S. No. Title Reference Book Page No.

UNIT I OPERATIONAL MODES OF BIOREACTORS (9)

1. Fed batch cultivation James E. Bailey & David F. Ollis, “Biochemical 600-602
BIO PROCESS ENGINEERING/BT6502 Engineering Fundamentals”, McGraw-Hill.
OBJECTIVES: 2. Cell recycle cultivation James E. Bailey & David F. Ollis, “Biochemical 602-603
To provide the students with the basics of bioreactor engineering. Engineering Fundamentals”, McGraw-Hill.
To develop bioengineering skills for the production of biochemical product using 3. Cell recycle cultivation in James E. Bailey & David F. Ollis, “Biochemical 603-605
integrated biochemical processes. waste water treatment Engineering Fundamentals”, McGraw-Hill.
4. Two stage cultivation. James E. Bailey & David F. Ollis, “Biochemical 605-609
UNIT I OPERATIONAL MODES OF BIOREACTORS 9 Engineering Fundamentals”, McGraw-Hill.
5. Packed bed reactor James E. Bailey & David F. Ollis, “Biochemical 609-610
Fed batch cultivation, Cell recycle cultivation, Cell recycle cultivation in waste water treatment,
two stage cultivation. Packed bed reactor, airlift reactor, fluidized bed reactor and bubble column Engineering Fundamentals”, McGraw-Hill.
reactor. 6. Airlift reactor James E. Bailey & David F. Ollis, “Biochemical 641-645

UNIT II BIOREACTOR SCALE – UP 8 Engineering Fundamentals”, McGraw-Hill.

7. Fluidized bed reactor James E. Bailey & David F. Ollis, “Biochemical 614-617
Regime analysis of bioreactor processes, oxygen mass transfer in bioreactors –microbial oxygen
demands; methods for the determination of mass transfer coefficients;mass transfer correlations. Engineering Fundamentals”, McGraw-Hill.
Scale up criteria for bioreactors based on oxygen transfer, power consumption and impeller tip 8. Bubble column reactor. James E. Bailey & David F. Ollis, “Biochemical 610-614
speed.
Engineering Fundamentals”, McGraw-Hill.
UNIT III BIOREACTOR CONSIDERATION IN ENZYME SYSTEMS 8 UNIT II BIOREACTOR SCALE – UP (8)
9. Regime analysis of bioreactor James M. Lee, “Biochemical Engineering”, PHI, USA. 240-247
Analysis of film and pore diffusion effects on kinetics of immobilized enzyme processes
reactions;formulation of dimensionless groups and calculation of effectiveness factors. Design of 10. Oxygen mass transfer in James M. Lee, “Biochemical Engineering”, PHI, USA. 241-242
immobilized enzyme reactors – packed bed, fluidized bed and membrane reactors. bioreactors
11. Microbial oxygen demands; James M. Lee, “Biochemical Engineering”, PHI, USA. 261-262
UNIT IV MODELLING AND SIMULATION OF BIOPROCESSES 11
12. Methods for the determination James M. Lee, “Biochemical Engineering”, PHI, USA. 264-265
Study of structured models for analysis of various bioprocess – compartmental models,models of of mass transfer coefficients;
cellular energetics and metabolism, single cell models, plasmid replication and plasmid stability 13. Mass transfer correlations. James M. Lee, “Biochemical Engineering”, PHI, USA. 248-250
model. Dynamic simulation of batch, fed batch, steady and transient culture metabolism.
14. Scale up criteria for bioreactors James M. Lee, “Biochemical Engineering”, PHI, USA. 272-274
based on oxygen transfer,
UNIT V RECOMBINANT CELL CULTIVATION 9
power consumption and
impeller tip speed.
Different host vector system for recombinant cell cultivation strategies and advantages. E.coli,
yeast Pichia pastoris/ Saccharomyces cereviseae, Animal cell cultivation, plant cell cultivation, UNIT III BIOREACTOR CONSIDERATION IN ENZYME SYSTEMS (8)
Insect cell cultivation. High cell density cultivation, process strategies, reactor considerations in 15. Analysis of film and pore James M. Lee, “Biochemical Engineering”, PHI, USA. 202,
the above system. diffusion effects on kinetics of 216
immobilized enzyme 208
TOTAL : 45 PERIODS reactions;

