Biosaintifika 9 (3) (2017) 572-578

Biosaintifika
Journal of Biology & Biology Education

http://journal.unnes.ac.id/nju/index.php/biosaintifika

Growth and Protein Content Establishment of Pleurotus ostreatus on

Liquid and Solid Medium

Aris Mumpuni, Nuraeni Ekowati, Purnomowati, Endang Sri Purwati

DOI: 10.15294/biosaintifika.v9i3.11660

Department of Microbiology, Faculty of Biology, Universitas Jenderal Soedirman, Indonesia

History Article Abstract

Received 24 September 2017 Pleurotus ostreatus cultivation is performed using solid medium to harvest fruit body
Approved 5 November 2017 and using liquid medium to harvest mycelia in submerged culture. Modifying nu-
Published 31 December 2017 trients in the medium to increase protein content of the fruitbody and mycelia
can be done through addition of nitrogen-containing materials. This study aims
Keywords to determine: the appropriate composition of the liquid medium for high myce-
Fruitbody; Mycelial biomass;
lial growth and protein content; and the exact composition of the solid medium to
Pleurotus ostreatus; Protein
obtain high fruitbody product and protein content. The method was experimental
with completely randomized design (CRD). The treatments were incubation of P.
ostreatus on three types of liquid medium and four types of solid medium. The re-
sults showed that the optimal liquid medium composition for mycelial growth was
Liquid Fermentation Medium 1 (FC1) with 10% corn fluor, and the highest protein
content was in Liquid Fermentation Medium 2 (FC2 = 29.76%). While the optimal
solid medium composition for fruitbody production was the medium with 3% corn
starch supplement (TJ3), and the highest protein content was obtained from the
medium without corn starch supplement (TJ0=24.69%). The increase of mycelial
and fruitbody weight from the medium with the addition of corn material indicated
a prospective in cultivation process, however effort to increase protein content of
the fruit body needs further research. Cultivating P. ostreatus in mycelial phase may
take shorter incubation time, may be produced in mass production with less space
consuming, and higher protein content than that by producing fruitbody.

How to Cite
Mumpuni, A., Ekowati, N., Purnomowati, P., & Purwati, E. S. (2017). Growth and
Protein Content Establishment of Pleurotus ostreatus on Liquid and Solid Medium.
Biosaintifika: Journal of Biology & Biology Education, 9(3), 572-578.

© 2017 Universitas Negeri Semarang


Correspondence Author: p-ISSN 2085-191X
Jl. HR Boenyamin 708, Grendeng, Purwokerto 53122 e-ISSN 2338-7610
E-mail: [email protected]
 Aris Mumpuni et al. / Biosaintifika 9 (3) (2017) 572-578

