Aug 2014 – Oct. 2014, Vol. 4, No. 4; 3413-3420.

E- ISSN: 2249 –1929

Journal of Chemical, Biological and Physical Sciences

An International Peer Review E-3 Journal of Sciences
Available online atwww.jcbsc.org

Section B: Biological Sciences

CODEN (USA): JCBPAT Review Article

Effect of Nutrient Sources and Environmental Factors

on the Biomass Production of Oyster Mushroom
(Pleurotus Ostreatus)
Mohamed F.M.Salem1; Khaled F.M.Salem2; Emily T.Hanna3; and Noura E.Nouh1
1
Environmental Biotechnology, GEBRI, Sadat University, Sadat city, PoBox79, Egypt
2
Plant Biotechnology, GEBRI, Sadat University, Sadat city, PoBox79, Egypt
3
National Nutrition Institute, Cairo University, GEBRI, Giza City, Egypt

Received: 23 September 2014; Revised: 01 October 2014; Accepted: 10 October 2014

Abstract: Studies were carried out on the growth condition for Pleurotus ostreatus
(oyster mushroom), an edible mushroom and the effect of nutrient Sources and
Environmental factors on the biomass production. This organism was able to grow
optimally at a temperature of 25 0C and pH of 6.5. It utilized various carbon sources such
as glucose, fructose, maltose, Galactose, sucrose and lactose, with glucose the most
utilized and this significantly enhanced mycelial growth closely followed by fructose,
sucrose and Galactose. The least utilized carbon sources were lactose. The organism
tested at different glucose concentration (20, 30, 40, 50 and 60 g/L),the maximum
glucose concentration for mycelial biomass production was achieved at 30 g/L. Oyster
mushroom (Pleurotus ostreatus) was cultivated on different carbohydrate substrates
(corn, rice and rye). Corn flour gave the highest mycelia yield while rye gave the lowest.
Keywords: Pleurotus ostreatus, mycelia growth, substrates, Growth requirements,
Biomass production.

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Evaluation… Salem et al.

INTRODUCTION

Oyster mushroom (Pleurotus ostreatus) is economically highly praised for its nutritional and
pharmacological value. Its fruit bodies are rich in carbohydrate, dietary fiber, proteins, essential amino
acids, vitamins and minerals1. Biologically active substances, especially polysaccharides and lectins,
isolated from Pleurotus ostreatus have hypocholesterolemic, antitumor, immunomodulating, antiviral and
antibacterial effects1, 2. Their mycelia also contain amino acids like glycine, valine, threonine, serine,
leucine, proline, methionine, asparagine, glutamine, lysine, arginine, histidine, cysteine and alanine3.
Mushroom cultivation serves as the most efficient and economically viable biotechnology for the
conversion of lignocellulose waste materials into high quality protein food and this will naturally open up
new job opportunities especially in rural areas4, 5.
Thus, the objectives of this study include: To determine the optimal growth condition: temperature, initial
pH, different carbon source and carbon source concentration which necessary for mycelial growth and
maximum biomass production of Pleurotus ostreatus, and to evaluate the suitability of carbohydrate
source for the cultivation of Pleurotus ostreatus.

MATERIALS AND METHODS

Sources of oyster mushroom: The cultivated mushroom was collected from sawdust from by serial
dilution method. The sample was aseptically collected from sawdust dumpsite at Somosta village, Assiut
Western Desert Road and a hand trowel was used to collect about 10 g each of the sample into a plastic
bag.
Cultivation of mushroom: Serial dilution method was used; about 0.1ml from 10-5 dilution was spread
on potato dextrose agar (PDA) plates using a glass spreader, sterilized by dipping in 95% ethanol and
flaming. The plates were incubated at room temperature for 4 days6.
Effects of different carbon sources on mycelia growth: The basal medium consisted of (g/L); yeast
extract 3g, peptone 2g, KH2PO4 0.6g, MgSO4.7H2O 0.4 g, vitamin B1 0.01 g ,0.50 g ampicillin7 . medium
supplemented with carbon sources; maltose, fructose, glucose, lactose, sucrose and galactose 3% (w/v)
The pH of the basal medium was adjusted to pH 6.5 with 1M NaOH. These were then sterilized 1.5 lbs
pressure for 15 minutes at 121oC and growth of the organism was assessed using the dry weight method.
Effects of different glucose source concentration on mycelia growth: For Pleurotus ostreatus, glucose
proved to be the best carbon source. Different concentrations of glucose 2-6 g were dissolved in 100ml of
the basal medium at pH of 6.5.
Effect of Temperature of substrate medium on mycelia growth:100ml of basal medium consisted of
(g/L); glucose 30g, yeast extract 3g, peptone 2g, KH 2PO4 0.6g, MgSO4 · 7H2O 0.4g, vitamin B1 0.01 g
,0.50 g ampicillin. The initial pH of the medium was adjusted to 6.5 was prepared in different conical
flasks of 250ml volume7. The flasks were autoclave for sterilization, then incubated under static condition
at different temperatures (20 oC, 25 oC, 30 oC, 35 oC, 40 oC) for 4 days. Flasks were inoculated with
Pleurotus isolates culture and incubated for 10 days. The organism growth was assessed using the dry
weight method.

