Study On Ethanol Production From Sugar Cane Molasses by Using Irradiated Saccharomyces Cervisiae

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Third International Conference on

Radiation Sciences and Applications


12 – 16 November 2012/ Hurghada, Egypt

Study on Ethanol Production from Sugar Cane Molasses


by Using Irradiated Saccharomyces cervisiae
Botros, H.W.; Ahmed, A.S.; Farag, S.S. and Hassan, I.A.
Atomic Energy Authority, Nuclear Research Center, Plant Research Department, Cairo,
Egypt.

ABSTRACT
In commercial ethanol production procedures often use sugar cane
molasses as a raw material due to their abundance and low costs. The most
employed microorganisms used for fermentation is Saccharomyces cerevisiae
yeasts due to its ability to hydrolyze sucrose from sugar cane molasses into
glucose and fructose, two easily assimilable hexoses.
The aim of this study was to evaluate the effect of gamma irradiation on
the activity of S. cerevisiae in the ethanol production yeast cells exposed to
different doses of gamma rays (0.05, 0.10, 0.2, 0.4, 0.6, 0.8 and 1.0 kGy. The
sugar cane substrate was optimized after maintaining deferent levels of sugar
concentrations (12-21%), medium pH (4.0-5.5), incubation temperature (25-
40oC) and rate of fermentation (24-168) h.
Data showed that rate of ethanol production was maximum by using the
irradiated S. cerevisiae cells at 0.1 kGy. dose at fermentation conditions as 15%
sugar concentration, initial pH 4.5, incubation temperature 30oC, fermentation
time 96 h at a fermentation medium volume 250 ml found in 500ml erlenmyer
flasks.
Keywords, Saccharomyces cerevisiae, Irradiation, Sugar cane molasses, ethanol.

INTRODUCTION
Ethanol is one of the most advanced liquid fuels because it is
environmental friendly. It is a clear, colorless liquid with a characteristic,
agreeable odor. In dilute aqueous solution, it has a sweet flavor, but in more
concentrated solutions it has a burning taste (Patil 1991). It is an alcohol, a
group of chemical compounds whose molecules contain an OH group, bonded
to a carbon atom. It melts at -114.1°C, boils at 78.5°C and has a density of
0.789 g/ml at 20°C (Kaur and Kocher 2002). Ethanol is produced by
fermentation: when certain species of yeast (notably Saccharomyces cerevisiae)
154 Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012)

metabolize sugar in the absence of oxygen, they produce ethanol and carbon
dioxide. Ethanol is particularly useful in industrial applications because of its
relatively high affinity for both water and organic compounds. The composition
of other alcohols limits their flexibility as compared to ethanol (Anxo et al.,
2008).
The molasses is obtained from different sources such as cane, beet and
citrus etc. It is a syrupy material left after the removal of sugar from the mother
syrup. The viscous material is composed of sucrose, glucose and fructose at
total carbohydrate concentration of 45-60% (w/v). The molasses is of three
types, the black strap, refinery and invert or high test molasses. Cane molasses
has less sucrose and more invert sugar, and lower content of nitrogen and
raffinose, more intense color and more buffer capacity (Wang et al., 1985;
Borzani et al., 1993; Borzani, 2001).
The production of raw cane sugar from sugarcane juice results in the
formation of molasses, a byproduct that contains minerals regarded as
impurities in raw sugar (Hugot and Jenkins 1986). However, this purification
process results in the loss of some high value disaccharides and
monosaccharides from the final raw sugar product that end up in the molasses.
The fermentable sugar content of molasses varies inversely with the purity of
the raw sugar produced at the factory. Molasses is a low value product that is
used as a cattle feed supplement, in specialized yeast propagation or as a
flavoring agent in some foods (Troiani, 2009a). Although the fermentable
sugars in the molasses cannot be further upgraded to raw sugar, they can be
converted to ethanol in a distillery. Hence, integrated sugarcane factories that
have sugar manufacturing co-located with an ethanol distillery can use molasses
as a feedstock for ethanol in addition to raw cane juice directly from the mill. A
significant number of sugarcane factories in Brazil and several hundreds of
others around the world are of this type (Szwarc, 2009).
Molasses is used in the baker’s yeast production, in the fermentation
technology for ethanol, citric, lactic and gluconic acids production, as well as
glycerol, butanol and acetone production, as an ingredient of mixed feeds or in
the production of amino acids (Belitz et al., 2009; Satyanarayana and Kunze,
2009). The fermentative yeast Saccharomyces cerevisiae is largely used in
ethanol production using such renewable biomass as sugar cane or sugar beet
molasses as the main carbon source (Echegaray et al., 2000; Sanchez and
Cardona, 2008). These types of S. cerevisiae were selected as production
Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012) 155

