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2. Draw a graph to calculate the initial rate of reaction due to enzyme X in absorbance units/min/100μl of extract.

2. Draw a graph to calculate the initial rate of reaction due to enzyme X in absorbance units/min/100μl of extract.

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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A baby fails to thrive after 8 weeks of life. She wakes frequently and is very irritable. The parents notice that the baby appears to be better after feeding but that her abdomen is protruding more and more. After her condition worsens, she is admitted to hospital where an examination indicates that his liver and kidneys are enlarged. A liver biopsy was performed and a liver extract was prepared by homogenising 200mg of liver in 1ml of 0.3M sucrose, 10mM Tris-HCl, pH 7.5, 1mM dithiothreitol, 1mM EGTA and 100μM phenylmethylsulphonyl fluoride (PMSF). The activity of enzyme X was measured in the liver sample by adding 100μl of the liver extract to 0.9 ml of 120mM Imidazole buffer (pH 7.0) containing 2.4mM NAD+, 30mM glucose-6-phosphate, 6 units/ml of glucose dehydrogenase in a cuvette with a 1cm light path. The increase in absorbance at 340nm was followed over a period of 10 minutes and compared with a blank sample containing everything but the glucose-6- phosphate. The results are shown in the table below: Time (mins) 0 2 4 6 8 10 Absorbance at 340nm With G6P 0 0.12 0.23 0.34 0.44 0.53 Without G6P 0 0.03 0.062 0.082 0.087 0.092
Transcribed Image Text:A baby fails to thrive after 8 weeks of life. She wakes frequently and is very irritable. The parents notice that the baby appears to be better after feeding but that her abdomen is protruding more and more. After her condition worsens, she is admitted to hospital where an examination indicates that his liver and kidneys are enlarged. A liver biopsy was performed and a liver extract was prepared by homogenising 200mg of liver in 1ml of 0.3M sucrose, 10mM Tris-HCl, pH 7.5, 1mM dithiothreitol, 1mM EGTA and 100μM phenylmethylsulphonyl fluoride (PMSF). The activity of enzyme X was measured in the liver sample by adding 100μl of the liver extract to 0.9 ml of 120mM Imidazole buffer (pH 7.0) containing 2.4mM NAD+, 30mM glucose-6-phosphate, 6 units/ml of glucose dehydrogenase in a cuvette with a 1cm light path. The increase in absorbance at 340nm was followed over a period of 10 minutes and compared with a blank sample containing everything but the glucose-6- phosphate. The results are shown in the table below: Time (mins) 0 2 4 6 8 10 Absorbance at 340nm With G6P 0 0.12 0.23 0.34 0.44 0.53 Without G6P 0 0.03 0.062 0.082 0.087 0.092
2. Draw a graph to calculate the initial rate of reaction due to enzyme X in absorbance units/min/100μl of extract.
Transcribed Image Text:2. Draw a graph to calculate the initial rate of reaction due to enzyme X in absorbance units/min/100μl of extract.
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