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Students collected Beta-fructosidase enzyme activity data in the presence and absence of an unknown inhibitor. P-nitrophenol...

Students collected Beta-fructosidase enzyme activity data in the presence and absence of an unknown inhibitor. P-nitrophenol was used as a read out for enzyme activity. Quantity of product was determined from the absorbance of each sample after 5 minutes of reaction. Using the information, complete the table below and create a properly labeled Lineweaver burke plot with the appropriate axes and best fit lines for the data, in excel (graph is worth 6pts). Using this plot, calculate the approximate Vmax and Km values for the reactions with and without the inhibitor. A picture of your graph can be inserted into this document as a picture.

		nmol product	nmol/min (1pt)	1/V (1pt)	Substrate Concentration (mM)	1/[S] (1pt)
Normal	1	150			0.036
	2	294			0.090
	3	423			0.180
	4	684			0.270
Inhibitor	5	73.0			0.036
	6	142			0.090
	7	217			0.180
	8	346			0.270

Normal Reaction

Vmax: _________________ (1pt)

Km: _________________(1pt)

Inhibited Reaction

Vmax: _________________(1pt)

Km: _________________(1pt)

What type of inhibitor is this? Be sure to justify your answer by discussing the Km AND Vmax values you calculated (2pts).

Expert Solution

Question:

Answer:

Lineweaver Burk plot equation:

1V= KmVmax S + 1Vmax

Slope = Km/Vmax

Intercept = 1/Vmax

So, Vmax = 1/intercept

Km= Slope / intercept

In absent of inhibitor:

Please find values from attached graph

Slope: 0.20629;

Intercept: 0.99035

Vmax = 1/0.99035 = 1.0097

Km = 0.20629/0.99035 = 0.2083

In presence of inhibitor:

Please find values from attached graph

Slope: 0.43088;

Intercept: 2.06855

Vmax= 1/0.43088 = 0.483429484

Km = 0.43088/2.06855= 0.2083

Vmax has been decrease in presence of inhibitor but km is still same. So, it is a non-competitive inhibition.

gladiator answered 2 years ago

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