Biju Patnaik University of Technology 2008-8th Sem B.Tech Protein Engineering - Question Paper

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Total number of printed pages - 7 B.Tech

PEBT 8405

Eighth Semester Examination - 2008 PROTEIN ENGINEERING Full Marks-70

Time: 3 Hours

Answer Question No. 1 which is compulsory and any five from the rest.

The figures in the right-hand margin indicate marks.

Total number of printed pages - 7 B.Tech

PEBT 8405

2x10

1. Answer the following questions :

(a) What is the contribution of conformational entropy of a polypeptide chain of 100 amino acids residues to its free energy ?

(b) Name the four different bond interactions to confer stability to the engineered protein.

(c) What do you mean by Molecular Chaperons ? What role it plays in protein architecture ?

(d) What do you mean by cotton effect in Circular Dichroism ?

(e) How many numbers of primers were used for PCR based site directed mutagenesis to insert a mutation in the DNA ?

(f) What is the approximate frequency of amide-1 band of peptide linkage in IR spectroscopy and what it infers ?

(g) What is anti-idiotopes ? What is its significance in enzyme engineering ?

(h) How many number of a-helices and (3-sheets are present in NAD binding domain ?

(i) The first three rotational Raman lines of linear tri-atomic molecules are at 4.86, 8.14 and 11.36 cm^-1 from the exciting Raman line. Estimate the rotational constant and the moment of inertia of the molecule.

(j) The structure of protein has been determined by X-ray diffraction of a protein crystal. It is found to contain 31% a-heli-ces, 58% (3-sheets and 11% random coils.

From the Circular Dichroism analysis the values are 60 % a-helices, 35% (3-sheets and 5% random coils. What inference will you draw about the structure ?

2. What are the protein engineering targets relating with biosensor research and development ? Briefly explain the strategies of genetically fused protein with suitable examples.

Total number of printed pages - 7 B.Tech

PEBT 8405

4+6

3. What do you mean by site directed Mutagenesis ? Briefly explain the various methods of site directed mutagenesis used for genetic engineering of novel protein. Add a note on the strategies of selection of mutants. 2+5+3

4. (a) Briefly explain the principle, Instrumenta

tion and applications of Mass Spectroscopy for protein analysis. 7

(b) The mass spectra of a constitutional isomer is shown below. It became gas at room temperature. The molecular ion is the small peak at m/z = 102 amu. Name the isomer, which will provide this spectrum. 3

Rel Abund

5. (a) Briefly explain the rational approaches of enzyme engineering for stabilization with reference to protease isolated from Bacillus subtilis ? 7

(b) Briefly explain the designing of lysozyme with reference to its thermo stability. 3

6. (a) Briefly explain the methods used for the

determination of the covalent structure of protein. 6

(b) What is the role of Edmanns reagent in Amino acid sequencing ? 4

7. Write down short notes on any two of the following: 5x2

(a) Ramachandran Plot

(b) Protein Engineering through covalent modifications

(c) Characteristic of IR bands of peptide linkage.

8. What are the various hierarchical level of protein structure ? Briefly explain the various kinds of bonds and interactions at various level of structure. Add a note on thermodynamics of polypeptide chain folding. 4+3+3