models with laccase. Bioorg Med Chem
tion of O2 to water at 0.6 V (NHE) at pH 7 on the ‘wired’ Pleurotus 1998;6:2161–9.
ostreatus Laccase Cathode. Biosens Bioelectron 2002;17:1071–4. D'Annibale A, Stazi SR, Vinciguerra V, Di Mattia E, Giovannozzi SG.
Call HP, Mücke I. History, overview and applications of mediated Characterization of immobilized laccase from Lentinula edodes
lignolytic systems, especially laccase-mediator systems (Ligno- and its use in olive-mill wastewater treatment. Process Biochem
zymR process). J Biotechnol 1997;53:163–202. 1999;34:697–706.
Calvo AM, Copa-Patiño JL, Alonso O, González AE. Studies of the D'Annibale A, Stazi SR, Vinciguerra V, Giovannozzi SG. Oxirane-
production and characterization of laccase activity in the immobilized Lentinula edodes laccase: stability and phenolics removal
basidiomycete Coriolopsis gallica, an efficient decolorizer of efficiency in olive mill wastewater. J Biotechnol 2000;77: 265–73.
alkaline effluents. Arch Microbiol 1998;171:31–6. D'Annibale A, Ricci M, Quaratino D, Federic F, Fenice M. Panus
Camarero S, Garcia O, Vidal T, Colom J, del Rio JC, Gutierrez A, et al. tigrinus efficiently removes phenols, color and organic load from
Efficient bleaching of non-wood high-quality paper pulp using olive-mill wastewater. Res Microbiol 2004;155:596–603.
laccase-mediator system. Enzyme Microb Technol 2004;35:113–20. Decher G. Fuzzy nanoassemblies: toward layered polymeric multi-
Campos R, Kandelbauer A, Robra KH, Cavaco-Paulo A, Gübitz GM. composites. Science 1997;277:1232–7.
Indigo degradation with purified laccases from Trametes hirsuta Dodor DE, Hwang HM, Ekunwe SIN. Oxidation of anthracene and
and Sclerotium rolfsii. J Biotechnol 2001;89:131–9. benzo[a] pyrene by immobilized laccase from Trametes versicolor.
Cantarella G, Galli C, Gentili P. Free radical versus electron-transfer routes Enzyme Microb Technol 2004;35:210–7.
of oxidation of hydrocarbons by laccase/mediator systems. Catalytic or Domínguez A, Rodríguez Couto S, Sanromán MA. Dye decolour-
stoichiometric procedures. J Mol Catal B Enzym 2003;22:135–44. ization by Trametes hirsuta immobilised into alginate beads.
Carter DN, McKenzie DG, Johnson AP, Idner K. Performance para- World Journal of Industrial Microbiology and Biotechnology
meters of oxygen delignification. Tappi J 1997;80:111–7. 2005;21:405–9.
Carunchio F, Crescenzi C, Girelli AM, Messina A, Tarola AM. Oxidation Donath E, Sukhorukov GB, Caruso F, Davis SA, Mohwald H. Novel
of ferulic acid by laccase: identification of the products and inhibitory hollow polymer shells by colloid-templated assembly of polyelec-
effects of some dipeptides. Talanta 2001;55:189–200. trolytes. Angew Chem Int Ed Engl 1998;37:2202–5.
Caruso F, Trau D, Möhwald H, Renneberg R. Enzyme encapsulation in Durán N, Esposito E. Potential applications of oxidative enzymes and
layer-by-layer engineered polymer multilayer capsules. Langmuir phenoloxidase-like compounds in wastewater and soil treatment: a
2000;16:1485–8. review. Appl Catal B Environ 2000;28:83–99.
Casa R, D'Annibale A, Pieruccetti F, Stazi SR, Giovannozzi SG, Lo Durante D, Casadio R, Martelli L, Tasco G, Portaccio M, De Luca P, et al.
Cascio B. Reduction of the phenolic components in olive-mill Isothermal and non-isothermal bioreactors in the detoxification of
510 S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513

waste waters polluted by aromatic compounds by means of assembled monolayers as a template. Macromolecules 1995;28:
immobilised laccase from Rhus vernicifera. J Mol Catal B Enzym 7569–71.