OUTCOMES:
16. Formulation of dimensionless Harvey W. Blanch, Douglas S. Clark, “Biochemical 128,
Upon completion of Bioprocess Engineering course graduates will be able to groups and calculation of 122
Engineering”, Marcel Decker Inc.
Select appropriate bioreactor configurations and operation modes based upon the nature of effectiveness factors.
bioproducts and cell lines and other process criteria. 17. Design of immobilized enzyme James E. Bailey & David F. Ollis, “Biochemical 609-611
reactors – packed bed
 Engineering Fundamentals”, McGraw-Hill. 33. High cell density cultivation, James E. Bailey & David F. Ollis, “Biochemical 269-271
process strategies, reactor Engineering Fundamentals”, McGraw-Hill.
18. Fluidized bed James E. Bailey & David F. Ollis, “Biochemical 614-617
considerations in the above
Engineering Fundamentals”, McGraw-Hill. system.
19. Membrane reactors. James E. Bailey & David F. Ollis, “Biochemical 610-614
Engineering Fundamentals”, McGraw-Hill.
UNIT IV MODELLING AND SIMULATION OF BIOPROCESSES (11)
20. Study of structured models for James M. Lee, “Biochemical Engineering”, PHI, USA. 174-176
analysis of various bioprocess -
compartmental models
21. Models of cellular energetics Harvey W. Blanch, Douglas S. Clark, “Biochemical 230-244
and metabolism,
Engineering”, Marcel Decker Inc.
22. Single cell models, Harvey W. Blanch, Douglas S. Clark, “Biochemical 244-246
Engineering”, Marcel Decker Inc.
B.E./B.Tech. DEGREE EXAMINATION, NOVEMBER/DECEMBER 2017
23. Plasmid replication Harvey W. Blanch, Douglas S. Clark, “Biochemical 251-257 FIFTH SEMESTER
Engineering”, Marcel Decker Inc. BIOTECHNOLOGY
BT6502-BIOPROCESS ENGINEERING
24. Plasmid stability model. Harvey W. Blanch, Douglas S. Clark, “Biochemical 251-257 (Regulations 2013)
Engineering”, Marcel Decker Inc. Time: Three Hours Maximum: 100 Marks
Answer ALL questions
25. Dynamic simulation of batch, Harvey W. Blanch, Douglas S. Clark, “Biochemical 277-280 PART A (10X2=20)
Engineering”, Marcel Decker Inc. 1. Give the advantages of Fed Batch culture.
(i) Production of highest possible amount of PHA by balanced cell growth and PHA
26. Fed batch, Harvey W. Blanch, Douglas S. Clark, “Biochemical 305-308 accumulation.
Engineering”, Marcel Decker Inc. (ii) N o n i t r o g e n l i m i t a t i o n d u r i n g t h e a c t i v e g r o w t h phase,
but it is introduced later to enhance PHA production.
27. Steady and transient culture Harvey W. Blanch, Douglas S. Clark, “Biochemical 280-296 (iii) R e s u lt s : m a x i m u m s p e c i f i c g r o w t h r a t e a c h i e v e d c ompared to the
metabolism. 309-322 max specific growth rate in batch cultivation.
Engineering”, Marcel Decker Inc.
UNIT V RECOMBINANT CELL CULTIVATION (9) 2. Draw and label the parts of bubble column reactor.

28. Different host vector system for James E. Bailey & David F. Ollis, “Biochemical 257-259
recombinant cell cultivation Engineering Fundamentals”, McGraw-Hill.
strategies and advantages
29. E.coli, yeast Pichia pastoris/ James E. Bailey & David F. Ollis, “Biochemical 259-261
Saccharomyces cereviseae, Engineering Fundamentals”, McGraw-Hill.
30. Animal cell cultivation, James E. Bailey & David F. Ollis, “Biochemical 261-265
Engineering Fundamentals”, McGraw-Hill.
31. Plant cell cultivation, James E. Bailey & David F. Ollis, “Biochemical 265-267
Engineering Fundamentals”, McGraw-Hill.
32. Insect cell cultivation. James E. Bailey & David F. Ollis, “Biochemical 267-269
Engineering Fundamentals”, McGraw-Hill.
 8. What is idiophase?
The idiophase is the phase in the growth of a culture during which secondary metabolites
are produced.
9. What is elicitor?
Elicitors in plant biology are extrinsic or foreign molecules often associated with plant
pests, diseases or synergistic organisms. Elicitor molecules can attach to special receptor
proteins located on plant cell membranes.
10. What is the difference between transduction and transformation when discussing gene
transfer to bacteria?

