African Journal of Biotechnology Vol. 7 (12), pp.

1983-1990, 17 June, 2008

Available online at http://www.academicjournals.org/AJB
ISSN 16845315 2008 Academic Journals

Full Length Research Paper

Autochthonous white rot fungi from the tropical forest:

Potential of Cuban strains for dyes and textile industrial
effluents decolourisation
Maria. I. Snchez-Lpez 1, Sophie F. Vanhulle2, Valrie Mertens2, Gilda Guerra1, Sara Herrera
Figueroa4, Cony Decock3, Anne-Marie Corbisier2 and Michel J. Penninckx5*
1

Departamento de Microbiologa, Facultad de Biologa, Universidad de la Habana, Calle 25 #455, CP:10400, Ciudad
Habana, Cuba.
2
Unit de Microbiologie, Facult dIngnierie Biologique, Agronomique et Environnementale, Universit catholique de
Louvain, Croix du Sud 3 bote 6, B-1348 Louvain-la-Neuve, Belgium.
3
Mycothque de lUniversit catholique de Louvain (MUCL, MBLA), Facult dIngnierie Biologique, Agronomique et
Environnementale, Universit catholique de Louvain, Croix du Sud 3, B-1348 Louvain-la-Neuve, Belgium.
4
Instituto de Ecologia y Systematica, Division de Micologia Carretera de Varona Km 3 , Capdevilla, Ciudad Habana,
Cuba.
5
Laboratoire de Physiologie et Ecologie Microbienne, Ecole Interfacultaire des Bioingnieurs, Universit Libre de
Bruxelles c/o Institut Pasteur, Rue Engeland 642, B-1180, Brussels, Belgium.
Accepted 26 March, 2008

Nineteen strains of wood-inhabiting pores fungi, representing thirteen species and ten genera,
collected from both natural and anthropic (urban) ecosystems in Cuba were tested for their ability to
decolourise the industrial anthraquinonic dye Acid Blue 62 (AB 62) in laboratory conditions, in both
solid and liquid media. On the basis of their decolourisation rate and growth inhibition, seven strains
viz. Tinctoporellus epimiltinus, Trametes maxima, Perenniporia tephropora, Coriolopsis rigida,
Hexagonia tenuis, Pachykytospora alabamae and Hexagonia hydnoides were selected for further
studies. All the strains were able to decolourise partially or completely the AB62 dye added to Malt
extract. Almost total decolourisation was obtained with T. maxima. Decolourising activity was also
shown with an industrial textile effluent, containing Remazol Yellow RGB, Remazol Red RR, and
Remazol Black B 133. Production of laccase, a ligninolytic enzyme possibly involved in decolourisation,
was stimulated by AB 62 for all the strains tested; T. maxima showing the highest production. Lignin
peroxidase and manganese peroxidase were not produced under the experimental conditions used. T.
maxima could represent a potential candidate for biotechnological applications. The exploitation of
local biodiversity in tropical area appears as a potentially productive approach for identifying promising
microbial strains for industrial use.
Key words: Anthraquinones, dyes, bioremediation, decolourisation, laccase, textile effluent, Trametes maxima,
white rot fungi, Acid Blue 62.
INTRODUCTION
Wastewaters from dyes and textile industries contain a
variety of pollutants, including dyes which are hardly
removed by conventional activated sludge treatment. A
successful management of these effluents often necessitates expensive physico-chemical treatment using, for
*Corresponding author. E-mail: upemulb@resulb.ulb.ac.be. Tel:
+32 2 373 33 03. Fax: + 32 2 3733309

instance, ozone (Khraisheh, 2003). Microbial decolourisation has been proposed as a less expensive and less
environmentally intrusive alternative (Selvam et al.,
2003). In the last ten years, so-called White Rot Fungi
(WRF) emerged as a promising group for biotechnological applications, especially in bioremediation (Reddy
and Mathew, 2001.). The oxidative enzymes of WRF
have broad substrate specificity and have been proposed
for their use in the transformation and mineralization of

1984

Afr. J. Biotechnol.