microorganisms on account of their commercial availability and an extensive


application in food industry.
MATERIALS AND METHODS
Materials:
Molasses were obtained from Egyptian sugar cane and integrated
industries company El-Hawamdia, Giza, Egypt and were used as a fermentation
medium for ethanol production. Table (1) shows the compositions of sugar cane
molasses used as a substrate fermentation.
Saccharomyces cerevisiae strain was obtained from the available stock
culture of Microbiology Department, Faculty of Science, Al-Azhar University.
Table (1): Composition of the sugar cane molasses used for experiment.
Molasses Water Soluble solids Total Total Mineral pH
type content, content, sugar, nitrogen substances
% % % content, % content,
%
Sugar cane 80.7 19.3 49.2 0.6 6.1 7.4
molasses

Methods:
Organism and culture maintenance:
Saccharomyces cervisiae maintained on yeast extract peptone glucose
(YPG) agar medium (pH 4.5), containing yeast extract (3.0 g/l), peptone (5.0
g/l), glucose (10.0 g/l), agar (20.0 g/l), The slants were incubated at 30°C for 1-
2 days for maximum growth.
Preparation of yeast cell suspension:
Sterilized distilled water (10 ml) was added to a 24-36 h old slant
culture of S. cerevisiae. The cells were scratched with a sterilized inoculating
needle and the tubes were shaken gently to form a homogeneous suspension.
The cell count was made using a Haemocytometer.
Development of inoculum:
Fifty milliliter of YPG medium was transferred to the individual 250ml
Erlenmeyer flasks. The flasks were cotton plugged, autoclaved and allowed to
cool at room temperature. One milliliter of cell suspension (2.74×106 CFU) was
added to each flask aseptically. The flasks were incubated in a rotary shaker
(160 rpm) at 30°C for 24h. Pre-grown culture of S. cerevisiae was centrifuged at
6000rpm for 15 min and yeast cells were separated out. The supernatant was
156 Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012)

discarded and the pellet was washed with saline water. It was re-centrifuged for
another 5 min to obtain the final pellet that was washed and then air-dried and
weighed. These were the free cells.
Pretreatment of molasses:
Pretreatment of molasses to reduce harmful compounds to the used
microorganisms (e.g. coloring substances, amino acids and heavy metals) that
may as inhibitors during fermentation. Sulfuric acid and activated carbon are
considered the best method for this purpose according to (Lazaridou et al.,
2002) by using molasses solution 20% total sugars were adjusted to pH 3 with
0.5M H2SO4. The solution was allowed to stand for 24 h and then centrifuged at
5000 g for 15min. The supernatant was also treated with 3% activated carbon by
stirred the mixture for 1h, then filtered through a Whatman no. 1 filter paper.
The pretreatment was repeated three the four times until the solution was almost
colorless and gave an absorbance reading at 490 nm.
Fermentation procedure and critical phases:
Three hundred milliliters of treated cane molasses with 15% (w/v) sugar
(initial pH 4.5) were taken into individual 500ml Erlenmeyer flasks. The flasks
were cotton plugged and steamed at 90°C in a water bath for 15-20 min. After
cooling to an ambient temperature, 2 g yeast cells were added to one flask and
placed in an incubator at 30°C for 120h. After the required incubation period,
the cells were separated out and stored for use of more experiments. The
fermented medium was used for estimation of ethanol and residual sugar
contents.
Assay methods:
The estimation of total reducing sugar was based on the dinitrosalicylic
acid (DNS) method. A double beam UV/VIS-scanning spectrophotometer was
used for measuring absorbance. Sugar contents in the supernatant were
determined by taking 1.0 ml of supernatant along with 2.0 ml of dinitrosalicylic
acid (DNS) reagent in a test tube. Blank containing 1.0 ml distilled water and
2.0 ml of DNS was run parallel. The tubes were heated in a boiling water bath
for 15 min. After cooling the tubes at room temperature, added 8 ml of distilled
water in each and absorbance was noted at 546nm using spectrophotometer.
Sugar concentration was determined from the standard curve of glucose.
Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012) 157

