LACCASE: MACRO AND M ICROBIAL SOURCES, PR ODUCTION, [626754]

LACCASE: MACRO AND M ICROBIAL SOURCES, PR ODUCTION,
PURIFICATION, AND BI OTECHNOLOGICAL APPLI CATIONS

Cristina Valentina ALBU (PROCA), Raluca Stefania ENCEA, Camelia DIGUTA,
Florentina MATEI, Calina Petruta CORNEA

University of Agronomic Sciences and Veterinary Medicine of Bucharest, Faculty of
Biotechnology, 59 Marasti Blvd, District 1, Bucharest, Romania
Corresponding author email: florentina.matei@bio tehnologii.usamv.ro

Laccase belongs to the blue multicopper oxidases and participates in cross -linking of monomers, degradation of
polymers, and ring cleavage of aromatic compounds. It is widely distributed in higher plants and fungi. It is present
in Asc omycetes, Deuteromycetes , and Basidiomycetes and abundant in lignin -degrading white -rot fungi. Lacasse
has been reported to be produced by different mushrooms (Trametes, Ganoderma, Pleurotus) and by filamentous
bacteria (Streptomyces) or fungi (Aspergillus). The article proposes a comparative analysis o f the optimal
conditions for laccase production in the case of mushrooms and some microorganisms, like fungi or filamentous
bacteria. Meanwhile will be described the isolation, purification , and c haracterization of the laccase produce d by
such organisms. All these issues will be approached through the biotechnological application of these enzymes (dye
decoloration, bioremediation, etc).

Keywords: laccase, macromycetes (mushrooms), micromycetes (ba cteria, fungi )

1. INTRODUCTION

Laccase was first detected in the Japanese lac
tree Toxicodendron verniciflua . Later, it was
found in certain other plants, in many insects,
and in a variety of fungi, including yeasts
(e.g., Cryptococcus ), molds (e.g.,
Penicillium ), mushrooms (e.g., Agaricus ),
and white -rot fungi (e.g., Pleurotus ) [1].
Laccases play an important role in the food
industry, paper and pulp industry, textile
industry, synthetic chemistry, cosmetics, soil
bioremediation and biodegradation of
environmental phenolic pollutant and
removal of endocrine disruptors [2]. These
enzymes are used for pulp delignification,
pesticide or insecticide degradation, organic
synthesis [4], waste detoxification, textile
dye transformation, food technological uses,
and biosensor and analytical applications.
Recently laccases have been efficiently
applied to nanobiotechnology due to their
ability to catalyze electron transfer reactions
without additional cofactor. The technique
for the immobilizat ion of biomolecule such
as layer -by-layer, micropatterning, and self –
assembled monolayer technique can be used for preserving the enzymatic activity of
laccases [5].

2. LACCASE – BIOCHEMIST RY
In recent years, enzymes have gained great
importance in Industries; laccases are one
among them which are widely present in
nature. Laccases are the oldest and most
studied enzymatic systems. These enzymes
contain 15 – 30 % carbohydrate and have a
molecul ar mass of 60 –90 kDa.
These are copper -containing 1,4 –
benzenediol: oxygen oxidoreductases (EC
1.10.3.2) found in higher plants and
microorganisms. These are glycosylated
polyphenol oxidases that contain 4 copper
ions per molecule that carry out 1 electron
oxidation of phenolic and its related
compound and reduce oxygen to water [2, 3].
When the substrate is oxidized by a laccase,
it loses a single electron and usually forms a
free radical which may undergo further
oxidation or non -enzymatic reactions
including hydration, disproportionation, and
polymerization [4] . These enzymes are
polymeric and generally contain 1 each of
type 1, type 2, and type 3 copper

center/subunit where the type 2 and type 3 are
close together forming a trinuclear copper
cluster.
Laccases are divided into “low -redox
potential” and “high -redox potential”
laccases depending on the structure and
properties of the copper center. The high –
redox potential laccases occur mainly in
basidiomycetes, especially white -rot fungi
(Gutierrez et al. 2006; Rebrikov et al. 2006;
Quaratino et al. 2007; Cherkas hin et al. 2007;
Hernandez -Luna et al. 2008), the low -redox
potential laccases seem to be widely
distributed in molds (Jung et al. 2002),
bacteria, insects, and plants. However,
detailed studies by Klonowska et al. (2002,
2005) have shown that the white -rot fungus
Trametes sp. C30 possesses not only the
high-redox potential laccase (LAC1) but also
two additional low -redox potential laccases
(LAC2 and LAC3) with minor activity.
Given their versatility and broad substrate
specificity, laccases as a family of copper –
containing oxidases catalyzing a variety of
oxidations could become among the most
important biocatalysts in fungal
biotechnology. Because of this, their
biochemical properties and molecular
evolution are of considerable interest and
have been summa rized in several reviews
(Xu 1999; Claus 2004; Nakamura and Go
2005; Baldrian 2006; Zhuklistova et al.
2008). [ 1]

