Answer Happy

QUESTION 1 (26 MARKS) a) Briefly discuss three (3) types of interactions typically introduced into the structure of a protein when engineering an enzyme for improved thermostability. Predict the type(5) of stabilization that might occur with the introduction of each type of interaction. (6 marks) b) Discuss three (3) advantages of immobilizing enzymes onto a solid support. How does these aspects improve the cost efficiency of manufacturing process? (6 marks) c) The following Scheme 1 depicts the elution profile of a crude enzyme mixture achieved by anion exchange chromatographic technique. Scheme i. State the region(5) containing the largest amount of unwanted protein contaminants. Explain your answer. ii. Identify the region of the onset of eluting the trapped protein. Explain the choice of this spot for introducing the elution buffer. iii. Explain the characteristics of the elution buffer to enable elution of trapped target proteins. (8 marks) iv. Explain the significance of region VI. Based on the above profile, provide justification in terms of quality of the purification process. (2 marks) v. If the sodium dodecyl polyacrylamide gel (SDS-PAGE) electrophoresis show's the eluted protein contains another another protein contaminant, propose and describe the principles of two (2) types of purification methods that may be used to further purify your target enzyme.