3. Define microbial oxygen demand.

Biochemical oxygen demand is a measure of the quantity of oxygen used by microorganisms
(e.g., aerobic bacteria) in the oxidation of organic matter.
4. What happens to the value of KLa when there is an increase in temperature?
KLa increases when there is an increase in temperature. The solubility of oxygen in the
liquid phase of a bioreactor was changed by a ramp change of temperature, and k La was
determined from the resulting return to equilibrium of dissolved oxygen activity.
5. State down the different methods available for immobilizing bio molecules.
(i) Self Assembly
(ii) Self Modification PART B (5X13=65)
(iii) Surface Modification 11. (a) Explain in detail the design and operation of
(iv) Photochemical Immobilization (i) Packed Bed Reactor
(v) Polymer Chemistry (ii) Airlift Reactor
6. What is effectiveness factor? Ans: Refer Reference Book 5 – Pg. No: 609, 610, 641
The effectiveness factor is defined as the ratio of the reaction rate actually observed to the (OR)
reaction rate calculated if the surface reactant concentration persisted throughout the (b) Explain in detail the kinetics of cell cycle cultivation.
Ans: Refer Reference Book 5 – Pg. No: 603-605
interior of the particle, Le., no reactant concentration gradient within the particle.
12. (a) (i) Explain the static method for the determination of mass transfer coefficient in
7. What is epigenetic system and metabolic system in a saturated model?
aerated bioreactor.
Epigenetics is the study of heritable changes in gene function that do not involve changes
Ans: Refer Reference Book 6 – Pg. No: 264-265
in the DNA sequence. The Greek prefix epi- (ἐπι- "over, outside of, around")
(ii) Calculate the Reynolds number and power required to agitate a 10,000 litre tank
in epigenetics implies features that are "on top of" or "in addition to" the
filled with water. The diameter of the tank is 2.0 m and is agitated at 100 rpm by a
traditional genetic basis for inheritance. Epigenetics most often denotes changes in
6-blade turbine type agitator. The agitator is half the tank diameter. (Np=4).
a chromosome that affect gene activity and expression, but can also be used to describe
(OR)
any heritable phenotypic change that does not derive from a modification of the genome,
(b) Explain in detail the regime analysis of a bioreactor process.
such as prions.
Ans: Refer Reference Book 5 – Pg. No: 240-244
Catabolism is the set of metabolic processes that break down large molecules. These
13. (a) Explain the effect of inhibitors, temperature and pH on immobilized enzyme catalytic
include breaking down and oxidizing food molecules. The purpose of the catabolic
activity.
reactions is to provide the energy and components needed by anabolic reactions which
Ans: Refer Reference Book 6 – Pg. No: 209-211
build molecules. (OR)
 (b) Explain the characteristics of the following bioreactors 4. What is the role of draft tube? (APR/MAY 2015)
(i) Membrane Reactors Gas hold-up and mass transfer were examined in a column with and without a draft tube. It was
(ii) Fluidized Bed Reactor. found that the introduction of a draft tube increases the gas hold-up but decreases the volumetric
mass transfer coefficient in Newtonian fluid systems.
Ans: Refer Reference Book 5 – Pg. No: 603-605
14. (a) Explain in detail the compartment model of William to illustrate the properties of cell 5. Write the applications of plug flow reactor.
growth kinetics. Large-scale reactions
Ans: Refer Reference Book 5 – Pg. No: 174-176 Fast reactions
(OR) Homogeneous or heterogeneous reactions
(b) Explain the kinetics of plasmid replication using a generalized model. Continuous production
Ans: Refer Text Book 2 – Pg. No: 251-254 High-temperature reactions
15. (a) Explain the strategies for High-cell density cultivation of E.coli.
6. Write the disadvantages of plug flow reactors. (APR/MAY 2014, APR/MAY 2013)
Ans: Refer text Book 2 – Pg. No: 277-280 Disadvantages of plug flow reactors are that temperatures are hard to control and can result in
(OR) undesirable temperature gradients. PFR maintenance is also more expensive than CSTR
(b) Explain in detail the guidelines for selecting a host vector system. maintenance.
Ans: Refer Reference Book 6 – Pg. No: 257-259
PART C (1X15=15) 7. Write the advantages of plug flow reactors(APR/MAY 2014)
16. (a) Explain in detail with example, how insect cells are used for protein production. Plug flow reactors have a high volumetric unit conversion, run for long periods of time without
Ans: Refer Reference Book 6 – Pg. No: 267-269 maintenance, and the heat transfer rate can be optimized by using more, thinner tubes or fewer,
thicker tubes in parallel.
(OR)
(b) Write notes on the medical and analytical applications of immobilized enzymes.
Ans: Refer Reference Book 5 – Pg. No: 208,216

8. Define Polymerase chain reaction? (APR/MAY 2013)

UNIT 1 OPERATIONAL MODES OF BIOREACTORS The polymerase chain reaction (PCR), a common laboratory technique, employs artificial
TWO MARKS synthesis in a cyclic manner to rapidly and specifically amplify a target DNA fragment from a
pool of DNA.
1. Define residence time distribution.(APR/MAY 2017)
Residence time distribution (RTD) describes the distribution of times required for fluid elements 9. Define fluidization.
to pass through a continuous-flow system. RTD function and associated flow models have been Fluidization is a process similar to liquefaction whereby a granular material is converted from a
widely used to characterize hydrodynamic behavior of a variety of systems encountered in static solid-like state to a dynamic fluid-like state. This process occurs when a fluid (liquid or
agricultural, food, and biological engineering. gas) is passed up through the granular material.