2004;27:191–206. Han S, Fang G, Deng Y, Li P, Asther M, Sigoillot J-C. Efficient effects
Edwards W, Leukes WD, Bezuidenhout J. Ultrafiltration of petro- on enhancement bleachabilities of soda-AQ wheat-straw pulps by
chemical industrial wastewater using immobilised manganese laccase treatment. Proceedings of the international symposium on
peroxidase and laccase: application in the defouling of poly- emerging technologies of pulping and papermaking 2nd, Guang
sulphone membranes. J Desalin 2002;149:275–8. zhou, China, Oct. 9–11; 2002.
Eggen T. Application of fungal substrate from commercial mushroom Hou H, Zhou J, Wang J, Du C, Yan B. Enhancement of laccase
production Pleuorotus ostreatus for bioremediation of creosote production by Pleurotus ostreatus and its use for the decolorization
contaminated soil. Int Biodeterior Biodegrad 1999;44:117–26. of anthraquinone dye. Process Biochem 2004;39:1415–9.
Fabbrini M, Galli C, Gentili P, Macchitella D. An oxidation of alcohols Huang H, Cai R, Du Y, Zeng Y. Flow-injection stopped-flow
by oxygen with the enzyme laccase and mediation by TEMPO. spectrofluorimetric kinetic determination of total ascorbic acid
Tetrahedron Lett 2001;42:7551–3. based on an enzyme-linked coupled reaction. Anal Chim Acta
Felby C, Pedersen LS, Nielsen BR. Enhanced auto adhesion of wood 1995;309:271–5.
fibers using phenol oxidases. Holzforschung 1997;51:281–6. Hublik G, Schinner F. Characterization and immobilization of the
Ferry Y, Leech D. Amperometric detection of catecholamine laccase from Pleurotus ostreatus and its use for the continuous
neurotransmitters using electrocatalytic substrate recycling at a elimination of phenolic pollutants. Enzyme Microb Technol
laccase electrode. Electroanalysis 2005;17:2113–9. 2000;27:330–6.
Fischlechner M, Zschörnig O, Hofmann J, Donath E. Engineering Hüttermann A, Mai C, Kharazipour A. Modification of lignin for the
virus functionalities on colloidal polyelectrolyte lipid composites. production of new compounded materials. Appl Microbiol
Angew Chem Int Ed Engl 2005;44:2892–5. Biotechnol 2001;55:387–94.
Freire RS, Durán N, Kubota LT. Effects of fungal laccase Itoh K, Fujita M, Kumano K, Suyama K, Yamamoto H. Phenolic acids
immobilization procedures for the development of a biosensor affect transformations of chlorophenols by a Coriolus versicolor
for phenol compounds. Talanta 2001;54:681–6. laccase. Soil Biol Biochem 2000;32:85–91.
Freire RS, Durán N, Kubota LT. Development of a laccase-based flow Jacob H, Del Grosso M, Kuever A, Nimmerfroh N, Suess HU.
injection electrochemical biosensor for the determination of phenolic Delignification of chemical pulp with laccase and mediators. A
compounds and its application for monitoring remediation of kraft E1 concept with a future? Papier 1999;53:85–95.
paper mill effluent. Anal Chim Acta 2002;463:229–38. Jaouani A, Guillen F, Penninckx MJ, Martinez AT, Martinez MJ. Role
Fritz-Langhals E, Kunath B. Synthesis of aromatic aldehydes by laccase- of Pycnoporus coccineus laccase in the degradation of aromatic
mediator assisted oxidation. Tetrahedron Lett 1998;39:5955–6. compounds in olive oil mill wastewater. Enzyme Microb Technol
Fukuda T, Uchida H, Takashima Y, Uwajima T, Kawabata T, Suzuki 2005;36:478–86.
M. Degradation of bisphenol a by purified laccase from Trametes Jarosz-Wilkołazka A, Ruzgas T, Gorton L. Use of laccase-modified
villosa. Biochem Biophys Res Commun 2001;284:704–6. electrode for amperometric detection of plant flavonoids. Enzyme
Gardiol AE, Hernandez RJ, Reinhammar B, Harte BR. Development Microb Technol 2004;35:238–41.
of a gas-phase oxygen biosensor using a blue copper-containing Jarosz-Wilkołazka A, Ruzgas T, Gorton L. Amperometric detection of
oxidase. Enzyme Microb Technol 1996;18:347–52. mono- and diphenols at Cerrena unicolor laccase-modified
Georis J, Lomascolo A, Camarero S, Dorgeo V, Herpoel I, Asther M, et graphite electrode: correlation between sensitivity and substrate
al. Pycnoporus cinnabarinus laccases: an interesting tool for food structure. Talanta 2005;66:1219–24.
or non-food applications. Meded ­ Fac Landbouwkd Toegep Biol Johannes C, Majcherczyk A. Natural mediators in the oxidation of
Wet 2003;68:263–6. polycyclic aromatic hydrocarbons by laccase mediator systems.
