Reg. No.

b. Lactase, also known as p-galactosidase, catalyses the hydrolysis of lactose to produce

glucose and galactose from milk and whey. Experiments are carried out to determine the
Kinetic parameters for the enzyme. From the given data, evaluate Vr* and Km B.Tech. DEGREE EXAMINATION, NOVEMBER 2016
Third Semester
Lactose concentration 2.50 2.27 1.84 1.35 t.25 0.730 0.460 0.204
(mol.1-1 x 102 I5BT2 lOE _ ENZYME ENGINEERING AND TECHNOLOGY
Reaction Velocity t.94 1.91 1.85 1.80 1.78 1.46 t.t7 0.779 (For the candidates admitted during the academic year 2015 - 2016 onwards)
(mol.l-rmin-r x 103) Note:
(D Part - A should be answered in OMR sheet within first 45 minutes and OMR sheet should be handed
over to hall invigilator at the end of 456 minute.
30. a. Analyze the internal mass tansfer effects in enzymes immobilized with porous support
(ii) Part - B and Part - C should be answered in answer booklet.
materials.
Time: Three Hours Max. Marks: 100
(oR)
b. Discuss the reactor configuration for immobilized enzymes and its applications. PART-A(20x 1:20Marks)
Answer ALL Questions
31. a. Elaborate the techniques available for determining the molecular weight of an enqFme- 1. Which of the following are coenzrymes?
(A) NAD, NADP, FAD, FMN (B) Vitamin, Fe, Cu
(oR) (C) NADPH2, Fe, CO (D) NAD, K CoA
b. Explain the following.
i. Nature of the extraction medium ii. Extraction of membrane bound en-rymes 2. Enzyme X requires Z* n order to caralyze the conversion of substrate Y. The Zn2* is best
identifred as.
32. a. Mention the applications of en-rymes in various industries. (A) Coen-zyme (B) Activator
(C) Substrate (D) Product
(oR)
b. Explain the applications of en-rymes in pharmaceutical and medicine. 3. Which of the following statement is incorrect?
(A) Enzymes hasten the completion of a @) The two terms 'substrate' and 'product'
****,t
reaction signify the starting and ending materials of
a reacti on
(C) Enqymes are effected by the (D) Enzyme exhibit specif,rcity for the reactions
reactions they catalyze they catalyze

4. An example of a substrate specific enq,rne is

(A) Hexokinase @) Thiokinase
(C) Lactase (D) Aminopeptidase

5. Uncompetitive inhibition process is described by

(A) Ko, increases, V-o decreases (B) Kn-, increases, V.* increases
(C) K- decreases, V-* decreases (D) K. remains unchanged V** remains
unchanged

6. An allosteric enryme has

(A) One active site (B) One active site and one allosteric site
(C) Active site and two types of (D) Two types of active sites
allosteric sites
7. The enzyme hexokinase which catalyzes glucose to Glucose - t - phosphate in glycolysis is
inhibited by Glucose - t - phosphate. This is an example of
(A) Feedback allosteric inhibition (B) Positive feedback
(C) Competitiveinhibition (D) Non - competitive inhibition

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 8. If K- : 30mM and V,o : 60mM min-t. What is the initial velocity at a substrate
concentration of 0. lmM? 19. The reaction catalyzed by glucose oxidase is used for
(A) 0.23 (B) 0.1e (A) Analysis of ATP (B) Diagnosis of diabetes mellitus
(c) 0.27 (D) 0.13 (C) Diagnosis of tumor (D) Aralysis of blood clots

9. Which one of the following reaction is used for the purpose of recycling enzymes in 20. The enryme used in confectionary to produce "Soft Corner" is
bioprocess? (A) Amylase (B) Cellulase
(A) Isomerization (B) Immobilization (C) Protease (D) Invertase
(C) Phosphorylation (D) Polymerization

10. The immobilization technique involving physical method is PART -B (5 x 4=20 Marks)
(A) Covalent bond formation dependent (B) Non - covalent bond formation dependent Answer ANY FM Questions
(C) Both A and B (D) Ionic bond formation dependent
21. Explore the effect of enryme and substrate concentration on en-ryme activity.
11. The most commonly employed cross - linked polymer is the
(A) Polyacrylamide gel (B) Collagen 22. An enzyme has a K. of 6.7 x 10-5M. If the V,,o of the preparation is 26 pmoles. lirr.min-r,
(C) Celluloses (D) Cation exchange resin what velocity would be observed in the presence of 6 x I 0a M substrate and 7 x 104 M of
Non - competitive inhibitors with Kr: 5 x 10-a M.
t2. An immobilized enzyme used in CSTR exhibits an effectiveness factor (r1) of 1.5. This
indicates. 23. Describe allosteric enzrymes and explain their characteristics,
(A) The enzymes follows substrate (B) The erlryrne experiences extemal frtm
inhibited Kinetics diffusion limitation 24. Explain the various methods of enzymes immobilization
(C) The en-q/me follows competitive (D) The immobilized enryme is operationally
inhibited Kinetics unstable 25. Illustrate the comprehensive process flow sheet for enryme purification.

13. Which of the following is not considered as the physical method for the cells rupturing? 26. How the molecular weight of an enzyme can be analyzed by Ultracentrifugation?
(A) Milling (B) Homogenization
(C) Ultrasonication (D) Alkali Treatment 27. Highlight the applications of en-zymes in IIFCS production.
14. Diethioteritol is added in extraction media to prevent
(A) Oxidation of sulphahydral PART-C(5x 12=60 Marks)
groups (B) Loss of enryme activity Answer ALL Questions
(C) Stabilization of solubilized enzyme (D) A, B and C
28. a. With illustrations, describe the mode of action of enzrymes, their structural of enzymes, their
15. The molec.ular weight of en4,mes can be determined by using
structural components and characteristics.
(A) SEC (B) rEC
(c) rEF (D) UV - Spectrophotometer (oR)
16. Enzyme TPA or Tissue plasminogen activator is used for
b. Derive the Michaelts-Menten kinetics equation for enryme substrate reactions.
(A) Dissolving blood clots (B) Maintaining plasma contents 29. a. i. Write short notes on the following.
(C) Clearing Eurbidity ofjuices (D) Stimulating thromboplastin production i. Feedback inhibiton
17. Enrymes are often used as Diagnostic tools, because
ii. Turn over number of an en-q.rne
(A) Damaged cells release en-4/mes into (B) The liver releases enzymes that accumulate ii. A marine microorganism contains an enryme that hydrolyzes. 6 - sulfate(s).
the blood so that plasma levels of the in the urine
Glucose -
The assay is based on the rate of glucose formation the en4rme in a cell - free extract has
enrymes are noticeably elevated
(c)
Enzymes destroy damaged cells, so (D) Enzymes destroy pathogenic bacteria so
KineticconstantsofKm:6.lx70aMandV,ou*:3OOpmoleslit'r.minlGalactose-6-
sulfate is acompetitive inhibitor(I). At 10-5 Mgalactose-t-sulfate and 2 x l0-5.glucose
X-rays reveals smaller body organs blood levels of bacteria deuline
- 6- sulfate, V was 1.5 pmol lit-t.min-r. Calculate K1 for galactose - 6- sulfate.
18. Size Exclusion chromatography, separates molecules on the basis of
(A) Size (B) Electricalcharge (oR)
(C) Molecular weight (D) AffiniU
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