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Biochemical Investigations of Different Mushroom Species for Their Biotechnological Potential

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Abstract

Among bioactive constituents occurring in mushrooms, phenolic compounds focus attention due to their antioxidant activity. Also, a special attention is given to mushrooms secreting extra cellular enzymes such as lignin peroxidases (LiP), manganese peroxidase (MnP) and laccase (Lac), enzymes which can be used in biodegradation processes. The aim of the present study was to investigate ten mushroom species for total phenolic compounds, lignocellulolytic enzymes and for their synthetic dyes decolourisation potential. For this purpose, 70% ethanol extracts of ten dried mushrooms were analysed using spectrophotometric methods. The results revealed that total phenolic compounds in the extracts were the highest in A. campestris, P. ostreatus var. Florida and T. versicolor. Laccase activity showed high values in extracts from P. ostreatus var. Florida, A. campestris, L. edodes, and G. applanatum. Lignin peroxidases (LiP) activity showed high values in extracts from A. campestris, F. velutipes, P. ostreatus var. Florida and T. versicolor, whereas manganese peroxidase (MnP) activity was highest in extracts from P. ostreatus var. Florida, A. campestris and G. applanatum. It was found that some of the fungal extracts showed high activities in decolorizing of synthetic dyes.
DOI: 10.2478/alife-2018-0088
562
BIOCHEMICAL INVESTIGATIONS OF DIFFERENT MUSHROOM
SPECIES FOR THEIR BIOTECHNOLOGICAL POTENTIAL
Mihai Bogdan NICOLCIOIU, Gabriela POPA, Florentina MATEI
University of Agronomic Sciences and Veterinary Medicine of Bucharest,
Faculty of Biotechnologies, 59 Mărăşti Blvd., District 1, 011464, Bucharest, Romania
Corresponding author email: popagabiro@yahoo.com
Abstract
Among bioactive constituents occurring in mushrooms, phenolic compounds focus attention due to their antioxidant
activity. Also, a special attention is given to mushrooms secreting extra cellular enzymes such as lignin peroxidases
(LiP), manganese peroxidase (MnP) and laccase (Lac), enzymes which can be used in biodegradation processes. The
aim of the present study was to investigate ten mushroom species for total phenolic compounds, lignocellulolytic
enzymes and for their synthetic dyes decolourisation potential. For this purpose, 70% ethanol extracts of ten dried
mushrooms were analysed using spectrophotometric methods. The results revealed that total phenolic compounds in the
extracts were the highest in A. campestris, P. ostreatus var. Florida and T. versicolor. Laccase activity showed high
values in extracts from P. ostreatus var. Florida, A. campestris, L. edodes, and G. applanatum. Lignin peroxidases
(LiP) activity showed high values in extracts from A. campestris, F. velutipes, P. ostreatus var. Florida and T.
versicolor, whereas manganese peroxidase (MnP) activity was highest in extracts from P. ostreatus var. Florida, A.
campestris and G. applanatum. It was found that some of the fungal extracts showed high activities in decolorizing of
synthetic dyes.
Keywords: dyes discoloration, lignocellulolytic enzymes, mushroom extracts, phenolic compounds.
INTRODUCTION
Mushrooms possess great potential for both
nutrition and therapeutic use. It has been found
that edible and medicinal mushrooms contain
many biologically active compounds with anti-
inflammatory, antitumor, antibacterial,
antiviral, cardiovascular, anti-
hypercholesterolemia and antioxidant activities
(Wani et al., 2010; Patel et al., 2012; Khatua et
al., 2013; Wasser, 2014). Amongst bioactive
constituents occurring in mushrooms, phenolic
compounds focus much attention due to their
antioxidant activity (Bubueanu et al., 2017).
