Bioresource Technology 96 (2005) 537544

Comparative culturing of Pleurotus spp. on coee pulp and

wheat straw: biomass production and substrate biodegradation
a,*
Dulce Salmones , Gerardo Mata a, Krzysztof N. Waliszewski b

a
Laboratorio de Hongos Comestibles, Departamento de Hongos, Instituto de Ecologa, P.O. Box 63, 91070 Xalapa, Mexico
b
Departamento de Ingeniera Qumica y Bioqumica, Instituto Tecnologico de Veracruz, P.O. Box 1420, 91808 Veracruz, Mexico

Received 16 September 2003; received in revised form 16 June 2004; accepted 17 June 2004
Available online 15 September 2004

Abstract

The results of the cultivation of six strains of Pleurotus (P. djamor (2), P. ostreatus (2) and P. pulmonarius (2)) on coee pulp and
wheat straw are presented. Metabolic activity associated with biomass of each strain was determined, as well as changes in lignin and
polysaccharides (cellulose and hemicellulose), phenolic and caeine contents in substrate samples colonized for a period of up to 36
days. Analysis were made of changes during the mycelium incubation period (16 days) and throughout dierent stages of fructi-
cation. Greater metabolic activity was observed in the wheat straw samples, with a signicant increase between 4 and 12 days of
incubation. The degradation of polysaccharide compounds was associated with the fruiting stage, while the reduction in phenolic
contents was detected in both substrates samples during the rst eight days of incubation. A decrease was observed in caeine con-
tent of the coee pulp samples during fruiting stage, which could mean that some caeine accumulates in the fruiting bodies.

2004 Elsevier Ltd. All rights reserved.

Keywords: Pleurotus; Coee pulp; Wheat straw; Biomass; Biodegradation

1. Introduction conversion of an agricultural waste to human food.

Coee pulp, one of the principal byproducts of wet-
processed coee (Coea arabica L.), which constitutes
almost 40% of the wet weight of the coee berry, is rich
in carbohydrates, proteins, minerals, and appreciable
quantities of tannins, caeine and potassium (Bresanni,
1979). Approximately 100,000 tons of coee pulp are
generated each year in Mexico, and the majority of this
waste has no further economic use; instead, coee grow-
ers generally spread it in the eld where it is allowed to
decompose.
Diverse technologies have been proposed for utilizing
the byproducts generated by the coee industry (Pandey
et al., 2000). Culturing edible mushrooms on coee pulp
seems especially attractive, since it represents a direct
*
Corresponding author: Fax: +52 228 8187809.
E-mail address: dulce@ecologia.edu.mx (D. Salmones).
Among edible mushrooms evaluated for this commer-
cial activity, Pleurotus strains appear promising, prima-
rily because their biological eciencies can exceed 100%
(wet base) (Martnez-Carrera et al., 1985; Martnez-
Carrera, 1989).
In the present study, two strains each of Pleurotus
ostreatus, Pleurotus pulmonarius and Pleurotus djamor
were cultured on coee pulp and wheat straw. The pur-
pose was to characterize dierences in biomass produc-
tion by the vegetative stage and fruiting bodies
production of the strains, as well as changes in lignin
and polysaccharide contents (cellulose and hemicellu-
lose) of the substrates attributable to dierential use
by strains. The degradation of toxic components, partic-
ularly phenols and caeine, was also studied. The prac-
tical goal of this investigation was to identify possible
advantages for substituting the commercially popular
wheat straw substrate with the agroindustrial waste,