from a cotton dyer company. It contained inter alia acid and reactive
dyes, salts and intermediates.

organo-pollutants structurally similar to lignin (Pointing,

2001). It has also been shown that the lignin degrading
system of WRF is exploitable for dye decolourisation
(Chander et al., 2004; Shah and Nerud, 2002;
Wesenberg et al., 2003). Among the ligninolytic enzymes
produced by WRF, laccases appear as particularly active
in dye degradation (Baldrian, 2004; Rodriguez et al.,
1999).
Many studies dedicated to bioremediation competence
of WRF assess the properties of strains deposited in
public collections (Jaouani et al., 2003). There have been
in contrast fewer contributions attempting to exploit directly local biodiversity, especially in African and Latin
America tropical area (Levin et al., 2004; Pointing et al.,
2000a; Tekere et al., 2001). However, this approach
appears to be potentially productive for identifying new,
promising strains for biotechnological applications
(Pointing et al., 2003).
During several surveys of wood-inhabiting fungi on
Cuba, in both natural and anthropic ecosystems, in different provinces, 200 strains were isolated in pure culture,
and later sorted according to their type of rot (viz. brown
or white rot), ecology, and habitat. On the basis of their
type of rot, ecology, and habitat, nineteen strains, representing thirteen species and ten different genera (Table
1) were then selected to evaluate their ability to decolourise dyes, either as raw material or mixed in a textile
effluent, in relation with laccase production. The results of
this screening are presented and discussed below.

The culture broth was filtered and biomass was washed and dried
at 105C for 2 days. The fungal biomass was determined as dry
mycelial weight per litre of culture broth.

MATERIAL AND METHODS

Laccase activity

Organisms and culture conditions

Laccase activity was determined by monitoring the A414 change

related to the rate of oxidation of 25 mM 2,2 -azino-bis-[3ethylbenzothiazoline-6-sulfonic acid] (ABTS) in 100 mM Na-tartrate
buffer (pH 4.5) (Lonergan and Baker, 1995). The value used for
molar extinction coefficient of ABTS ( 414) was 34450 M-1 cm-1.
Assays were performed in 1-ml cuvette at ambient temperature (2225C) with 50 l of adequately diluted culture liquid. One unit
activity (U) was defined as the amount of enzyme, which oxidized 1
mol of ABTS per minute and the activities were expressed in U l-1.

The strains used in the present study are listed in Table 1. After
isolation in pure culture and identification, strains were deposited
and stored at the BCCMTM/MUCL and at the CRGF (Colleccion de
Recursos Geneticos Fungicos) at the Institute of Ecology and
Systematic, Cuba. The strains were preserved by cryopreservation
at 130C (at BCCMTM/MUCL) or alternatively on agar slopes
under mineral oil or water in both collections. Pycnoporus sanguineus (MUCL 41582) was used as a reference strain (Vanhulle et
al., 2001).
The strains were grown first on 2% (w/v) malt extract agar (MA2)
incubated at 25C for 7 days. One plug of agar was sampled at the
margin of the colony with a hollow-punch of 4 mm diameter to serve
as inoculum for subsequent MA2 plates or MA2 plates added with
1.75 mM Acid Blue 62. The diameter of growth and decolourisation
were measured each day. Averages of radial growth rates
(mm/day) were calculated.
For growing in liquid medium, seventeen plugs of agar were
sampled with a hollow-punch of 4 mm diameter at the margin of the
colony of a MA2 7 days old pre-culture to serve as inoculums.
Erlenmeyer of 250 ml contained either 100 ml of a 2% (w/v) malt
extract broth (ML2), ML2 supplemented with 1.75 mM Acid Blue 62,
or ML2 supplemented with 0.5% (v/v) of the industrial dye effluent.
The cultures were incubated a 25C for 14 days in a rotary shaker
at 125 rpm [stroke 50 mm, and shaking plate dimensions 760 x 600
(W x D) mm], with daily sampling. Acid Blue 62 was courteously
supplied by Yorkshire Europe (Tertre, Belgium).
The industrial effluent was obtained as an anonymous sample

