pbl13_ppt (final hanisah)
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lab com 2TRANSCRIPT
PowerPoint Presentation
KKEK 3171: Laboratory and Communication IISemester 1, Session 2014/15Lecturer: Dr. Ang Teck NamAssoc. Prof. Dr. Ngoh
Group:Noor Hanisah Abdul Hamid (KEK120038)Ng Sau Wei (KEK120037)Ong Kien Yeap (KEK120046)PBL 13: Batch Fermentation
CONTENTSIntroduction to Fermentation
Problem Statement & Objectives
Literature Review
Methodology
Results & Analysis
Conclusion
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3Background of FermentationEthanol / Lactic acid fermentation
Metabolic process that converts sugar into acids or alcohol
Anaerobic process carried out by microorganisms
Liquid medium contains of specific nutrients for growth
GlucoseCO2 + EthanolPyruvic Acid
NAD+NADHYeast cellsIntroduction to FermentationBackgroundApplication
en.wikipedia.org/wiki/Fermentation
Liquid medium for fermentationYeast Extract a form of processed yeast products have the same characteristic of yeast cells
Glucose serves as a source of energy for growth under anaerobic condition
4Ammonium Sulphate, (NH4)2SO4 acts as a pH buffer
Magnesium sulphate, MgSO4Source of nutrients
Dipotassium phosphate, K2HPO4Source of nutrientsLiterature ReviewLiquid MediumFactors for Fermentation
Factors affecting alcoholic fermentationI) pHOptimum: 4 < pH < 8II) Inoculum Dosage
III) TemperatureOptimum: 30oC < T < 38oCBeyond 45oC: yeast cells are destroyed.Low temperature: dormant stage -> slow fermentation rate
5Literature ReviewLiquid MediumFactors for Fermentation
Applications of Fermentation6Introduction to FermentationBackgroundApplication
http://science.jrank.org/pages/2677/Fermentation-Uses.html
Alcoholic beverages
Antibiotics and drugs
Activated sludge process
PROBLEM STATEMENTSACTIVATOR OF FERMENTATION: Yeast -> convert glucose in liquid medium to ethanol.
PROBLEM: Yeast suspension added to medium PRIOR to autoclaving.
OBSERVATION: No yeast growth and alcohol production.
HYPOTHESIS: The high temperature & high pressure autoclave treatment destroyed the yeast cells.7Problem StatementObjectives
ObjectivesTo investigate the correct sequence of alcoholic fermentation by Bakers yeast.
To verify the process of alcoholic fermentation by conducting a series of qualitative and quantitative analyses
8Problem StatementObjectives
METHODOLOGYNegative Control (Problem Statement)Actual experimentChemical test for carbon dioxide Oxidation test for ethanolCell mass analysisGlucose assay (DNS method)9MethodologyNegative ControlActual ExperimentCO2 Test
Students experiment (negative control)10MethodologyNegative ControlActual ExperimentCO2 Test
Yeast extractGlucose(NH4)2SO4 MgSO4K2HPO4
Bakers yeast
11MethodologyNegative ControlActual ExperimentCO2 Test
Yeast extractGlucose(NH4)2SO4 MgSO4K2HPO4
Bakers yeast
Actual Experiment
Chemical test for carbon dioxide12MethodologyNegative ControlActual ExperimentCO2 Test
Acidified saline(25% NaCl, 3% H2SO4)
5ml of K2CrO7Few drops of 0.1M H2SO4
Warm13MethodologyOxidation Test for EthanolDNS assayCell Mass Analysis
Oxidation Test for ethanol
Heat in water bath
3mL of DNS solutionSodium Metabisulfide(Rochelle salts)Potassium Sodium Tartrate Tetrahydrate14MethodologyOxidation Test for EthanolDNS assayCell Mass Analysis
Glucose Assay ( DNS Method )
Cell Mass AnalysisThe dried filter paper is weighed.
The fermented mixture is filtered using Buchner funnel.
After filtration, the filtrate is dried together with the filter paper for 24hours.
The final weight is measured.15
MethodologyOxidation Test for EthanolDNS assayCell Mass Analysis
16RESULTS & ANALYSISNegative Control (Problem Statement)Actual experimentChemical test for carbon dioxide Oxidation test for ethanolCell mass analysisGlucose assay (DNS method)
Physical ObservationsAlcohol scent is detected.Bubbles floating Yeast is still growing 17Results & AnalysisPhysical ObservationsCO2 volumeOxidation test for Ethanol
18No significant level change in saline solution!Results & AnalysisPhysical ObservationsCO2 volumeOxidation test for Ethanol
Oxidation Test for EthanolINITIALFINAL19Results & AnalysisPhysical ObservationsCO2 volumeOxidation test for Ethanol
Initial colour of acidified K2Cr2O7solution20Negative controlActual experimentResults & AnalysisPhysical ObservationsCO2 volumeOxidation test for Ethanol
Spectrometer result: Absorbance plot 21Run 1Run 2Results & AnalysisPhysical ObservationsCO2 volumeOxidation test for Ethanol
Calculation for glucose concentration22NEGATIVE CONTROL0.672= 0.2504*x 0.0264 x = 2.9329 mg/mL
ACTUAL EXPERIMENT 0.604 = 0.2504*x 0.0264 x = 2.6613 mg/mL
Results & AnalysisGlucose contentCell mass analysis
Run 1NEGATIVE CONTROL 0.462=0.1752*x-0.0346 x = 2.8345 mg/mL x(negative)= 10* 2.8345 mg/mL = 28.345 mg/mL
ACTUAL EXPERIMENT 0.022=0.1752*x-0.0346 x = 0.3231 mg/mL
x(actual)= 10*0.3231 mg/mL = 3.231 mg/mL
Run 2
Mass of dry yeast at the end of experiment(g)Trial 1Actual0.9798Negative0.3251Trial 2Actual0.9906Negative0.1838
Initial mass of Bakers yeast: 1 g23Cell Mass AnalysisResults & AnalysisGlucose contentCell mass analysis
CONCLUSIONSSequence of alcoholic fermentation is significant.->Yeast must be added after autoclaving medium.
The qualitative and quantitative analysis were done and verified. Presence of alcohol scentBubbling in saline solutionK2Cr2O7 turns from orange to greenGlucose is consumedIncreasing final mass of yeast 24
Fermentation occurs!Conclusion
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25Q & A SessionTHANK YOU!