Turkish Journal of Biology

Volume 36 Number 3 Article 5

1-1-2012

Bioactive compounds from discarded mushroom beds

SVSSSL HIMA BINDU NIDADAVOLU

RAM PRASAD METUKU

SAMATHA BURRA

RAJITHA BYRAM

SINGARA CHARYA MARINGANTI

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NIDADAVOLU, SVSSSL HIMA BINDU; METUKU, RAM PRASAD; BURRA, SAMATHA; BYRAM, RAJITHA; and
MARINGANTI, SINGARA CHARYA (2012) "Bioactive compounds from discarded mushroom beds," Turkish
Journal of Biology: Vol. 36: No. 3, Article 5. https://doi.org/10.3906/biy-1011-168
Available at: https://journals.tubitak.gov.tr/biology/vol36/iss3/5

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 S.V.S.S.S.L. H. B. NIDADAVOLU, R. P. METUKU, S. BURRA, R. BYRAM, S. C. MARINGANTI

Turk J Biol
36 (2012) 275-284
© TÜBİTAK
doi:10.3906/biy-1011-168

Bioactive compounds from discarded mushroom beds

S.V.S.S.S.L. Hima Bindu NIDADAVOLU*, Ram Prasad METUKU, Samatha BURRA,

Rajitha BYRAM, Singara Charya MARINGANTI
Department of Microbiology, Kakatiya University, Warangal 506 009 - INDIA

Received: 26.11.2010

Abstract: Discarded dried button, dried oyster, and fresh button mushroom beds were examined for their bioactive
compounds and industrial enzymes. Levels of reducing sugars and total sugars were high in all mushroom beds.
Carboxymethyl cellulase activity was higher (920 μg/mL) in the dried oyster mushroom bed extract. Schizophyllum
commune, Fomitopsis feei, Trametes gibbosa, and Trametes elegans were grown on direct mushroom bed extract, 0.2%
glucose-containing mushroom bed extract, and common production medium for determination of growth and
lignolytic activities. Since lignolytic activities were high with the direct mushroom bed extract, this medium was further
diluted to 1:1 and 1:2 ratios and again tested for growth and lignolytic activities with F. feei, T. gibbosa, and T. elegans.
It was determined that the 1:1 ratio gave good results for all these organisms. The button mushroom bed extract was
concentrated using a rotary evaporator and compared for growth and lignolytic enzymes with dried powder extract
using F. feei, T. gibbosa, and T. elegans. Growth was high with these 3 organisms in the dried powder extract, but
lignin peroxidase activity was the highest with the rotary evaporator extract using T. gibbosa. The results indicate that
these waste mushroom bed extracts can be used as cost-effective media for the growth of microorganisms and for the
production of bioactive compounds and industrial enzymes.

Key words: Discarded mushroom beds, bioactive compounds, industrial enzymes, white rot fungi

Introduction time and even in small extra spaces in their homes.

Today mushrooms are becoming more and Hence, this has developed as a village industry and
more popular amongst people as a continental is earning respectable amounts of money to support
or Chinese delicacy. As the number of diabetic rural families. It is becoming one of the important
patients is increasing steadily in India, mushrooms activities of the Development of Women and Children
can supplement a good diet for these patients with in Rural Areas groups formed in the villages to help
low calories and high protein value. Due to their themselves. This program has been revolutionary
fast growth and simple cultivation without need of in all districts of the state of Andhra Pradesh and is
any chemical fertilizers or pesticides, mushroom becoming very popular in other states of India.
farming is becoming a very popular cottage industry. This mushroom production technology is
The government is encouraging it as a small-scale considered a very attractive and useful program
industry and giving subsidiaries for mushroom because it not only recycles agricultural wastes such
cultivation. Using agricultural waste materials, rural as paddy/jowar/maize straw, cotton waste, groundnut
women are cultivating mushrooms in their spare waste, and oil cake but also produces highly nutritious