Many Basidiomycete fungi such as Pleurotus
spp., Lentinula edodes, Hericium coralloides,
Trametes versicolor, Laetiporus sulphureus,
Ganoderma spp. can enzymatically degrade
various substrates containing lignin,
hemicellulose, and cellulose into soluble
compounds with low molecular weight (Lim et
al., 2013). The wood-rotting Basidiomycetes
are the most efficient degraders of
lignocellulose substrates. The enzymes
involved in lignin degradation are mainly lignin
peroxidase (LiP), which catalyses the oxidation
of both phenolic and non-phenolic units,
manganese peroxidase (MnP) and laccase (Lac)
which oxidizes phenolic compounds (Jhadav et
al., 2009). These enzymes have been shown to
degrade not only lignocellulose but also some
recalcitrant environmental pollutants such as
crude oil wastes (Popa et al., 2013), textile dyes
discoloration (Nagai et al., 2002; Iordache et
al., 2016), delignification and pulp bleaching
(Palonen and Viikari, 2004), and
bioremediation (Jaouani et al., 2005). Because
most of biologically active compounds are
involved in antioxidant activity and different
biotechnological processes the objectives of
this study were to investigate ten mushroom
isolates for total phenolic compounds,
lignocellulolytic enzymes activity and for their
synthetic dyes decolourisation potential.
MATERIALS AND METHODS
Mushroom samples
The fresh fruiting bodies of ten medicinal
and/or edible mushroom species of Laetiporus
563
sulphureus, Pleurotus eryngii, Agaricus
campestris, Flammulina velutipes, Lentinus
edodes, Pleurotus ostreatus var. Florida,
Ganoderma lucidum, Ganoderma applanatum,
Trametes versicolor and Hericium coralloides
were collected from different Romanian
geographical locations (Fundata, Sibiu, Sinaia,
Chitila). The fungal materials were kindly
authenticated by Dr. Ioana Tudor and Eng. Paul
Covic. The fresh fruiting bodies were left to
completely dry at the room temperature (25 -
26oC) for a week.
Extracts preparation
Fine powder of each dried fruiting bodies was
subject of hydro ethanol (70%) extraction (10
ml/g). The mixture was macerated for 24 hours
at room temperature, and then the homogenate
was centrifuged at 10.000 rpm, for 15 min. The
obtained supernatant was used for total
phenolic content determination, enzyme assays
and synthetic dyes discoloration analysis.
Total phenolic assay
Total phenolic content was determined by the
use of Folin Ciocalteu reagent (Singleton et
al., 1999) and expressed as Gallic acid
equivalents. An aliquot (1 ml) of extract or
standard solution of Gallic acid was added to
25 ml of volumetric flask, containing 9 ml
distilled water and 1 ml Folin Ciocalteu
reagent. After 5 minutes, 10 ml of 5% (w/v)
sodium carbonate solution was added to the
mixture. After 90 minutes at room temperature,
the absorbance of the samples was determined
at 760 nm with a UV/VIS spectrophotometer.
Total phenolic content was expressed as mg
GAE/l.
Meanwhile, phenolic content has been analysed
by TLC. Thin-layer chromatography (TLC)
was performed using silica gel TLC plates (60
F254 10 cm x 10 cm, Merck) and pre-coated
silica gel TLC plates (POLYGRAM SIL
G/UV254-Merck, 40 x 80 mm).
About 5 µl of mushroom extracts was loaded
on TLC plates. The plates were developed in
hexane: methanol: acetic acid (3:9:2) as mobile
phase.
The developed plates were air dried and
observed under UV (365 nm). After this
examination, for detection of phenolic
compounds, the TLC plates were sprayed
with 10 ml Folin - Ciocalteu reagent and then
evaluated in visible light. The appearance of
the blue spots indicates the presence of
phenolic compounds. The retention factor (Rf)
value of the spots was also determined by
calculating a ratio between the distance from
the starting point to the center of the spot on the
TLC plate and the distance from the starting
point to the solvent front (Kagan and Flythe,
2014).