Decolourisation in liquid medium

Dye decolourisation was determined spectrophotometrically by
monitoring of the absorbance at 500 and 595 nm for the dye Acid
Blue 62. For the industrial effluent a spectrophotometric integrative
method (SIM) was adapted from a method described by the
"American Public Health Association" (APHA). APHA describes the
determination of the colour standard in Hazen units, wherein 1 mg/l
Pt equals one Hazen. The stock solution contained 2.49 g K2PtCl6
and 2.02 g CoCl26H2O dissolved in 200 ml of concentrated analytical grade HCl (d =1.19) and diluted to 1 litre with distilled water.
A445 of this solution represented 1000 Hazen. This method was able
to measure coloration of the water in the visible spectrum. The
spectrophotometric integrative method (SIM) used here monitored
the absorbance of the sample in the visible region from 380 to 740
nm. The integration of the curve absorbance/wavelength gave a
numerical result in area units. To convert this result in colour units
(APHA), five dilutions of the stock solution of the standard (containing Pt) described above were measured by both APHA and SIM
methods. A graph area/hazen unit was constructed for the standard. A conversion factor was calculated, and the result obtained
for the coloured sample was converted in colour units (APHA).

Biomass

Statistics
All the experiments were performed at least two times using three
replicates. The data presented correspond to mean values with a
standard error less than 10%.

Cluster analysis
Cluster analysis of hierarchical order, based on Euclidian distances,
using the percentages of decolourisation of the dye and the industrial
effluent reached by the different strains, was performed by UPGMA
(Unweighted Pair-Group Method, Arithmetic Average) method, using
the statistical package NTSys-pc version 2.02i (1998), which presented
the more adequate cophenetic values. The robustness of the tree
topology was assessed by 2000 times resampling using a bootstrap
analysis (Yap and Nelson, 1996).

Snchez-Lpez et al.

1985

Table 1. Origin of the fungi strains and growth parameters on solid medium.

Strains (*)(**)
Coriolopsis rigida MUCL44156 (CRGF121)
Coriolopsis sp MUCL 41639
Flavodon flavus MUCL 44097(CRGF 175)
Hexagonia hydnoides MUCL 41636 (CRGF
241)
Hexagonia hydnoides MUCL 43543 (CRGF
242)
Hexagonia tenuis MUCL 43553 (CRGF 248)
Hexagonia tenuis MUCL 43554 (CRGF 251)
Lentinus crinitus MUCL 41620 ( CRGF 273)
Megasporoporia setulosa MUCL 43550 ( CRGF
268)
Pachykytospora alabamae
MUCL 44100 (
CRGF 296)
Pachykytospora alabamae MUCL 44126 (CRGF
298)
Perenniporia cubensis MUCL 41656 (CRGF
313)
Perenniporia cubensis MUCL 43209 (CRGF
314)
Perenniporia tephropora MUCL 43535 (CRGF
323)
Perenniporia tephropora MUCL 43536 (CRGF
325)
Pycnoporus sanguineus MUCL 41627 (CRGF
437)
Tinctoporellus epimiltinus MUCL 43523 (CRGF
465)
Trametes maxima MUCL 44155 (CRGF467)
Trametes maxima MUCL 44167(CRGF 468)
Pycnoporus sanguineus MUCL 41582

Growth rate
MA2 (mm/d)

Growth rate Acid

Blue 62 (mm/d)

Decolourisation rate
(mm/d)

Yield (***)
(mm decolourisation/
mm growth)

Inhibition
(%) (****)

5.3
4.2
5.1
4.6

3.3
3.2
2.6
2.0

3.7
2.8
2.6
2.9

1.13
0.88
1.00
1.4

38.2
23.7
48.8
56.3

dead wood, Prov. Pinar del Ro

3.4

1.8

2.5

1.4

46.5

dead branch, Prov. Pinar del Ro

dead branch, Prov. Pinar del Ro
dead wood, Prov. Cienfuegos
dead hanging branch, Prov. Pinar
del Ro
dead branch, Prov. Pinar del Ro