Gianfreda L, Xu F, Bollag J-M. Laccases: a useful group of Appl Environ Microbiol 2000;66:524–8.
oxidoreductive enzymes. Bioremediat J 1999;3:1­25. Johannes C, Majcherczyk A, Huttermann A. Oxidation of ace-
Golz-Berner K. Walzel B. Zastrow L. Doucet O. (04.03.2004). naphthene and acenaphthylene by laccase of Trametes versicolor
Cosmetic and dermatological preparation containing copper- in a laccase-mediator system. J Biotechnol 1998;61:151–6.
binding proteins for skin lightening. Int Pat Appl WO2004017931. Jolivalt C, Brenon S, Caminade E, Mougin C, Pontié M. Immobili-
Gomes SASS, Rebelo MJF. A new laccase biosensor for polyphenols zation of laccase from Trametes versicolor on a modified PVDF
determination. Sensors 2003;3:1­25. microfiltration membrane: characterization of the grafted support
Gomes SASS, Nogueira JMF, Rebelo MJF. Amperometric biosensor and application in removing a phenylurea pesticide in wastewater.
for polyphenolic compounds in red wine. Biosens Bioelectron J Membr Sci 2000;180:103–13.
2004;20:1211–6. Juang RS, Tseng RL, Wu FC, Lin SJ. Use of chitin and chitosan
Gómez J, Pazos M, Rodríguez Couto S, Sanromán MA. Chestnut in lobster shell wastes for colour removal from aqueous so-
shell and barley bran as potential substrates for laccase production lutions. J Environ Sci Health Part A Environ Sci Eng 1996;31:
by Coriolopsis rigida under solid-state conditions. J Food Eng 325–38.
2005;68:315–9. Jung H, Hyun K, Park Ch. Production of laccase and bioremediation of
Gupta G, Rajendran V, Atanassov P. Laccase biosensor on monolayer- pentachlorophenol by wood-degrading fungus Trichophyton sp.
modified gold electrode. Electroanalysis 2003;15:1577–83. LKY-7 immobilized in Ca-alginate beads. Polpu Chongi Gisul
Haghighi B, Gorton L, Ruzgas T, Jönsson LJ. Characterization of graphite 2003;35:80–6.
electrodes modified with laccase from Trametes versicolor and their Kandioller G, Christov L. Evaluation of the delignification and bleaching
use for bioelectrochemical monitoring of phenolic compounds in flow abilities of selected laccases with HBT on different pulps. In:
injection analysis. Anal Chim Acta 2003;487:3­14. Argyropoulos DS, editor. Oxidative delignification chemistry funda-
Hammond PT, Whitesides GM. Formation of polymer microstructures mentals and catalysis. ACS symposium series, Vol. 785. USA: Oxford
by selective deposition of polyion multilayers using patterned self- University Press; 2001. p. 427–43.
S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513 511

Kang KH, Dec J, Park H, Bollag JM. Transformation of the fungicide Lu R, Xia L-M. Advances in research and application of laccase
cyprodinil by a laccase of Trametes villosa in the presence of mediator system. Xianweisu Kexue Yu Jishu 2004;12:37–44.
phenolic mediators and humic acid. Water Res 2002;36:4907–15. Lucas M, De La Rubia T, Martinez J. Oxidation of low molecular
Karamyshev AV, Shleev SV, Koroleva OV, Yaropolov AI, Sakharov weight aromatic components of olive-mill wastewaters by a
IY. Laccase-catalyzed synthesis of conducting polyaniline. En- Trametes versicolor laccase. Polyphenols Actual 2003;23:36–7.
zyme Microb Technol 2003;33:556–64. Lund M, Ragauskas AJ. Enzymatic modification of kraft lignin
Kasinath A, Novotny C, Svobodova K, Patel KC, Šašek V. Decoloriza- through oxidative coupling with water-soluble phenols. Appl
tion of synthetic dyes by Irpex lacteus in liquid cultures and packed- Microbiol Biotechnol 2001;55:699–703.