0960-8524/$ - see front matter
2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2004.06.019
538 D. Salmones et al. / Bioresource Technology 96 (2005) 537544
coee pulp. In addition, this study permitted the identi-
cation of those strains most capable of developing in
coee pulp. This research is part of a series of investiga-
tions that are focused on optimizing the use of coee
pulp as a medium for mushroom cultures.
2. Methods
2.1. Strains
The following six strains of Pleurotus were studied:
IE-38 and IE-49 of P. ostreatus (Jacq.: Fr.) Kumm
and IE-137 of P. pulmonarius (Fr.) Quel. were obtained
from commercial strains available in Europe and Asia;
IE-121 of P. djamor (Fr.) Boedjin was isolated from a
wild specimen in Mexico and nally, IE-218 of P. dja-
mor and IE-225 of P. pulmonarius were obtained from
genetic crosses of monosporic cultures in our labora-
tory. All cultures were deposited in the culture collection
of the Instituto de Ecologa (Xalapa, Mexico).
2.2. Culture conditions
Spawn was prepared from sorghum seeds that had
been hydrated to a moisture content of 55% by weight.
Two hundred grams of hydrated grains (wet weight)
were placed in polypropylene bags, and then sterilized
at 121 C for 1 h (Gaitan-Hernandez et al., 2002). Once
cooled, each bag of sorghum seeds was inoculated with
mycelia from one of the six pre-cultured Pleurotus
strains. Inoculated bags were incubated in darkness at
27 C until mycelia had completely covered the sorghum
grains. This process took approximately 23 weeks to
complete.
Coee pulp was collected at a coee processing plant
(located in Coatepec, Veracruz, Mexico), air-dried to a
moisture content of 1820%, and then stored at ambient
temperature. Dry wheat straw was broken into 13 cm
fragments with an electric thresher and also stored un-
der identical conditions as the coee pulp. In all experi-
ments, each of these substrates was rehydrated in water
for 12 h, and then excess moisture was allowed to run o
until a moisture content of 60% (5%) was reached. Fol-
lowing this treatment, substrates were placed in polypro-
pylene bags and sterilized at 121 C for 1 h. Two
hundred gram samples (wet weight) of wheat straw or
coee pulp were established for (1) estimating biomass
in the vegetative stage, (2) analyzing ber components,
(3) measuring phenolic contents, and (4) assessing caf-
feine concentrations.
Each 200 g sample was seeded with 10 g (5% inocula-
tion) of spawn and incubated at 27 C in total darkness.
Eight sample replicates 6 strains 2 substrates were
prepared (N = 96 total samples) in order to undertake
the analyses. In parallel, three samples per substrate,
without mycelia, were prepared and these served as con-
trol groups. The time required for the complete coloni-
zation of samples of either substrate was established as
16 days of incubation at 28 1 C, in total darkness
(Velazquez-Cedeno et al., 2002). Later, samples colo-
nized were placed, without their plastic bags, in a con-
trolled environment and under conditions favorable
for the induction of fruiting; i.e., with an average ambi-
ent temperature of 24 5 C, an relative humidity of
84 6%, 400 m3 of air circulating per hour, and
11 1 h of natural illumination daily. Samples were cul-
tured under these conditions for 36 days. During the
experiment, the samples in the following stages (S) were
collected and analyzed: 4, 8, 12, and 16 days of incuba-
tion (S2, S3, S4, and S5, respectively); rst appearance
of primordia (S6); beginning of rst harvest (S7); and
one week after nishing rst harvest (S8). Control sam-
ples were collected at 0, 16, and 36 days of incubation
(S1, S5, and S9, respectively). All collections were frozen
at 20 C, lyophilized, pulverized in a domestic blender,
and then stored at 4 C.
2.3. Estimation of biomass production during the vegeta-
tive stage of the culture
Biomass production was estimated from mycelial
metabolism during the substrate colonization phase
and was determined using the method of uorescein
diacetate hydrolysis (FDA; Sigma) (Inbar et al., 1991;
Mata et al., 2002). This determination requires sterile
culture conditions, and for this reason, only sample
stages under active incubation and covered with plastic
lids were analyzed (i.e., S1S5, inclusive). This experi-
ment was undertaken by placing 0.2 g of lyophilized
material per strain, per substrate, per collection stages
(i.e., S2S5) in each of ve, sterile test tubes (N = 6
strains 2 substrates 5 tubes 4 collection stages =
300 total tubes). In three tubes, 3 ml of FDA (10 mg/l)
were added, and in the two remaining tubes (considered
as controls) 3 ml of phosphate buer solution (60 mM,
pH 7.6) were added. Test tubes were incubated at 30
C for 10 min, after which time the reaction was halted
by the addition of 3 ml of acetone. The suspension was
ltered (Wattman lter, No. 1) and the remaining solu-
tion was detected having a spectrophotometric absorb-
ency of 490 nm (Spectronic Genesys 5, Macedar, NY).
One unit of metabolic activity was dened as 1 lmol
of FDA hydrolyzed min 1 g 1 (dry weight). Control
samples were analyzed for the stages S1 and S5 (N = 2
substrates 5 tubes 2 collection stages = 20 tubes).
2.4. Analysis of ber components
The ber component of all colonized substrates was
determined with a ber analyzer (Ankom, Model 200/
220, Macedar, NY). Using the detergent analysis
 D. Salmones et al. / Bioresource Technology 96 (2005) 537544
method (Van Soest and Wine, 1967), both the neutral
detergent ber (NDF) and the acid detergent ber
(ADF) were extracted. Lignin fractions were determined
with sulfuric acid (72%), and the cellulose content was
estimated directly from ADF-lignin. Hemicellulose was
arithmetically calculated as NDF ADF. Material ana-
lyzed corresponded to S1, S5, and S8 stages and S9 for
control samples.
2.5. Variation in phenolic content and caeine
To determine the concentration of phenolic com-
pounds, 1 g of lyophilized substrate was mixed with 10
ml of distilled sterile water and the solution agitated
with a rotor at 45 rpm for 30 min. The resulting suspen-
sions were passed through an inert nylon cloth and cen-
trifuged twice at 10,016g, for 15 min each time.
Supernatants were maintained at 4 C. Phenols present
in the substrates were determined by adding Folin and
Ciocalteu reagent to the supernatant (Box, 1983). After
1 h of incubation at 37 C, absorbency was read to 750
nm and results were expressed in mM of water-soluble
phenols per gram of substrate. Material corresponded
to the S2S8 stages and was analyzed in three replicates
(N = 6 strains 2 substrates 7 collection stages 3
repetitions = 252 vials). Control samples were analyzed
for the stages of S1, S2, S5, and S9 (N = 2 substrates 3