INTRODUCTION conditions. Furthermore, according to Hoa et

al. (2015) difference in mineral content of mush-
Pleurotus ostreatus (Jacq. Ex.Fr) Kummer room not only depended on mushroom species
or white oyster mushroom can grow on lignin but also depended on substrates used. That was
and cellulose containing medium and so is eas- due to mineral concentration of substrate formu-
ily found in weathered woods. People have been las. On the other hand, the electro conductivity
widely cultivating P. ostreatus as edible mushroom. of substrate is one of factors affected on mineral
It is charactized by high nutritional contents of uptake of mushroom. Ekowati et al. (2011) stated
10.5-30.4% protein, 1.6-2.2% fat, 57.6-81.8% that with the addition of minerals to the liquid
carbohydrates, 7.5-8,7% fiber, and 6.1-9.8% ash fermentation medium, the yield of L. edodes bioa-
(Rashad et al. 2009). P. ostreatus also has medici- ctive compounds is higher than the yield from the
nal properties through its bioactive compounds standard medium. Amuneke et al. (2011) stated
isolated from fruitbody that showed good activ- that the use of different growth substrates strong-
ity in inhibiting the proliferation of cervix cancer ly affect the protein, carbohydrate and vitamin
cells (HeLa) (Ekowati et al., 2017). The nutrient content of P. ostreatus.
content of fruitbody and mushroom mycelia is P. ostreatus cultivation is usually performed
influenced by the constituent material of growth using solid medium by harvesting the fruit body.
media, which is an important aspect to determine Besides using solid medium, P. ostreatus can also
the quality of mushroom fruit body. be cultured using liquid medium by harvesting its
P. ostreatus is the second most cultivated ed- mycelia. The mycelia are produced by different
ible mushroom worldwide after Agaricus bisporus. techniques with that of the fruit body. Mycelia
It has economic and ecological values and me- production technique that has been developed re-
dicinal properties (Sánchez, 2010). The top level cently is using a liquid medium with submerged
mushroom exporting country is China which culture. Good mycelia biomass growth in liquid
reach 87%, while mushroom export from Indo- medium should be supported by the availability
nesia is only 4%. Indonesia began to export ed- of nutrients on the medium and enrichment of
ible mushroom in 2003, but to date there has been the medium with the addition of organic nitrogen
no rapid progress (Marshall and Nair, 2009). Up sources, carbon sources and mineral additions
to 2008, the production of mushroom in China which include macro and micro elements. The
was still the highest in the world, reaching 70% addition of nutrients to the culture medium will
of world production, while Indonesia was still be- increase nutrient content in the mycelia, so that
low the top 10 (Vostrovský and Jablonská, 2011). the mycelia can be used as spawn to obtain good
Studies to increase mushroom production quality and highly nutritious fruitbody.
needs to keep on going as Indonesia’s mushroom Lomberh et al. (2002) reported that sub-
production considerably low. According to Misa- merged cultivation in liquid media of mushroom
chi (2017) it ranked 14th in the world. Other data mycelium is a promising method which can be
from Wakchaure (2014) China’s mushrooms pro- used in novel biotechnological processes for ob-
duction was 1,568,523 tons in 2007, while Indo- taining pharmaceutical substances of anticancer,
nesia only produced 48,247 tons in the same year. antiviral, immunomodulating, and antisclerotic
While in 2011, (Misachi, 2017), China’s mush- action from fungal biomass and cultural liquids
rooms production increased to 5,008,850 tons, and also for the production of liquid spawn. As a
on the other hand Indonesia sloped down to only result of the recent studies new data was obtained
produced 45,851 tons. Indonesia’s ability in pro- on the cultivation conditions which provide for
ducing mushroom including that of P. ostreatus is fast growth and high productivity of the investi-
still very low compared to other countries. gated strains in liquid media. The culture media
The low production of Indonesian mush- including glucose, peptone, yeast and corn steep
rooms, especially P. ostreatus is due to inadequa- extract was proposed to be the best for most of
te capability of cultivation technology, as it is the species investigated.
stated by Iriantinah (2014) that the inadequacy The addition of nutrients to the mushroom
of technological performance influencing yield growth medium as an effort to increase protein
productivity; and nutrient-poor raw materials of content in the fruitbody can be done through ad-
growth media used on mushroom cultivation, as dition of nitrogen-containing materials such as
it is stated by Curvetto et al. (2002) that in fungus corn (Zea mays). Acording to Enyisi et al. (2014)
growth and development, both quality and quan- the proximate composition of maize and maize
tity aspects (biological productivity and efficien- products are of 44.8 – 69.6% carbohydrate; 11.6
cy), are closely linked to nutrient type and growth – 20% moisture contents; 4.5 – 9.87; 2.17 – 4.43

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fat; 2.10 – 26.77 fibre; and 1.10 – 2.95% ash. Corn In phase 2 (solid medium) the treatments
starch is applied as supplement sources of carbo- were as follows :
hydrates, fats and proteins. 0% corn starch supplement on solid
Based on the above background, this re- TJ 0 :
growth medium
search was performed to: 1) obtain liquid medi-
1% corn starch supplement on solid
um composition which can support the growth TJ 1 :
growth medium
of P. ostreatus mycelia with high protein content;
and 2) obtain solid medium composition which 2% corn starch supplement on solid
TJ 2 :
can support the formation of P. ostreatus fruitbody growth medium
with high protein content. The result of this rese- 3% corn starch supplement on solid
TJ 3 :
arch may contribute ideas and practical technique growth medium
on the mushroom cultivation that emphasize on Observation held on the first harvest.
gaining high yield of protein content either from
mycelia or from the fruitbody.