Enzymatic assays
Laccase activity. Laccase activity was
determined using guaiacol as substrate (Kalra
et al., 2013). The reaction mixture contained 3
ml sodium acetate buffer (10 mM, pH 5.0), 1ml
guaiacol (2 mM) and 1 ml enzyme source. The
mixture was incubated at 30oC for 15 min. The
changes in absorbance due the oxidation of
guaiacol in the reaction mixture were recorded
by spectrophotometer at 450 nm, with a molar
extinction coefficient for guaiacol (Ɛ450 =6740
M-1 cm-1). One unit of enzyme activity (U/ml)
is defined as the amount of enzyme that
oxidized 1 μmol of guaiacol per minute.
Lignin peroxidase activity. Lignin peroxidase
(LiP) activity was determined
spectrophotometrically at 310 nm (Ɛ310 =9300
M-1cm-1) using the method of Takamiya et al.,
2008). The reaction mixture contained veratryl
alcohol (8 mM), 0.3 M citrate/ 0.4 M phosphate
buffer (pH 4.5), 60 µl supernatant, and distilled
water. The mixtures were incubated for 2
minute at 30oC. The reaction was initiated by
addition of 5 mM H2O2. One unit of LiP
activity was defined as the amount of enzyme
that oxidized 1 µmole of veratryl alcohol per
minute.
Manganese peroxidase activity. Manganese
peroxidase (MnP) activity was determined by
monitoring the oxidation of guaiacol as a
substrate at 465 nm with extinction coefficient,
Ɛ465 = 12100 M-1cm-1 (Mabrouk et al., 2010).
The reaction mixture contained sodium
succinate buffer (0.5 M, pH 4.5 at 30oC),
guaiacol (4mM), manganese sulphate (1mM),
300 µl supernatant, and distilled water. The
mixtures were incubated for 2 minute at 30oC.
The reaction was initiated by addition of 1 mM
H2O2. One unit of MnP activity was defined as
the amount of enzyme that oxidized 1 µmole of
guaiacol per minute.
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Dyes discoloration. Four reactive dyes,
Bemacid Rot N-TF (BR), Bemacid Gelb N-TF
(BG), Bemacid Blau N-TF (BB) (Bezema AG)
provided from The National Research and
Development Institute for Textiles and Leather
(INCDTP) Bucharest, Romania, and Congo red
(CR) were used for monitoring the dye
discoloration capacity of fungal extracts. An
aliquot of fungal extract (50 μl) was incubated
for three weeks with 2 mL of 20 mM sodium
acetate buffer (pH 4.0) containing 0.05% of
each dye, at 25oC in the dark. The control
experiments were performed under identical
conditions without addition of fungal extracts.
Dyes discoloration was measured
spectrophotometrically every week at the
corresponding absorption wavelength for each
compound: BR-λ340 nm, BG- λ440 nm, BB- λ640
and CR- λ595 nm. The discoloration percentage
(D %) was calculated as follows: D (%) = (A0
A)/A0 × 100, where A0 is the dye absorbance of
the control, and A is the dye absorbance of the
test sample.
RESULTS AND DISCUSSIONS
Total phenolic content in mushroom extracts
Polyphenols are a large group of secondary
plant metabolites with a significant role in the
protection of oxidation processes (Croft, 1999).
The contents of total phenolic compounds (mg
GAE/l) in the ten mushroom 70 % ethanol
extracts are shown in Figure 1.
Figure 1. Total phenolic content in the 70% ethanol
extracts of ten mushroom isolates
1-Laetiporus sulphureus; 2-Pleurotus eryngii;
3-Agaricus campestris; 4-Flammulina velutipes;
5-Lentinus edodes; 6-Pleurotus ostreatus var. Florida;
7-Ganoderma lucidum; 8-Ganoderma applanatum;
9-Trametes versicolor; 10-Hericium coralloides.
The results show that the amount of phenolic
compounds was the highest in the extracts from
the Agaricus campestris, Pleurotus ostreatus
var. Florida and Trametes versicolor (489.18;
339.37 and 264.64 mg GAE/l) respectively.
