Indian Institute of Technology Kharagpur (IIT-K) 2007 B.Tech Biotechnology Down Stream Processing - Question Paper
INDIAN INSTITUTE OF TECHNOLOGY KHARAGPUR DEPARTMENT OF BIOTECHNOLOGY Spring Mid Semester Examination 2007 Date - 20.2. 2007 Time-2hrs Full Marks-30 No. of students-23 Subject: Down Stream Processing Sub No. BT41012
ATTEMPT ALL THE QUESTIONS
1. a)Enumerate the design equation for protein release during disruption of microbial cell in a high pressure homogenizer. 2
J||t*)What are major problems( five points) in cell disruption by sonication? 2
2.What is the order of relative effectiveness of cations and anions for salting out of proteins? Why (NH4 )2S04 is chosen for salt induced protein precipitation?
From an experiment it was observed that the solubility of albumin is 11.5 and 0.005 g/1 at ( NH4 )2S04 concentration of 2.5M and 3.5M respectively. What is the solubility of albumin at 3.0M ( NH4 )2S04 ? 2+2+3
3. Describe inclusion bodies and how will you isolate inclusion bodies from E.coli and solubilize it. Describe the chromatographic refolding of the solubilize protein from inclusion bodies. 1+1+1 +2
4. Define sedementation coefficient ( S) and what is its unit? Describe the equation related to S and molecular weight of the molecule with all abbreviations.
Calculate the sedimentation coefficient S) of a particle centrifuged at 30,000 rpm in a swinging out rotor mid 'movement of the panicle iToirPthe meniscus of the tube with respect to time is given table below. 1 + 1+3
|
X (cm) |
Time ( sec) x 104 |
|
4 |
0 |
|
5 |
1.04 |
|
6 |
2.16 |
|
7 |
3.24 |
|
8 |
4.32 |
/5.a) You want to isolate peripheral blood mononulclear cells from blood. Which centrifugation( Differential or Density gradient)) you will choose and why? What type of rotor( swinging bucket or near vertical) you will use for this purpose and why? 1+1
b) Describe four important differences between sedimentation velocity and sedimentation equilibrium experiments in ultracentrifuge 2
c)Deduce the equation related to feed flow rate ( Q), settling velocity under gravity ( vg) and sigma factor( ) in case tubular bowl centrifuge. 3
6.Schematically describe the isolation of lactic dehydrogenase from a crude enzyme solution by using two phase aquous system ( PEG3400 derivatized with ligand porcian yellow, and Dextran). 2
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| Earning: Approval pending. |