bed bioreactor. Enzyme Microb Technol 2003;32:167–73. Maceiras R, Rodríguez Couto S, Sanromán A. Influence of several
Keum YS, Li QX. Fungal laccase-catalyzed degradation of hydroxy inducers on the synthesis of extracellular laccase and in vivo
polychlorinated biphenyls. Chemosphere 2004;56:23–30. decolourisation of Poly R-478 by semi-solid-state cultures of
Knutson K, Ragauskas A. Laccase-mediator biobleaching applied to a Trametes versicolor. Acta Biotechnol 2001;21:255–64.
direct yellow dyed paper. Biotechnol Prog 2004;20:1893–6. Mai C, Majcherczyk A, Hütterman A. Chemo-enzymatic synthesis and
Kuhad RC, Singh A, Eriksson KEL. Microorganisms and enzymes characterization of graft copolymers from lignin and acrylic
involved in the degradation of plant fiber cell wall. In: Eriksson compounds. Enzyme Microb Technol 2000;27:167–75.
KEL, editor. Biotechnology in the Pulp and paper industry. Majcherczyk A, Johannes C. Radical mediated indirect oxidation of a
Advances in biochemical engineering biotechnology. Berlin: PEG-coupled polycyclic aromatic hydrocarbon (PAH) model
Springer Verlag; 1997. Chapter 2. compound by fungal laccase. Biochim Biophys Acta 2000;1474:
Kulys J, Vidziunaite R. Amperometric biosensors based on recombi- 157–62.
nant laccases for phenols determination. Biosens Bioelectron Majcherczyk A, Johannes C, Hüttermann A. Oxidation of polycyclic
2003;18:319–25. aromatic hydrocarbons (PAH) by laccase of Trametes versicolor.
Kulys J, Drungiliene A, Wollenberger U, Krikstopaitis K, Scheller F. Enzyme Microb Technol 1998;22:335–41.
Electroanalytical determination of peroxidases and laccases on Marko-Varga G, EmnCus J, Gotton L, Ruzgas T. Development of
carbon paste electrodes. Electroanalysis 1997;9:213–8. enzyme-based amperometric sensors for the determination of
Kulys J, Vidziunaite R, Schneider P. Laccase-catalyzed oxidation of phenolic compounds. Trends Anal Chem 1995;14:319–28.
naphthol in the presence of soluble polymers. Enzyme Microb Martele Y, Callewaerta K, Naessens K, Van Daeleb P, Baetsb R, Schacht
Technol 2003;32:455–63. E. Controlled patterning of biomolecules on solid surfaces. Mater Sci
Kurisawa M, Chung JE, Uyama H, Kobayashi S. Enzymatic synthesis Eng C Biomim Mater Sens Syst 2003;23:341–5.
and antioxidant properties of poly(rutin). Biomacromolecules Mathiasen TE. Laccase for improved beer storage. Trends Food Sci
2003;4:1394–9. Technol 1996;7:272.
Kuuva T, Lantto R, Reinikainen T, Buchert J, Autio K. Rheological Mccarthy JT, Levy VC, Lonergan GT, Fecondo JV. Development of
properties of laccase-induced sugar beet pectin gels. Food optimal conditions for the decolourisation of a range of industrial
Hydrocoll 2003;17:679–84. dyes using Pycnoporus cinnabarinus laccase. Hazard Ind Waste
Kuznetsov BA, Shumakovich GP, Koroleva OV, Yaropolov AI. On 1999;31:489–98.
applicability of laccase as label in the mediated and mediatorless McKay G. Waste colour removal from textile effluents. Am Dyest
electroimmunoassay: effect of distance on the direct electron transfer Report 1979;68:29–36.
between laccase and electrode. Biosens Bioelectron 2001;16:73–84. Messerschmidt A. Blue copper oxidases. Adv Inorg Chem 1993;40:
Lang G, Cotteret J, (22.07.1999). Hair dye composition containing a 121–85.
laccase. (L'Oreal, Fr.). Int Pat Appl WO9936036. Messerschmidt A. Multi-copper oxidases. Singapore: World Scientif-
Lante A, Crapisi A, Krastanov A, Spettoli P. Biodegradation of ic; 1997.
phenols by laccase immobilised in a membrane reactor. Process Meyer U. Biodegradation of synthetic organic colorants. Microbial
Biochem 2000;36:51–8. degradation of xenobiotic and recalcitrant compounds. FEMS
Leech D, Daigle F. Optimisation of a reagentless laccase electrode for symposium, Vol. 12. London: Academic Press; 1981. p. 371–85.
the detection of the inhibitor azide. Analyst 1998;123:1971–4. Micard V, Thibault JF. Oxidative gelation of sugar-beet pectins: use of
Leite OD, Lupetti KO, Fatibello-Filho O, Vieira IC, de Barbosa AM. laccases and hydration properties of the cross-linked pectins.