Except the fungal isolates that showed the
lowest content of total phenol (Laetiporus
sulphureus,Lentinus edodes and Hericium
coralloides), Ganoderma applanatum,
Pleurotus eryngii, Flammulina velutipes and
Ganoderma lucidum isolates had a relatively
similar phenolic content (Figure 1). The higher
contents of total phenol found in the mushroom
extracts could be considered as indicators for
higher antioxidant ability (Boonsong et al.,
2016; Popa et al., 2016).
Phenolic compounds detection by TLC
analysis. Detection of phenolic compounds
was also performed by TLC assay. The TLC
plates were developed in hexane: methanol:
acetic acid (3:9:2) and sprayed with 10 ml
Folin - Ciocalteu reagent. Under UV light (365
nm), the eluted compounds showed a more
pronounced greenish yellow fluorescence for
Agaricus campestris (3), Flammulina velutipes
(4), Ganoderma lucidum (7), Ganoderma
applanatum (8) and Trametes versicolor (9)
extracts (Figure 2 -A).
Figure 2. TLC analysis for phenolic compounds:A- plate
under UV (365 nm); B- samples under visible light
1-Laetiporus sulphureus; 2-Pleurotus eryngii;
3-Agaricus campestris; 4-Flammulina velutipes;
5-Lentinus edodes; 6-Pleurotus ostreatus var. Florida;
7-Ganoderma lucidum; 8-Ganoderma applanatum;
9-Trametes versicolor; 10-Hericium coralloides.
565
After treatment with Folin - Ciocalteu reagent
and examination under visible light it was
observed that all samples of fungal extracts
showed blue spots corresponding to the
presence of phenolic compounds (Figure 2-B).
According to obtained TLC analysis,
respectively to the retention factors (Rf)
obtained and summarized in Table 1, it can be
noticed that the majority of the mushroom
extracts had detected two spots indicating
phenolic compounds and these samples are
Laetiporus sulphureus, Flammulina velutipes,
Lentinus edodes, Ganoderma lucidum,
Ganoderma applanatum, Trametes versicolor.
There is another group of three detected spots
indicating phenolic compounds, respectively
Pleurotus eryngii, Agaricus campestris and
Pleurotus ostreatus var. Florida. The probe
with four spots indicating phenolic compounds
is represented by Hericium coralloides. In what
concerns the similarities of the phenolic
compounds Rf value within samples we
observed that there are some compounds that
are common to some samples. The phenolic
compounds with Rf value 0.75 were common
to Agaricus campestris and Trametes
versicolor. The phenolic compounds with Rf
value 0.83 were common to Flammulina
velutipes, Pleurotus ostreatus var. Florida,
Hericium coralloides.
The phenolic compunds with Rf value 0.90
were common to Pleurotus eryngii, Agaricus
campestris. The phenolic compounds with Rf
value 0.92 were common to Laetiporus
sulphureus, Lentinus edodes, Pleurotus
ostreatus var. Florida, Ganoderma lucidum and
these compounds had the biggest number of
samples in which they could be observed.
The phenolic compounds with Rf value 0.97
were common to Pleurotus eryngii, Agaricus
campestris and Ganoderma applanatum
(Table 1).
Table1. Phenolic profiles of different mushrooms extracts obtained by TLC analysis
Legend: 1 - Laetiporus sulphureus; 2 - Pleurotus eryngii; 3 - Agaricus campestris;
4 - Flammulina velutipes; 5 - Lentinus edodes; 6 - Pleurotus ostreatus var. Florida;
7 - Ganoderma lucidum; 8 - Ganoderma applanatum; 9 - Trametes versicolor; 10 - Hericium coralloides.
Quantitative estimation of the enzymes. All
collected fungal isolates were examined for the
production of lignocellulolytic enzymes.