3414 J. Chem. Bio. Phy. Sci. Sec. B, Aug. 2014 – Oct. 2014; Vol.4, No.4; 3414-3420
Evaluation… Salem et al.

Effect of initial pH of substrate medium on mycelia growth: The ingredients of the basal medium
were weighed and dissolved in distilled water.100 ml of medium were distributed in each conical flask.
The pH of the each medium was adjusted as 3, 4, 5, 5.5, 6, 6.5, 7, 7.5, and 8. Then the conical flasks with
different pH level as mentioned above were autoclaved for sterilization, then incubated under static
condition. The flasks were then incubated at 25 °C for 10 days. The organism growth was assessed using
the dry weight method.
Effects of different carbohydrates sources: The effects of different carbohydrate sources were
investigated by water extracts method. The different sources used were corn flour, rice powder and rye at
0.1 mg/mL concentrations each in basel medium After 10 days of incubation the different dry weights
were obtained and used to assay against each carbohydrate source. Medium without carbohydrates
sources serve as control.

RESULTS

Effect of carbon compounds on the growth of Pleurotus isolates: The results of the study carried out
on carbon source type of Pleurotus ostreatus showed that dextrose was the best carbon source. This was
closely followed by fructose, sucrose and galactose. The utilization of dextrose by tropical edible
mushroom has not been reported while glucose utilization has been reported 8-11. Dextrose, which is an
isomer of glucose, can be transformed to glucose during metabolism12.

Figure 1: Effect of carbon sources on the mycelial biomass production in Pleuotus ostreatus.

Effect of glucose concentration on the growth of Pleurotus isolates: The study suggested that the rate
of production of oyster mushroom from its mycelia stage was significantly influenced by the glucose
concentration .The effect of different carbon sources concentration on the mycelia growth is shown in
(Figure 2). The highest mycelia growth was achieved at 30 g/L glucose in the basal medium.

3415 J. Chem. Bio. Phy. Sci. Sec. B, Aug. 2014 – Oct. 2014; Vol.4, No.4; 3414-3420.
Evaluation… Salem et al.

Figure 2: Effect of glucose concentration on the mycelial biomass production in Pleuotus ostreatus.

Effect of Temperature of substrate medium on mycelia growth: After inoculation of Pleurotus

ostreatus in conical flasks and incubated at different temperatures (20- 40 oC) for 10 days, maximum
growth was observed at temperatures of 250C for Pleurotus ostreatus whereas minimal growth was
observed till temperatures 35 oC and while the heat tolerance reached its Maximum at 40 0C, no growth
above 40 oC. Intensity decreased by decreasing of temperature and stopped at 10 oC. The effect of
different Temperature on the mycelia growth is shown in (Figure 3).

Figure 3: Effect of Temperatures on the mycelial biomass production in Pleuotus ostreatus

Effect of initial pH of substrate medium on mycelia growth: Observations on basal medium with
different pH (3- 8) suggest that there was no growth at pH 3. The maximum growth was observed at pH
6.5 and the moderate growth was observed at pH 7.0 and 7.5 whereas minimal growth at pH 8. These

3416 J. Chem. Bio. Phy. Sci. Sec. B, Aug. 2014 – Oct. 2014; Vol.4, No.4; 3414-3420
Evaluation… Salem et al.

results indicate that the optimum pH for growth of Pleuotus ostreatus at these conditions was 6.5 and
moderate growth in between 7 to 7.5.

Figure 4: Effect of initial pH on the mycelial biomass production in Pleuotus ostreatus.

Effects of different carbohydrates sources: The effect of different carbohydrate sources on the mycelia
growth is shown in (Figure 5). The highest mycelia growth (1.36425 g/100 mL) was achieved using corn
flour while rye gave the least mycelia growth (1.111g/100 mL).

Figure 5: Growth yield of Pleuotus ostreatus on different carbohydrates sources.