Synergic effect studies of the bi-enzymatic system laccase- Carbohydr Polym 1999;39:265–73.
peroxidase in a voltammetric biosensor for catecholamines. Michniewicz A, Ledakowicz S, Jamroz T, Jarosz-Wilkolazka A,
Talanta 2003;59:889–96. Leonowicz A. Decolorization of aqueous solution of dyes by the
Levin L, Forchiassin F, Viale A. Ligninolytic enzyme production and laccase complex from Cerrena unicolor. Biotechnologia
dye decolorization by Trametes trogii: application of the Plackett– 2003;4:194–203.
Burman experimental design to evaluate nutritional requirements. Mikolasch A, Hammer E, Jonas U, Popowski K, Stielow A, Schauer F.
Process Biochem 2005;40:1381–7. Synthesis of 3-(3,4-dihydroxyphenyl)-propionic acid derivatives
Li K, Xu F, Eriksson K-E. Comparison of fungal laccases and redox by N-coupling of amines using laccase. Tetrahedron 2002;58:
mediators in oxidation of a nonphenolic lignin model compound. 7589–93.
Appl Environ Microbiol 1999;65:2654–60. Minussi RC, Pastore GM, Durán N. Potential applications of laccase in
Lisdat F, Wollenberger U, Makower A, Hortnagl H, Pfeiffer D, the food industry. Trends Food Sci Technol 2002;13:205–16.
Scheller FW. Catecholamine detection using enzymatic amplifi- Mishra G, Tripathy M. A critical review of the treatments for
cation. Biosens Bioelectron 1997;12:1199–211. decolourization of textile effluent. Colourage 1993;40:35–8.
Lorenzo M, Moldes D, Rodríguez Couto S, Sanromán A. Improving Moeder M, Martin C, Koeller G. Degradation of hydroxylated
laccase production by employing different lignocellulosic wastes compounds using laccase and horseradish peroxidase immobilized
in submerged cultures of Trametes versicolor. Bioresour Technol on microporous polypropylene hollow fiber membranes. J Membr
2002;82:109–13. Sci 2004;245:183–90.
512 S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513

Moldes D, Gallego PP, Rodríguez Couto S, Sanromán A. Grape seeds: mentals and catalysis. ACS symposium series, Vol. 785. USA: Oxford
the best lignocellulosic waste to produce laccase by solid state University Press; 2001. p. 358–72.
cultures of Trametes hirsuta. Biotechnol Lett 2003;25:491–5. Potin O, Veignie E, Rafin C. Biodegradation of polycyclic aromatic
Mougin C, Jolivalt C, Malosse C, Chaplain V, Sigoillot JC, Asther M. hydrocarbons (PAHs) by Cladosporium sphaerospermum isolated
Interference of soil contaminants with laccase activity during the from an aged PAH contaminated soil. FEMS Microbiol Ecol
transformation of complex mixtures of polycyclic aromatic 2004;51:71–8.
hydrocarbons in liquid media. Polycycl Aromat Compd Rehorek A, Tauber M, Gubitz G. Application of power ultrasound for azo
2002;22:673–88. dye degradation. Ultrason Sonochem 2004;11:177–82.
Murugesan K. Bioremediation of paper and pulp mill effluents. Indian Reyes P, Pickard MA, Vazquez-Duhalt R. Hydroxybenzotriazole
J Exp Biol 2003;41:1239–48. increases the range of textile dyes decolorized by immobilized
Mustafa R, Muniglia L, Rovel B, Girardin M. Phenolic colorants laccase. Biotechnol Lett 1999;21:875–80.
obtained by enzymatic synthesis using a fungal laccase in a hydro- Riu J, Schönsee I, Barcelo D. Determination of sulfonated azo dyes in
organic biphasic system. Food Res Int 2005;38:995­1000. groundwater and industrial effluents by automated solid-phase
Nicotra S, Cramarossa MR, Mucci A, Pagnoni UM, Riva S, Forti L. extraction followed by capillary electrophoresis/mass spectrome-
Biotransformation of resveratrol: synthesis of trans-dehydrodimers try. J Mass Spectrom 1998;33:653–63.