3.2
2.8
5.5
1.8

2.4
2.4
3.3
0.9

2.5
2.7
3.5
0.9

1.06
1.10
1.08
1.00

25.9
12.1
40.3
50.0

2.1

1.2

2.4

1.96

43.3

dead branch, Prov. Pinar del Ro

2.4

1.2

2.3

1.88

50.5

dead stump, Prov. Cienfuegos

3.4

1.8

2.2

1.22

47.9

dead stump, Prov. Cienfuegos

3.5

1.9

2.0

1.09

46.6

dead wood, Prov. Pinar del Ro

5.1

3.4

3.6

1.05

33.5

dead wood, Prov. Pinar del Ro

5.4

3.1

3.3

1.07

43.7

dead trunk, Prov. Cienfuegos

6.0

3.1

3.3

1.06

48.0

decaying stump, Prov. Pinar del

Ro
dead stump, Prov. Pinar del Ro
dead trunk, Prov. Cienfuegos
Reference strain

6.0

6.0

6.0

1.00

31.1

6.0
3.8
5.7

4.0
2.8
2.9

4.1
3.1
3.0

1.02
1.12
1.04

34.1
26.4
48.5

Origin
dead branch, Prov. Pinar del Ro
dead trunk, Prov. Cienfuegos,
dead wood, Prov. Pinar del Ro
dead branch, Prov. Cienfuegos

(*)The strains were identified by their deposit number at the Belgian Coordinated Collections of Microorganisms/Mycothque de lUniversit Catholique de Louvain (BCCM/MUCL).
(**) In brackets, the strains deposit number at the CRGF (Colleccion de Recursos Geneticos Fungicos) at the Institute of Ecology and Systematic.
(***) The decolourisation yield was expressed as the ratio between the diameter of decolourisation and diameter of growth
(****)Percentage of growth inhibition was calculated as follows: % Inhibition = 100 X (Diameter of growth on MA2 - Diameter of growth on MA2 added with AB 62)/ Diameter of growth on MA2.

1986

Afr. J. Biotechnol.

Table 2. Maximal biomass and laccase activity obtained in medium ML2 without and with the dye Acid Blue 62.

Strain
Coriolopsis rigida 44156
Hexagonia hydnoides 41636
Pycnoporus sanguineus 41582
Pachykytospora alabamae 44100
Tinctoporellus epimiltinus 43523
Perenniporia tephropora 43535
Hexagonia tenuis 43554
Trametes maxima 44155

Medium ML2
Biomass at
Maximal Laccase
14 days (g/l)
activity (U/L)
7.987
42.4 (11)*
4.803
70.7 (8)
6.067
28.3 (9)
8.000
267.5 (11)
5.060
26.6 (8)
5.070
68.4 (8)
7.450
17.8 (11)
7.420
68.4 (8)

Medium ML2 + Acid Blue 62

Biomass at 14 Maximal Laccase
days (g/l)
activity (U/L)
4.270
261.6 (10)
3.580
600.4 (11)
4.067
43.6 (9)
6.313
592.5 (11)
3.513
451.7 (7)
4.073
134.9 (8)
6.127
541.4 (10)
5.813
758.1 (8)

*Values reported in parentheses are cultivation time (days) at which maximal laccase activity was obtained.

RESULTS

The growth inhibition in presence of AB 62 and the

capacity to decolourise the dye on solid medium were
chosen as criteria for a preliminary sorting of fungal
strains. Growth and decolourisation rate attained for the
nineteen selected strains and the P.sanguineus reference
strain are shown in Table 1.
Addition of AB 62 to the media affected the growth rate
of all the strains tested, but to different extent. Hexagonia
tenuis (MUCL 43554) was the less affected, with a
growth reduction of 12% (Table 1) while Hexagonia
hydnoides (MUCL 41636), Megasporoporia setulosa
(MUCL 43550), and Pachykytospora alabamae (MUCL
44126) were the most severely affected, with a growth
rate reduction attaining about 50%.
Dye decolourisation was observed for all the strains,
including the reference P. sanguineus (MUCL 41582).
Tinctoporellus epimiltinus (MUCL 43523), T. maxima
(MUCL 44155), P. tephropora (MUCL 43535) and C.
rigida (MUCL 44156) were found to have the highest
decolourization rate in the presence of the AB 62 dye
incorporated in MA2. However, in terms of decolourisation yield (Table 1), the isolates of P. alabamae and of
H. hydnoides appeared as the most performing.
Taking into account all these results, seven strains,
namely, T. epimiltinus (MUCL 43523), T. maxima (MUCL
44155), P. tephropora (MUCL 43535), C. rigida (MUCL
44156), H. tenuis (MUCL 43554), P. alabamae (MUCL
44100) and H. hydnoides (MUCL 41636), were selected
for further studies in liquid medium.