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Bioactive compounds from discarded mushroom beds

mushrooms for human consumption. It also gained was extracted with 40 mL of distilled water using a
significance because it is low-tech and manageable pestle and mortar. After homogenization, the extracts
in normal conditions. With a simple 2-day training were filtered and centrifuged. The supernatant
process, illiterate woman can easily practice the obtained was used for further studies and compared
preparation of mushroom beds with agricultural with fresh button mushroom bed extract.
wastes, their sterilization methods, spawning, casing, A total of 950 g of fresh button mushroom bed
cropping, and so on. Previously, villagers were afraid was soaked in 4.5 L of distilled water for 1 day.
of the mushrooms and never ventured to touch them.
After filtration and centrifugation, the obtained
Their reasons were, first, that they did not know
supernatant was used for further studies. This extract
how to distinguish between edible and poisonous
was used as broth for the production of lignin
mushrooms, and, second, that the appearance of
peroxidase and laccase by Schizophyllum commune,
poisonous mushrooms (colored or spotted) in the
Fomitopsis feei, Trametes gibbosa, and Trametes
house or agricultural fields was considered a bad
elegans, comparing it with common production
omen. Now in the changed conditions, people are
medium as control (1 g peptone, 2 g yeast extract, 1
fascinated with the production and are cultivating
g dipotassium hydrogen phosphate, 0.2 g magnesium
mushrooms, for which they are receiving attractive
sulfate heptahydrate, 5 g ammonium sulfate, 20 g
payment.
glucose, and 1000 mL distilled water at pH 6.0) and
The use of mushrooms for their bioactive the same extract containing 2% glucose after 7 and 14
compounds has been in practice for a long time, since days of incubation.
these mushrooms can provide compounds that have
nutritional, medicinal, and biological importance (1- Since lignin peroxidase activity was high in the
4). However, the isolation of bioactive compounds direct mushroom bed extract, it was further diluted
from discarded mushroom beds is not in common with distilled water at 1:1 and 1:2 ratios and was
practice. When our scientific group visited a village, tested for the production of lignin peroxidase and
we found huge piles of discarded mushroom beds on laccase by inoculation with F. feei, T. gibbosa, and T.
the outskirts of the village and saw that their disposal elegans after 7 days of incubation.
was becoming a problem. To solve this disposal Using a rotary evaporator, 500 mL of the fresh
problem and to get “wealth out of waste” (WOW), button mushroom extract was concentrated under
the present investigation was undertaken. Collection vacuum. It was then weighed (approximately 4 g)
of these discarded beds from many villages and and stored. The fresh button mushroom bed was
extraction of industrially important enzymes and dried under shade and powdered. Two grams of this
bioactive compounds is the topic of this paper. powder was extracted with 0.2% glucose solution by
boiling for 30 min. After filtration, the supernatant
obtained was used as broth. Quantitative estimation
Material and methods
of the laccase and lignin peroxidase activities of
Collection of waste mushrooms beds this extract was done and compared with the rotary
After the completion of mushroom harvesting evaporator extract from F. feei, T. gibbosa, and T.
(button and oyster), the discarded beds were collected elegans that were incubated for 7 days.
from the villages in and around the city of Warangal pH
and were used in the present investigation.
The pH levels of both the button and oyster
Preparation of extracts mushroom bed extracts were determined by using
The preparation of the extract from the discarded pH papers (BDH Chemicals Ltd., UK).
beds was done by following the methods of earlier Bioactive compounds
reports, slightly modified in the present study (5,6).
From the discarded dried mushroom bed, 3 g of solid Total soluble sugars
fermented material was collected in the zone where The amount of total soluble sugars was estimated
spawn had spread and fruit bodies had grown. This using the phenol-sulfuric acid method (7). The

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 S.V.S.S.S.L. H. B. NIDADAVOLU, R. P. METUKU, S. BURRA, R. BYRAM, S. C. MARINGANTI