3. MACROBIAL SOURCES OF
LACCASE

The attention of researchers in recent decades
has been focused on the comprehensive study
of different mush room species, which belong
particularly to Macrofungi (Macromycetes).
Ascomycetes and Basidiomycetes are two the
most diverse and exceedingly numerous
group of Macrofungi that have been
intensive ly investigated in various aspects.
Exploration of the enzymatic activity of these
mushrooms is one of the important trend for
understanding their physiological,
biochemical features and in order to clear up the considerable promising potential for
industria l and biotechnological applications.
Laccase is one of the most studied and key
enzyme s. According to the Journal of
Microbiology, Biotechnology and Food
Sciences positive reaction was detected in 21
species mostly in 2 -4 days after inoculation
and formed a reddish -brown zone around the
mycelium. The most active producers of
laccase were Lentinus edodes (Fig 1B), L.
luscina and Coprinus comatus (the latter two
mentioned species are referred to the soil
saprotrophic). We should note the fact of
laccase prese nce in two ( Agrocybe aegerita
and Fomitopsis pinicola ) out of four brown –
rot species. Souza et al. (2008) found in the
genome of brown -rot silencing genes of
laccase. Investigations showed laccase
activity of species L. luscina, Crinipellus
schevczenkovi, A. aurea, Hypsizygus
marmoreus, L. schimeji, Oxyporus obducens ,
and S. litschaueri for the first time. The
existence of laccase has been detected in
earlier studies in similar species: A. agerita
(Saparrat et al., 2000; Erden et al., 2009; Jang
et al., 200 9), C. comatus (Kalmiș et al.,
2008), Fomes fomentarius (Elisashvili et al.,
2009), Fomitopsis pinicola, Flamulina
velutipes, Ganoderma lucidum (Jarosz –
Wilkołazka et al., 2002), G. applanatum
(Jarosz -Wilkołazka et al., 2002; Elisashvili et
al., 2009), Grifola frondosa (Jang et al.,
2009; Buchalo et al., 2011a), L. edodes
(Jarosz -Wilkołazka et al., 2002; Kalmiș et al.,
2008; Buchalo et al., 2011a; Seshikava and
Singara, 2012), P. eryngii (Kalmiș et al.,
2008; Jang et al., 2009; Seshikava and
Singara, 2012) a nd P. ostreatus (Erden et al.,
2009; Jang et al., 2009; Elisashvili et al.,
2002, 2008, 2009; Shraddha et al., 2011;
Seshikava and Singara, 2012). This enzyme
has been found in P. djamor (Kalmiș et al.,
2008), H. myxotricha (Jang et al., 2009), L.
sulphure us (Shraddha et al., 2011; Seshikava
and Singara, 2012), and T. versicolor (Jarosz –
Wilkołazka et al., 2002; Elisashvili et al.,
2008, 2009; Erden et al., 2009; Jang et al.,
2009; Shraddha et al., 2011; Seshikava and
Singara, 2012). [6]

4. MICROBIAL SOURCES OF
LACCASE

Considering the microbial sources, laccase is
produced by a large variety of bacteria (Table
1) and filamentous fungi (Table 2). In lower
fungi, as Zygomycetes or Chytridiomycetes ,
the production of laccase was not
demonstrated [9].
There are also laccase -producing yeasts, like
Kluyveromyces dobzhanskii, Pichia
manshurica (Wakil et al., 2017) and
Cryptococcus neoformans (Eisenman et al.,
2007), but they have far fewer known
representants than the other classes of
microorganisms.

