Shake Flask Fermentation

3192 WordsMay 2, 201313 Pages
Table of Contents 1.0 Abstract 2 2.0 Introduction 2 3.0 Aims 3 4.0 Theory 3 5.0 Apparatus 5 6.0 Procedure 5 7.0 Result 7 8.0 Calculation 10 9.0 Discussion 11 10.0 Conclusion 13 11.0 Recommendation 13 12.0 References 13 13.0 Appendix 14 1.0 Abstract In this experiment, Escherichia coli is used as a sample to study the growth kinetic of microorganism in shake flask. A different volume of E.coli was transferred into 250ml Erlenmeyer/shake flask containing media for the nutrient of microorganism. The different volume of microorganism transferred will give the different effect of reading on the constant volume of media used. There are three ways to test the growth kinetic rate of microorganism on shake flask, which are by optical…show more content…
5) More absorbance means higher number of cell detected. 6) The sampling is repeated 12 times so that we have 12 samples in a time of 24 hours with samples taken in every 2 hours. 6.3 Glucose analysis 1) 2 ml of inoculums is transferred into micro centrifuge tube. 2) Sample is then centrifuge at 10000 rpm with temperature 4 0C for 10 minutes. 3) After centrifuge, cell (solid) will deposited at the bottom of the centrifuge tube and supernatant (liquid) is separated from the cell. 4) This supernatant is transfer into another centrifuge tube for glucose analysis; the cell will then followed the next test which is cell dry weight. 5) Centrifuge tube containing supernatant is then slotted in the slot of the YTI turntable Biochemical Analyzer in order to analyzer the glucose existed inside the supernatant. 6) After 24 hours, the inoculums is transferred back into 135 ml medium prepared making a total of 150 ml of inoculums prepared thus finishing the steps of preparation of inoculums. 6.4 Sampling for cell dry weight 1) All 12 centrifuge tube containing cell (solid) from procedure for glucose analysis is arranged on the centrifuge tube rack. 2) The cap is opened inserted into the oven. 3) Centrifuge tube rack containing centrifuge is put inside the oven at 60 0C for 24 hours. 4) The dried biomass is then being placed inside

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