quantity of total sugars was expressed as μg/mL run to find the standard deduction in titer value.
of extract using a standard graph (100 mg/100 mL The activity was expressed as the amount of enzyme
glucose). required to liberate one micromole equivalent of fatty
Reducing sugars acid per milliliter per minute.
Estimation of reducing sugars was carried out (8) and Cellulase (EC 3.2.1.4)
the amount of reducing sugar was calculated using a Cellulase activity was assayed by determination of
standard curve prepared from glucose (100 mg/100 the reducing sugars released from carboxymethyl
mL). The quantity of reducing sugar was expressed as cellulase (CMC) (16,17). A volume of 0.5 mL of the
μg/mL of extract. culture supernatant fluid was incubated with 1 mL
Total lipids of 2% CMC in 0.05 M sodium acetate buffer (pH
The total lipid content was assayed (9) using a mixture 4.8) at 50 °C for 10 min. The reduced sugar product
of chloroform and methyl alcohol (2:1). was assayed using the dinitrosalicylic acid method
using glucose as the sugar standard. The activity was
Proteins expressed in μg/mL of extract.
Total protein content was estimated (10) and the Protease (EC 3.4.21.99)
amount of protein in the sample was calculated using
a standard graph drawn with bovine serum albumin Protease enzyme activity was assayed by following
(10 mg/100 mL). a spectrophotometric method (18,19). The activity
of the reaction mixture, which was prepared by
Enzyme assays incubating the enzyme extract with casein, was
Laccase (EC 1.10.3.2), lignin peroxidase (EC terminated by using 110 mM trichloroacetic acid.
1.11.1.14), and manganese peroxidase (EC Next, the filtrate obtained after filtration through
1.11.1.13) Whatman No. 1 filter paper was added to sodium
Laccase activity was measured (11) by taking the carbonate and Folin-Ciocalteu reagent and incubated
optical density of the reaction mixture prepared by at 37 °C for 30 min, and its optical density was taken
mixing 0.5 mL of distilled water, 1 mL of sodium at 660 nm. A standard curve was prepared using
acetate buffer (pH 4.5), and 0.5 mL of substrate tyrosine and expressed as the quantity of the enzyme
solution (46 mM guaiacol) to 0.5 mL of crude enzyme that releases soluble fragments of trichloroacetic acid
extract at 440 nm for up to 90 s with time intervals giving a blue color equivalent to 0.5 mg/mL tyrosine
of 30 s. Lignin peroxidase activity was evaluated (12) under the conditions of the assay.
by following the same procedure as for laccase, but Alpha amylase (EC 3.2.1.1)
0.5 mL of hydrogen peroxide was also added to the Amylase activity was calculated by the following
mixture. Manganese peroxidase activity was assayed formula (20,21), and 1 unit of alpha amylase is the
(13) using 0.5 mL of sodium tartrate buffer (pH 5), amount of protein that will hydrolyze 10 mL of starch/
0.5 mL of 100 μM guaiacol, 1 mL of distilled water, min under specific conditions. The reaction mixture
0.1 mL of culture filtrate, and 0.5 mL of hydrogen (3 mL), containing 1 mL of 0.1 M acetate buffer (pH
peroxide (30% w/v) containing a reaction mixture 4.8), 0.5 mL of extract, and 1 mL of starch solution,
and by reading its optical density at 465 nm. For these was incubated for 10 min and then the reaction was
3 enzymes, 1 activity unit was defined as the amount stopped by the addition of 1 mL of iodine reagent and
of enzyme necessary to oxidize 1 μmol substrate/min. 3 mL of 0.05 N hydrochloric acid. Absorbance was
Lipase (EC 3.1.1.3) recorded at 620 nm. A decrease in absorbance was a
Lipase activity was measured using the universal measure of amylase activity.
titrimetric method (14,15). The oil-water emulsion
and enzyme extract (0.1 mL: 9.9 mL: 1 mL) was mg of starch Abc – Abd
titrated against 0.1 N sodium hydroxide using a
phenolphthalein indicator. A blank (9.9 mL water : 0.1 hydrolyzed = ----------- × mg of starch
mL Tween 20 : 1 mL sterilized broth) was previously Abc initially present,

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Bioactive compounds from discarded mushroom beds

where Abc = absorbance of control and Abd = extract in a total volume of 3 mL. The absorbance of
absorbance of digested sample. H2O2 was taken at 240 nm and the activity of the
Beta amylase (EC 3.2.1.2) enzyme was expressed in units/mL of extract.