Table 1. Bacterial laccase sources (after
Desai and Nityanand, 2011)
Sources References
Bacteria
Azospirillium
lipoferum Givaudan et al.,
1993; Faure et al.,
1994
Bacillus subtilis Martins et al., 2002
S. maltophilia
AAP56 Galai et al., 2009
Streptomyces
coelicolor Dube et al., 2008

Table 2. Fungal laccase sources (after
Baldrian, 2006)
Sources References
Fungi
Botrytis cinerea Slomczynski et al.,
1995
Aspergillus
nidulans Scherer & Fischer,
1998
Chetomium
thermophilum Chefetz et al., 1998
Chalara paradoxa Robles et al., 2002
Magnaporthe
grisea Iyer & Chattoo, 2003
Mauginiella sp. Palonen et al., 2003
Melanocarpus
albomyces Kiiskinen et al., 2002
Monocillium
indicum Thakker et al., 1992 Myrothecium
verrucaria Sulistyaningodyah et
al., 2004
Neurospora
crassa Froehner & Eriksson,
1974
Ophiostoma novo –
ulmi Binz & Canerascini,
1997
Podospora
anserina Durrens, 1981
Rhizoctonia solani Wasaki et al., 1967
Trichoderma
giganteum Wang & Ng, 2004

Properties of laccase: extracellular
localization; molecular weight of
approximatively 60 to 70 kDa (typical in
fungi); isoelectric pH point between 3,0 and
7,0; pH optima in fungi is between 3,6 and
5,2 (highly dependent on the substrate); its
structure i s usually monomeric
(homodimeric laccases were found too, in
Gaeumannomyces graminis, Monocillium
indicum, Podospora anserina ); optimal
temperature around 55 -60°C (for B.cinerea).
Purified laccase has a blue appearance
around 600nm, in spectrophotometry.
Bacterial laccases have higher
thermostability and halotolerance than fungal
laccases and are thought to be more valuable
in dye decolorization, biofuel production and
biobleaching (Fang et al.; Martins et al.,
2014).
There are some lignolytic -activity
actinomycetes, like Trichoderma sp. or
Botryosphaeria sp. , which displayed laccase
production.
The laccase produced by Monocillium
indicum was the first Ascomycetae laccase,
characterized with similar immunological
features with Coriolus versicolor and
Agaricus bisporus laccases and with lignin
peroxidase activity similar to Phanerochaete
chrysosporium laccase (Thakker et al., 1992;
Shraddha et al., 2011).
Pseudomonas putida was revealed to be a
laccase -producing bacteria, with the attribute
of decolori zing synt hetic dyes and industrial
effluents.
Streptomyces cyaneus and Streptomyces
ipomoea showed laccase activity, the former
having it tens of times stronger [10].

Although it is known that Streptomyces
griseus is also a producer of laccase, in the
cited assessment it did not show this kind of
enzymatic activity – the localization of the
laccase could be responsible for the absence
of activity detected.
A fungal laccase, produced by and extracted
from the Ascomycete Chaetomium sp. , was
characterized with the ability to decolorize
different dyes, even in the presence of high
concentrations of sodium chloride [11]. The
purified laccase from this fungus is able to
degrade or transform various synthetic dyes,
as Acid Orange, Direct Red, Direct Blue and
RBBR (Remazol Brilliant Blue R), with and
without mediators.
Another laccase, able to decolorize synthetic
dyes, is the one produced by Spirulina
platensis , a cyanobacterium [12] purified and
characterized the laccase from this Spirulina
genus, and evaluated its decolorization
property on Reactive Blue 4: almost
complete decolorization, without any
mediators.
A psychrotolerant bacterial strain of Serratia
marescens was studied for its laccase
production [13]. It was proved to synthesize
laccase even under ext reme conditions,
which is likely to be beneficial for
biotechnological applications.
The laccase obtained from Ascomycetae
Myceliophtora thermophila is suitable for
industrial pulp bleaching and delignification
[14]

5. ISOLATION, PURIFICAT ION
AND CHARACTE RIZATION OF
LACCASE

5.1. Isolation of laccase
When the enzyme is immobilized, it
becomes more vigorous and resistant to
alteration in the environment which allows
easy recovery and reuse of enzyme for
multiple purposes. That is why researchers
are moving towards the efficient methods of
immobilization which influence the
properties of the biocatalyst. Laccase
immobilization has been studied with a wide range of different immobilization methods
and substrates. [17]
Immobilization techniques can improve the
process economy of laccase -based
biotechniques. A variety of different
methods have been shown to be useful.
These include immobilization o n polyamide
matrices (Silva et al. 2007), on glass supports
(Cho et al. 2008), on epoxy -activated carriers
(Berrio et al. 2007; Kunamneni et al. 2008),
on magnetically separable silica spheres
(Zhu et al. 2007), on magnetic chitosan
microspheres (Jiang et al. 2005), on
nanoparticles, and on kaolinite (Hu et al.
2007). Furthermore, laccase may be
immobilized by entrapment in alginate –
chitosan microcapsules (Lu et al. 2007) or in
Cu–Al and Cu -alginate beads (Teerapatsakul
et al. 2008; Niladevi and Prema 2008) . In
most cases, immobilization leads to
increased enzyme stability and improved
resistance to changes in pH and temperature.
[1]