catalyzed by laccases from Myceliophtora thermophyla and from Rodríguez Couto S, Sanromán MA. Coconut flesh: a novel raw
Trametes pubescens. Tetrahedron 2004;60:595–600. material for laccase production by Trametes hirsuta under solid-
Niku-Paavola M-L, Viikari L. Enzymatic oxidation of alkenes. J Mol state conditions. Application to Lissamine Green B decolouriza-
Catal B Enzym 2000;10:435–44. tion. J Food Eng 2005;71:208–13.
Norsker M, Jensen M, Adler-Nissen J. Enzymatic gelation of sugar Rodríguez Couto S, Sanromán MA. Effect of two wastes from
beet pectin in food products. Food Hydrocoll 2000;14:237–43. groundnut processing on laccase production and dye decolouriza-
Nyanhongo GS, Gomes J, Gübitz G, Zvauya R, Read JS, Steiner W. tion ability. J Food Eng 2006;73:388–93.
Production of laccase by a newly isolated strain of Trametes Rodríguez Couto S, Gundín M, Lorenzo M, Sanromán A. Screening of
modesta. Bioresour Technol 2002;84:259–63. supports for laccase production by Trametes versicolor in semi-
Nyanhongo GS, Rodríguez Couto, S, Gübitz GM. Coupling of 2,4,6- solid-state conditions. Determination of optimal operation condi-
trinitrotoluene (TNT) metabolites onto humic monomers by a new tions. Process Biochem 2002;38:249–55.
laccase from Trametes modesta Chemosphere In Press. Rodríguez Couto S, Hofer D, Sanromán MA, Gübitz GM. Production
Okazaki S-Y, Michizoe J, Goto M, Furusaki S, Wariishi H, Tanaka H. of laccase by Trametes hirsuta grown in an immersion bioreactor.
Oxidation of bisphenol A catalyzed by laccase hosted in reversed Application to decolourisation of dyes from a leather factory. Eng
micelles in organic media. Enzyme Microb Technol 2002;31:227–32. Life Sci 2004a;4:233–8.
O'Neill C, Hawkes FR, Hawkes DL, Lourenco ND, Pinheiro HM, Rodríguez Couto S, Rosales E, Gundín M, Sanromán MA.
Delee W. Colour in textile effluents — sources, measurement, Exploitation of a waste from the brewing industry for laccase
discharge consents and simulation: a review. J Chem Technol production by two Trametes sp. J Food Eng 2004b;64:423–8.
Biotechnol 1999;74:1009–18. Rodríguez Couto S, Sanromán MA, Hofer D, Gübitz GM. Stainless
Paice MG, Bourbonnais R, Reid ID. Bleaching kraft pulps with steel sponge: a novel carrier for the immobilisation of the white-rot
oxidative enzymes and alkaline hydrogen peroxide. Tappi J fungus Trametes hirsuta for decolourisation of textile dyes.
1995;78:161–9. Bioresour Technol 2004c;95:67–72.
Palmieri G, Cennamo G, Sannia G. Remazol brilliant blue R Rodríguez Couto S, Sanromán MA, Gübitz GM. Influence of redox
decolourisation by the fungus Pleurotus ostreatus and its oxidative mediators and metal ions on synthetic acid dye decolourization by
enzymatic system. Enzyme Microb Technol 2005;36:17–24. crude laccase from Trametes hirsuta. Chemosphere 2005;58:417–22.
Palmore GTR, Kim H-H. Electro-enzymatic reduction of dioxygen to Rodríguez Couto S, López E, Sanromán MA. Utilisation of grape
water in the cathode compartment of a biofuel cell. J Electroanal seeds for laccase production in solid-state fermentors. J Food Eng
Chem 1999;565:110–7. 2006;74:263–7.
Pazarlıoglu NK, Sariişik M, Telefoncu A. Laccase: production by Roure M, Delattre P, Froger H, (03.03.1992). Composition for an
Trametes versicolor and application to denim washing. Process enzymic coloration of keratin fibres, especially for hair and its use
Biochem 2005;40:1673–8. in a dyeing process. Eur Pat Appl EP0504005.