The beta amylase activity was assayed (8) using a

reaction mixture contained 200 μL of soluble starch Results
in a phosphate buffer (0.5 M, pH 7.5), 200 μL of Of the 3 mushroom bed extracts, the first and second
enzyme extract, and 300 μL of phosphate buffer. The were button and oyster mushroom beds collected
reaction was incubated for 15 min at 30 °C, then 300 15 days after their harvest, while the third was a
μL of dinitrosalicylic acid solution was added and the button mushroom bed collected immediately after its
mixture was boiled for 15 min. Before cooling, 100 μL harvest. These mushroom bed extracts were assayed
of Rochelle salt (40% sodium potassium tartrate) was for bioactive compounds and enzymes and the results
added and the color was measured at 575 nm. One are shown in Table 1. The pH levels of the dried
unit of amylase activity was defined as the amount of button, dried oyster, and fresh button mushroom
enzyme that releases one milligram of reducing sugar bed extracts were 5, 4.5, and 4, respectively. The
as glucose per milliliter per minute under the assay amounts of total sugars were 830, 390, and 720 μg/
conditions. mL from the dried button mushroom bed extract,
Cellobiohydrolase (EC 3.2.1.91) the dried oyster mushroom bed extract, and the fresh
mushroom bed extract, respectively. The reducing
Exoglucanase or cellobiohydrolase activity was sugar content of the dried button, dried oyster, and
measured (22) using the reaction mixture for a fresh button mushroom extracts was 720, 630, and
reducing sugar assay containing 0.5 mL of enzyme 1330 μg/mL, respectively. The total lipid content of
extract, 1.5 mL of 0.05 M citric acid buffer (pH 4.8), the dried button mushroom bed extract was 20 mg/g,
and 0.05 g of cellulose substrate. After incubation while that of the oyster mushroom bed extract was 10
for 1 h at 50 °C, the reaction was stopped by the mg/g and the fresh button mushroom bed extract’s
addition of 2 mL of 3,5-dinitrosalicylic acid reagent. was 70 mg/g of sample. The total protein content
The resulting mixture was boiled for 15 min and the of the dried button, dried oyster, and fresh button
reduced content was measured by absorbance at 575 mushroom bed extracts was 180, 131, and 375 μg/mL
nm and expressed in terms of milligrams of reducing of extract, respectively.
groups (as μg glucose/mL) liberated using a 100
In terms of cellobiohydrolase activity, 20 μg/mL
mg/100 mL glucose standard graph.
of activity was noticed in the dried button mushroom
Xylanase (EC 3.2.1.8) bed extract, but there was no activity in the oyster
Xylanase activity was assayed (23) by preparing a mushroom bed extracts, and 790 μg/mL was shown
reaction mixture containing 0.5 mL of 0.1% (w/v) in the fresh mushroom bed extract. Catalase activity
substrate in a 0.1 M sodium acetate buffer (pH 5.0) was recorded from the dried button, dried oyster,
and 0.1 mL of enzyme extract. The mixture was and fresh button mushroom bed extracts (0.15, 0.12,
incubated at 40 °C in a water bath with agitation for 30 and 0.5 U respectively). The oyster mushroom bed
min. The reducing sugar released was measured using extracts did not show a positive result for protease
3,5-dinitrosalicylic acid. The color was developed by and beta amylase activity, but 340 and 310 μg/mL of
boiling in a water bath for 5 min. Absorbance was beta amylase activity was recorded from the dried
read at 540 nm and 10 μM xylose was used to prepare button and fresh button mushroom bed extracts,
a standard graph. The enzyme activity was expressed respectively. Neither the dried button nor the oyster
in units/mL. mushroom bed extract showed a positive result for
the lignolytic enzyme assays, such as laccase, lignin
Catalase (EC 1.11.1.6) peroxidase, and manganese peroxidase, but 48 U of
Catalase activity was determined (24) using a laccase activity was recorded from the fresh button
reaction mixture containing 40 mM H2O2 in a 50 mushroom bed extract. In terms of xylanase activity,
mM phosphate buffer (pH 7.0) and 0.1 mL of enzyme 8.4 U/mL was shown in the dried button mushroom

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 S.V.S.S.S.L. H. B. NIDADAVOLU, R. P. METUKU, S. BURRA, R. BYRAM, S. C. MARINGANTI

Table 1. Bioactive compounds and enzymatic activities of 3 mushroom bed extracts.

Dried button Dried oyster Fresh button

Parameters mushroom mushroom mushroom
bed extract bed extract bed extract
pH 5 4.5 4
Total sugars (μg/mL) 830 390 720
Reducing sugars (μg/mL) 720 630 1330
Total lipids (mg/g) 20 10 70
Lipase (μm) 0.5 1.5 0.5
Proteins (μg/mL) 180 131 375
Cellobiohydrolase (μg/mL) 20 - 790
Xylanase (U/mL) 8.4 5.7 24.3
Carboxymethyl cellulase (μg/mL) 200 920 100
Protease (μg/mL) 40 - 47
Catalase (U/mL) 0.15 0.12 0.5
α-amylase (U) 1.18 9.0 6.37
β-amylase (μg/mL) 340 - 310
Manganese peroxidase (U/mL) - - -
Laccase (U/mL) - - 48
Lignin peroxidase (U/mL) - - -
- = not detected