Ethanol estimation
Dichromate method:
Ethanol was also determined with good precision by oxidation with acid
dichromate solution (Kiransree et al., 2000). The ethanol in the known masses
of the solution was oxidized to acetic acid using a known mass of standard
potassium dichromate (0.1N) in the presence of sulfuric acid.
Irradiation of organism:
Slants of S. cerevisiae cells were exposed for doses of 0.05, 0.1, 0.2,
0.4, 0.6, 0.8 and 1.0 kGy of gamma rays using cobalt-60 gamma radiation cell at
the Nuclear Research Center, Inshas, Egypt. The dose rate at the time of
radiation treatment was 3.1 kGy/hr.
RESULTS AND DISCUSSION
Effect of gamma irradiation on growth of Saccharomyces cerevisiae cells:

The irradiated and non-irradiated S. serevisiae cells maintained on yeast


extract peptone glucose (YPG) agar medium (pH 4.5) in sterilized slants and
incubated at 30oC for 1-2 days for maximum growth then sterilized distilled
water was add to the 1-2 days old slants culture of S. serevisiae. The cells were
scratched with a sterilized inoculating needle and the tubes were shaken gently
to form a homogeneous suspension. The cell count was made using
haemocytometer. Spore suspension of yeast cells were prepared at initial
concentration 1.75x106/ml, then 10 milliter of the spore suspension was added
to each 500 ml Erlenmeyer flask containing 250 ml of liquid (YPG) medium.
Irradiated and non-irradiated samples of yeast cells incubated at 30oC
for 72 hours. Then the biomass concentration /1ml of the tested samples of yeast
were counted by haemocytometer and the dry weight of cells in 250 ml media
were determined.
Data in Table (2) showed that there was an increase the biomass
concentration /ml in the non-irradiated yeast samples by increasing the
incubation time, whereas the initial concentration of the yeast cells
(1.75x106)/1ml increased gradually to 3.25x106/ ml at 24 h incubation time,
4.65x106/1ml at 48h and 6.25x106/ml at 72h incubation time.
158 Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012)

Table (2): Effect of gamma irradiation on growth of S. serevisiae cells.

Incubation time (h)


Dose 24 48 72
kGy Biomass cell Biomass Biomass Biomass Biomass Biomass
count /1 ml dry weight cell count dry weight cell count dry weight
g/250 ml /1 ml g/250 ml /1 ml g/250 ml
0.00 3.25x106 0.85 4.65 x106 1.75 6.25 x106 2.15
0.05 3.60x106 1.15 5.95 x106 1.88 7.38 x106 2.85
0.10 3.95x106 1.82 6.78 x106 2.64 9.39 x106 3.12
0.20 1.75x106 0.92 2.48 x106 1.92 3.52 x106 2.01
0.40 1.25x106 1.68 1.65 x106 1.10 2.24 x106 1.23
0.60 0.83x106 0.42 0.89 x106 0.51 1.11 x106 0.60
0.80 0.32x106 0.31 0.35 x106 0.33 0.39 x106 0.39
1.00 0.02x106 0.04 0.04 x106 0.06 0.06 x106 0.09
Sugar conc. 15%, incubation temperature 30oC, initial pH 4.5 and incubation time 24-72h.