2. Define pasteurization. (APR/MAY 2017, APR/MAY 2016) 10. Define batch cultivation
The treatment of foods or beverages with mild heat, irradiation, or chemical agents to improve A technique used to grow microorganisms or cells. A limited supply of nutrients for growth is
keeping quality or to inactivate disease-causing microorganisms. Originally, Louis Pasteur provided; when these are used up, or some other factor becomes limiting, the culture declines.
observed that spoilage of wine and beer could be prevented by heating them a few minutes at Cells, or products that the organisms have made, can then be harvested from the culture.
122–140°F (50–60°C).
11. Define fed batch cultivation
3. Define sterilization(APR/MAY 2016, APR/MAY 2015) Fed-batch culture is, in the broadest sense, defined as an operational technique in
A form of monetary action in which a central bank or federal reserve attempts to insulate itself biotechnological processes where one or more nutrients (substrates) are fed (supplied) to the
from the foreign exchange market to counteract the effects of a changing monetary base. The bioreactor during cultivation and in which the product(s) remain in the bioreactor until the end of
sterilization process is used to manipulate the value of one domestic currency relative to another, the run.
and is initiated in the forex market.
12. Define cell recycle cultivation 2. Good uniformity of temperature
Continuous cultivation with cell recycling is used to obtain high cell concentration. Knowing that 3. Catalyst can be continuously regenerated with the use of an auxiliary loop
secondary metabolite formation rate is proportional to cell concentration, continuous cultivations
with cell recycling is an alternative to increase the productivity
20. Advantages of fluidized bed reactor
13. What are the two stages of cultivation?
 Uniform Particle Mixing: Due to the intrinsic fluid-like behavior of the solid material,
Autotrophic
Mixotrophic fluidized beds do not experience poor mixing as in packed beds. This complete mixing
allows for a uniform product that can often be hard to achieve in other reactor designs. The
14. Define packed bed reactor
elimination of radial and axial concentration gradients also allows for better fluid-solid
In packed bed reactors, cells are immobilized on large particles. These particles do not move
with the liquid. Packed bed reactors are simple to construct and operate but can suffer from contact, which is essential for reaction efficiency and quality.
blockages and from poor oxygen transfer.  Uniform Temperature Gradients: Many chemical reactions require the addition or
removal of heat. Local hot or cold spots within the reaction bed, often a problem in packed
beds, are avoided in a fluidized situation such as an FBR. In other reactor types, these local
temperature differences, especially hotspots, can result in product degradation. Thus FBRs
are well suited to exothermic reactions. Researchers have also learned that the bed-to-
surface heat transfer coefficients for FBRs are high.
 Ability to Operate Reactor in Continuous State: The fluidized bed nature of these
15. Define airlift reactor
Air-lift bioreactors are similar to bubble column reactors, but differ by the fact that they contain reactors allows for the ability to continuously withdraw product and introduce new reactants
a draft tube. The draft tube is always an inner tube (this kind of air-lift bioreactor is called "air- into the reaction vessel. Operating at a continuous process state allows manufacturers to
lift bioreactor with an internal loop) or an external tube (this kind of air-lift bioreactor is called
"air-lift bioreactor with an external loop) which improves circulation and oxygen transfer and produce their various products more efficiently due to the removal of startup conditions
equalizes shear forces in the reactor. The figure below illustrates the basic structure of an air-lift in batch processes.
bioreactor with an internal loop

16. Define fluidized bed reactor 21. Advantages of bubble column reactor
A fluidized bed reactor (FBR) is a type of reactordevice that can be used to carry out a variety of Provide several advantages during operation and maintenance such as high heat and mass
multiphase chemical reactions. transfer rates, compactness and low operating and maintenance costs.