collection stages 3 repetitions = 18 vials).
The analysis of caeine was based on Newmans
method (1981), but modied as follows: 0.5 g of sample
(dehydrated and pulverized) was mixed with chloro-
form, sodium metabisulte, and an acetic acid/NaOH
buer (pH 6.5). This solution was then agitated in an
ice bath for 45 min, and the chloroform layer separated
with a Wattman No. 1 lter paper. The chloroform in
solution was evaporated over a hot water bath and the
caeine adhering to the walls of the receptacle was redis-
solved in 10 ml of 50% ethanol. Caeine concentrations
were read at an absorbency level of 294 nm. Collection
stages subjected to analysis corresponded to S1, S5,
and S8. This test was carried on three replicates per con-
dition (N = 6 strains 3 collection stages 3 tubes = 54
tubes). Three replicates of the control samples (S1, S5,
and S9 collection stages) were also analyzed. In addi-
tion, caeine content in the fruiting bodies of IE-225
strain were quantied. Results were recorded as percent-
age of caeine per gram of dry matter.
2.6. Determination of strain productivity
For the production of fruiting bodies, the sample size
was incremented to 2 kg of sterile substrate (500 g dry
weight). Each sample was placed in a plastic bag, homo-
geneously mixing the substrate with 100 g of spawn (5%
inoculation ratio) and incubating the mixture in dark-
ness at 28 C. Ten sample replicates 6 strains 2 sub-
539
strates were prepared (N = 120 total bags). Sixteen days
following inoculation (or earlier if primordia were
observed) the plastic covers were removed and sam-
ples were transferred to a room with adequate condi-
tions for the development of fruiting bodies (Section
2.2). Samples were maintained under these environmen-
tal conditions until the completion of the culture cycles
of 36 days. Strain productivity, reported as biologi-
cal eciency (BE), was calculated as percent fresh
weight of fruiting bodies with respect to dry weight of
substrate.
2.7. Statistical analysis
Means and standard deviations were calculated, and
signicant dierences were estimated using the Tukey
multiple-range test at p < 0.05. In addition, correlation
analysis were made between biological eciencies and
metabolic activity in order to nd out relations between
biomass production during vegetative and reproductive
stages.
3. Results and discussion
Metabolic activities for the six Pleurotus strains on
coee pulp and wheat straw substrates are shown in
Fig. 1. No activity was detected for the control samples
at any time, whereas metabolic activity was recorded for
colonized samples of both substrates starting at the
fourth day of incubation.
Statistically signicant increases in metabolic activity
on wheat straw samples were observed for all strains be-
tween S2 and S3, and between S3 and S4. Strains IE-
121, and IE-218 attained their maximum activities at
S3, whereas IE-38, IE-49, IE-137 and IE-225 reached
maximum activities at S4. The highest metabolic level
among all strains was noted from IE-225.
Metabolic activity on coee pulp samples increased
between S2 and S3, although this was only statistically
signicant for strains IE-49, IE-218, and IE-225. IE-
38, IE-49, IE-137 and IE-225 strains continued to in-
crease metabolic activities between S3 and S4; however,
this was only signicantly dierent for IE-38, IE-49 and
IE-225. Maximum metabolic activities were noted at S3
for P. djamor strains (IE-121 and IE-218), and S4 for the
other four strains. IE-225 showed the greatest metabolic
level of all strains tested on coee pulp, which was also
the case with wheat straw. Although it was not possible
to establish a correlation between values for metabolic
activity and the biological eciencies measured for each
strain, it is interesting to note that IE-225 showed high
metabolic activities and yields in both substrates. This
nding suggests that in Pleurotus species (as well as in
other mushroom species under culture) the ability of
strains to grow on a new substrate is related to their
540 D. Salmones et al. / Bioresource Technology 96 (2005) 537544
Fig. 1. Metabolic activities (uorescein diacetate hydrolysis) recorded for Pleurotus strains during the rst 16 days of incubation on wheat straw and
coee pulp substrates.
metabolic activity during early stages of mycelial coloni-
zation. Although high metabolic activity does not guar-
antee high sporophore production, this activity depends
on the ability of the strain to utilize nutrients available
in the substrate, some of which will be indispensable
to the morphogenesis of fruiting bodies.
In addition this study demonstrated that Pleurotus
strains cultured on wheat straw represented more bio-
logical activity than strains cultured on coee pulp. This
result suggests that the complex chemical composition
of the coee pulp impedes its ecient conversion to
mycelium. In parallel studies (Savoie et al., submitted
for publication), we found that laccase and Mn-peroxi-
dase activity of Pleurotus strains on coee pulp is greater
than that taking place on wheat straw; suggesting that it
is likely associated with a greater level of enzymatic
detoxication of the coee pulp substrate compared to
the less-complex, wheat straw substrate. Both results
are in agreement with previous studies of Lentinula
and Pleurotus (Mata and Perez-Merlo, 1999; Salmones
and Mata, 2002), in which mycelial metabolic activity
during the rst few days of colonizing coee pulp was
more related to the detoxication of this substrate than
to an increase in mycelial biomass.
Consumption of the main ber components of the
two substrates at S5, S8 and S9 are presented in Table 1.
By the end of 16-day incubation period, the polysaccha-
ride content of the substrates had been very little con-
sumed by the mycelia. However, consumption was
noticeable after the rst harvest. Although results dem-
onstrated that all six strains consumed lignocellulose,
degradation percentage varied with each strain. In par-
ticular, IE-49 (P. ostreatus) eciently degraded hemicel-
lulose on coee pulp and wheat straw and lignin on
wheat straw, whereas the two P. djamor strains (IE-
121 and IE-218) showed a preference for the cellulose
of coee pulp. The remaining strains, IE-38, IE-137
and IE-225, were distinguished by a high consumption
of lignin when grown on coee pulp. In general, a great-
er decrease in lignocellulosic compounds was observed
for wheat straw samples than for coee pulp.
Cellulose to lignin ratios varied for both substrates
during the experiment (Table 1). The wheat straw, con-
trol samples showed ratios from 4.55 to 4.75, whereas
 D. Salmones et al. / Bioresource Technology 96 (2005) 537544 541