Irradiation of the initial yeast suspension caused increasing in


the biomass count of yeast cells after 24 hours incubation. The biomass
count/1ml was 3.6x106 in 0.05 kGy irradiated samples and 3.95x106/1ml in 0.1
kGy irradiated samples. By increasing the irradiation dose (0.20 – 1.0 kGy),
there was a gradual decrease in the biomass cell count of the yeast, whereas it
was from (1.75x106 to 0.02x106)/1ml in 1.0 kGy irradiated yeast samples.
By increasing the incubation time up to 72 hours there was an
increase in the biomass cell count in all the treated sample and the maximum
biomass cell count was noticed in (0.10 kGy) irradiated yeast samples after 72
hours of incubation. Also irradiation affected on the biomass dry weight of the
yeast cells samples, where it was noted that the dry weight increased up to (0.10
kGy) where it reached 1.88 g/250 ml medium and then decreased gradually by
increasing the irradiation dose where it reached 0.04 g/250 ml in (1.0 kGy)
irradiated yeast samples. The maximum biomass dry weight of yeast cells was
in 0.1 kGy irradiated samples incubated up to 72 hours where it reached to 3.12
g/250 ml of medium. Reports have been published on the production of ethanol
under stationary conditions (Tyagi and Ghose 1982; Roukas, 1996). In the
present study, cane molasses was used as the basal fermentation medium.
Cachot and Marie-Nolelle (1991) treated cane molasses with sulphuric acid and
got encouraging results. Other workers have also reported cane molasses as the
best raw material for enhanced and consistent yields of ethanol (Hamdy et al.,
1992; Kiss, et al., 1999).
Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012) 159

Rate of ethanol production by irradiated S. cerevisiae exposed for 0.1 kGy


dose:
Rate of ethanol production by the irradiated S. cerevisiae cells was
investigated as shown in Table (3). The rate was studied from 24-168 hours
after inoculation. The sugar consumed by the non-irradiated and irradiated yeast
cells after 24 hours was 4.3% and 4.7% and ethanol yield was 0.8% and 1.1%.
maximum ethanol yield was 6.8% by the irradiated cells was obtained after 96
hours of incubation with a maximum sugar consumption 16.1%, followed by
ethanol yield 6.4% by the non-irradiated yeast cells obtained after 120 hours of
incubation with sugar consumption 15.8%. So irradiation caused increase in the
activity of the yeast cells and increase in both the ethanol yield and sugar
consumption. These results are in harmony with results reported by Tyagi and
Ghose 1982; Roukas, 1996; Kiss et al., 1999; Fregonesi et al., 2007).
Table (3): Rate of ethanol production and sugar consumption by irradiated S.
serevisiae cells for 0.1 kGy from cane molasses.
Fermentation Non-irradiated cells Irradiated cells
rate (h)
Ethanol Sugar Ethanol Sugar
production consumption production consumption
% %
% %
24 0.8 4.3 1.1 4.7
48 2.5 7.2 3.2 8.1
72 4.0 8.6 5.0 9.5
96 5.5 4.4 6.8 16.1
120 6.4 15.8 6.4 16.1
144 5.8 15.8 6.2 16.0
168 5.1 15.8 5.8 15.7
Sugar conc. 15%, incubation temperature 30oC, initial pH 4.5 and incubation time 24-72h.
Effect of the initial pH of cane molasses on the ethanol production % and
sugar consumption % by the irradiated S. cerevisiae (0.1 kGy):
Table (4) highlights ethanol production by irradiated yeast cells
at (0.1 kGy) dose of gamma ray at different initial pH. The pH ranged from 4.0-
5.5 and each fermentation was run from 24-168 h after inoculation. Ethanol
yield by the irradiated cells after 24 h incubation at 30oC and sugar content 15%
was 0.9% and consumed sugar was 3.1%, at pH 4.0 and increased to 1.4% for
160 Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012)

