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International Journal of Applied Environmental Sciences
ISSN 0973-6077 Volume 11, Number 1 (2016), pp. 27-36
© Research India Publications
http://www.ripublication.com
Absorption of Organic Compounds by Saccharomyces
cerevisiae on Industrial Waste Media
Endah Rita Sulistya Dewi
Doctoral Program on Environmental Science, Diponegoro University,
Jl. Imam Barjo, Semarang -50241, Indonesia.
Anang M. Legowo
1) Doctoral Program on Environmental Science, Diponegoro University,
Jl. Imam Barjo, Semarang -50241, Indonesia.
2) Department of Agricultural Sciences, Faculty of Animal and Agricultural Science,
Diponegoro University, Jl. Imam Barjo, Semarang -50241, Indonesia.
Munifatul Izzati
Jl. Prof. Soedarto SH Semarang Indonesia 50275
1) Doctoral Program on Environmental Science, Diponegoro University,
Jl. Imam Barjo, Semarang -50241, Indonesia.
2) Department of Biology, Faculty of Sains & Mathematics,
Diponegoro University, Jl. Prof. Soedarto SH, Semarang-50275, Indonesia.
Abstract
Abstract-Industrial wastewater non-dairy creamer can pollute the environment
if not done properly processing. Pollutants contained in liquid waste in the
form of organic compounds and anorganic either in a state of suspended and
dissolved able to degrade the quality of water bodies pose even smell that can
disrupt the environmental balance.Utilization of Saccharomyces cereviceae to
tackle environmental pollution continue to be developed . The removal of
heavy metals in soil and water environments can be performed by
Saccharomyces cerevisiae, while research capabilities in Saccharomyces
cerevisiae biosorption of organic compounds have not been done. The use of
Saccharomyces cerevisiae in wastewater treatment non-dairy creamer is
expected to have a high enough prospects in degrade organic compounds.
Based on this background , it is important to do research on liquid waste
management Non-dairy creamer media grows as Saccharomyces cerevisiae
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Endah Rita Sulistya Dewi et al
for single bioremediation function.Media effluent Non-dairy creamer (NDC)
contains organic compounds such as carb, fat and protein that can be used for
the growth of microorganisms. Saccharomyces cerevisiae grown in a variety
of media concentration waste (Non-dairy Creamer 100%, Non-dairy Creamer
75%, Non-dairy Creamer 50% , Non-dairy Creamer 25%), analyzed the
ability of these microorganisms utilize glucose and glycerol compounds.
Factorial design is a research, data analysis using Manova. Furthermore, based
on multivariate analysis , the odds observations media concentration with a pvalue of 0.001 , the p < 0.05, which showed that Ho is rejected so that it can
be interpreted that the concentration of media influence on the absorption
ability of organic compounds by Saccharomyces cerevisiae .The results
showed that levels of the substrate affect the absorption capacity. The highest
absorption of organic compounds by Saccharomyces cerevisiae is a
compound glycerol. The ability of Saccharomyces cerevisiae to utilize organic
compounds in the wastewater medium non-dairy creamer as bioremediation
agents can be recommended
Keyword: organic compounds, S.cerevisiae, Non-dairy Creamer (NDC)
Introduction
Non-dairy creamer is milk or cream substitute product that is a product of lipid
emulsion in water, made from vegetable oils such as coconut oil hydrogenated with
the addition of food additives are permitted, namely glucose syrup, sodium caseinate,
emulsifiers and stabilizers, salt and water. The main technology used in the product is
a spray-drying technology, the cutting-edge technology that is applied to transform
the liquid phase into a powder creamer ingredient. Furthermore, in the process of
making Non-dairy creamer, waste is produced which can be either solid or liquid
waste[1].
Non-dairy creamer effluent which flowed along the flow of waste disposal can lead to
contamination when not treated optimally. In addition to causing the quality of the
water body is lowered, the odor generated can disturb the environment. Odor
generated can be caused by excessive content of organic compounds contained in
wastewater. Associated with these conditions need to be a processing using
bioremediation techniques.