Table 1
Cell wall compositions for wheat straw and coee pulp substrates colonized with Pleurotus strains
Strain Mycelial growth stages NDF Hemicellulose Cellulose Lignin Cellulose:Lignin ratio
Wheat straw
Control S1 91.3 31.4 49.2 10.8 4.55
S5 90.1 32.1 47.5 10.0 4.75
S9 89.4 32.7 46.2 10.1 4.57
P. djamor
IE-121 S5 83.2 25.6 47.4 10.2 4.64
S8 65.6 20.6 37.4 7.6 4.92
IE-218 S5 78.2 26.4 42.6 9.1 4.68
S8 58.3 19.6 33.5 5.2 6.44
P. ostreatus
IE-38 S5 72.3 27.3 38.8 6.1 6.36
S8 68.4 22.3 40.5 5.6 7.23
IE-49 S5 72.6 22.3 40.7 8.8 4.62
S8 58 17.8 35.5 4.7 7.55
P. pulmonarius
IE-137 S5 78.8 28.3 42.7 7.8 5.47
S8 68.1 20.9 41.1 5.9 6.96
IE-225 S5 56.6 29.4 42.1 9.1 4.62
S8 58.9 20.9 33.4 4.6 7.26
Coee pulp
Control S1 67.7 17.1 24.5 26 0.94
S5 64.4 15.3 23.2 25.9 0.89
S9 63.5 15.1 23 25 0.92
P. djamor
IE-121 S5 59.3 14 23.5 21.8 1.07
S8 51.2 13.3 18.3 19.7 0.92
IE-218 S5 54 14.2 20.4 19.4 1.05
S8 51.9 13.4 18.9 19.6 0.96
P. ostreatus
IE-38 S5 55.9 14.2 22.3 19.4 1.14
S8 50.1 12.1 23 15 1.53
IE-49 S5 55.6 11.2 23.3 21.1 1.10
S8 53.6 11 22.3 20.3 1.09
P. pulmonarius
IE-137 S5 50.9 11.2 24.3 15.4 1.57
S8 49.5 11.5 24.1 13.9 1.73
IE-225 S5 57 14.3 22.9 19.8 1.15
S8 47.9 12.2 21.5 14.2 1.51
Values for each cell wall component were calculated as percent dry weights and were measured during the mycelial growth stages S1, S5 and S8 (or S9
for control samples).