bed extract, 5.7 U/mL was given by the oyster Since the mushroom bed extract gave remarkable
mushroom bed extract, and 24.3 U/mL was found in results for lignin peroxidase and laccase activity
the fresh button mushroom extract. Carboxymethyl compared to the glucose-containing extract and the
cellulase activity was 200, 920, and 100 μg/mL in common production medium, it was further diluted
the dried button, dried oyster, and fresh button to 1:1 and 1:2 ratios and tested again for the same
mushroom bed extracts, respectively. after 7 days of incubation with F. feei, T. gibbosa,
Table 2 shows the dry weight and the lignin and T. elegans. The results are given in Table 3. Dry
peroxidase and laccase activities of S. commune, F. weight, lignin peroxidase, and laccase activities were
feei, T. gibbosa, and T. elegans in direct mushroom high in the 1:1 ratio medium compared to the 1:2
bed extract, extract prepared by mixing with ratio medium.
(0.2%) glucose, and common medium after 7 and Lignin peroxidase and laccase activity from
14 days of incubation. Dry weight was high in both rotary evaporator and dried powder extracts
the glucose-containing extract for all organisms. containing broths inoculated with F. feei, T. gibbosa,
Lignin peroxidase activity (1668 U/mL) was high and T. elegans were assayed after 7 days (Table 4).
with the mushroom bed extract when compared The dry weight of all of the organisms was high in
to the glucose-containing extract and the common the dried powder extract medium (4, 3.2, and 2.4
production medium for T. gibbosa. Laccase activity g/L, respectively), but T. gibbosa showed high lignin
was high with the glucose-containing extract for T. peroxidase activity (376 U/mL) and laccase activity
gibbosa (1980 U/mL). (414 U/mL) in the rotary evaporator extract medium.

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Bioactive compounds from discarded mushroom beds

Table 2. Growth and lignolytic enzyme activity of 4 white rot fungi in 3 media.

Days of
Type of media Type of activity S. commune F. feei T. gibbosa T. elegans
incubation

Dry weight 7 12.8 4.8 5.6 5.2

(g/L) 14 5.2 4.8 4.8 4.6
Lignin 7 - - 1668 -
Direct
peroxidase
mushroom 14 324 - 444 -
(U/mL)
bed
extract 7 - - 1860 -
Laccase
(U/mL) 14 - - 602 600

Dry weight 7 12.8 13.6 12.8 5.6

(g/L) 14 18.4 10.8 14.8 6.8
Mushroom Lignin 7 - - 796 -
bed peroxidase
extract (U/mL) 14 - - 388 -
+
Laccase 7 - - 1980 -
0.2% glucose
(U/mL) 14 - - 1690 -

Dry weight 7 4.8 6.8 9.6 7.2

(g/L) 14 11.6 6.9 8.4 5.6
Lignin 7 - 56 12 -
Production
peroxidase
medium 14 - - - 32
(U/mL)
(control)
Laccase 7 - 96 - -
(U/mL) 14 - - 36 -

- = no activity

Table 3. Dry weight and lignolytic enzyme activity of 3 white rot fungi using 1:1 and 1:2 ratios of button mushroom bed extract after
7 days of incubation.

Dilution of media Lignin peroxidase Laccase

Organism Dry weight (g/L)
with distilled water (U/mL) (U/mL)
1:1 2.8 20 42
F. feei
1:2 1.2 6 -
1:1 2.4 588 602
T. gibbosa
1:2 1.2 262 360
1:1 2.0 - -
T. elegans
1:2 1.2 - -

- = No activity

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 S.V.S.S.S.L. H. B. NIDADAVOLU, R. P. METUKU, S. BURRA, R. BYRAM, S. C. MARINGANTI

Table 4. Comparison of rotary evaporator and dried powder extract media containing 3 white rot fungi for growth and lignolytic
enzymes after 7 days of incubation.