Peralta-Zamora P, Pereira CM, Tiburtius ERL, Moraes SG, Rosa MA, Roy JJ, Abraham TE, Abhijith KS, Sujith kumar PV, Thakur MS. Biosen-
Minussi RC, et al. Decolorization of reactive dyes by immobilized sor for the determination of phenols based on Cross-Linked Enzyme
laccase. Appl Catal B Environ 2003;42:131–44. Crystals (CLEC) of laccase. Biosens Bioelectron 2005;21:206–11.
Petersen BR, Mathiasen TE. Deoxygenation using laccase enzyme. Saito T, Kato K, Yokogawa Y, Nishida M, Yamashita N. Detoxification
Trends Food Sci Technol 1997;8:249. of bisphenol A and nonylphenol by purified extracellular laccase
Pickard MA, Roman R, Tinoco R, Vazquez-Duhalt R. Polycyclic aromatic from a fungus isolated from soil. J Biosci Bioeng 2004;98:64–6.
hydrocarbon metabolism by white rot fungi and oxidation by Schäfer A, Specht M, Hetzheim A, Francke W, Schauer F. Synthesis
Coriolopsis gallica UAMH 8260 laccase. Appl Environ Microbiol of substituted imidazoles and dimerization products using cells
1999;65:3805–9. and laccase from Trametes versicolor. Tetrahedron 2001;57:
Pointing SB. Feasibility of bioremediation by white-rot fungi. Appl 7693–9.
Microbiol Biotechnol 2001;57:20–33. Schliephake K, Mainwaring DE, Lonergan GT, Jones IK, Baker WL.
Poots VJP, McKay JJ. The removal of acid dye from effluent using Transformation and degradation of the disazo dye Chicago Sky
natural adsorbents — I Peat. Water Res 1976;10:1061–6. Blue by a purified laccase from Pycnoporus cinnabarinus. Enzyme
Poppius-Levlin K, Tamminen T, Kalliola A, Ohra-aho T. Characterization of Microb Technol 2000;27:100–7.
residual lignins in pulps delignified by laccase/N-hydroxyacetanilide. In: Sealey J, Ragauskas AJ, Elder TJ. Investigations into laccase-mediator
Argyropoulos DS, editor. Oxidative delignification chemistry funda- delignification of kraft pulps. Holzforschung 1999;53:498–502.
S. Rodríguez Couto, J.L. Toca Herrera / Biotechnology Advances 24 (2006) 500–513 513

Selinheimo E, Kruus K, Buchert J, Hopia A, Autio K. Effects of Tsioulpas A, Dimou D, Iconomou D, Aggelis G. Phenolic removal in
laccase, xylanase and their combination on the rheological olive oil mill wastewater by strains of Pleurotus spp. In respect to
properties of wheat doughs. J Cereal Sci 2006;43:152–9. their phenol oxidase (laccase) activity. Bioresour Technol
Semenov AN, Lomonosova IV, Berezin V, Titov I. Peroxidase and 2002;84:251–7.
laccase as catalysts for removal of phenylhydrazide protecting group Uchida H, Fukuda T, Miyamoto H, Kawabata T, Suzuki M, Uwajima T.
under mild conditions. Biotechnol Bioeng 1993;42: 1137–41. Polymerization of bisphenol A by purified laccase from Trametes
Setti L, Giuliani S, Spinozzi G, Pifferi PG. Laccase catalyzed- villosa. Biochem Biophys Res Commun 2001;287:355–8.
oxidative coupling of 3-methyl 2-benzothiazolinone hydrazone Ünyayar A, Mazmanci MA, Ataçağ H, Erkurt EA, Coral G. A
and methoxyphenols. Enzyme Microb Technol 1999;25:285–9. Drimaren Blue X3LR dye decolorizing enzyme from Funalia
Sigal GB, Mrksich M, Whitesides GM. Effect of surface wettability on trogii one step isolation and identification. Enzyme Microb
the adsorption of proteins of proteins and detergents. J Am Chem Technol 2005;36:10–6.
Soc 1998;120:3464–73. Uyama H, Kobayashi S. Enzyme-catalyzed polymerization to
Soares GMB, Costa-Ferreira M, Pessoa de Amorim MT. Decoloriza- functional polymers. J Mol Catal B Enzym 2002;19–20:117–27.
tion of an anthraquinone-type dye using a laccase formulation. Vandertol-Vanier HA, Vazquez-Duhalt R, Tinoco R, Pickard MA.