5.2. Purification of laccase
Ammonium sul fate is being commonly used
for the enzyme purification for many years.
But research ers have found much more
efficient methodologies such as protein
precipitation by ammonium sul fate, anion
exchange chromatography, desalt/buffer
exchange of protein, and gel filtration
chromatography. Single -step laccase
purification from Neurospora crassa takes
place by using celite chromatography and 54
fold purification was obtained with a specific
activity of 333 U mg−1 [ 7]. Laccase from
LLP13 was first purified with column
chromatography and then purified with gel
filtration [ 8, 15]. Laccase from T. versicolor
is purified by using ethanol precipitation,
DEAE -Sepharose, Phenyl -Sepharose and
Sephadex G -100 chromatography which is a
single monomeric laccase with a specific
activity of 91,443Umg−1 [ 16]. Laccase from
T. versicolor is purified with Ion Exch ange
chromatography followed by gel filtration
with a specific activity of 101UmL−1 and
34.8-fold purification [18]. Laccase from
Stereum ostrea is purified with ammonium

sulfate followed by Sephadex G -100 column
chromatography with 70 -fold purification
[19]. Laccase from fruiting bodies is purified
with ammonium sul fate precipitation with
40–70% saturation and DEAE cellulose
chromatography then 1.34 and 3.07 fold
purification is obtained respectively [ 17].
Laccase can be immobilized on different
pyrolytic graphite (best support), ceramics
supports and on a carbon fiber electrode
where it acts as a biosensor. At the 12th day,
maximum laccase activity 40,774.0UL−1
was achieved [20]. An optical biosensor is
fabricated by using stacked films for the
detection of phenolic compounds; 3 -methyl –
2-benzothiazolinone hydrazone (MBTH)
was immobilized on a silic ate film and
laccase on a chitosan film [21].

5.3. Characterization of laccase
According to Takao Saito et al. 2003 [22] , the
purified laccase produced one band on an
SDS-PAGE gel at the apparent molecular
mass of approximately 73 kDa (Fig. 2A).

Fig. 2. SDS-PAGE (A) and IEF (B) of the
purified strain I -4 laccase

This laccase was used in gel filtration
chromatography on a HiLoad 16/60
Superdex 200 pg column, the molecular mass
of the native enzyme being estimated at 80
kDa. The isoelectric point (pI), de termined by analytic isoelectric focusing, was 3.5 (Fig.
2B). This pI is similar to that of the laccase
from the basidiomycete PM1 [ 23], which
has an acidic isoelectric point. To determine
the state of its catalytic center, the laccase
was characterized sp ectroscopically. The
purified laccase had a blue color, typical of
copper -containing proteins. The UV-Vis
spectrum of the laccase showed a peak
absorption at about 611 nm, typical for the
type I Cu(II), that is responsible for the deep
blue color of the en zyme. A shoulder at about
333 nm suggests the presence of type III
binuclear Cu(II) pair [ 24]. The EPR spectrum
of the laccase showed the superimposed
signals from type I and type II Cu(II), each in
a different coordination environment. The
parameters of the type I Cu(II) signal were
gII = 2.21 and AII = 8.3 × 10−3 cm−1, and
those of the type II Cu(II) signal were gII =
2.25 and AII = 1.93×10−2 cm−1. These
spectral characteristics are similar to those of
other blue copper proteins that ha ve four
copper atoms [ 25, 26].

6. APPLICATIONS IN
BIOTECHNOLOGY

6.1. Food Processing I ndustry
In the food industry, laccase is used for the
elimination of undesirable phenolic
compound in baking, juice processing, wine
stabilization, and bioremediation of
wastewater. [17]
Wine browning, due, primarily, to enzymatic
and chemical oxidation of phenolic
compounds, represents one of the most
unwanted processes that can occur in wine –
making. During the crushing of the grapes,
the release of laccase from Botryt is cinerea
affected beans in the must could determine a
significant reduction of phenolic compounds.
Polyphenols, including their major classes,
important in wine (phenolic acids, catechins,
anthocyanins, tannins , and stilbenes), are
converted to the corre sponding quinones,
which pass into dark -colored polymers.
These polymers are insoluble in water and
aqueous solutions and precipitates from the
must and wine. Moreover, the oxidation of