Quantitative estimation of laccase (Lac), lignin
peroxidase (LiP) and manganese peroxidase
(MnP) produced by selected fungi, showed that
the highest activities of Lac (16.18 U/ml) and
MnP (2.248 U/ml) were obtained from the
Pleurotus ostreatus var. Florida ethanolic
extract (Figure 3).
The highest lignin peroxidase activity was
obtained from the extracts of Agaricus
campestris (5.048 U/ml), Pleurotus ostreatus
var. Florida (4.606 U/ml), Trametes versicolor
(3.332 U/ml) and Flammulina velutipes (3.235
U/ml) (Figure 3).
Previous studies showed that the enzymes
extracted from the mushrooms have industrial
value (Maciel et al., 2010, Nigam, 2013). The
production of lignocellulolytic enzymes can be
566
stimulated by the presence of a wide variety of
inducing substrates mainly aromatic or
phenolic compounds related to lignin or lignin
derivatives such as ferulic acid, 2,5-xylidine,
and veratryl alcohol (Barbosa et al., 1996).
Figure 3. Quantitative estimation of Laccase (Lac),
Lignin peroxidase (LiP) and Manganese peroxidase
(MnP) - produced by selected fungi
Discoloration dyes by fungal extracts
Four dyes, Bemacid Rot N-TF (BR), Bemacid
Gelb N-TF (BG), Bemacid Blau N-TF (BB)
(Bezema AG) and Congo red (CR) were used
for the evaluation of dye discoloration capacity
of the fungal extracts.
Bemacid dyes are acid dyes for dyeing
polyamide and wool for application in neutral
to slightly acid pH ranges. Bemacid Rot and
Bemacid Gelb are azo dyes, and Bemacid Blau
is an anthraquinonoid dye. Congo red is an
organic compound, the sodium salt of 3,3′-bis4-
aminonaphthalene-1-sulfonic acid.
It is an azo dye. In our experiment, the fungal
extracts had no discoloration activity against
Congo red, but some of them showed high
activities in decolorizing Bemacid Rot (BR)
and Bemacid Gelb (BG) dyes (Figure 4).
All fungal extracts had discoloration activities
against BR dye. L. sulphureus showed the
highest discoloration activity (90.8 %) followed
by L. edodes (76.82 %), P. eryngii (73.75 %),
T. versicolor (69.35 %) and P. ostreatus
Florida (64.37 %). T. versicolor and Hericium
coralloides showed the highest discoloration
activities on BG dye, exhibited similar
decolourization ability (82.72 %).
Among all tested samples, only Lentinus
edodes, P. ostreatus Florida and G. applanatum
showed discoloration abilities on all Bemacid
dyes (Figure 4).
Figure 4. Colour removal of the synthetic dyes Bemacid
Rot N-TF, Bemacid Gelb N-TF and Bemacid Blau N-TF,
during 3 weeks, in the presence of the fungal extracts
1 - Laetiporus sulphureus; 2-Pleurotus eryngii;
3 - Agaricus campestris; 4-Flammulina velutipes;
5 - Lentinus edodes; 6-Pleurotus ostreatus var. Florida;
7 - Ganoderma lucidum; 8-Ganoderma applanatum;
9 - Trametes versicolor; 10-Hericium coralloides
These results are in agreement with some of
previously reported. Many studies have shown
that fungi due to their enzymes are able to
decolorize and detoxify industrial dyes
(Michniewicz et al., 2008, Teixeira et al.,
2010).
CONCLUSIONS
Ten mushroom isolates were investigated for
total phenolic compounds, lignocellulolytic
567
enzymes activity and for their synthetic dyes
discoloration potential. It was found that all
samples of the fungal extracts showed the
presence of phenolic compounds. The highest
content of total phenols was observed in
Agaricus campestris, Pleurotus ostreatus var.
Florida and Trametes versicolor extracts.