17. Define bubble column reactor 22. Disadvantages of packed bed reactor
A bubble column reactor is an apparatus used to generate and control gas-liquid chemical 1. Undesired thermal gradients may exist
reactions. It consists of a vertically-arranged cylindrical column filled with liquid, at the bottom 2. Poor temperature control
of which gas is inserted 3. Channeling may occur
4. Unit may be difficult to service and clean
18. Advantages of packed bed reactor
1. High conversion per unit mass of catalyst 23. Disadvantages of airlift reactor
2. Low operating cost 1. Bed-fluid mechanics not well known
3. Continuous operation 2. Severe agitation can result in catalyst destruction and dust formation
3. Uncertain scale-up
19. Advantages of airlift reactor
1. Good mixing 24. Disadvantages of bubble column reactor
  considerable degree of back mixing in both the liquid and the gas phase 27. Define Recycle stream
 short gas phase residence time A process stream that returns material from downstream of a process unit back to the process
 higher pressure drop with respect to packed columns unit.
 rapid decreasing of interfacial area above values of the aspect ratio greater than, say
12, due to the increased rate of coalescence 28. Define bypass stream
It is a stream that skips one or more stages of the process and goes directly to another down
25. Disadvantages of fluidized bed reactor stream stage.

29. Define Purge Stream

 Increased Reactor Vessel Size: Because of the expansion of the bed materials in the Purge stream is a bled off to remove an accumulation of inerts or unwanted material that might
reactor, a larger vessel is often required than that for a packed bed reactor. This larger vessel otherwise buildup in the recycle stream.
means that more must be spent on initial capital costs.
30. Explain external biomass feedback.
 Pumping Requirements and Pressure Drop: The requirement for the fluid to suspend A cell separator such as a centrifuge or settling tank is used to concentrate biomass leaving the
the solid material necessitates that a higher fluid velocity is attained in the reactor. In order to reactor.Once biomass removed cell is recycled.
achieve this, more pumping power and thus higher energy costs are needed. In addition,
PART B
the pressure drop associated with deep beds also requires additional pumping power.
 Particle Entrainment: The high gas velocities present in this style of reactor often result 1. Explain packed bed reactor with applications and write down the design equations.
APR/MAY 2016,APR/MAY 2011,MAY/JUNE 2014
in fine particles becoming entrained in the fluid. These captured particles are then carried out
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
of the reactor with the fluid, where they must be separated. This can be a very difficult and McGraw-Hill.(Page No-609-610)
expensive problem to address depending on the design and function of the reactor. This may
often continue to be a problem even with other entrainment reducing technologies. 2. Explain fluidized bed reactor with applications and write down the design
 Lack of Current Understanding: Current understanding of the actual behavior of the equations. APR/MAY 2011
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
materials in a fluidized bed is rather limited. It is very difficult to predict and calculate the
McGraw-Hill.(Page No-614-617)
complex mass and heat flows within the bed. Due to this lack of understanding, a pilot
plant for new processes is required. Even with pilot plants, the scale-up can be very difficult 3. Explain in detail Airlift reactor, Bubble column reactor with their applications and
and may not reflect what was experienced in the pilot trial. write down the design equations. APR/MAY 2016,APR/MAY 2011,MAY/JUNE
 Erosion of Internal Components: The fluid-like behavior of the fine solid particles 2013,2014
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
within the bed eventually results in the wear of the reactor vessel. This can require expensive
McGraw-Hill.(Page No-641)
maintenance and upkeep for the reaction vessel and pipes.
 Pressure Loss Scenarios: If fluidization pressure is suddenly lost, the surface area of the 4. Describe in detail Fed batch cultivation with the design equations. APR/MAY 2008
bed may be suddenly reduced. This can either be an inconvenience (e.g. making bed restart Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
difficult), or may have more serious implications, such as runaway reactions (e.g. for McGraw-Hill.(Page No-600)
exothermic reactions in which heat transfer is suddenly restricted).
5. Explain in detail cell recycle cultivation.APR/MAY 2008
26. What are the reasons for non-ideality in a fluidized bed reactor? MAY/JUNE 2012 Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
Mass transfer, McGraw-Hill.(Page No-602)
Partitioning.
 6. How the cell recycle cultivation is used in waste water treatment and explain the Although scale up models and the use of characteristic time analysis are potentially attractive, a
stages of cell cultivation APR/MAY 2009 more immediate approach to the rational scaling of reactors is scale down.
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
9. Write the advantages of fed batch mode of fermentation. APR/MAY 2015
McGraw-Hill.(Page No-603)
production of high cell densities due to extension of working time (particularly important in the
production of growth-associated products)
controlled conditions in the provision of substrates during the fermentation, particularly
UNIT 2 BIOREACTOR SCALE UP regarding the concentration of specific substrates as for ex. the carbon source
TWO MARKS control over the production of by-products or catabolite repression effects due to limited
provision of substrates solely required for product formation
1. Define mass transfer coefficient.
Determinations of volumetric mass transfer coefficient were conducted in a three-phase internal- 10. What should be the value of impeller Reynolds number for laminar and turbulent
loop airlift reactor with an enlarged degassing zone. The effectof parameters such as the airflow regime in a bioreactor? MAY/JUNE 2012
rate (riser superficial gas velocities between 0.01 and 0.5 m/s), solids loading (up to 30%, v/v), For ratios that are above 10,000 we know that the mixing regime is fully turbulent. For numbers
solids density (1023 and 1048 kg/m3) and the liquid-phase properties on kLa was studied. between 1000 and 10,000 we know that the mixing environment is transitional. For numbers or
ratios below 1000, the mixing regime is partially laminar or fully laminar.
2. What is scale up? APR/MAY 2016
Generally, fermenters maintain a height to diameter ratio of 2 to 1. If The height to diameter ratio 11. List out the criteria to be followed for scaling up of a bioreactor. MAY/JUNE 2012
remains constant, and then the surface to volume ratio decreases during scale-up.
A. Constant power input per unit volume (P/V = constant).
3. What is scale down?
Although scale up models and the use of characteristic time analysis are potentially attractive, a B. Constant KLa
more immediate approach to the rational scaling of reactors is scale down.
C. Constant mixing quality
4. What are the effects of scale up? D. Constant momentum factor (MF = ND. NWL (D – W)) = constant)
(i) Decreases the relative contribution of surface aeration to oxygen supply.
(ii) Removal of dissolved carbon dioxide removal. E. Similar drop size distribution (ds = constant)