the colonized straw samples attained ratios from 4.62
(IE-121) to 7.55 (IE-49). Control samples for coee pulp
showed ratios of 0.890.94, whereas values in degraded
coee pulp ranged from 0.92 (IE-121) to 1.73 (IE-137).
These results conrm the importance of the cellulose
content of the substrate in the production of fruiting
bodies, as previously indicated by other authors (Geetha
and Sivaprakasam, 1998; Thomas et al., 1998). How-
ever, it was not possible to correlate these same data
for the coee pulp substrate because it is likely that, in
this substrate, the mushroom received nutrition and en-
ergy from the abundant free sugars that were present
(Elas, 1978), and thus made limited use of the cellulose
fraction. The preceding nding is also backed up by the
minimal variation in cellulose that was observed during
the culture cycle. Additionally, the hemicellulose frac-
tion, although less abundant than the cellulose fraction,
may have functioned as an important source of carbon
during developmental stages previous to primordia for-
mation, as has been proposed for Agaricus bisporus (Sa-
voie, 1998).
Variation in the phenolic content of substrates (Fig.
2) during the culture cycle is an important aspect in
the adaptation of the strain, especially since the capacity
of a substrate to resist degradation has been partially
attributed to its array of phenolic compounds (Fermor
and Macauley, 1991). In the present study, the wheat
straw cultures showed a reduction in phenolic contents
during the rst eight days of incubation (0.030.06
mM/l) from initial concentration of 0.93 mM/l. By S4
S7, these values were maintained between 0.42 and
0.62 mM/l. In the wheat straw control samples no de-
crease was observed for the duration of the experiment.
The coee pulp samples presented the same pattern: the
542 D. Salmones et al. / Bioresource Technology 96 (2005) 537544

