PHILIPP AGRIC SCIENTIST ISSN 0031-7454

Vol. 96 No. 2, 198–204

June 2013

Philippine Native and Exotic Species of Edible Mushrooms Grown on

Rice-Straw-Based Formulation Exhibit Nutraceutical Properties
Renato G. Reyes1, Sofronio P. Kalaw1, Rich Milton R. Dulay1, Hiroaki Yoshimoto2, Noriko
Miyazawa3, Tomoko Seyama4 and Fumio Eguchi4,*
1
Center for Tropical Mushroom Research and Development, Department of Biological Sciences, College of Arts and
Sciences, Central Luzon State University, Science City of Muñoz, Nueva Ecija, Philippines
2
Mushtech Co. Ltd., Japan
3
Takasaki University of Health and Welfare, Gunma, Japan
4
Tokyo University of Agriculture, Sakuragaoka, Setagayaku, Tokyo, Japan
*
Author for correspondence; e-mail: f1eguchi@nodai.ac.jp

To establish the nutraceutical potential of Philippine native and exotic species of edible mushrooms,
the amino acid profile and functional activities of Schizophyllum commune Fr., Lentinus tigrinus (Bull.)
Fr., Lentinus sajor caju Fr., Ganoderma lucidum (Curtis: Fr.) P. Karst. and Pleurotus florida (Mont.)
Singer were elucidated. Out of 21 standard amino acids present in eukaryotes, 20 were found in P.
florida and S. commune and 19 in L. sajor caju and L. tigrinus. Arginine was not detected in L. tigrinus
and L. sajor caju while methionine was not found in G. lucidum. Several kinds of nonstandard amino
acids, namely, phosphoserine, urea, α-aminobutyric acid, cystathionine, β-alanine, γ-aminobutyric acid,
monoethanolamine, ammonia and ornithine were detected in all species of mushrooms used. On the
other hand, phosphoethanolamine and α-aminoacidipic acid were found in P. florida only while 1-
methylhistidine was detected in S. commune, G. lucidum and P. florida. The antiplatelet aggregation
test was performed to determine the anticoagulant activity of the extract. The different species of
mushrooms demonstrated inhibition of platelet aggregation as mediated by aggregating factor,
arachidonic acid Na and adenosine diphosphate. P. florida produced the highest inhibition of activating
factor while S. commune exhibited the highest inhibition of arachidonic Na and adenosine diphosphate
activity. Finally, all the mushroom species demonstrated anti-inflammatory activities as indicated by the
inhibition of IL-8 gene expression.

Key Words: exotic mushrooms, medicinal mushrooms, mushroom nutraceutical, wild edible mushrooms

Abbreviations: AA – arachidonic acid, ADP – adenosine diphosphate, DMEM – Dulbecco’s modification Eagle’s
medium, DMSO – dimethylsulfoxide, FAO – Food and Agriculture Organization, IL-8 – interleukin-8, GAPDH –
glyceraldehyde-3-phosphate dehydrogenase, M-MLV – Moloney-Murine Leukemia Virus, PAF – platelet activating
factor, PPP – platelet poor plasma, PRP – platelet rich plasma, RT-PCR – reverse transcription-polymerase chain
reaction, TNF-α – tumor necrosis factor alpha

INTRODUCTION mushroom eaters, their consumption of edible

As a tropical country, the Philippines has very rich
mycological resources which unfortunately are still in the
wilderness growing on fallen logs, decomposing piles of
straws, lawns, meadows and gardens. Emergence of wild
mushrooms via the germination of spores commences in
May and lasts until September when there is sufficient
moisture and optimal temperature (Bulseco et al. 2005;
Mariano et al. 2005; Musngi et al. 2005; Paderes et al.
2004; Reyes et al. 2006; Reyes et al. 2003; Reyes and
Abella 2002; San Agustin et al. 2004). This situation
makes the collection of edible mushrooms from the wild
seasonal. Even though most Filipinos are known to be
mushrooms is limited by the unavailability of research-
based production technology and the lack of information
on the edibility and nutraceutical benefits of these
mushrooms (FAO Statistics 2007).
To popularize the production and consumption of
cultured mushrooms from the Philippines, we have
initiated in the last decade the development of mushroom
production technologies using locally available agro-
industrial residues in a more practical, economical yet
financially rewarding setup. Efforts have been done to
domesticate Lentinus tigrinus (Dulay et al. 2012a and b),
the more popular Volvariella volvacea (Reyes et al.
2004b; Reyes 2000), Collybia reinakeana which used to