5. Write few mass transfer correlations. APR/MAY 2016 F. Constant impeller tip speed (π NDi = constant)
Correlations for the volumetric mass transfer coefficient with the riser superficial gas velocity
and solids loading were determined for the two solids density and the two liquid-phases. A good G. Constant mixing rate number = (N/K)(Di/Dt)α constant
agreement between experimental data and the calculated values was obtained.
12. What is meant by flooding in bioreactor? APR/MAY 2011
6. Write few bioreactors. When air flow dominates the flow pattern flooding is occur in the bioreactor. Flooding of
Ideal (stirred) Batch reactor Ideal tank reactor, also named Continuous Stirred Tank Reactor fermenter occurs due to inappropriate combination of air flow rate and speed of agitation.
(CSTR) Ideal Plug Flow Reactor (PFR)
7. Write the features of scale up. 13. What are the different types of impeller used in bioreactors? APR/MAY 2011
Maintenance of constant power to volume ratios, constant kla, constant tip speed, a combination 1) High Head Closed Channel Impeller – high-efficiency design for pumping water and other
of mixing time and renolds number and the maintenance of a constant substrate or product level liquids at higher head pressures
(usually dissolved oxygen concentration). 2) Vortex Impeller – Used for pumping stringy solids and debris-laden liquids
3) Centrifugal Screw Impeller – Used for pumping oils and other viscous liquids
8. Write the difference between scale up and scale down. 4) Propeller – Used for pumping high volumes of water at low heads
Scale up 5) Shredder Impeller – Used for chopping solids to smaller pieces when they enter the pump
Generally, fermenters maintain a height to diameter ratio of 2 to 1. IfThe height to diameter ratio 6) Closed Channel Impeller – Used for pumping sewage and wastewater
remains constant, then the surface to volume ratio decreases during scale-up. 7) Mixed Flow Impeller – Used for high volume water pumping at low to medium heads
Scale down 8) Semi-Open Impeller – Used for trash and debris laden liquids
9) Hardened Sand/Slurry Impeller – Used for pumping abrasive liquids
 22. Scale up criteria for bioreactors based on power consumption
14. Write the equation for KLa. MAY/JUNE 2013 P/VαN3Di5/Di3=N3Di2
The increase in dissolved oxygen concentration is given by –
23. Scale up criteria for bioreactors based on impeller tip speed.
dCL / dt = KLa(C*-CL) (ii) NDi
Taking logarithm after Integration of equation (ii) we have
ln(C*-CL) = - KLa.t 24. What are the effects on scale up?
The objective of a successful scale up is a homogeneous environment (temperature, pH and pO2,
15. How will you calculate the power number? nutrients, metabolites, cell density, etc.). This is achieved using optimisations based on:
NP = Power Number = P/(n3Da5r)  Agitation based parameters (mixing times, power input, tip speed)
 Gas based parameters (gassing rates)
16. What are the factors affecting the value of KLa?
• A number of factors have been demonstrated to affect the KLa value. Such factors  Heat transfer
include
– the air-flow rate employed in vessels, 25. write down the benefit of scale down.
– the degree of agitation inside vessels ,  Production of data at much smaller effort and time
– the rheological properties of the culture broth and  Improved Safety ( Smaller is safer )
– the presence of antifoam agents.  Improve the knowledge of Process Characteristics

