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B.Tech./201S/ES/BIO-30S/2/4 NATIONAL INSTITUTE OF TECHNOLOGY (Established by Ministry of Human Resource Development, Yupia, District- Papum Pare, Arunachal Fax: 0360-2284972. Govt. Of India) Pradesh- 791112 Email: [email protected] End Semester Examination (July- Dec, 2015) BT 305: Enzyme Engineering Full Marks: 100 Time: 3 Hr Instructions: 1. Answer as many questions as you can but maximum score will be limited to 100. 2. Please read the instruction carefully (Vmax= Maximum velocity; Km=Michaelis constant; [s] = Substrate; Ve =Velocity) UNIT-I (i) Answer all questions l.(a) How sulfa drug acts as a competitive inhibitor? Prove ~ 2 (b)What is active site? 1 (c) How aspartyl protease can be use for HIV prevention? 2 2. Short note (ANY ONE) (a) Lyophilization (b) Enzyme immune-biosensor (ii) Answer any ONE question (5x1=5) (15x1=15) 1. (a) Mention enzyme specificity. 6 (b) Describe that free energy is a useful thermodynamics function for understanding enzyme. 5 (c) Explain Lineweaver - Bruk equation. 4 2. (a) Explain what are cofactors and coenzymes? 3 (b) Explain enzyme inhibition with all reactions and double reciprocal plots. 10 (c) What are the factors get influence the rate of enzyme action? 2 UNIT-II (i) 1. Answer all questions A solution contains initially 35.0xl0-4 moll," of peptide substrate and1.50ig chymotrypsin in 2.5rnL volume. After 10 minutes, 18.6xl0-4 moll," of peptide substrates remain. Molar mass of chymotrypsin is 45,000 g mol". Calculate rate of reaction, enzyme activity and specific activity and turn over number. 5 2. An enzyme-catalyzed reaction has a Km of ImM and a Vmax of 5nM· S-I. What the reaction velocity when the substrate concentration is (a) 0.35mM (b) 1.6mM. (Note: when units in the numerators and denominator cancel) (ii)Write short notes (ANY THREE) (a) Spectrophotometer (b) Chromatographic assay (3+2) (5x3=15) (c) Gel electrophoresis (d) Ultra centrifugation (e) Ultra filtration UNIT III (i) Answer all questions (a) Explain catabolism and anabolism with an example. 2 (b) At the time of enzyme isolation, cell lysis methods are required. Explain briefly about cell lysis methods. 8 (ii) Answer any ONE question 1. (a) Prove that V. - (15xl=15) 7 Vmax[S] Km+[S] o (b) Mention what are the working principals of biosensor? 7 (c) Explain "all enzymes are protein but all proteins are not enzyme". 1 2. (a) Briefly describe enzyme substrate interaction. 5 (b) Described how different enzymes are classified and named. 8 (c) Prove that Km=[S] When Vo= 1/2Vmax 2 (i) UNIT IV Answer all questions 1. What are the difference between NMR and X-ray crystallography? 10 (ii) Answer any ONE question 1. Short notes (ANY THREE) (5X3=15) Application of enzymes in (a) (b) (c) (d) (e) Biomedical industries Food industries Brewing industries Dairy and paper industries Drug industries 2. Briefly explain what are the enzyme based biosensors has been used for health care. 15