DISSCUTION

The results of the study carried out on carbon source type of Pleurotus ostreatus showed that dextrose was
the best carbon source. This was closely followed by fructose, sucrose and galactose. The utilization of
dextrose by tropical edible mushroom has not been reported while glucose utilization has been

3417 J. Chem. Bio. Phy. Sci. Sec. B, Aug. 2014 – Oct. 2014; Vol.4, No.4; 3414-3420.
Evaluation… Salem et al.

reported8, 10-12. Dextrose, which is an isomer of glucose, can be transformed to glucose during
metabolism6.
The results of these present findings are contradictory to the observation of Griftin13 who suggested that
mannitol and fructose are the most commonly utilizable sugars after glucose. The utilizable carbon
sources used during this study have significant effect on the growth of the fungi (Figure 1). The
preference of glucose over other carbon source by Pleurotus ostreatus may be due to the ease with which
it was metabolized to produce cellular energy for the growth of the organism. Kurtzman and Zadrazil14
mentioned that the ability of Pleurotus spp to use different carbon sources may be an expression of the
physiological differences in the species or of the isolates, since other isolates of the same species might
give different results. The highest mycelia growth was achieved at 30 g/l glucose concentration
(Figure 2).
The mycelium of Pleurotus ostreatus grew optimally at a temperature of 250C and pH of 6.5 (Figure 3
and 4). Thus the ability of the mycelia to growth at this temperature and pH range enabled them to
flourish in agro wastes in the tropics15. Mycelial development forms the vegetative growth phase of
mushroom growing and temperature is highly important since it affects the growth and adaptability as
well as the quantity of the quality of fruiting-bodies produced 12.
The temperature obtained was in agreement with that reported by Ayman et al for Pleurotus ostreatus,16
which also reported by Sridevi et al.17 in the studies on growth parameters of Pleurotus ostreatus and
Pleurotus florida . The temperature obtained was not in agreement with that reported by Nwokoye et al.18
for Pleurotus ostreatus, the organism was able to grow optimally at a temperature of 28oC, thus inferring
that the strain used for this study was different from that mentioned above. Chang and Miles 19 mentioned
that there were three strains of Pleurotus ostreatus, the high temperature, medium and low temperature
strains in the range of 25 oC -30 oC, 16 oC -22 oC and 12 oC -15 oC, respectively, inferring that the strain
used for this study belonged to the high temperature strain. Optimum pH for growth of Pleurotus
ostreatus at these conditions was 6.5 in agreement with Ayman et al.16 and Ibekwe et al.20 while Fasidi,21
reported an optimum pH of 6.0 for Volvariella Esculenta. Kuforiji and Fasidi,22 found that 35oC and 7
were optimal temperature and pH values for mycelial growth of V. volvacea. Tseng,23 reported that
optimum pH of V. volvacea varies from strain to strain as strain CT13 had an optimum pH of 5 and
another strain, a pH of 6.6. This further confirms that the strain used for this study is different from that of
Leeward et al.24.
The study suggested that the production of oyster mushroom from its mycelia stage was significantly
influenced by the carbohydrate source, the results gave corn flour as the best carbohydrates sources for
the mycelia growth and biomass production of Pleurotus ostreatus (Figure 5).This may be due to the
saprophytic ability of this fungus to produce extracellulase hydrolyzing enzymes such as cellulose,
lignase and laccase to hydrolyse cellulose, hemicelluloses and lignin present in these substrates to simple
sugars for growth25.

CONCLUSION

Successful utilization of agro-wastes for both mycelial and fruitbody formation of mushroom supply
nutrients needed by this fungus to convert them to protein-rich palatable foods. Thus it can be concluded
that maximal mycelial growth of Pleurotus ostreatus can be achieved by cultivating the fungus at
temperature of 25oC and pH of 6.5. The mycelial mass can also be enriched by growing in a medium

3418 J. Chem. Bio. Phy. Sci. Sec. B, Aug. 2014 – Oct. 2014; Vol.4, No.4; 3414-3420
Evaluation… Salem et al.

containing 3% glucose as carbon sources in this research dextrose (isomer of glucose) was the best carbon
source witch closely followed by fructose, sucrose and galactose.
Utilization of agro wastes helps in reducing the mountain range of wastes, converting them into
mushroom protein and vitamins. This represents one of the world’s untapped resources of tasteful food in
the future and because mushroom cultivation needs only a small space, land can be conserved. Thus the
use of wastes can provide more food, more jobs, better family income, and improved living standard, curb
global warming and clear up the crop residues on road sides and forest margins 26.

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* Corresponding author: Noura E.Nouh

Department of Environmental Biotechnology, GEBRI, Sadat University,
P.O. Box 79, Egypt
Email: noraeissa2020@gmail.com

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