Chemical and Biochemical Engineering 2290A/B Lecture Notes - Alkaline Phosphatase, Reaction Rate, Sodium Hydroxide
DepartmentChemical and Biochemical Engineering
Course CodeCBE 2290A/B
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Aim: To study the effect of temperature on enzyme reaction rate
Protocol: End point Assay method
3.9ml Tris HCl pH 9.0
1.0ml substrate solution
At T=0 , add 0.1ml alkaline phosphatase (enzyme)
At T=4min, add 2ml 2N NaOH
Read absorbance at end of 4min at 410nm.
Repeat 3 times at each temperature
Observation (End point assay method)
Observation (Absorbance @ 410nm) t= 4 min
a. For each treatment, plot product concentration vs. time.
b. For each treatment, determine reaction rate using the graphical method and put the
appropriate information on the figure as a text box.
c. For each treatment’s figure legend, in addition to the standard information, add the
resulting reaction rate and product concentration at equilibrium (based on absorbance at
the plateau; if available).
d. Plot temperature vs. reaction rate.
i) Effect of temperature on enzyme activity
ii) Graphical representation of Temperature vs Reaction rate
iii) Based on your knowledge of enzyme structure, function and means of regulation, what
effect(s) do/does temperature have on enzyme function (i.e. rate and saturation, if you
recorded saturation) and how do your data demonstrate that/those effect(s)?
Using your knowledge of enzyme structure and function, explain why a prolonged period of high fever is
dangerous to humans.
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