Code No: R05322302 Set No.

1
III B.Tech II Semester Regular Examinations, Apr/May 2008
ENZYME ENGINEERING
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
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1. Discuss in detail how the enzyme structure helps in catalysis? [16]

2. The Purification table of purification process is as follows.

SNo Procedure Total protein (mg) Activity(units)
i Crude extract 20,000 4,000,000
II Precipitation (salt) 5,000 3,000,000
iii Precipitation (pH) 4,000 1,000,000
iv Ion exchange chromatography 200 800,000
v Affinity 50 750,000
vi Size-exclusion 45 675,000

(a) From the above table, calculate specific activity of the enzyme solution after
each purification procedure.
(b) Which of the purification procedures used for this enzyme is most effective (i.e.,
gives the greatest relative increase in purity)? Which one is least effective?
(c) Is there any indication based on the results shown in the table that the enzyme
after step 6, is no pure?
(d) What else could be done to estimate the purity of the enzyme? [4+4+4+4]

3. What are the different types of mechanisms that the catalyst functional groups use
in catalysis? [16]

4. A single substrate enzyme -catalysed reaction was studied at fixed temperature pH,
and enzyme concentration, in the presence and absence of a fixed concentration of
an allosteric inhibitor, l. the following results were obtained.
Initial concentration of Initial velocity of reaction
(m mol. l−1 ) (µ mol 1−1 min−1 )
uninhibited inhibited
0.5 266 160
0.67 379 300
1.0 534 534
2.0 711 752
3.0 758 785
4.0 776 794
5.0 784 797

What can you deduce about the reaction mechanism given that enzyme was know
to be trimetric? [16]

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Code No: R05322302 Set No. 1
5. (a) Discuss the criteria to select an immobilization method.
(b) Discuss the advantages and disadvantages of covalent binding method of en-
zyme immobilization. [8+8]

6. Discuss the effect of intra particle difuusion on the reaction kinetics in immobilized
enzyme processes. [16]

7. (a) Discuss the kinetics of Enzyme bound membrane reactors.

(b) Write the essential components of an Enzyme reactor. [8+8]

8. Discuss how enzyme biosensors are superior to conventional biosensors. [16]

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Code No: R05322302 Set No. 2
III B.Tech II Semester Regular Examinations, Apr/May 2008
ENZYME ENGINEERING
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
⋆⋆⋆⋆⋆

1. Write about the one-dimensional and two-dimensional electrophoresis in protein

purification? [16]

2. Explain in detail about the isolation of enzymes from animal cells. [16]

3. What is an active site of an enzyme? How the active site involves in enzyme
substrate complex formation. Explain in detail about the determination of active
site concentration. [16]

4. Show pictorially the Dixon plot for competitive and non competitive inhibitions
and explain in detail. [16]

5. (a) What are the factors that determine the adsorption of enzyme to the supports
in the immobilization of enzymes by adsorption method.
(b) Describe the covalent method of enzyme immobilization on trialcoxysilanes.
[8+8]

6. Discuss the external diffusional effects on the immobilized enzyme kinetics on non-
porous supports. [16]

7. Discuss the use of enzyme bound membrane reactors in the reactions involving
organic solvents. [16]

8. Write an assay on the use of enzymes as biosensors. [16]

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Code No: R05322302 Set No. 3
III B.Tech II Semester Regular Examinations, Apr/May 2008
ENZYME ENGINEERING
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
⋆⋆⋆⋆⋆

1. Write an essay on production of enzymes from plant and animal sources? [16]

2. Explain in detail about the isolation of enzymes from animal cells. [16]

3. Are the following true or false? In each case give in detail explanation for your
conclusions.

(a) Enzymes are specific to only one compound and hence only one reaction.
(b) Substrate specificity is a consequence of the particular shape and charge dis-
tribution of the active site.
(c) Enzymes alter the equilibrium position of the reaction they speed up.
(d) Binding between an enzyme and by two groups on the substrate enables the
enzyme to distinguish between stereoisomers. [4+4+4+4]

4. How much model account for cooperativity and explain alloseric regulation. [16]

5. (a) Write a short notes on prevention of distortion of enzyme in the covalent

method of enzyme immobilization.
(b) What are the commonly used gels or fibre materials used in the entrapment
method of enzyme immobilization. [8+8]

6. Explain about internal mass transfer resistance and how to monitor it in bio-
processes involving immobilized enzyme systems. [16]

7. Derive the mathematical expression for an irreversible and reversible single sub-
strate reaction in a CSTR. [16]

8. Write an assay on the mediated enzyme electrodes with emphasis on metal complex
mediators. [16]

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Code No: R05322302 Set No. 4
III B.Tech II Semester Regular Examinations, Apr/May 2008
ENZYME ENGINEERING
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
⋆⋆⋆⋆⋆

1. What are the remarkable properties of enzymes that differ from other catalysis?
Explain in detail. [16]

2. (a) Explain in detail about the finishing operation of enzyme purification process.
(b) Discuss in detail about the “purification table”? [8+8]

3. Derive the Adiar equation for a tetrameric enzyme consisting of identical subunits,
and deduce the relationships between the apparent binding constants required for
the various types of co-operativity. [16]

4. Discuss allosteric enzymes in terms of cooperativity. [16]

5. Discuss the method,supports,advantages and disadvantages of enzyme immobiliza-

tion by adsorption technique. [16]

6. Discuss the effect of intra particle difuusion on the reaction kinetics in immobilized
enzyme processes. [16]

7. What are the operational and economic advantages with conventional chemical
reactors and enzyme reactors. [16]

8. Write an assay on the mediated enzyme electrodes with emphasis on metal complex
mediators. [16]

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