Bioremediation is the development of environmental biotechnology field by utilizing
the biological processes in controlling pollution. Microorganisms have been used to
reduce organic compounds and toxic substances which derive from household waste
industry[2]. The new thing is that bioremediation technique proved to be very
effective and inexpensive in terms of the economy to clean up contaminated soil and
water by toxic chemical compounds or toxic[3].
Chemical degradation by microbes in the environment is a very important process to
reduce the levels of hazardous substances in the environment, which takes place
through a series of chemical reactions that are quite complex. In the process of
Absorption of Organic Compounds by Saccharomyces cerevisiae
29
degradation microorganisms use chemical compounds for growth and reproduction
through the process of oxidation [4].
Based on this condition important to do research on the management of waste liquid
non- dairy creamer as a growing medium in Saccharomyces cerevisiae bioremediation
function, so that the environment is also protected from contamination by waste liquid
non-dairy creamer.
Material and Methods
Preparing tool and ingredient pure culture of Saccharomyces cerevisiae, the waste
liquid non-dairy creamer, yeast extract, peptone, dextrose, distilled water, 0,5N
NaOH, H2O, Na2CO3, CuSO4.
Creating medium YEPD.YEPD medium made of a mixture of yeast extract,
peptone, dextrose, and distilled water. All materials included in the glass beaker and
stirred until homogeneous. The mixture is then put in erlemeyer or jam jar to a
volume of 150 mL. Jar covered with foam and aluminum foil, and then sterilized by
autoclaving at 121oC for 15 minutes. Furthermore, the medium is cooled and after
cold ready inoculated.
Making the medium of liquid waste Non-dairy creamer. Medium is made from
liquid waste non-dairy creamer with a concentration of 100%, 25%, 50% and 75%.
Furthermore, the material is stirred until homogeneous, filtered, and put in a jam jar.
Jam jar and covered with foam and sterilized by autoclaving at 121oC for 15 minutes,
then cooled and after a cold medium ready inoculated
Growing S. cerevisiae on medium YEPD and medium liquid waste Non-dairy
creamer.Growth of S. cerevisiae in YEPD medium and liquid waste based on the
method of [5]In the medium and medium YEPD liquid waste non-dairy creamer,
S.cerevisiae suspended in sterile distilled water with a density of 10 6 cells / mL. Each
medium in the bottle hours inoculated with 50 mL of the suspension. Furthermore,
both the medium and the medium YEPD liquid waste non-dairy creamer incubated in
an incubator at a temperature of 26 ° C for 96 hours. Then the pH of the medium is
measured and calculated cell Saccharomycescerevisiae population. cerevisiae every
24 hours for 6 days.
Measuring pH medium. pH measurement and wastewater mediumYEPD Non-dairy
creamer done at the 24th hour, 48, 72, and 96 hours by using a pH meter.
Counting the number of cells S. cerevisiae.Counting the number of cells / mL done
every 24 hours based on the method of [6]with a hemocytometer on a light
microscope or through analysis kerapatn optical / OD (Optical Density) using a
photometer at a wavelength of 550 nm
Calculation of potential biosorption S.cerevisiae. Analyzing the glucose content,
nitrogen and glycerol in the medium at 0 hours, 48 hours, 96 hours, 144 hours (each
2x24 hours). Factorial design is a research, and data analysis using Manova
Endah Rita Sulistya Dewi et al
30
The ability of Saccharomyces cerevisiae biopsorbsi to use the formula:
Absorption capacity(mg/g) =
Legend :
Co = initial concentration (mg/L)
Ce = final concentration (mg/L)
m = biosorben mass (gram)
V = Volume solvent (L) [7].