Fig. 2. Phenolic concentrations of wheat straw and coee pulp substrates measured during the growth and fruiting stages of dierent Pleurotus
strains.

substrate initially registered 2.10 mM/l of phenolic com-
pounds, but this amount was drastically reduced during
S2 and S3 from 0.10 to 0.58 mM/l. In previous similar
studies of Pleurotus developing on coee pulp (Murrieta
et al., 2002; Salmones and Mata, 2002), there was a sig-
nicant negative relationship between phenolic concen-
tration in the substrate and laccase activity during the
rst week of culturing. These authors demonstrated that
enzymatic activity not only contributed to the nutrition
of the vegetative stages of Pleurotus strains (as observed
for other edible mushrooms, including Lentinula edodes
and Agaricus bisporus (Mata and Savoie, 1998; Savoie,
1998)), but that laccase also actively participated in
detoxifying the coee pulp. This is particularly impor-
tant for the use of coee pulp as a growth medium
because a rapid reduction in phenolic concentration
would accelerate colonization, and consequently, de-
crease the risks of invasion by mold contaminants, such
as Trichoderma and Monilia, species frequently found in
decomposing coee pulp.
In our experiment, the caeine content of the coee
pulp substrate decreased during mycelial growth (espe-
cially during the development of fruiting bodies), such
that by the end of the rst harvest it was as much as
50% lower than the control (Fig. 3). However, caeine
was not actually lost during this process, part of this
compound was transferred to the sporophores during
their development, since the fruiting bodies analyzed
showed concentrations between 0.17% and 0.22% of caf-
feine. Our results are in accordance with previous stud-