ethanol production and 4.7% for sugar consume and the these data decreased by
increasing the pH value. Also, the value of ethanol production and sugar
consume increased by increasing the rate time of fermentation in all initial pH
values. Maximum production of ethanol yield was in yeast samples irradiated
for 0.1 kGy and the sugar consume was 16.1% at 96 h incubation at pH value
4.5 in initial cane molasses medium. These results showed that irradiation of
yeast cells at dose 0.1 kGy increased its activity and increase the yield of
ethanol production during fermentation process of cane molasses. So, data
showed that the rate of ethanol production by yeast cells is highly affected by
the pH of the fermentation medium. S. cerevisiae showed maximum growth
under acidic conditions and exhibited the maximum yield of ethanol (7.1%) at
pH 4.5 after 96h incubation time as similar to Rapeanu et al. (2000a). More
acidic and basic conditions, both retard the yeast metabolic pathways and hence
the growth of cells (Willaert and Viktor 2006). Productivity was decreased by
increase and decrease in pH due to the growth of other microbes of other
microbes as the fermentation was carried out without sterilization (Amutha and
Paramasamy 2001; Kourkoutas et al., 2004). In addition, pH of the surrounding
medium change the configuration and permeability of the cell membrane thus
reduced the rate of sugar fermented enzymes.
Table (4): Effect of the initial pH of cane molasses on ethanol production by
irradiated S. serevisiae cells (at 0.1 kGy) dose.
pH
Rate of 4.0 4.5 5.0 5.5
fermentation Ethanol Sugar Ethanol Sugar Ethanol Sugar Ethanol Sugar
(h) production consump- production consump- production consump- production consump-

% tion % % tion % % tion % % tion %


24 0.9 3.1 1.4 4.7 1.1 3.2 0.6 2.8
48 2.5 7.3 2.9 8.1 2.7 6.7 1.3 5.2
72 3.7 7.9 4.4 9.5 3.9 7.9 2.1 5.9
96 5.3 12.2 7.1 16.1 5.8 16.3 4.7 11.1
20 5.3 12.2 6.8 16.1 5.6 16.3 4.7 11.0
44 5.1 12.2 6.8 16.1 5.1 16.3 4.1 11.1
168 4.8 12.2 6.4 16.1 4.7 16.3 3.5 10.8
o
Sugar conc. 15%, incubation temperature 30 C and fermentation rate (24-168) h.
Effect of incubation temperature on ethanol production by irradiated S.cerevisiae
cells for (0.1 kGy) dose:

Table (5) is depicted the effect of different incubation temperature on


Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012) 161

ethanol production with irradiated (0.1 kGy) yeast cells. The temperature ranged
from 25-40oC and caused of fermentation was studied from 24-168 hours. Data
showed that ethanol yield increased with increasing the incubation period in
parallel with increasing in sugar consume. The optimal ethanol production was
obtained at 30oC by the irradiated cells giving ethanol yield 6.8% with 13.1%
sugar consumption at 96 hours of incubation. Changing in the incubation
temperature caused change in ethanol yield and sugar consumption, where at
slightly higher temperature growth rate may be adversely affected. Some strains
of S. cerevisiae have also been reported, capable of growing and fermenting
cane molasses at 40-45oC under batch conditions (Wang et al., 1985; Cachot
and Marie-Noelle 1991; Amutha and Paramasamy 2001).
Table (5): Effect of incubation temperature on ethanol production by irradiated S.
serevisiae cells (at 0.1 kGy) dose.
Incubation temperature oC
Rate of 25oC 30oC 35oC 40oC
fermentation Ethanol Sugar Ethanol Sugar Ethanol Sugar Ethanol Sugar
(h) production consump- production consump- production consump- production consump-

% tion % % tion % % tion % % tion %


24 1.4 7.0 1.2 7.4 0.0 7.2 0.7 5.6
48 2.8 7.3 2.6 8.1 2.2 7.8 1.8 5.9
72 4.6 8.3 4.5 9.5 4.0 8.1 2.7 7.8
96 6.8 13.8 6.1 14.3 5.3 12.7 3.4 13.9
20 5.9 14.1 6.9 15.3 5.0 14.3 3.2 14.2
44 5.9 15.3 5.5 16.1 4.7 15.0 3.0 14.1
168 5.0 15.3 5.0 16.1 4.3 15.7 2.8 14.2
Sugar conc. 15%, initial pH 4.5 and fermentation rate (24-168) h.
Effect of different initial sugar concentration on ethanol production by
irradiated S. cerevisiae cells for (0.1 kGy) dose:
Effect different initial sugar concentrations on ethanol
production was investigated in Table (6). Sugar concentration ranged from 12-
21% and fermentation period was ranged from 24-168 hours. The ethanol yield
and sugar consumption increased by increasing the incubation periods in all the
different sugar concentration. Maximum production was obtained at 15% sugar
level by the irradiated yeast cells where the produced ethanol reached to 7.1%
and sugar consumption 14.4% at 96 hours incubation period, while the initial
sugar concentration less than or more than 15% showed decreases in sugar
consumption and ethanol production in agreement with (Irfana et al., 2009),
they reported that maximum production was obtained at 15% sugar level by S.
cervisiae which gave 6.49% ethanol with 14.9% sugar consumption.
162 Botros, H.W et al., J. Rad. Res. Appl. Sci., Vol. 5, No. 2(2012)