Experimental research conducted at the Laboratory of Chemistry, Faculty of Science
and Mathematics Christian Satya Wacana University
Result and Discussion
Absorption of organic compounds by the cells in the medium used for the growth of
these cells. Batch system used in this study, a fermentation system without the
addition of the substrate at a certain time. Saccharomyces cerevisiae fermentation of a
way to get energy for metabolism processes in the form of ATP that is characterized
by a phosphorylation substrate that produces the synthesis of the final product.
Media effluent non-dairy creamer derived from vegetable oil feedstocks contain
carbohydrate, protein, and fat that can be used for microbial growth. Non-dairy
creamer is a product substitute for milk or cream that is a product of lipid emulsion in
water, made from vegetable oils such as coconut oil hydrogenated with the addition of
food additives are permitted, namely glucose syrup, sodium caseinate, emulsifiers and
stabilizers , salt and water [1].
The Absorption of Glucose Compound
Carbohydrates are the main substrates were broken in the process of fermentation.
Polysaccharides first be broken down into simple sugars, for example hydrolysis of
starch into glucose units. The results showed the uptake of glucose by cells of
Saccharomycescerevisiaeis presented in Fig 1, Fig 2, Fig 3 and Fig 4.
Figure 1: Glucose Absorbtion Capacity of Waste NDC Media Concentration
of 100% (A)
Absorption of Organic Compounds by Saccharomyces cerevisiae
31
Description :
A5 = concentration of cell 105 cells/ml
A6 = concentration of cell 106 cells/ml
A7 = concentration of cell 107 cells/ml
A8 = concentration of cell 108 cells/ml
Figure 2: Glucose Absorbtion Capacity of Waste NDC Media Concentration
of 75% (B)
Description :
B5 = concentration of cell 105 cells/ml
B6 = concentration of cell 106 cells/ml
B7 = concentration of cell 107 cells/ml
B8 = concentration of cell 108 cells/ml
Figure 3: Glucose Absorbtion Capacity of Waste NDC Media Concentration
of 50% (C)
Description :
C5 = concentration of cell 105 cells/ml
C6 = concentration of cell 106 cells/ml
C7 = concentration of cell 107 cells/ml
C8 = concentration of cell 108 cells/ml
Endah Rita Sulistya Dewi et al
32
Figure 4: Glucose Absorbtion Capacity of Waste NDC Media Concentration
of 25% (D)
Description :
D5 = concentration of cell 105 cells/ml
D6 = concentration of cell 106 cells/ml
D7 = concentration of cell 107 cells/ml
D8 = concentration of cell 108 cells/ml
The results showed the uptake of glucose by cells of Saccharomyces cerevisiae high
of 3.823 mg/g on a substrate concentration of 75%, with a concentration cell count
106 cells / ml, it may be possible because the nutrient content in the concentration of
75%, still relatively fulfilled for growth, [8]starting that the nutrient-containing sugar
will provide energy for metabolic processes Saccharomyces cerevisiae, while in the
manufacture of non-dairy creamer products are sugar content.
According [9] glucose in the fermentation process will be broken down into other
compounds. Glucose fermentation pathways in microbes can be through lanes
Embden-Meyerhoff-Parnas (EMP) which is characterized by the formation of fructose
diphosphate, followed by a breakdown of fructose diphosphate into two molecules of
glyceraldehyde phosphate. This reaction is catalyzed by the enzyme aldolase. Then
there griseraldehida phosphate dehydrogenation reaction which is the oxidation
reactions that produce energy in the form of ATP. The hydrogen atom is released will
be captured by NAD, forming NADH2. The fermentation process can continue if
NADH2 can be oxidized back to the second stage of fermentation thus releasing
hydrogen atoms back. Thus, the NAD serves as a hydrogen carrier in the fermentation
process. The whole reaction as follows:
Glucose + 2 (ADP + 2 NAD+ + Pi) → 2 piruvat + 2 ATP + 2 (NADH + H+)
When the existence of reduced oxygen or no oxygen, glucose can still be converted
into pyruvic acid via glycolysis [10]. The absorption of glucose into the cells and the
solution is presented in Fig.5.