17. Define oxygen mass transfer in bioreactors. 26. Define dilution rate?
• The majority of fermentation processes are aerobic At steady state the growth rate (µ) of the micro-organism is equal to the dilution rate
• Therefore oxygen is an important requirement in aerobic processes like (D). The dilution rate is defined as the rate of flow of medium over the volume of culture in the
bioreactor.
27. Define Maximal growth rate?
• Therefore, 192 grams of oxygen are required for the complete oxidation of 180 grams of Each microorganism has a maximal growth rate (µMAX) (the rate of growth observed if
glucose. none of the nutrients are limiting). If a dilution rate is chosen that is higher than the maximal
growth rate, the culture will not be able to sustain itself in the bioreactor, and will wash out.
18. Define microbial oxygen demands.
Definition. Standard method for indirect measurement of the amount of organic pollution (that 28. What are the types of bioreactors?
can be oxidized biologically) in a sample of water. BOD test procedure is based on the activities A bioreactor may be classified as batch, fed batch or continuous (e.g. continuous stirredtank
of bacteria and other aerobic microorganisms (microbes), which feed on organic matter in reactor model). An example of a continuous bioreactor is the chemostat
presence of oxygen.
29. Write the advantages of fed batch mode of fermentation.
19. List out the methods for the determination of mass transfer coefficients. production of high cell densities due to extension of working time (particularly important in
 Static the production of growth-associated products)
 Dynamic controlled conditions in the provision of substrates during the fermentation, particularly
 Sulphite oxidation regarding the concentration of specific substrates as for ex. the carbon source
control over the production of by-products or catabolite repression effects due to limited
20. Define mass transfer correlations. provision of substrates solely required for product formation
In engineering, the mass transfer coefficient is a diffusion rate constant that relates the mass
transfer rate, mass transfer area, and concentration change as driving force: Where: kc is the mass 30.Mension theReuse potential of MBR effluent
transfer coefficient [mol/(s. · m2)/(mol/m3)], or m/s.  The UMBR system can provide Good Quality effluent that is completely acceptable for
reuse.
21. Scale up criteria for bioreactors based on oxygen transfer.  The reclaimed water can be directly reused for municipal watering, toilet flushing and car
Q/VαNDi3/Di3=N washing. After the softening treatment, the reclaimed water can be used as a cooling
water supply or processing water.
  Therefore, lots of urban waste water can be effectively harnessed, and moreover, large variables, such as the dimensions of the reactor, can be estimated. PFRs are also sometimes
quantities of water can be saved. As a result, the water industry would move towards a called as Continuous Tubular Reactors (CTRs).
more sustainable future.
3.Define diffusion.

PART B

1. Explain the resistance involved in transport of oxygen from a bubble to biochemical

reaction site. Explain clearly the assumptions made and explain the importance of
oxygen mass transfer determination for aerobic fermentation with suitable
examples. MAY/JUNE 2012
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-240-247)

2. Discuss the methods used for finding kLa in bioreactors. APR/MAY Diffusion is the movement of molecules from a high concentration to a low concentration.
2011,MAY/JUNE 2013 Many molecules diffuse across cell membranes.
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-264-265)
4. Define kinetics. APR/MAY 2015
3. Scale up criteria for bioreactors based on oxygen transfer, power consumption and That part of classical mechanics which deals with the relation between the motions of material
bodies and the forces acting upon them. It is synonymous with dynamics of material bodies.
impeller tip speed. APR/MAY 2011
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-272-274) 5. Define enzyme.
4. Derive the expression for power requirement in an aerated bioreactor and in an Enzymes are biological catalysts, or chemicals that speed up the rate of reaction between
ungassed bioreactors. MAY/JUNE 2012 substances without themselves being consumed in the reaction.
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-274)
6. What is dimensionless number?
A ratio of various physical properties (such as density or heat capacity) and conditions (such as
5. What are the various factors which were followed while scaling up process in the
flow rate or weight) of such nature that the resulting number has no defining units of weight,
bioreactor? And comment on its significance. MAY/JUNE 2013 rate, and so on. Also known as nondimensional parameter.
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-255)
7. What is Damkohler numbers? APR/MAY 2015
6. Problem based on finding kLa in bioreactors. APR/MAY 2015, NOV/DEC 2015 The Damköhler numbers (Da) are dimensionless numbers used in chemical engineering to relate
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-267) chemical reaction timescale to other phenomena occurring in a system.

