Bioethanol production from cheese whey.pptx

It deals about production of bioethanol from cheese whey which is not sterilized and other characters of the ethanol producing yeasts were also covered

1. Bioethanol production from cheese whey using yeast, a non -Saccharomyces, Kluyveromyces marxianus Asmamaw Tesfaw 1

2. Introduction Bioethanol Yeast Starch Lignocelluloses Wastes Simple sugars Sorting Hydrolysis Chemical Physical Biological Agricultural residues 2

3. natural ecosystems rich in sugar 3

4. Growth factors temperature pH oxygen initial sugar concentrations Optimization of multiple variables: single factor at a time Limitations Which one comes first? You can’t predict for future? Solution Response surface methodology (RSM) 4

5. RSM employ Statistical and Mathematical technique RSM enables to design experiments build model evaluate interactions look for optimum conditions for responses reduce the number of experiments 5

6. Cheese whey Liquid waste 130millions tons word wide 50% water bodies High BOD and COD increasing at a rate of 3% per year Ethiopia 20, 000 tons Increasing Objective to evaluate ethanol production capability of K. marxianus ETP87 from crude whey. 6

7. MATERIALS AND METHODS Optimizing growth variables Temperature: 30, 35, 40 pH: 4, 5, 6 Time:24, 48, 72 Where, Y = ethanol produced in g/L (dependent output)  = intercept β 1 , β 2 , and β 3 = Linear, quadratic and interaction regression coefficients for temperature, pH and time respectively X 1 , X 2 , and X 3 = independent variable for temperature (degree centigrade), pH, and time (hours) respectively ε = random experimental error 7

8. Factor 1 (Temperature, o C ) Factor 2 (pH) Factor 3 (time, hours) Response (Ethanol produced, g/L) 35.00 5.00 48.00 30.00 6.00 24.00 30.00 6.00 72.00 26.59 5.00 48.00 35.00 5.00 48.00 35.00 5.00 48.00 40.00 4.00 72.00 35.00 5.00 48.00 35.00 5.00 88.36 30.00 4.00 24.00 43.41 5.00 48.00 40.00 4.00 24.00 35.00 5.00 48.00 40.00 6.00 72.00 35.00 3.32 48.00 40.00 6.00 24.00 30.00 4.00 72.00 35.00 6.68 48.00 35.00 5.00 7.64 35.00 5.00 48.00 8 Experimental design

9. Crude whey Yeast: K. marxianus ETP87 Experimental condition : non-deproteinized and deproteinized by heating Supplementation of whey with molasses and nitrogen sources extract (0.55%), peptone (1%), and ammonium sulfate (0.33%). Molasses 100% molasses alone 25% whey and 75% molasses 50% whey and 50% molasses 75% whey and 25% molasses 100% whey 9

10. Effect of pH and temperature pH: crude3.9 and adjusted 5.0 Temperature:25, 30, 35, 40, 45, and 50 o C Lactose determination : reducing sugar Ethanol determination : GC-MS and Pycnometer methods 10

11. RESULTS AND DISCUSSION 11

12. Model diagnostics 12 Normality test No need to transform

13. 13 DFFITS

14. Diagnostics Case Statistics Report Internally Externally Influence on Run Actual Predicted Studentized Studentized Cook's Fitted Value Standard Order Value Value Residual Leverage Residual Residual Distance DFFITS Order 1 5.43 10.43 - 5.00 0.607 -2.240 -3.009 0.776 -3.743 1 11 2 9.30 8.04 1.26 0.670 0.616 0.596 0.077 0.849 2 3 19.25 14.68 4.57 0.670 2.232 2.990 1.011 1 4.258 1 6 4 15.52 15.29 0.23 0.166 0.070 0.067 0.000 0.030 16 5 15.02 12.07 2.95 0.670 1.440 1.535 0.421 2.186 1 7 6 18.46 15.07 3.39 0.670 1.655 1.842 0.555 2.623 1 5 7 9.84 12.91 -3.07 0.607 -1.376 -1.450 0.293 -1.803 9 8 9.47 12.35 -2.88 0.607 -1.291 -1.342 0.258 -1.669 10 9 8.56 7.74 0.82 0.670 0.402 0.384 0.033 0.547 4 10 6.70 7.65 -0.95 0.607 -0.427 -0.409 0.028 -0.509 12 11 14.90 15.29 -0.39 0.166 -0.120 -0.114 0.000 -0.051 20 12 15.78 15.29 0.49 0.166 0.150 0.143 0.000 0.064 15 13 8.13 13.54 -5.41 0.607 -2.424 -3.579 0.908 -4.451 1 14 14 15.63 15.29 0.34 0.166 0.104 0.099 0.000 0.044 17 15 7.65 8.01 -0.36 0.670 -0.176 -0.167 0.006 -0.238 3 16 10.22 8.61 1.61 0.670 0.785 0.769 0.125 1.095 8 17 4.00 4.54 -0.54 0.607 -0.243 -0.232 0.009 -0.288 13 18 15.11 15.29 -0.18 0.166 -0.056 -0.053 0.000 -0.024 18 19 7.85 5.25 2.60 0.670 1.272 1.318 0.328 1.876 1 20 15.83 15.29 0.54 0.166 0.166 0.157 0.001 0.070 19 14