In the present study, on the basis of ethanol yield 96 hrs was found to be
optimal for maximal production by the irradiated cells. It might be due to the
fact that the time necessary to complete fermentation of sugar cane molasses to
ethanol is correlated with the initial sugar concentration and yeast cells (Kaur
and Kocher, 2002; Willaert and Viktor, 2006).
Table (6): Effect of different initial sugar concentration on ethanol production by
irradiated S. serevisiae cells (at 0.1 kGy) dose.
Sugar content %
Rate of 12% 15% 18% 21%
fermentation Ethanol Sugar Ethanol Sugar Ethanol Sugar Ethanol Sugar
(h) production consump- production consump- production consump- production consump-

% tion % % tion % % tion % % tion %


24 1.0 5.2 1.5 6.4 1.5 7.7 1.3 6.5
48 1.2 6.0 2.8 8.1 2.7 7.8 2.1 5.9
72 2.0 7.4 4.4 9.5 4.0 8.7 3.0 7.4
96 4.1 12.8 7.1 14.4 6.5 15.0 6.4 16.5
20 4.9 13.3 6.7 15.3 6.6 15.5 6.5 16.9
44 5.1 13.3 6.6 16.1 6.6 16.5 6.5 16.9
168 5.1 13.3 6.5 16.1 6.5 16.5 6.5 16.9
Incubation temperature 30oC, initial pH 4.5 and fermentation rate (24-168) h.

CONCLUSION
The study showed that Saccharomy cerevisiae was able to produce
ethanol from cane molasses and fermentation process depends on sugar
concentration 15% and pH 4.5 and incubation temperature 30oC. Exposing S.
cerevisiae for (0.1 kGy) dose of gamma ray increased the ability of the yeast
cells for producing ethanol during the fermentation process of sugar cane at the
above conditions of fermentation.
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‫اﻟﻤﺆﺗﻤﺮ اﻟﺪوﻟﻲ اﻟﺜﺎﻟﺚ‬
‫ﻟﻠﻌﻠﻮم اﻹﺷﻌﺎﻋﯿﺔ وﺗﻄﺒﯿﻘﺎﺗﮭﺎ‬
‫‪ ١٦ – ١٢‬ﻧﻮﻓﻤﺒﺮ ‪ – ٢٠١٢‬اﻟﻐﺮدﻗﺔ ‪ -‬ﻣﺼﺮ‬

‫دراﺳﺔ ﻋﻠﻰ إﻧﺘﺎج اﻷﯾﺜﺎﻧﻮل ﻣﻦ ﻣﻮﻻس ﻗﺼﺐ اﻟﺴﻜﺮ ﺑﺎﺳﺘﺨﺪام ﺧﻼﯾﺎ اﻟﺨﻤﯿﺮة‬
‫اﻟﻤﺸﻌﻌﺔ‬
‫ھﺎﻧﺊ وھﯿﺐ ﺑﻄﺮس – أﺷﺮف ﺻﺒﺮى أﺣﻤﺪ – ﺳﻮزى ﺻﺒﺤﻰ ﻓﺮج – إﺳﻤﺎﻋﯿﻞ ﻋﺒﺪ اﻟﺤﻠﯿﻢ ﺣﺴﻦ‬