METHODS

This research was held from April to Oc-

tober 2014 in the Laboratory of Mycology and
Plant Pathology Faculty of Biology and the Lab-
oratory of Organic Chemistry Faculty of Mathe- Figure 1. Incubation of P. ostreatus in liquid fer-
matics and Natural Sciences, Jenderal Soedirman mentation media (A); and solid media (B)
University, Purwokerto, Indonesia. Elaborating
P. ostreatus isolate (collection of Laboratory of In phase 1 the experiment was conducted
Mycology and Plant Pathology), wood sawdust, as follows : preparation of culture media for P.
rice bran¸ corn fluor, CaCO3, CaSO4, cotton, al- ostreatus mycelia, preparation of liquid culture
uminium foil, alcohol 70%, polypropylena plastic media, inoculation of the isolate, incubation, har-
bag (25x15x0.05 cm), baglog collar, nose masker, vesting, drying, and protein analysis (Figure 1A).
hand glove, corn seed, Potato Dextrose Agar While the experiment in phase 2 were as follows
(PDA) medium, glucose, pepton, yeast extract, : preparation of growth medium for P. ostreatus
and malt extract. fruit body, inoculation of spawn to the solid treat-
Experimental method with Completely ment media, incubation, harvesting of fruitbody,
Randomized Design (CRD) and 4 replicates was and protein analysis (Figure 1B). Media composi-
used. tion for phase 2 experiment were as follows (Tab-
In phase 1 (liquid medium) the treatments le 1).
were as follows : Data obtained were then analyzed using
anova followed by Duncan Multiple Range Test
Liquid fermentation medium 1
(DMRT) at 5% error rate.
(composition: (g/l) potato 200.0;
FC 1 :
dextrose 20.0; corn seed extract
10.0). RESULTS AND DISCUSSION
Liquid fermentation medium 2 The cultivation of P. ostreatus in three types
(composition: (g/l) malt extract of liquid fermentation medium shows that on
FC 2 :
10.0; yeast extract 2.0; CaSO4 1.0; the 28th day the fungus has entered the station-
rice bran extract 5.0; dextrose 10.0). ary phase, so that secondary metabolites can be
Liquid fermentation medium 3 harvested. As the three types of medium used
(composition: (g/l) glucose 40.0; contain different nutrients, it is possible that the
FC 3 : peptone 1.0; yeast exract 2.0; stationary phase were not be achieved at the same
KH2PO4 1.0; MgSO4.7H2O 0.2; time. This resulted in difference of weight of har-
(NH4)SO4 5.0). vested mycelia, either fresh or dry (Table 2 and
Observation was conducted at 28th day of incuba- Figure 2 A and B)
tion The highest weight of mycelia was ob-

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Tabel 1. Composition of solid medium material of P. ostreatus

Composition
Treatment Wood saw- Rice bran Gypsum CaCO3 TSP Corn
dust (kg) (kg) (kg) (kg) (kg) flour (kg)
TJ0 12.5 1.87 0.125 0.125 0.625 0
TJ1 12.5 1.87 0.125 0.125 0.625 0.125
TJ2 12.5 1.87 0.125 0.125 0.625 0.250
TJ3 12.5 1.87 0.125 0.125 0.625 0.375