15. 15 Central Composite design Factor 1 Factor 2 Factor 3 Response 1 Std Run A:Temperature B:pH C:Time Ethanol Produced 11 1 35.00 3.32 48.00 5.43 2 2 40.00 4.00 24.00 9.3 6 3 40.00 4.00 72.00 19.25 16 4 35.00 5.00 48.00 15.52 7 5 30.00 6.00 72.00 15.02 5 6 30.00 4.00 72.00 18.46 9 7 26.59 5.00 48.00 9.84 10 8 43.41 5.00 48.00 9.47 4 9 40.00 6.00 24.00 8.56 12 10 35.00 6.68 48.00 6.7 20 11 35.00 5.00 48.00 14.9 15 12 35.00 5.00 48.00 15.78 14 13 35.00 5.00 88.36 8.13 17 14 35.00 5.00 48.00 15.63 3 15 30.00 6.00 24.00 7.65 8 16 40.00 6.00 72.00 10.22 13 17 35.00 5.00 7.64 4 18 18 35.00 5.00 48.00 15.11 1 19 30.00 4.00 24.00 7.85 19 20 35.00 5.00 48.00 15.83

16. ANOVA: Model validation Source Sum of Squares df Mean Square F Value p-value Prob > F Model 258.14 9 28.68 12.81 0.0002 A-Temperature 0.49 1 0.49 0.22 0.6493 B-pH 22.63 1 22.63 10.10 0.0098 C-Time 113.49 1 113.49 50.67 < 0.0001 AB 1.13 1 1.13 0.51 0.4933 AC 5.59 1 5.59 2.50 0.1451 BC 8.51 1 8.51 3.80 0.0799 A 2 32.89 1 32.89 14.68 0.0033 B 2 45.27 1 45.27 20.21 0.0012 C 2 48.95 1 48.95 21.85 0.0009 Residual 22.40 10 2.24 Lack of Fit 21.69 5 4.34 30.64 0.0009 Pure Error 0.71 5 0.14 Cor Total 280.53 19 Std. Dev. 1.50 R-Squared 0.9202 Mean 11.88 Adjusted R-Squared 0.8483 C.V. % 12.60 Predicted R-Squared 0.4102 16

17. 17 Where X 1 temperature ( o C ) X 2 pH X 3 incubation time(hours) Optimum temperature (34.2 o C), pH (4.43), and incubation time (71.93 hours)

18. 18 (B) (A) (D) (C) K. marxianus ETP87 Response surface

19. Whey samples pH Lactose (g/L) Ethanol (g/L) by Efficiency (%) Biomass (g/L) Sampling 2 week later microflora on time of sampling microflora 2 weeks in refrigerator Ethanol by K. marxianus ETP87 Tigist dairy 4.5 34.4±1.3 27.3±1.1 1.2±0.09 11.62±0.7 66.23 7.67±0.5 Amanual dairy 3.8 28.6±1.2 21.5±0.9 0.6±0.08 1.7±0.08 10.54±0.8 72.26 6.20±0.4 Household 3.1 18.7±0.8 14.4±0.7 2.3±0.1 1.4±0.07 6.43±0.5 67.42 5.48±0.4 Shola dairy 4.2 27.8±1.2 26.1±1.1 0.8±0.08 12.49±0.9 88.09 7.12±0.7 19 Ethanol production from different whey

20. Ethanol production from non-sterilized and sterilized whey Refrigerator days Non Sterilized whey Sterilized whey Ethanol produced % of reduction Ethanol produced % of reduction 11.71 11.68 1 11.46 2.13 11.69 +0.09 2 10.34 11.7 11.48 1.71 3 10.97 6.3 10.77 7.79 4 10.01 14.51 11.12 4.79 5 9.22 21.26 10.56 9.59 6 8.61 26.47 10.21 12.59 7 8.18 30.15 9.63 17.55 8 7.54 35.61 9.44 19.18 14 5.67 51.58 8.37 28.34 Average 9.11 10.36 20

21. Effect of whey pH 21

22. Molasses supplementation to whey 22

23. Effects of external nutrient additions to whey 23

24. Effect of Temperature 24

25. CONCLUSIONS AND RECOMMENDATIONS Putting whey sample in refrigerator doesn’t guarantee the existence of lactose for more than 5 days without decreasing in its amount Ethanol could be produced from crude non-sterilized whey using K. marxianus 25

26. Thank You 26