Bioresour Technol 2001a;79:171–7. Enhanced activity by poly(ethylene glycol) modification of Cor-
Soares GMB, Pessoa de Amorim MT, Costa-Ferreira M. Use of iolopsis gallica laccase. J Ind Microbiol Biotechnol 2002;29:214–20.
laccase together with redox mediators to decolourize Remazol Vianello F, Cambria A, Ragusa S, Cambria MT, Zennaro L, Rigo A. A
Brilliant Blue R. J Biotechnol 2001b;89:123–9. high sensitivity amperometric biosensor using a monomolecular
Soares GMB, Pessoa Amorim MT, Hrdina R, Costa-Ferreira M. layer of laccase as biorecognition element. Biosens Bioelectron
Studies on the biotransformation of novel disazo dyes by laccase. 2004;20:315–21.
Process Biochem 2002;37:581–7. Vinod S. Enzymatic decolourisation of denims: a novel approach.
Solomon EI, Sundaram UM, Machonkin TE. Multicopper oxidases Colourage 2001;48:25–6.
and oxygenases. Chem Rev 1996;96:2563–605. Xiao Y, Zhang S, Hu Q, Jiang W, Pu Ch, Shi Y. Immobilization of
Stephen JA. Electrooxidation of dyestuffs in waste waters. J Chem fungal laccase on chitosan and its use in phenolic effluents
Technol Biotechnol 1995;62:11­117. treatment. Weishengwu Xuebao 2003;43:245–50.
Tamminen T, Kleen M, Ohra-aho T, Poppius-levlin K. Chemistry of Xu F. Recent progress in laccase study: properties, enzimology, production
mediated-laccase delignification analyzed by pyrolysis-GC/MS. and applications. In: Flickinger MC, Drew SW, editors. The
J Pulp Pap Sci 2003;29:319–24. encyclopedia of bioprocessing technology: fermentation, biocatalysis
Tanaka T, Tonosaki T, Nose M, Tomidokoro N, Kadomura N, Fujii T, and bioseparation. New York: John Wiley & Sons; 1999. p. 1545–54.
et al. Treatment of model soils contaminated with phenolic Yaropolov AI, Skorobogat'ko OV, Vartanov SS, Varfolomeyev SD.
endocrine-disrupting chemicals with lactase from Trametes sp. in a Laccase. Properties, catalytic mechanism, and applicability. Appl
rotating reactor. J Biosci Bioeng 2001;92:312–6. Biochem Biotechnol 1994;49:257–80.
Tanaka T, Nose M, Endo A, Fujii T, Taniguchi M. Treatment of Zavarzina AG, Leontievsky AA, Golovleva LA, Trofimov SY. Bio-
nonylphenol with laccase in a rotating reactor. J Biosci Bioeng transformation of soil humic acids by blue laccase of Panus tigrinus
2003;96:541–6. 8/18: an in vitro study. Soil Biol Biochem 2004;36:359–69.
Tavares APM, Gamelas JAF, Gaspar AR, Evtuguin DV, Xavier Zhang X, Eigendorf G, Stebbing DW, Mansfield SD, Saddler JN.
AMRB. A novel approach for the oxidative catalysis employing Degradation of trilinolein by laccase enzymes. Arch Biochem
polyoxometalate–laccase system: application to the oxygen Biophys 2002;405:44–54.
bleaching of kraft pulp. Catal Commun 2004;5:485–9. Zille A, Tzanov T, Guebitz GM, Cavaco-Paulo A. Immobilized laccase
Timur S, Pazarlıoglu N, Pilloton R, Telefoncu A. Thick film sensors for decolourization of Reactive Black 5 dyeing effluent. Biotech-
based on laccases from different sources immobilized in polyani- nol Lett 2003;25:1473–7.
line matrix. Sens Actuators B Chem 2004;97:132–6. Zollinger H. Synthesis, properties and applications of organic dyes and
Toca-Herrera JL, Krastev R, Bosio V, Küpcü S, Pum D, Fery A, et al. pigments. Colour chemistry. New York: John Wiley-VCH Publish-
Recrystallization of bacterial S-layers on flat polyelectrolyte surfaces ers; 2002:92­100.
and hollow polyelectrolyte capsules. Small 2005;1:339–48.
Trudeau F, Daigle F, Leech D. Reagentless mediated laccase electrode for
the detection of enzyme modulators. Anal Chem 1997;69:882–6.