Code No: 7A203/MT

NR
M.Tech. – II Semester Regular Examinations, September, 2008

DOWNSTREAM PROCESSING
(Biotechnology)

Time: 3hours Max. Marks:60

Answer any FIVE questions

All questions carry equal marks
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1. Explain in detail about the characteristics of protein and various
steps to be adopted for recombinant protein purification.

2. Write about different types of centrifuges and how they are scaled
up. Compare the advantages disadvantages centrifugation with
microfiltration.

3.a) Explain the chromatography equipment.

b) Write briefly about two methods of affinity chromatography.

4. Write short notes on the following:

a) Aqueous two phase extraction
b) Ammonium sulphate precipitation.

5. Write notes on the following:

a) Ultra filtration
b) Elution methods in ion exchange chromatography
c) Super critical extraction.

6. 10g of BSA is eluted from an 80 litre Sephadex column which has

a void fraction of 0.4. The concentration in the column peaks after
470 litres are eluted and the maximum concentration is 1.8% of
that originally in the column. Estimate
a) The equilibrium constant for binding the albumin to the
sephadex
b) The number of stages in the column
c) The concentration profile in the column.

7.a) For disruption of baker’s yeast, a homogenizer was found to give a

yield of 49% when operated at 55 Mpa and 85% at 105 Mpa.
Determine the number of passes required to obtain 85% disruption
at 55 Mpa.
Contd….2.,
Code No: 7A203/MT ::2::

b) Pichia pastoris fermentation broth has to be clarified for

separating cells and clear liquid using microfiltration. In
laboratory microfiltration of membrane are 120 sq.cm, 10 ml/min
permeate flux was obtained. Calculate the flux in LMH and
membrane area required for processing 10,000 litres in one shift of
8 hrs.

8. Pencillin F is to be extracted from clarified fermentation broth

using amyl acetate as solvent at pH 4.0. The distribution
coefficient of the system was found to be 32. The initial
concentration of pencillin in the feed is 400 mg/l. The flow rates of
feed and solvent stram was 500 l/h and 30 l/h respectively.
a) How many ideal stages (Counter current contact) are required
to recover 97% pencillin in the feed.
b) If you use 3 counter current stages what will be the percent
recovery?
c) If you use 3 cross current stages with equal solvent flow rate of
10 l/h in each, what will be recover percent?

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