addition of AB 62 to the medium affected the growth of

the fungi, and consequently the biomass yield. For all the
strains tested, the percentage of growth inhibition was
smaller than 50%. Strains of H. tenuis, P. tephropora and
P. alabamae were the less affected, although linear
growth of P. alabamae on the solid medium was the most
severely reduced (Table 1). In the absence of dye,
biomass ranged from 4.7 8.0 g/l. In the presence of AB
62, biomass production was reduced to 3.5 - 6 g/l
depending on the strain studied.
All the selected fungi were able to decolourise, partially
or completely, the dye AB 62. The A 595 nm value decreased to a large extent for all the strains during the first
five days of growth (Figure 1A). This reduction corresponded to a shift in the coloration of the medium from
blue to red. The appearance of the red colour resulted
from an increase of A 500nm (Figure 1 B).
Later the red colour disappeared gradually, resulting for
example into almost total decolourisation by T. maxima
after 18 days of culture (not shown).
Except for H. tenuis and T. epimiltinus, production of
laccase in medium ML2 (without dye) was higher in the
tested strains than in the reference P. sanguineus MUCL
41582 (Table 2). The higher level of activity in our
cultivation conditions was obtained by P. alabamae
(267.5 U/L), followed by H. hydnoides MUCL 41636 (70.7
U/L), P. tephropora MUCL 43535 (68.4 U/L) and T.
maxima (68.4 U/L).
However, for all strains tested, the production of
laccase was stimulated by the presence of the dye (Table
2). In presence of the dye, the greatest levels of laccase
activity were obtained by T. maxima (758.1 U/L), followed
by H. hydnoides (600.4 U/L), P. alabamae (592.5 U/L)
and H. tenuis (541.4 U/L).

Growth and decolourisation in liquid medium

Decolourisation of the textile effluent

Biomass growth as well as decolourisation was attained

for all strains after about 14 days of cultivation in liquid
medium (Table 2). As for cultivation in solid media,

The industrial effluent used contained mainly three dyes,

Remazol Yellow RGB, Remazol Red RR and Remazol
Black B 133, at a total concentration of about 70 g/l. The

Growth and
medium

decolourisation

capacity

on

solid

Snchez-Lpez et al.

1987

A
18

16

14

Absorbance 595nm

12

10

0
0

10

12

14

16

time (days)

Absorbance 500nm

0
0

10

12

14

16

time (days)

Figure 1. Time course of decolourisation of the ML2 medium supplemented with Acid Blue 62. (A) values of
A595nm and (B) values of A500nm. ( ) Coriolopsis rigida 44156; ( ) Hexagonia hydnoides 41636; ( ) Pycnoporus
sanguineus 41582; () Pachykytospora alabamae 44100; (*)Tinctoporellus epimiltinus 43523; ( ) Perenniporia
tephropora 43535; (+) Hexagonia tenuis 43554 and ( ) Trametes maxima 44155.

effluent contained also other additives like molan,

fumexol among others. Figure 2 shows the absorbance
spectrum of the ML2 medium added with the textile
effluent after 14 days of incubation with the fungal strains.
All the strains were able to decolourise to more or less

extent the industrial effluent. A noticeable collapse of the

whole visible spectrum, as compared to the spectrum of
the uninoculated control, was obtained with five strains;
T. maxima being the most effective with a 94% value of
decolourisation. The decolourisation performance of P.

1988

Afr. J. Biotechnol.

