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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 28 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. 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