In this study we evaluated the optimal physical requirements such as the shaking rate and incubation time for applying in liquid media malt extract broth to obtain a high emestrin compound and its toxicity.
Effect of shaking rate on the production of bio-active components are not fixed, It depends on the environmental factors. You perform a study with and without agitation so that you will conclude whether agitation is required or not. If it is required, then you try with shaking at low and high speeds.
You may use placket burman design, it is one of statistical approach to study and evaluate statistically the main effect of each parameter in your studies. It is also one of the approach to study/large large parameters by conducting small number of experiments.
Shaking rate is crucial for aerobes to provide an enabling condition for organism's metabolisms. However, since your study evaluated the optimal physical requirements such as the shaking rate, it is pertinent to optimized the shaking rate with others using response surface methodology, a statistical tool of Design Expert to determine the optimized conditions.
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