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The glutamate dehydrogenase (GDH) catalyses the following reaction: +H3N- H с CH₂ CH₂ COO •COO™ + NAD+ Time (min) A340 + H₂O GDH COO CIO CH₂ CH₂ - Vo= 1.1.107 M.s¯¹ EA in 0.1 mL of GDH = 0.33 nkat COO acide glutamique The activity of GDH is monitored in the sense of the formation of glutamate using the following conditions: 0.2 mL of 5 M ammonium sulphate 2.4 mL of buffer at pH 8 0.1 mL of NADH at 6.15 mg.mL-¹ (M = 709 g.mol-¹) 0.2 mL of 1 M a-ketoglutarate solution Warm mixture at 25 °C for 5 min Add 0.1 mL of GDH solution containing 1.6 mg.mL-¹protein to start the reaction. + NH₂+ acide α-cétoglutarique The change in absorbance at 340 nm is monitored, in a 1-cm cuvette, every minute for 10 min. Results are given in the table below: Data: ENADH at 340 nm = 6220 M¹.cm1 4 5 6 1 2 3 1.760 1.718 1.675 1.635 1.595 1.550 + NADH + H* 7 8 10 9 1.510 1.489 1.476 1.451 · Calculate the initial rate Vo of the reaction in M.s¹. · Calculate the enzyme activity of the volume of GDH introduced in the test (in nkat). The answer:

The glutamate dehydrogenase (GDH) catalyses the following reaction: +H3N- H с CH₂ CH₂ COO •COO™ + NAD+ Time (min) A340 + H₂O GDH COO CIO CH₂ CH₂ - Vo= 1.1.107 M.s¯¹ EA in 0.1 mL of GDH = 0.33 nkat COO acide glutamique The activity of GDH is monitored in the sense of the formation of glutamate using the following conditions: 0.2 mL of 5 M ammonium sulphate 2.4 mL of buffer at pH 8 0.1 mL of NADH at 6.15 mg.mL-¹ (M = 709 g.mol-¹) 0.2 mL of 1 M a-ketoglutarate solution Warm mixture at 25 °C for 5 min Add 0.1 mL of GDH solution containing 1.6 mg.mL-¹protein to start the reaction. + NH₂+ acide α-cétoglutarique The change in absorbance at 340 nm is monitored, in a 1-cm cuvette, every minute for 10 min. Results are given in the table below: Data: ENADH at 340 nm = 6220 M¹.cm1 4 5 6 1 2 3 1.760 1.718 1.675 1.635 1.595 1.550 + NADH + H* 7 8 10 9 1.510 1.489 1.476 1.451 · Calculate the initial rate Vo of the reaction in M.s¹. · Calculate the enzyme activity of the volume of GDH introduced in the test (in nkat). The answer:

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
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The glutamate dehydrogenase (GDH) catalyses the following reaction: *H3N- C H - CH₂ - CH₂ COO™ acide glutamique -COO + NAD+ + H₂O Time (min) A340 GDH COO™ с 5 1 2 3 4 1.760 1.718 1.675 1.635 1.595 CH₂ The answer: -1 - Vo 1.1.107 M.s-¹ - EA in 0.1 mL of GDH = 0.33 nkat CH₂ The activity of GDH is monitored in the sense of the formation of glutamate using the following conditions: 0.2 mL of 5 M ammonium sulphate 2.4 mL of buffer at pH 8 0.1 mL of NADH at 6.15 mg.mL-¹ (M = 709 g.mol-¹) 0.2 mL of 1 M a-ketoglutarate solution Warm mixture at 25 °C for 5 min Add 0.1 mL of GDH solution containing 1.6 mg.mL protein to start the reaction. COO CO acide a-cétoglutarique The change in absorbance at 340 nm is monitored, in a 1-cm cuvette, every minute for 10 min. Results are given in the table below: Data ENADH at 340 nm = 6220 M¹.cm¹ -1 + NH4+ NADH + H+ 6 1.550 7 8 10 9 1.510 1.489 1.476 1.451 Calculate the initial rate Vo of the reaction in M.s¹. - Calculate the enzyme activity of the volume of GDH introduced in the test (in nkat).
Transcribed Image Text:The glutamate dehydrogenase (GDH) catalyses the following reaction: *H3N- C H - CH₂ - CH₂ COO™ acide glutamique -COO + NAD+ + H₂O Time (min) A340 GDH COO™ с 5 1 2 3 4 1.760 1.718 1.675 1.635 1.595 CH₂ The answer: -1 - Vo 1.1.107 M.s-¹ - EA in 0.1 mL of GDH = 0.33 nkat CH₂ The activity of GDH is monitored in the sense of the formation of glutamate using the following conditions: 0.2 mL of 5 M ammonium sulphate 2.4 mL of buffer at pH 8 0.1 mL of NADH at 6.15 mg.mL-¹ (M = 709 g.mol-¹) 0.2 mL of 1 M a-ketoglutarate solution Warm mixture at 25 °C for 5 min Add 0.1 mL of GDH solution containing 1.6 mg.mL protein to start the reaction. COO CO acide a-cétoglutarique The change in absorbance at 340 nm is monitored, in a 1-cm cuvette, every minute for 10 min. Results are given in the table below: Data ENADH at 340 nm = 6220 M¹.cm¹ -1 + NH4+ NADH + H+ 6 1.550 7 8 10 9 1.510 1.489 1.476 1.451 Calculate the initial rate Vo of the reaction in M.s¹. - Calculate the enzyme activity of the volume of GDH introduced in the test (in nkat).
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