Absorption of Organic Compounds by Saccharomyces cerevisiae
33
Figure 5: Chronology of Establishment of ATP in the Krebs Cycle, adapted from
Campbell, et al[16].
In all types of cells, and regardless of the mechanism used to extract energy, the
reaction is accompanied by the formation of Adenosine triphosphate (ATP). ATP is a
common intermediate (reactant) in both the reactions that produce energy and
reactions that require energy, and its formation requires a mechanism by which the
available energy can be channeled into the biosynthesis of cell reactions that require
energy.
The Absorption of Glycerol Compound
The results showed the uptake of glycerol by cells of Saccharomyces cerevisiaeis
presented in Fig. 6, Fig.7, Fig.8 and Fig.9.
Figure 6: Glicerol Absorbtion Capacity of Waste NDC Media Concentration
of 100% (A)
Description :
A5 = concentration of cell 105 cells/ml
A6 = concentration of cell 106 cells/ml
A7 = concentration of cell 107 cells/ml
A8 = concentration of cell 108 cells/ml
Endah Rita Sulistya Dewi et al
34
Figure 7: Glicerol Absorbtion Capacity of Waste NDC Media Concentration
of 75% (B)
Description :
B5 = concentration of cell 105 cells/ml
B6 = concentration of cell 106 cells/ml
B7 = concentration of cell 107 cells/ml
B8 = concentration of cell 108 cells/ml
Figure 8: Glicerol Absorbtion Capacity of Waste NDC Media Concentration
of 50% (C)
Description :
C5 = concentration of cell 105 cells/ml
C6 = concentration of cell 106 cells/ml
C7 = concentration of cell 107 cells/ml
C8 = concentration of cell 108 cells/ml
Figure 9: Glicerol Absorbtion Capacity of Waste NDC Media Concentration
of 25% (D)
Absorption of Organic Compounds by Saccharomyces cerevisiae
35
Description :
D5 = concentration of cell 105 cells/ml
D6 = concentration of cell 106 cells/ml
D7 = concentration of cell 107 cells/ml
D8 = concentration of cell 108 cells/ml
The absorption capacity of the highest glycerol 6.594 mg / g at a concentration of 106
cells / ml, on a substrate concentration of 75%, higher than the capacity of absorption
of glucose at a concentration of 106 cells / ml in medium liquid waste non-dairy
creamer. Non-dairy creamer is a vegetable oil based products is coconut oil. When in
the growth medium widely available fatty compounds are glycerol, then the microbes
in this case Saccharomyces cerevisiae will be utilizing the presence of glycerol to
growth compared to glucose.
According to research conducted by Bleve, et al[11], Saccharomyces cerevisiae
capable of using olive oil mill wastewaters (OMW) olive oil or liquid waste as a
source of nutrients for growth. Furthermore [12], states Saccharomyces cerevisiae is
the most adaptive microbe that can live in conditions of minimal nutritional
environment including waste inexpensive media.
Glycerol plays an important role in maintaining osmotic balance and redox balance in
yeast cells Saccharomyces cervisiae. These compounds can be formed during the
fermentation process or made available to the media [13].
The existence of glycerol by microbes would be converted into Phosphoglyceric acid,
pyruvic acid, asetyl-CoA which would then be included in the Krebs cycle and
generate ATP.
The ability to degrade chemical compounds by microbes in the environment is a very
important process to reduce the levels of hazardous substances in the environment,
which takes place through a series of chemical reactions that are quite complex, so
with these conditions microbes have the potential as a bioremediation agent.
Saccharomyces cerevisiae one microbe species of fungi that can serve as a
bioremediation agent [14],[15]
Conclusion
The ability of Saccharomyces cerevisiae to utilize organic compounds in the
wastewater medium Non-dairy creamer as bioremediation agents can be
recommended. The highest absorption of organic compounds by Saccharomyces
cerevisiae is a compound glycerol.The absorption capacity of the highest glycerol
6.594 mg/g at a concentration of 106 cells/ml, on a substrate concentration of 75%,
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