Figure 2. Absorbance spectra in the visible range of the medium ML2 supplemented
with 0.5 % (v/v) of the industrial textile effluent. Data were recorded after 14 days
incubation in the presence of the different WRF strains. Decolourisation values for each
strain, reported as percentage in bracket, was calculated using the integrated values of
the absorbance between 380 740 nm. (1) Uninoculated control; (2) Perenniporia
tephropora [41]; (3) Pachykytospora alabamae [32]; (4) Tinctoporellus epimiltinus [43];
(5) Pycnoporus sanguineus [67]; (6) Hexagonia tenuis [78]; (7) Hexagonia hydnoides
[83]; (8) Coriolopsis rigida [88] and (9) Trametes maxima [94].

sanguineus was lower than for C. rigida, H. hydnoides, H.

tenuis and T. maxima.
Laccase activity was detected in all cultures; P. alabamae and T. maxima being the best enzyme producers.
DISCUSSION
Collection of strains characterized here for their decolourisation capabilities were realized in both natural and
anthropic ecosystems in Cuba, mostly in Pinar del Rio
Province. The Sierra Del Rosario Biosphere Reserve is
characterized by several vegetation types, in particular
evergreen forest presenting some similarities with
neotropical rain forest (Capote et al., 1988), and was
reported as an important reservoir of fungal biodiversity
(Minter et al., 2001). A significant part of the selected
strains nevertheless came from anthropic environments,
including urban areas. The presence in the urban zone of
many dead fallen trunks or stump, consequence of
numerous tropical storms, constitute an interesting ecosystem characterized by non buffered conditions, due to
an open exposition with very high diary and seasonal
variation in both surface temperature and water content.
Several polypores species are commonly observed on
this substrate in Cuba, as well as in the entirety of tropical
areas. They include P. sanguineus, H. hydnoides, T.
maxima, C. rigida, P. tephropora, Earliella scabrosa, or
H. tenuis. These species might constitute the primary

colonizers of freshly fallen dead trunk. Previous studies in

other tropical area, e.g. French Guyana have also shown
that polypores isolated from similar environment (freshly
fallen dead trunk), produced higher enzyme activity,
either linked to white or brown rotting (Decock et al.,
unpublished). However, the link between the ecological
conditions of such substrate and the physiological
properties of the fungus, of which enzyme production,
remain to be obviously elucidated. Primary colonisers
should have efficient enzymatic systems to penetrate and
invade quickly the fresh unaltered wood in order to
establish a viable colony within deeper zones of the
material, less affected by drastic changes of T and water
content. They should preserve a volume of wood as large
as possible from colonisation by other species, and in
consequence, their enzymatic activities should also be
efficient for intra or inner species competition. These
species should also be able to react quickly when the
conditions are optimal, although these being highly
variable.
In 2003, the world textile dye consumption was 680 000
tons, with an annual turnover of 4.49 billion euros
(Sunfaith China Ltd, 2004). Among these, anthraquinonic
acid dyes, like Acid Blue 62, represent 166 000 tons in
the World and 40 000 tons in Europe. This class forms
the basis of many of the modern synthetic dyes and is
reported in the Colour Index as one of the most important
group of organic dyes (Hobson and Wales, 1998).
Moreover, anthraquinonic structures have been isolated

Snchez-Lpez et al.

1989

Figure 3. Cluster analysis of the white-rot fungi strains, obtained from decolourisation
analysis of the dye AB 62 and the industrial effluent. Ther strains are Coriolopsis rigida
(MUCL 44156), Hexagonia hydnoides (MUCL 41636), Pycnoporus sanguineus (MUCL
41582), Pachykytospora alabamae (MUCL 44100), Tinctoporellus epimiltinus (MUCL
43523), Perenniporia tephropora (MUCL 43535), Hexagonia tenuis (MUCL 43554) and
Trametes maxima (MUCL 44155).