the phenolic compounds can adversely affect
the sensorial and nutritio nal properties of the
wine.
The storage of beer depends on various
factors, such as the haziness, the oxygen
content, the temperature. The haziness is
caused by small amounts of
proanthocyanidins, which are naturally
occurring polyphenols and proteins that
could cause precipitation. This type of
complex is known as „cold haze” and occurs
during chilling – can be re -dissolved at room
temperature or at higher temperatures. Even
the products that do not have this disorder
when packaged, could form it during lo ng
storage. The usage of laccase for the
oxidation of polyphenols as an alternative to
traditional therapy has been tested many
times. Also, laccase is used to eliminate the
oxygen at the end of the production process
of beer (Osma et al., 2010).
Suga r beet pectin is a functional aliment,
which can form thermo -irreversible gelatins.
These types of gelatin are very interesting
and can be used in the food industry because
they can be warming while retaining the gel
structure. Compared with peroxidase, which
is used as a food additive, laccase proved to
be more effective and safer for consumption.
One of the biggest problems in the fruit juice
processing is the enzymatic and the chemical
browning. The color and the taste of the fruit
depends on the phenolic com pounds, which
should be selectively removed from the
composition, in order to prevent any
alteration of taste, flavor , and color –
attributed to oxidation of polyphenols. To
prevent the discoloration of the fruit
beverages, by replacing the chemical
adsorb ents, enzymes are being used, such as
laccase. It has the potential to eliminate
unwanted phenols responsible for browning
and disorder in many beverages, such as fruit
juice, beer , and wine.
In the bread -making process , it is known that
additives are adde d to improve bread and
bread dough, from which results improved
texture and flavor , a larger volume and longer
freshness. In recent years, enzymes have
been increasingly used as enha ncing agents, including laccase. Even if the laccase used in
the preparati on of doughs could be of any
origin, inclusively vegetal, it is preferable to
be of microbial origi n, because it is easier to
handle and produce on a larger scale (Si,
2001) [27].

6.2. Dye Decolorization
Textile industry utilizes a large volume of
water and chemicals for wet processing.
These chemicals range from inorganic
compounds to organic compounds. The
chemical structure of dyes provides
resistance to fading when exposed to light,
water, and other chemicals.
Laccase degrades dye; that is why laccase –
based processes have been developed which
include synthetic dyes and are being used in
the industry nowadays [30, 31].
Bl´anquez et al. [32] used T. versicolor in the
form of pellets to treat a black liquors
discharge for detoxifying and reducing the
color, aromatic compounds, and chemical
oxygen demand (COD). They found that
color and aromatic compounds were reduced
up to 70 –80% and COD Enzyme Research 5
was reduced up to 60%. They concluded that
T. versicolor is able to produce laccase. T.
versicolor completely decolorizes the
Amaranth, Tropaeolin O, Reactive Blue 15,
Congo Red, and Reactive Black 5 with no
dye sorption while it partially decolorizes
Brilliant Red 3G -P, Brilliant Yellow 3B -A
and Remazol Brilliant Blue R with some dye
sorption. They found that after
decolorization, the toxicity of few dyes
remained the same while some became non –
toxic [33]. Laccase -based hair dyes are less
irritant and easier to handle than conventional
hair dyes because laccases replace H2O2 in
the dye formulation [34]. La ccase is also used
in the dechlorination process.
Xylidine is a laccase inducer which increases
dichlorination activity due to which
dissolved oxygen concentration is reduced
[35]. Romero et al. found that bacteria S.
maltophilia decolorizes some synthetic dyes
(methylene blue, methyl green, toluidine
blue, Congo red, methyl orange, and pink) as
well as the industrial effluent. [17]

TEMPO oxidation by laccase is used in the
production of hydrophobic cotton fibers and
hydrophobic jute fibers (with dodecyl ga llate
in this case).

