Apr_02_237 - Name: _ Student Number: _ _ THE UNIVERSITY OF...

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Name: ________________________________ Student Number: __________________ _______________________________________________________________________________________ THE UNIVERSITY OF MANITOBA April 25, 2002, 9:00 AM - 12:00 PM Page 1 (of 4) Paper # 557/558 Biochemistry II Lab Section Final Examination Microbiology/Chemistry 60.237/2.237 Examiner: Dr. A. Scoot _______________________________________________________________________________________ 1. Answer ALL questions in the space provided. . 2. The back side of each page provides space for preliminary work. _______________________________________________________________________________________ Marks 6 1. Two different methods of column chromatography were used to isolate hexokinase from a liver homogenate. A 5 mL aliquot of homogenate was applied to column A and another 5 mL applied to column B. Several 10 mL fractions were collected from each column. The enzyme activity in 0.1 mL of a 1 in 50 dilution of the homogenate was found to be 4.0 mmoles glucose-6-phosphate (G6P) produced per min. The protein content of 0.5 mL of a 1 in 50 dilution of the homogenate was found by biuret assay to be 4.0 mg. For column A the enzyme activity was found in fraction 2 where 0.1 mL of a 1 in 10 dilution was found to produce 5.0 mmoles G6P per min and the protein content of 1.0 mL of a 1 in 10 dilution was found to be 2.5 mg. For column B the enzyme activity was in fraction 6 where 0.1 mL was found to produce 20.0 mmoles G6P/min and the protein content of 1.0 mL was found to be 5.0 mg. Which of the two columns would you choose for purification of the enzyme for each of the following purposes? (a) A more pure enzyme or (b) A higher yield of enzyme. Show calculations and cite values including the degrees of purification to support your answer.
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THE UNIVERSITY OF MANITOBA April 25, 2002, 9:00 AM - 12:00 PM Page 2 (of 4) Paper # 557/558 Biochemistry II Lab Section Final Examination Microbiology/Chemistry 60.237/2.237 Examiner: Dr. A. Scoot _______________________________________________________________________________________ Marks 2. A tissue extract containing six proteins, PK, PL, PM, PN, PO, and PP was dissolved in 0.15 M phosphate
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Apr_02_237 - Name: _ Student Number: _ _ THE UNIVERSITY OF...

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