from a number of fungi including Drechslera S. Ito,

Trichoderma Pers., Aspergillus Link and Curvularia
Boedijn strains, and could serve as basis for development of a future generation of new natural and/or semisynthetic dyes (Alvi and Rabenstein, 2004). Acid blue 62
was chosen as a model for evaluating the Cuban strains
and all strains studied were able to decolourise this dye.
The action of the seven selected strains toward other
types of colorants (e.g. azo dyes), remain to be evaluated. However, the noteworthy decolourisation activity of
the strain towards a textile effluent containing different
dyes and some additives indicate that they could
potentially have a positive action against complex
pollution situations.
Considering decolourisation analysis of the dye AB 62
at two different optical densities (500 and 595 nm) and
the industrial effluent, it is possible to determine several
highly related strains (Figure 3). The four strains of
subgroup A (T. maxima MUCL 44155, C. rigida MUCL
44156, H. hydnoides MUCL 41636 and H. tenuis MUCL
43554) which produced the highest decolourisation,
behave quite similarly to the strain P. sanguineus MUCL
41582, used as reference for dye decolourisation [Van
Hulle et al., 2001]. A strain of P. sanguineus was also

described as a very efficient laccase producer (Pointing

et al., 2000b).
No correlation was found between the levels of laccase
activity and the decolourisation rate in the strains tested.
T. maxima MUCL 44155, H. hydnoides MUCL 41636 and
H. tenuis MUCL 43554 demonstrated high level of both
laccase activity and decolourisation. C. rigida MUCL
44156 obtained a high level of decolourisation with a
lower level of laccase. P. alabamae produced one of the
highest levels of laccase activity but notwithstanding, the
lowest level of decolourisation. Nevertheless, for all the
strains, the laccase activity was stimulated by the
presence of dye. T. maxima MUCL 44155 showed the
highest decolourisation capacity and also the highest
laccase activity (Table 2) in the culture conditions tested.
For that reason we consider this strain as very promising
for future studies. Members of the genus Trametes, in
particular T. versicolor, have been reported as very
efficient in dye decolourisation (Amaral et al., 2004; Libra
et al., 2003; Liu et al., 2004). The genus Trametes
emerged also as a potent decolouriser from recent fungal
screening of natural environment, for example from the
Chirinda and Chimanimani sub-tropical hardwood forests
in Zimbabwe (Tekere et al., 2001). Several members of

1990

Afr. J. Biotechnol.

the genus Trametes, in particular T. versicolor (L.: Fr.)

Pilt, T. hirsuta (Fr.) Pilt and T. ochracea (Pers.) Gilb.
and Ryvarden were reported as potent laccase producers
(Tomsovsky and Homolka, 2003). However, to our
knowledge, this is the first report of decolourisation and
laccase production by T. maxima. Laccase has been
identified as the principal enzyme active in dye decolourisation by several WRF strains (Baldrian, 2004; Levin et
al., 2004; Liu et al., 2004; Wesenberg et al., 2003)
although manganese peroxidase (Boer et al., 2004) or
other intracellular enzymes were reported as the main
activities in some strains (Blanquez et al., 2004). Lignin
and Mn peroxidases were not detected in the cultivation
conditions we used in the present investigation (data not
shown). This indicated a plausible involvement of laccase
in decolourisation by T. maxima. The noticeable increase
in laccase activity in the presence of AB 62 or the textile
effluent supported also this hypothesis.
In conclusion, T. maxima seems thus to constitute a
promising strain for biotechnological applications. Its
description highlighted the interest of screening of
environments still little exploited. This type of approach
which has clear connexions with white biotechnology
(Frazzetto, 2003) could also constitute a good incentive
for better protection of fungal biodiversity.
ACKNOWLEDGMENTS
This work was supported by the Elite Internationale
(CUBACHAM) and FIRST Objectif 3 (CHAMBOIS)
programmes from the Walloon Region, the CIUF (Conseil
Inter-Universitaire de la Communaut Franaise de
Belgique) -CUD (Commission Universitaire pour le
Dveloppement) cooperation programme (project CIUFCUD-MUCL-Cuba) and the EU Sixth Framework
Program (NMP2-CT2004-505899 - SOPHIED). Cony
Decock gratefully acknowledges the financial support
received from the Belgian Federal Science Policy
Officeontract BCCM C3/10/003).
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