Fig. 3. Caeine loss measured in coee pulp samples during the growth and development of dierent Pleurotus strains.
 D. Salmones et al. / Bioresource Technology 96 (2005) 537544
ies reported by Leifa et al. (2000) and Fan et al. (2003),
who documented that Pleurotus as did not have the abil-
ity to degrade caeine when cultured on coee pulp and
coee husks. Even so, it is necessary to establish through
further studies, if the afore mentioned characteristic is
attributable to all cultivated species of Pleurotus, as well
as its implications in the nutritional quality of the mush-
rooms harvested.
Data on the production of fruiting bodies are pre-
sented in Table 2. Each of the six Pleurotus strains
showed optimal mycelial growth on both substrates,
and all samples were completely colonized during the
rst week of culturing. Strains cultured on wheat straw
developed fruiting bodies earlier than strains cultured
on coee pulp. P. djamor strains were the most preco-
cious of all species and strains tested, forming their rst
primordia after 11 days of incubation on wheat straw
and after 13 days of incubation on coee pulp. One to
two harvests were obtained during the 20 days in which
samples were maintained in the production area. Aver-
age biological eciencies (expressed as percentages)
were recorded as 30.5 (18.5) to 77.1 (43.8) on wheat
straw and as 40.2 (4.7) to 86.5 (40.9) on coee pulp.
The greatest average biological eciencies corresponded
to IE-225 on wheat straw and IE-49 on coee pulp. In
spite of the early development of primordia in P. dja-
mor, productivities of these strains were low, especially
for IE-121. In addition, four of the strains studied
(i.e., IE-38, IE-49, IE-218, and IE-225) were able to in-
crease their productivities when cultured on coee pulp.
Comparing the results of this study on biological e-
ciencies with previous reports for some of these strains,
a decrease in values was clearly seen in the present study.
For example, Salmones et al. (1997) and Mestizo-Valdes
(1997) reported biological eciencies of 72.4% and
93.3% for the IE-121 and IE-218 strains, respectively,
when cultured on wheat straw; whereas Velazquez-
Cedeno et al. (2002) recorded a biological eciency of
Table 2
Productivity of Pleurotus strains, expressed as biological eciencies, during 36 days of culturing
Strains Wheat straw
Primordia initiation (days) Biological eciencies (%)
P. djamor
IE-121 1112 40.4(10.5)bca
IE-218 1114 30.5(18.5)c
P. ostreatus
IE-38 1819 50.2(15.8)abc
IE-49 1719 54.2(11.8)abc
P. pulmonarius
IE-137 1819 66.4(24.7)abc
IE-225 1632 77.1(43.8)ab
a
Values for biological eciencies per strain, per substrate, correspond to averages for 10 replicates and are expressed as percentages. Standard
deviations appear in parentheses. Dierent small-case letters following the data entries indicate signicant dierences among strains p < 0.05 (Tukeys
multiple range test).
543
125% for IE-38 when cultured on coee pulp. Neverthe-
less, it is important to note that the lower biological e-
ciencies reported in this study were derived from
basidiocarp weights from only one or two harvests
(70% of the expected total production), whereas the
higher eciencies reported in the earlier studies were de-
rived from 3 to 5 harvests (representing approximately
more than 95% of the total expected production). Thus,
the dierences in biological eciencies observed between
this study and earlier works might likely be explained by
dierences in the number and weights of harvests.
Therefore, the suitability of a mushroom-culturing
substrate depends, in great part, on its composition,
availability, and cost. In this regard, the advantages of
coee pulp are (1) it is an adequately nutritive substrate
for mushroom growth and development, (2) it is cur-
rently available as a no-cost, agricultural waste product,
and (3) it can be dehydrated and stored for long periods
of time with no apparent eect on mushroom yields, a
factor osetting its seasonal availability as fresh material
(Martnez-Carrera et al., 1996). Knowledge gained to
date about culturing Pleurotus on coee pulp shows that
this substrate has advantages over alternative waste sub-
strates and might serve as a substitute for wheat straw.
However, more studies will be necessary to nd strains
able to eciently degrade coee pulp, particularly phen-
ols and caeine, since many factors may aect this sub-
strates composition and the availability of its nutrients
for culturing mushrooms. Such factors might include,
but are not limited to (1) dierences in methods used
to separate the coee beans from the pulp, (2) phyto-
chemical dierences in pulp that can be attributed to dif-
ferent bean varieties, and (3) dierences in cropping
systems used to grow coee in the eld. The conse-
quences of these factors on mushroom yields are already
well known for straw-based substrate (Chalaux et al.,
1995; Labuschange et al., 2000). In addition, mushroom
resistance to attack by antagonistic molds might be
Coee pulp
Primordia initiation (days) Biological eciencies (%)
1320 40.2(4.7)bc
1523 61.8(31.4)abc
1832 71.6(28.1)ab
1731 86.5(40.9)a
2232 50.3(27.9)abc
2331 80.5(34.4)ab
544 D. Salmones et al. / Bioresource Technology 96 (2005) 537544
increased by inducing the production of lignocellulolytic
enzymes during the early stages of substrate coloni-
zation (Murrieta et al., 2002). Finally, coee pulp
degraded by mushroom cultures might have further
post-production value in the elaboration of forages, ver-
micomposts, and organic fertilizers. This possibility
deserves further attention and experimentation.
Acknowledgments
The authors are indebted to Ms. C. Rosa Elena
Caballero and Ms. C. Aracelly Vega, from Universidad
de Chiriqui (Panama), and Dr. Jean-Michel Savoie,
from INRA (France), for their collaboration in caeine
and metabolic activities techniques. This work was sup-
ported in part with nancial assistance from CONA-
CYT (project no. 28530-N).
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