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S.cerevisiae fermentation in shake flask experiment
- 1. BIOTECHNOLOGY ENGINEERING LAB IV BTE3211 I, 2017/2018 Mohammad Firdaus Bin Abu Bakar 1416451 Mohd Haffizi Bin Hasni 1421995 Abdul Hadi Bin Annuar 1412769 Abdullah Umar Bin Shahrin 1414575 Raja Muhammad Norfarres bin Raja Nordin 1413989 Osama Abdelhadi 1314409 S.cerevisiae Fermentation in Shake Flask
- 2. Introduction Shake Flask Easy to manage Preparation of Culture
- 3. Introduction Saccharomyces cerevisiae fungi (Feldmann, 2010) withstand a wide range of pH(Ortiz-Muñiz, 2010) withstand ethanol far better (Prasertwasu et al., 2014)
- 4. Introduction Fermentation (Moran et al., 2014) C6H12O6 + O2 H2O + CO2 + Energy+ biomass C6H12O6 CO2+ C2H6O +Energy Ethanol as a Secondary metabolite
- 5. Introduction Media (Eddleman, 1999) 20 g/L Peptone source of nitrogen 10 g/L yeast extract Source of nutrient 50 g/L Glucose
- 6. Objectives To study growth kinetics of baker’s yeast To familiarise with batch culturing in flask To practice proper aseptic techniques
- 7. Procedure Yeast Culture 0.2% w/w yeast added into media Shake for 90 min Every 15 min sample taken for analysis
- 8. Procedure Analysis I Optical density measure at 620nm Media use as blank Graph is plotted
- 9. Procedure Analysis II Boat from aluminium foil prepared Dry at 80C Centrifuge sample (1mL, 10min, 10000rpm) Supernatant removed, pellet resuspended Content poured in boat, dry overnight at 80C, weight the boat
- 10. Results Boat time (min) Absorban ce transmittance (%) weight before drying (g) Weight after drying (g) dry cell weight (g) 0a 15 0.552 28.1 0.7152 0.7153 0.0001 0b 15 0.594 28.4 0.8186 0.8194 0.0008 1a 30 0.383 41.4 0.5357 0.5375 0.0018 1b 30 0.384 41.3 0.6624 0.6641 0.0017 2a 45 0.274 53.2 0.4388 0.4405 0.0017 2b 45 0.286 51.7 0.8199 0.8215 0.0016 3a 60 0.469 33.9 0.4119 0.4131 0.0012 3b 60 0.306 49.4 0.5418 0.544 0.0022 4a 75 0.391 40.7 0.53 0.5312 0.0012 4b 75 0.469 33.9 0.8077 0.8112 0.0035 5a 90 0.496 31.9 0.6884 0.6902 0.0018 5b 90 0.717 19.2 0.7512 0.7566 0.0054 6a 105 0.8 15.9 0.7435 0.7456 0.0021 6b 105 1.169 6.78 0.6101 0.6125 0.0024
- 11. Results 0 0.2 0.4 0.6 0.8 1 1.2 1.4 0 20 40 60 80 100 120 Absorbance Vs time A B
- 12. Results 0 0.001 0.002 0.003 0.004 0.005 0.006 0 0.2 0.4 0.6 0.8 1 1.2 1.4 Cell dry weight vs absorbance a b
- 13. Results 0 0.001 0.002 0.003 0.004 0.005 0.006 0 20 40 60 80 100 120 Cell dry weight vs time a b
- 14. DISCUSSION Hypothesis As number of cells increase the absorbance should increase Error o Use of the same micropipette tip o The yeast and media are not entirely homogenized o Contamination in certain vials due to poor aseptic techniques o Error in weighing the boats o Some sample spilled during resuspension o The wrong cuvette may had been used as a blank
- 15. CONCLUSION Practice the aseptic techniques Observe the growth of yeast through optical density measure the dry weight of the cell precaution steps must be taken