6.3. Bioremediation and Biodegradation
Laccase is used in bioremediation, bio –
solubilization , and desulfurization, in the
production of biosensors and biofuels and
also in the production of fiberboards.
Because of her oxidizing ability towards
phenolic and non -phenolic compounds,
laccase can and is used for degradation of
industrial wastes, like paper, oil, leather,
pharmaceutical, pesticides [29].
The presence of phenols in the industrial
wastewater attracted interest in the
application of bioremediation processes and
their treatment with laccase. The presence of
phenolic compounds in drinking water is a
real danger. The distillery wastewater is
generated during the production of ethanol by
the fermentation of sugar cane molasses . This
produces a major environmental impact, due
to the high content of soluble organic matter
and due to its dark brown color. Fungal
laccase showed better properties in the
reduction of total phenolic compounds in
color than the laccase from other sourc es.
Due to rapid industrialization and extensive
use of pesticides for better agricultural
productivity, contamination of soil, water,
and air take place which is a serious
environmental problem of today.
Polychlorinated biphenyls (PCB), benzene,
toluene, ethylbenzene, xylene (BTEX),
polycyclic aromatic hydrocarbons (PAH),
pentachlorophenol (PCP), 1,1,1 -trichloro –
2,2-bis (4 -chlorophenyl) ethane (DDT), and
trinitrotoluene (TNT) are the substances
which are known for their carcinogenic as
well as mutagenic ef fect and are persistent in
the environment. Fungi renovate a wide
variety of hazardous chemicals; that is why
the researcher’s interest is generated in them
[36].
T. versicolor is used for the bioremediation of
atrazine in soil with low moisture and organi c
contents that are normally found in semiarid
and Mediterranean -like ecosystems [37]. Keum and Li [38] obtained laccase from
T. versicolor and Pleurotus ostreatus for the
degradation of PCBs as well as phenol and
found as chlorination increases, degradati on
rate decreases and concluded that 3 -hydroxy
biphenyl was more resistant to laccase
degradation than 2 – or 4-hydroxy analogs.
After five days of incubation, when glucose
and fructose were used as a cosubstrate than
71% of p -hydroxy benzoic acid and 56% o f
protocatechuic acid were degraded [39].
Laccase obtained from T. villosa remediates
the soil by degrading 2,4 -DCP (2,4 –
dichlorophenol). An experiment was
performed by Ahm in which he took 2 types
of soil: in soil 1, both free and immobilized
laccase remo ve 100% of 2,4 -DCP (without
regard of moisture content). In soil 2,
immobilized laccase removed more 2,4 -DCP
(about 95%) than free enzyme (55%, 75%,
and 90%, at 30%, 55%, 100% maximum
water holding capacity) [40]. Cerrena
unicolor produces laccase in the l ow nitrogen
medium which has the capability of reducing
lignin content from sugarcane bagasse up to
36% within 24 h at 30◦C [41].

6.4. Paper and Pulp Industry
Chlorine and oxygen -based chemical
oxidants are used for the separation and
degradation of ligni n which is required for
the preparation of paper at an industrial level.
But some problems such as recycling, cost,
and toxicity remain unsolved. However, in
the existing bleaching process, LMS could be
easily implemented because it leads to a
partial repl acement of ClO2 in pulp mills
[42].

6.5. Medical applications
The biosensors used in biomedical
engineering are also produced with laccase.
In the determinations of chemical
compounds, in nanobiotechnology and
biomedicine and cosmetics is used laccase
too.
The effect of poison ivy dermatitis, which is
caused by urushiol, can be reduced with
laccase treatment. It has been shown that

laccase can oxidize the urushiol to a quinone
derivative, that is innocuous [28].
Some important drugs, like anti -cancer
drugs , antioxidants, hormones , and hormone
derivatives are prepared with the help of
laccase (by oxidation) and are added to some
cosmetics [29].
Laccase can also oxidize iodide to iodine,
which is used as a disinfectant.

7. CONCLUSIONS

Laccases are the versatile enzymes which
catalyze oxidation reactions coupled to four –
electron reduction of molecular oxygen to
water. They are multicopper enzymes which
are widely distributed in higher plants and
fungi. They are capable of degrading lignin
and are present abundantly in many white -rot
fungi. They decolorize and detoxify the
industrial effluents and help in wastewater
treatment. They act on both phenolic and
nonphenolic lignin -related compounds as
well as highly recalcitrant environmental
pollutants which he lp researchers to put them
in various biotechnological applications.
They can be effectively used in paper and
pulp industries, textile industries, xenobiotic
degradation, and bioremediation and act as a
biosensor. Laccase has been applied to
nanobiotechno logy which is an increasing
research field and catalyzes electron transfer
reactions without additional cofactors.
Recently several techniques have been
developed for the immobilization of
biomolecule s such as micropatterning, self –
assembled monolayer, and layer -by-layer
technique which immobilize laccase and
preserve their enzymatic activity. [17]

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