Table 2. The mycelial weight and protein content of P. ostreatus mycelium cultured on three types of
liquid fermentation medium
Average of mycelial Average of mycelial dry Average of mycelial pro-
Treatment
fresh weight (g) weight (g) tein content (%)
Medium
11.71 ± 0.24 b 1.01 ± 0.30 b 14.31 ± 1.23 a
FC1
Medium
7.76 ± 1.96 a 0.67 ± 0.11 a 29.76 ± 1.88 b
FC2
Medium
6.70 ± 1.89 a 0.71 ± 0.14 a 27.06 ± 1.40 b
FC3
Note : numbers with the same alphabetic font in the same collumn are not significantly different
(DMRT 5%)
tained from the medium of FC1 which was 11.71 ents and less suitable condition of the medium
g, the weight was obtained from the liquid medi- and results in low protein content in the mycelia.
um with the initial volume of 100 ml and the final Meanwhile FC2 and FC3 media contained me-
volume of 80 ml. If the medium is converted to 1 asurable minerals, which have very important
L, it is estimated that the weight of mycelia will function in metabolisms, especially as cofactors
be 110,71 g / L. Medium FC1 is a natural medi- so that the formation of primary and secondary
um because it uses potato extract and corn seeds metabolites were optimal. Among the other two
extract and yet enriched with dextrose, in this me- treatments, protein content in FC2 was the high-
dium the sources of carbon, nitrogen and mineral est. This could be due to the medium cointains
are highly available. Medium FC2 contains car- rice bran. It is conformed by Donini et al. (2009)
bon, nitrogen, macronutrient and micronutrient that among the most used cultivation supple-
sources, while FC3 medium contains high car- ments, cereal brans are sources of organic nitro-
bon source, little nitrogen source, and incomplete gen (N2), necessary to the growth of the myce-
mineral content. This experiment showed that lium mass, which may interfere in productiveness
cultivation in natural medium spurred the growth and biological efficiency of the fungus.
of mycelium. Protein content analysis was also per-
Protein content of mycelia biomass ranged formed on the fruit body cultured on four types
14.31-29.76% (Table 2.). According to Chang of solid medium with different amount of corn
and Miles (2004) the protein content of P.ostreatus flour supplement. It was observed that protein
range is 10.5 - 30.4%. This revealed that the pro- content in the fruitbody of P. ostreatus was not
tein content in mycelial biomass of P. ostreatus straightly proportional to the weight of the fruit-
cultivated in the three types of media were good. body obtained either. The percentage of protein
However, mycelial biomass weight on FC1 content in the fruitbody ranged from 18.85 to
medium was not in line with its protein content 24.69%, this showed that fruitbody protein was
as the capacity of mycelial growth in this medium slightly lower when compared with the protein
was not proportional to its mycelial protein. Pro- content in mycelial biomass.
tein content of mycelium biomass was obtained Furthermore, the analysis of protein con-
high in FC2 and FC3 media. The data showed tent was held to determine whether the high
disadvantageous of using natural medium (FC1) weight of fruitbody was also followed by high
of which the less measurable chemical composi- protein content. The analysis of fruitbody weight
tion of the medium affected the balance of nutri- and its protein content is presented in Table 3.