198 The Philippine Agricultural Scientist Vol. 96 No. 2 (June 2013)

Philippine Native and Exotic Species of Edible Mushrooms
be a wild edible species from Puncan, Carranglan, Nueva
Ecija in the Philippines (Reyes et al. 2004a), Coprinus
comatus (Reyes et al. 2009a) and S. commune, a wild
edible mushroom known to the Bicolanos as kurakding,
to the Visayans as kudopdop, to the Ilonggos as Kudyadyi
and to the Ilocanos as kudit (Gisala et al. 2005; Garcia et
al. 2004). Exotic species of ligninolytic edible
mushrooms such as Pleurotus sajor-caju and P. florida
have already been successfully grown in recent years on
rice-straw-based formulation in the rural areas of the
country (Reyes et al. 2009b). The Center for Tropical
Mushroom Research and Development at the Central
Luzon State University has developed a technology on
the efficient utilization of rice straw as substrate for the
cultivation of ligninolytic edible mushrooms (Reyes et al.
2006; Villaceran et al. 2006) which conventionally are
produced on enriched sawdust-based formulation.
At the turn of the 21st century, the mushroom-eating
habit of Filipinos and their perception of mushrooms
have changed. Imported mushroom-based nutraceutical
products such as Ganoderma and Agaricus blazei coffee,
Ganoderma-based dietary supplements in addition to
dried Lentinula edodes and Auricularia from China and
canned Agaricus bisporus from the US were introduced
in the local market. As such, mushrooms are no longer
regarded these days by Filipinos as simply gourmet food
but as nutraceutical commodities as well.
Nutraceuticals are substances that may be considered
as food or parts of a food that exhibit medical or health
benefits (Khatun et al. 2012). Mushrooms are considered
as one of the nutraceutical commodities because they
possess functionalities such as antitumor, anti-
inflammatory, immune enhancers (Rogers 2006) and
prevent diabetes, hyperlipidemia, arteriosclerosis and
chronic hepatitis (Takaku et al. 2001). Being nutritious
food sources, they are richly endowed with all essential
and commonly occurring non-essential amino acids
(Quimio 2004), which indeed elucidated their nutritional
role in the human diet. With this premise, our research
team has initiated an intensive effort of domesticating
wild edible mushrooms from the Philippines with
potential in the nutraceutical industry.
Our study was conducted to elucidate the amino acid
profile and functionalities of native and exotic species of
edible mushrooms. Inhibition of platelet aggregation and
IL-8 gene expression in vitro were used to establish the
anticoagulation and anti-inflammatory activities of these
mushrooms.
MATERIALS AND METHODS
Cultivation of Fruit Bodies
Three Philippine strains of edible mushrooms, namely, S.
commune, L. sajor-caju, L. tigrinus and G. lucidum and
one exotic species (P. florida) were grown on 7 parts
The Philippine Agricultural Scientist Vol. 96 No. 2 (June 2013)
Renato G. Reyes et al.
composted rice straw (soaked in water tank overnight,
washed, hauled out from the tank, piled, and covered
with sacks) and 3 parts of coconut sawdust (v/v)
contained in 17.78 cm x 35.56 cm polypropylene plastic
bags. Bags were incubated at room temperature (30 °C).
To allow the emergence of fruit bodies, fully ramified
fruit spawn bags were transferred to a mushroom-
growing house (28 °C) and the usual protocol in the
cultivation of edible mushroom in the tropics was
adopted (Reyes et al. 2009c).
Preparation of the Mushroom Samples for Assay
The fruit bodies were dried at 45–60 °C in a mushroom
convection dryer. The dried fruit bodies were ground
using a Wonder Blender WB-1 (Osaka Chemical Co.,
Ltd., Osaka, Japan) and subsequently sieved through a
1000-µm mesh and the resultant powder was used. For
the platelet aggregation inhibition test and IL-8 gene
expression inhibition test, the powder was extracted in 10
volumes of hot water (80 °C) for 1 h, and the hot water
extract was concentrated under reduced pressure after
filtration (Advantec No. 2 filter paper, Toyo Roshi
Kaisha, Ltd. Tokyo, Japan). For amino acid analysis, the
hot water extraction procedure of Eguchi et al. (1999)
was followed. The active components of the milled
mushroom sample (20 g) were extracted in 600 mL hot
water at 80–90 °C in a water bath for 2 h.
Amino Acid Profile
The determination of the nutritional content of the
mushroom samples was limited to their free amino acid
profile. Standard and nonstandard amino acid contents
(McKee and McKee 1999) were calculated in an L-8900
amino acid analyzer (Hitachi Co. Ltd. Japan).
Platelet Aggregation Inhibition Test
The platelet aggregation inhibitory effect, induced by
platelet activating factor (PAF) and arachidonic acid Na
(AA), was evaluated. Human peripheral blood was
collected from the median cubital vein of a medication-
free healthy adult for at least 2 wk. The blood was
centrifuged (1100 rpm, 20 min, room temperature) and
the upper layer was collected as the platelet rich plasma
(PRP). Subsequently, the lower layer was centrifuged
(3000 rpm, 5 min, at room temperature) and the platelet
poor plasma (PPP) was collected.
PRP and PPP (223 µL of each) were preheated at 37
°C and the methanol extract of each sample, dissolved in
2 µL of a 2% dimethylsulfoxide (DMSO) solution, was
added to PRP and PPP. These experiments were
incubated for 3 min at 37 °C. After incubation, 25 µL of
PAF or an aqueous solution of arachidonic acid (500 nM)
was added and platelet aggregation induced. Ion
exchanged water was used as control. Induced
aggregation was measured using an aggregometer (MCM
Hema Tracer 313M, MC Medical Co., Ltd. Tokyo,
199