Quantitative estimation of lignocellulolytic
enzymes produced by selected fungi, showed
that the highest activities of laccase and
manganese peroxidase were obtained from the
Pleurotus ostreatus var. “Florida”. The highest
lignin peroxidase activity was obtained from
the extracts of Agaricus campestris, Pleurotus
ostreatus var. Florida, Trametes versicolor
and Flammulina velutipes. The ten fungal
extracts had no discoloration activity against
Congo red, but some of them showed high
activities in decolorizing of synthetic azo dyes.
Among all tested samples, only Lentinus
edodes, P. ostreatus Florida and G. applanatum
showed discoloration abilities on all Bemacid
dyes.
REFERENCES
Barbosa A.M., Dekker R.F.H., St. Hardy G.E, 1996. Veratryl
alcohol as an inducer of laccase by an ascomycete,
Botryosphaeria sp., when screened on the polymeric dye
Poly R-478. Letters in Applied Microbiology, 23(2): 93-96.
Boonsong S., Klaypradit W., Wilgggaipun P., 2016.
Antioxidant activities of extracts from five edible
mushrooms using different extractants. Agriculture and
Natural Resources, 50(2): 89-97.
Bubueanu C., Grigore A., Șerban E., Popa G., Cornea C.P,
2017. HPTLC identification of bioactive compounds and
antioxidant activity of Pleurotus ostreatus and Lentinus
edodes extracts. Scientific Bulletin. Series F.
Biotechnologies, Vol. XXI, ISSN 2285-1364, 343-348.
Croft K.C., 1999. Antioxidant effects of plant phenolic
compounds. In T. K. Basu, N. J. Temple, M. L. Garg
(Eds.), Antioxidants in Human Health, CABI Publishing,
Wallingford Oxon, New York, 109-121.
Iordache O., Cornea C. P., Popa G., Diguţă C., Dumitrescu I.,
Vărzaru E., Rodino S., Ionescu I., Matei A., 2016.
Evaluation of decolourization abilities of some textile dyes
by fungal isolates. Revista Industria textilă, ISSN 1222–
5347, vol. 67(3), 181-188.
Kagan I.A., Flythe M.D, 2014. Thin-layer Chromatographic
(TLC) Separations and Bioassays of Plant Extracts to
Identify Antimicrobial Compounds. J. Vis. Exp. (85),
e51411, 1-8.
Jaouani A., Guillén F., Penninckx M.J., Martínez A.T.,
Martínez M.J., 2005. Role of Pycnoporus coccineus
laccase in the degradation of aromatic compounds in olive
oil mill wastewater. Enzyme Microb. Technol., 36(4):
478486.
Jhadav A., Vamsi K.K., Khairnar Y., Boraste A., Gupta N.,
Trivedi S., Patil P., Gupta G., Gupta M., Mujapara A.K.,
Joshi B., Mishra d., 2009. Optimization of production and
partial purification of laccase by Phanerochaete
chrysosporium using submerged fermentation. International
Journal of Microbiology Research, 1(2): 9-12.
Kalra K., Chauhan R., Shavez M., Sachdeva S., 2013. Isolation
of laccase producing Trichoderma spp. and effect of pH
and temperature on its activity. Int. J. Chem. Environ.
Technol., 5(5): 2229 2235.
Khatua S., Paul, S., Acharya K., 2013. Mushroom as the
potential source of new generation of antioxidant: a
review. Res J Pharm Technol, 6(5): 496-505.
Lim S-H., Lee Y-H., Kang H-W., 2013. Efficient recovery of
lignocellulolytic enzymes of spend mushroom compost
from oyster mushrooms, Pleurotus spp., and potential use
in dye decolorization. Mycobiology, 41(4): 214-220.
Mabrouk A.M., Kheiralla Z.H., Hamed E.R., Youssry A.A.,
Abd El Aty A.A., 2010. Screening of some marine-
derived fungal isolates for lignin degrading enzymes
(LDEs) production. Agriculture and Biology Journal of
North America, 1(4): 591-599.
Maciel M. J. M., Castro e Silva A., Ribeiro H. C. T, 2010.