‫ﯾﺴﺘﺨﺪم ﻣﻮﻻس ﺳﻜﺮ اﻟﻘﺼﺐ ﻏﺎﻟﺒﺎً ﻓﻰ اﻹﻧﺘﺎج اﻟﺘﺠﺎرى ﻟﻜﺤﻮل اﻷﯾﺜﺎﻧﻮل ﻛﻤﺎدة ﺧﺎم ﻧﻈﺮًا‬
‫ﻟﺮﺧﺺ ﺛﻤﻨﮫ وﺗﻮاﻓﺮه‪ .‬ﺧﻼﯾﺎ ﺧﻤﯿﺮة ‪ Saccharomyces cervisiae‬ﺗﺴﺘﺨﺪم أﺳﺎﺳﺎً ﻓﻰ إﻧﺘﺎج اﻷﯾﺜﺎﻧﻮل‬
‫ﻧﻈﺮاً ﻟﻘﺪرﺗﮭﺎ ﻋﻠﻰ ﺗﺤﻠﯿﻞ ﺳﻜﺮ اﻟﺴﻜﺮوز اﻟﻤﻮﺟﻮد ﻓﻰ ﻣﻮﻻس ﻗﺼﺐ اﻟﺴﻜﺮ وﺗﺤﻮﯾﻠﮫ إﻟﻰ ﺳﻜﺮ اﻟﺠﻠﻮﻛﻮز‬
‫واﻟﻔﺮﻛﺘﻮز ﺣﯿﺚ أﻧﮭﻤﺎ ﺳﻜﺮﯾﺎت ﺳﺪاﺳﯿﺔ ﺑﺴﯿﻄﺔ‪ .‬ﺗﮭﺪف ھﺬه اﻟﺪراﺳﺔ ﻟﺘﻘﯿﯿﻢ إﺳﺘﺨﺪام أﺷﻌﺔ ﺟﺎﻣﺎ ﻋﻠﻰ ﻗﺪرة‬
‫ﺧﻼﯾﺎ اﻟﺨﻤﯿﺮة ﻓﻰ ﻋﻤﻠﯿﺔ إﻧﺘﺎج اﻷﯾﺜﺎﻧﻮل ﺣﯿﺚ ﺗﻢ ﺗﻌﺮﯾﻀﮭﺎ ﻟﺠﺮﻋﺎت )‪،٠٫٦ ،٠٫٤ ،٠٫٢ ،٠٫١ ،٠٫٠٥‬‬
‫‪ (١٫٠ ،٠٫٨‬ﻛﯿﻠﻮﺟﺮاى ﻣﻦ أﺷﻌﺔ ﺟﺎﻣﺎ‪ .‬ﺗﻤﺖ ﻋﻤﻠﯿﺔ ﺗﺨﻤﯿﺮ اﻟﻤﻮﻻس ﺗﺤﺖ ﻇﺮوف ﻣﺤﺘﻮى ﺳﻜﺮ )‪-١٢‬‬
‫‪ ،(%٢١‬وﺳﻂ ﺣﺎﻣﺾ )‪ ،(٥٫٥-٤‬ﺣﺮارة اﻟﺘﺤﻀﯿﻦ )‪ ( ٤٠-٢٥‬درﺟﺔ ﻣﺌﻮﯾﺔ وﻓﺘﺮات اﻟﺘﺤﻀﯿﻦ ﺗﺘﺮاوح‬
‫ﺑﯿﻦ )‪ (١٦٨-٢٤‬ﺳﺎﻋﺔ‪ .‬أﻇﮭﺮت اﻟﻨﺘﺎﺋﺞ أن أﻋﻠﻰ ﻣﻌﺪل ﻹﻧﺘﺎج اﻷﯾﺜﺎﻧﻮل ﻛﺎن ﻋﻨﺪ ﺗﻌﺮﯾﺾ ﺧﻼﯾﺎ اﻟﺨﻤﯿﺮة‬
‫ﻟﺠﺮﻋﺔ ‪ ٠٫١‬ﻛﯿﻠﻮﺟﺮاى ﻣﻦ أﺷﻌﺔ ﺟﺎﻣﺎ ﻋﻨﺪ ﻧﺴﺒﺔ ﺳﻜﺮ ‪ ،%١٥‬درﺟﺔ ﺣﺎﻣﻀﯿﺔ ‪ ٤٫٥‬ﻋﻨﺪ درﺟﺔ ﺣﺮارة ‪٣٠‬‬
‫درﺟﺔ ﻣﺌﻮﯾﺔ وﻣﺪة ﺗﺤﻀﯿﻦ ‪ ٩٦‬ﯾﻮﻣﺎً‪.‬‬

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