Shake Flask Fermentation
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 density (OD), glucose analysis and cell dry weight (CDW). Then, from these test the result then used to plot the growth curve graph which next use to compared and analyze the growth curve of microorganism.
2.0 Introduction
Shake flask fermentation is one of the fermentation method which are widely used for screening of high producing strains. The shake flask fermentation is the simplest way to do the fermentation using small amount volume of nutrient broth in laboratory. A nutritionally rich medium, Lysogeny broth or luria bertani (LB) is used for the growth of bacteria. Usually the shake flask used is between 250ml to 500ml range. From the previous research, it is shown that smaller volume of shake flask give better oxygen transfer rate but it is only suitable for short term fermentation, otherwise the medium will be evaporate. The shake flask have several design which are standard shake flask or Erlenmeyer flask, flying saucer shake flask, shake flask with baffles and flat bed Thompson bottle. In this experiment, the standard shake flask is used.
Erlenmeyer flask which equipped with cotton wool stoppers and autoclaved including the nutrient broth inside the flask. Then, the flask is allowed to cool to the
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 density (OD), glucose analysis and cell dry weight (CDW). Then, from these test the result then used to plot the growth curve graph which next use to compared and analyze the growth curve of microorganism.
2.0 Introduction
Shake flask fermentation is one of the fermentation method which are widely used for screening of high producing strains. The shake flask fermentation is the simplest way to do the fermentation using small amount volume of nutrient broth in laboratory. A nutritionally rich medium, Lysogeny broth or luria bertani (LB) is used for the growth of bacteria. Usually the shake flask used is between 250ml to 500ml range. From the previous research, it is shown that smaller volume of shake flask give better oxygen transfer rate but it is only suitable for short term fermentation, otherwise the medium will be evaporate. The shake flask have several design which are standard shake flask or Erlenmeyer flask, flying saucer shake flask, shake flask with baffles and flat bed Thompson bottle. In this experiment, the standard shake flask is used.
Erlenmeyer flask which equipped with cotton wool stoppers and autoclaved including the nutrient broth inside the flask. Then, the flask is allowed to cool to the
References: 1. Growth and Vulturing Bactering, 24.3.2012 from url http://www.mansfield.ohio-state.edu/~sabedon/black06.htm 2. Microbial Growth Curve Part 2:Stationary PhaseFacts andFallacies,24.3.2012 from url http://fermentationtechnology.blogspot.com/2010/01/stationary-phase-facts-and-fallacies.html 3. Talaat E. Shehata and Allen G. Marr, Effect of Nutrient Concentration on the Growth of Escherichia coli (1971) Department of Bacteriology, University of California, Davis, California 95616 4. Roger G. Harrison,Paul todd,Bioseparation science and engineering(2003),oxford university press. 5. Christie john geankoplis, transport processes and separation process principles (fourth ed),pearson prentice hall (2003) 13.0 Appendix