Organism Type of media pH Dry weight (g/L) Lignin peroxidase (U/mL) Laccase (U/mL)

F. feei RE 3.5 1.6 6 -

DPE 3.5 4 - -

RE 5.0 2.4 376 414

T. gibbosa
DPE 5.5 3.2 284 282

RE 5.0 0.8 - -
T. elegans
DPE 5.0 2.4 - -

RE = media prepared from mushroom bed extract using rotary evaporator

DPE = media prepared from dried mushroom bed powder
- = no activity

The growth of these 3 organisms was better on the sustain the growth of Salvia officinalis by improving
dried powder extract than on the rotary evaporator the air porosity and mineral content of the soil.
extract when grown on agar plates (Figure). Earlier research reported that mushroom compost
waste can be used as an alternative fertilizer to farm
yard manure in strawberry growing (30). Many
Discussion reports have shown that mushroom compost waste
Similar to our present observations, the wastes can be a good culture substrate when it is mixed
produced by mushroom forms for making compost with soil as a farm yard fertilizer or used alone in
have previously been studied (6). Furthermore, order to replenish the physical condition of the soil
Pleurotus spp. have been analyzed for possible (31-33). The results obtained in the present study
utilization as valuable bioactive compounds. The clearly indicate that improvement in the physical
separation of sugars, lipids, and diversified enzymes and chemical status of the soil is closely associated
from mushroom bed extracts is a novel concept with the compounds released by spent mushrooms
for the creation of WOW. Reutilization of spent and the subsequent discharge of a variety of enzymes
mushroom substrate (SMS) and the separation of during the decomposition process. Hence, SMS is an
bioactive compounds with soil properties like bulk added advantage for the improvement of soil fertility.
density, stability, surface crust, temperature changes, Recycling of Pleurotus waste from the cultivation
infiltration rate, aeration, and water retention capacity of Pleurotus sajor-caju gave a significant yield of
have been utilized (25,26). It was also noticed in our Pleurotus sajor-caju on starch-, peptone-, and wheat
study that temperature and water content played an bran-supplemented SMS (34).
important role in the extraction of compounds and Recycling of waste mushroom substrate for
enzymes. mushroom cultivation has been carried out using the
The addition of straw to the soil caused an increase sawdust from waste shiitake bed log for the cultivation
in the number of total bacteria, actinomycetes, and of Pleurotus cornucopiae (35). Pretreatment of waste
fungi of the rhizosphere (27). The yield of green mushroom beds and methods for converting the same
gram increased in plots previously supplied with to yield sugars and ethanol has been patented (36). In
mushroom spent rice straw (28). Recently, a report accordance with prior work (37), the determination
(29) stated that Pleurotus waste was adequate to of laccase enzymes from mushroom bed extracts is

281
Bioactive compounds from discarded mushroom beds

Media prepared by using rotavapor extract

Media prepared by using dried powder extract

Figure. Growth pattern of 3 white rot fungi on agar plates containing media prepared using rotary
evaporator and dried powder extracts.

an innovative technology for the separation of sugars biogas. Since these waste beds are ecofriendly, very
from lignocellulosic materials (which are not suitable cheap, easily extractable, and proven to be good
for food or fodder) for the large scale production media, these extracts can be used directly as media
of bioethanol to improve petrol. A change in the for the growth of microorganisms in comparison to
physicochemical properties of recycled spent cost-effective synthetic media.
mushroom compost through vermicomposting by
epigeic earthworms Eisenia foetida and E. andrei was
reported (38). Thermal treatment technologies were Acknowledgments
compared to determine an appropriate method of The authors are very grateful to the University Grants
recovering energy from 2 wastes, spent mushroom Commission, New Delhi, for providing financial
compost and coal tailings (39). assistance.
As the ingredients for growth media for
microorganisms are enormously increased by
SMS, different bioactive compounds can effectively Correspondence author:
substitute for costly commercial media. Our present S.V.S.S.S.L. Hima Bindu NIDADAVOLU
efforts were successful in the cultivation of 4 white rot
fungi and establishment of production efficiencies of Department of Microbiology,
lignolytic enzymes during 7 and 14 days of incubation Kakatiya University,
time. It has been concluded that certain bioactive Warangal 506 009 - INDIA
compounds and industrial enzymes can be extracted
from these waste beds. These waste beds can be used E-mail:*[email protected]
for the production of bioethanol, bioenergy, and

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 S.V.S.S.S.L. H. B. NIDADAVOLU, R. P. METUKU, S. BURRA, R. BYRAM, S. C. MARINGANTI

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