Industrial and biotechnological applications of ligninolytic
enzymes of the basidiomycota: a review. Electronic Journal
of Biotechnology, 13(6): 1415.
Michniewicz A., Ledakowicz S., Ullrich R., Hofrichter M.,
2008. Kinetics of the enzymatic decolorization of textile
dyes by laccase from Cerrena unicolor, Dyes and
Pigments, 77(2): 295302.
Nagai M., Sato T., Watanabe H., Saito K., Kawata M., Enei
H., 2002. Purification and characterization of an
extracellular laccase from the edible mushroom Lentinula
edodes, and decolorization of chemically different dyes.
Appl. Environ. Microbiol., 60(3): 327335.
Nigam P. S., 2013. Microbial enzymes with special
characteristics for biotechnological applications.
Biomolecules, 3(3):597611.
Palonen H., Viikari L., 2004. Role of oxidative enzymatic
treatments on enzymatic hydrolysis of
softwood. Biotechnol. Bioeng., 86(5): 550557.
Patel Y., Naraian R. Singh V. K., 2012. Medicinal properties of
Pleurotus species (oyster mushroom): a review. World
Journal of Fungal and Plant Biology, 3(1): 1-12.
Popa G., Nicolcioiu M. B., Cornea C.P., 2013, In vitro
evaluation of crude oil degradation potential of some
Pleurotus ostreatus isolates. Scientific Bulletin. Series F.
Biotechnologies, Vol. XVII, ISSN 2285-1364, 143-146.
Popa G., Cornea C.P., Luta G., Gherghina E., Israel Roming F.,
Bubueanu C., Toma R., 2016. Antioxidant and
antimicrobial properties of Laetiporus sulphureus (Bull.)
Murrill. AgroLife Scientific Journal - Volume 5 (1): 168
173.
Singleton V. L., Orthofer R., Lamuela-Raventos R. M., Lester
P., 1999. Analysis of total phenols and other oxidation
substrates and antioxidants by means of Folin-Ciocalteu
reagent. Meth. Enzymol., 299: 152-178.
Takamiya M., Magan N., Warner P.J., 2008. Impact assessment
of bisphenol A on lignin-modifying enzymes by
basidiomycete Trametes versicolor. Journal of Hazardous
Materials, 154(1-3): 33-37.
Teixeira R.S., Pereira P.M., Ferreira-Leitao V., 2010.
Extraction and Application of Laccases from Shimeji
Mushrooms (Pleurotus ostreatus) Residues in
Decolourisation of Reactive Dyes and a Comparative Study
Using Commercial Laccase from Aspergillus oryzae.
Enzyme Research, 2010: 1-8.
Wani, B. A., Bodha, R. H. and Wani, A. H., 2010. Nutritional
and medicinal importance of mushrooms. Journal of
Medicinal Plants Research, 4(24): 2598-2604.
Wasser S.P., 2014. Medicinal mushroom science: Current
perspectives, advances, evidences, and challenges. Biomed.
Journal, 37(6):345-56.
... "Fungal polyethylene degraders," BioResources 16(4), 7056-7070. 7057 Nicolcioiu et al. 2018;Kumar and Chandra 2020). Among these enzymes, laccases produced by white-rot basidiomycetes have high redox potential and diverse catalytic properties compared with the laccases from plants and bacteria (Rodgers et al. 2010). ...
... A subset of 20 isolates representing mostly basidiomycetes were selected for qualitative assay of laccase production. PDA plates amended with 0.01% guaiacol (G5502, Sigma, St Louis, MO, USA) were prepared (G-PDA) and inoculated with an actively growing mycelial disk (5 mm diam.) of each fungal culture (Swe 2011; Nicolcioiu et al. 2018). Plates were incubated at room temperature for 5 to 7 days, and a color change from colorless to reddish brown was recorded. ...
... Reddish brown color development in the PDA medium containing guaiacol (G-PDA) was used as a qualitative test for laccase production (Nicolcioiu et al. 2018). ...
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