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Table 3. Fresh weight and protein content of P. ostreatus fruitbody grown on solid medium.
Average of fresh weight of Average of protein content
Treatment
P. ostreatus fruitbody (g) of P. ostreatus fruitbody (%)
0% corn starch supplement
67.33 ± 9.07 a 24.69 ± 0.72 c
on solid growth medium
1% corn starch supplement
85.33 ± 11.93 ab 18.85 ± 0.73 a
on solid growth medium
2% corn starch supplement
90.33 ± 15.30 b 20.82 ± 0.18 ab
on solid growth medium
3% corn starch supplement
94.00 ± 8.88 b 23.44 ± 3.13 bc
on solid growth medium
Note : numbers with the same alphabetic font in the same collumn are not significantly different
(DMRT 5%)
P. ostreatus cultured on four different solid in the fruitbody.
medium showed that the supplementation of Protein obtained from this experiment was
corn flour was able to increase fruitbody produc- a little bit lower than the protein content from
tion, the more corn flour given, the higher the fresh fruitbody (30,20%) of P. florida grown on
fruitbody yielded. Fruitbody weight ranged from solid-mixed medium of cardboard and banana
67.33 - 94.00 g, this weight was obtained from leaves, as it was reported by Tirkey (2017). The
the first harvest only (Figure 2B). This result was discrepancy maybe due to, as stated by Singh et
in consonant with the report of Jeznabadi et al. al. (2008), several parameters that affect the en-
(2016) that the quality of P. eryngii was signifi- zyme production; however, its nitrogen source is
cantly affected by substrate ingredients. The type a major factor.
of substrate as well as the type and quantity of The protein content in the fruitbody was
supplement appeared to have a substantial ef- not much different to the results of research re-
fect on prolonging the delayed-release nutrients. ported by Patil et al. (2010) and Bonginkhosi et al.
Carrasco-Gonz´alez et al. (2017) reviewed that (2012) who states that the protein content of P.
the protein quality of P. ostreatus is one of its ma- ostreatus range from 20.33 to 24.66%. The amino
jor strengths because it has high contents of all acid content analysis showed that the mushroom
essential amino acids and excellent protein di- contains high value of 19 kinds of amino acid
gestibility. both essential and non-essential, especially glu-
tamic acid i.e. 64.02 mg/100 g fruit body. This
high glutamic acid content that causes delicious
taste in mushrooms.
Muhsrooms such as Agaricus bisporus, Pleu-
rotus florida, Russula delica and Lyophyllum decastes
are protein and fiber rich with low fat content.
The ash content and carbohydrate content was
Figure 2. A twenty eighth-day mycelial incuba- less than other food from plant and animal ori-
tion of P. ostreatus in liquid medium (A); harvest- gin (Teklit, 2015). This research revealed that the
ed fresh mycelia of P. ostreatus (B); and ready- increase of fruitbody weight from the medium
to-harvest P. ostreatus fruitbody from solid media with the addition of corn flour indicated a pro-
(C). spective in cultivation process, as the increase in
mushroom production is a common problem that
The weight of the fruitbody was not direct- is still difficult to overcome until now, the results
ly proportional to the content of protein in the of this study also provide great benefits for the
fruitbody (Table 3.). The highest protein content development of mushroom cultivation industry,
was obtained from the addition of 0% corn flour however effort to increase protein content of
or without corn fluor and from 3% corn fluor the fruit body needs further research, such as by
treatment. The results showed that the addition modifying nutrient sources in the medium due to
of corn flour spur growth, the existing nutrients the results of this study were not yet consistent.
were used for the growth of mycelium until the As a nutriceutical foods, protein content in mush-
formation of fruitbody, however, the nutrients rooms is an ultimate parameter.
were not enough to increase the protein content Proteins of Pleurotus sp. mushroom have

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superior quality because some of the members of mushroom from agro base waste products.
this genus contains complete proteins with the Journal of Microbiology and Biotechnology Re-
well distribution of essential amino acids, as well search, 1(3), 1-14.
as non-essential amino acids particularly gamma- Bonginkhosi, E. D., Diana, M. E., & Masarirambi,
M. T. (2012). Growth and Yield Response of
aminobutyric acid (GABA), that act as neurot-
Oyster Mushroom (Pleurotus ostreatus) Grown
ransmitter and ornithine which is a precursor on Different Locally Available Substrates. Cur-
in the synthesis of arginine (Deepalakshmi and rent Research Journal of Biological Sciences, 4(5),
Mirunalini, 2014). In term of obtaining protein 623-629.
from P. ostreatus, cultivating the mushroom in Carrasco-Gonz´alez, J. A., Serna-Sald´ıvar’, S. O.,
mycelial phase in liquid media is a prospective & Guti´errez-Uribe, J. A. (2017). Nutritional
effort to elaborate as it may take shorter incuba- composition and nutraceutical properties of
tion time, may be produced in mass production the Pleurotus fruiting bodies: Potencial use as
with less space consuming, and higher protein food ingredient. Journal of Food Composition and
Analysis, 58, 69-81.
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Chang, S. T., & Miles, P. G. (2004). Edible Mushrooms
lial biomass can be preserved in certain forms and Their Cultivation. CRC Press, Inc, Florida:
such as powder for further utilization as it was Boca Raton.
reported by Tupamahu and Budiarso (2017) that Curvetto, N. R., Figlas, D., Devalis, R., Delmastro, S.
the addition of 1% P. ostreatus mushroom powder (2002). Growth and productivity of different
resulting in increased yoghurt quality, and the Pleurotus ostreatus strains on sunflower seed
preferred yoghurt product by most of the pane- hulls supplemented with N-NH4+ and/or Mn
lists. It was then proven that the addition of the (II). Bioresource Technology, 84(2), 171-176.
mushroom powder increase yoghurt quality and Deepalakshmi, K & Mirunalini, S. (2014). Pleurotus
ostreatus: an oyster mushroom with nutritional
public acceptance.
and medicinal properties. Journal of Biochemical
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CONCLUSION Donini, L. P., Bernardi, E., Minotto, E., & Nascimen-
to, J. S. (2009). Growing Shimeji on elephant
Different composition of liquid and solid grass substrate supplemented with different
medium influenced growth and protein content types of sharps. Scientia Agraria, 1, 67-74
of P. ostreatus. The optimal liquid medium for Ekowati N., Kasiamdari, R. S., Pusposendjojo, N., &
P.ostreatus mycelial growth (11.71g) was Liquid Soegihardjo, C. J. (2011). Daya Antimikroba
Metabolit Bioaktif Jamur Shiitake (Lentinula
Fermentation Medium 1, while the highest pro-
edodes (Berk.) Pegler) yang Dikultur pada Tiga
tein content (29.76%) was observed in Liquid Jenis Medium Fermentasi. Majalah Obat Tradis-
Fermentation Medium 2. The optimal solid me- ional, 16(3), 132-137.
dium for P. ostreatus fruitbody production was the Ekowati, N., Mumpuni, A., & Muljowati, J. S. (2017).
medium with 3% corn starch supplement, while Effectiveness of Pleurotus ostreatus Extract
the highest protein content was obtained from Through Cytotoxic Test and Apoptosis Mecha-
the medium without corn starch supplement nism of Cervical Carcer Cells. Biosaintifika:
(24.69%). Journal of Biology & Biology Education, 9(1), 148-
155.
Enyisi, I. S., Umoh, V. J., Whong, C. M. Z., Abdullahi,
ACKNOWLEDGEMENT I. O., & Alabi, O. (2014). Chemical and nutri-
tional value of maize and maize products ob-
Thanks to Kemendikbud Dikti who has tained from selected markets in Kaduna State,
provided research fund of BOPTN Higher Edu- Nigeria. African Journal of Food Science and Tech-
cation Research of Fiscal Year 2014 (Year II) nology, 5(4), 100-104.
through LPPM Unsoed Contract Number DIPA Hoa, H. T., Wang, C. L., & Wang, C. H., (2015). The
023.04.2.189899/2014. Acknowledgments also Effects of Different Substrates on the Growth,
to the Head of LPPM Unsoed and staff who have Yield, and Nutritional Composition of Two
Oyster Mushrooms (Pleurotus ostreatus and Pleu-
provided good facilities, information and coop-
rotus cystidiosus). Mycobiology, 43(4), 423–434.
eration, and the Dean of the Faculty of Biology Iriantinah, C. (2014). Strategi Pengembangan Komodi-
Unsoed who has given permission and support to tas Jamur Tiram Putih (Pleurotus florida) Di Ka-
the implementation of this research. bupaten Nganjuk. Jurnal Manajemen Agribisnis,
14(2), 161-172.
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