8. What is Data acquisition?

UNIT 3 BIOREACTOR CONSIDERATION IN ENZYME SYSTEMS Data acquisition is the sampling of the real world to generate data that can be manipulated by a
TWO MARKS computer. Sometimes abbreviated DAQ or DAS, data acquisition typically involves acquisition
of signals and waveforms and processing the signals to obtain desired information.
1. Define metabolism. APR/MAY 2016
The term metabolism refers to all of the chemical reactions by which complex molecules taken 9. Define thiele modulus. APR/MAY 2014, APR/MAY 2011
into an organism are broken down to produce energy and by which energy is used to build up The Thiele modulus was developed by E.W. Thiele in his paper 'Relation between catalytic
complex molecules. activity and size of particle' in 1939. Thiele reasoned that with a large enough particle, the
reaction rate is so rapid that diffusion forces are only able to carry product away from the surface
2. What is PFR? APR/MAY 2016 of the catalyst particle.
The plug flow reactor (PFR) model is used to describe chemical reactions in continuous, flowing
systems. The PFR model is used to predict the behavior of chemical reactors, so that key reactor 10. What is known as internal mass transfer resistance? APR/MAY 2011
The internal mass transfer resistance to either the products or reactants that occurs between the Examples of commonly used matrixes for entrapment are:
external pellet surface and the interior of the pellet.
(1). Polyacrylamide gels
11. Write down the disadvantage of sulphite oxidation method? (2). Cellulose triacetate
Disadvantages i) slow, (3). Agar
ii) effected by surface active agents (4). Gelatin
iii) Rheology of soln not like media
(5). Carrageenan
12. Name any four polymers used in enzyme immobilization. MAY/JUNE 2012 (6). Alginate
(a). Alginate: A natural polymer derived from the cell wall of some algae. Calcium or
15. Write down the factors which influence the pore diffusion of immobilized enzyme.
magnesium alginate is the most commonly used matrix. They are inert and have good water Analysis of the effect of internal diffusion is complicated by such factors as the shape of the
holding capacity. particles, the distribution in size and shape of the pores, the total volume of the pores with
(b). Chitosan and chitin: They are structural polysaccharides occurring naturally in the cell wall respect to the particle volume (porosity, h), the depth to which the pores penetrate the particles
(e.g. pellicular particles)
of fungi and the exoskeleton of Arthropods. The various functional groups in enzymes can bind
to the – OH group of chitin and can form covalent bonds. 16. Compare the role of enzymes and cells in the manufacture of a biochemical product.
(c). Collagen: It is the protenaceous support with good porosity and water holding capacity. The The basic function of an enzyme is to increase the rate of a reaction. Most cellular reactions
side chains of the amino acids in the collagen and that of enzyme can form covalent bonds to occur about a million times faster than they would in the absence of an enzyme. Second,
most enzymes act specifically with only one reactant (called a substrate) to produce products.
permanently hold the enzyme to the support.
(d). Carrageenan: It is a sulfated polysaccharide obtained from some red algae. Their good Enzymes are what make all the chemical reactions in the cell possible. The human body is made
gelling properties together with its high protein holding capacity makes it good support for up of trillions of cells, and there are different cells for differentfunctions. Cells are little bundles
immobilizing enzymes. of chemical reactions. They reproduce, they create energy, and they break molecules down and
build them up.
13. What are the distinct advantages of membrane reactors?
Making a gaseous product in a membrane reactor generally affects the way that pressure affects 17. Define packed bed reactor
the extent of reaction at thermodynamic pseudo-equilibrium. In an ordinary flow reactor, the
A packed bed is a hollow tube, pipe, or other vessel that is filled with a packing material. The
composition of the exhaust gas is determined by the composition of the feed gas and the extent
of reaction. As a result, at pseudo-equilibrium, the extent of reaction is entirely determined by packing can be randomly filled with small objects like Raschig rings or else it can be a
the feed composition and the exhaust equilibrium constant, the latter being determined by the specifically designed structured packing. Packed beds may also contain catalyst particles or
temperature and pressure of the exhaust. In a membrane reactor, the partial pressure of the
components at psueudo-equilibrium are not uniquely determined by the total pressure, exit adsorbents such as zeolite pellets, granular activated carbon, etc.
temperature, and feed composition. There is also a significant (and beneficial) effect that derives The purpose of a packed bed is typically to improve contact between two phases in a chemical or
from the controlled removal of a product or addition of reactant.
similar process. Packed beds can be used in a chemical reactor, a distillation process, or
a scrubber, but packed beds have also been used to store heat in chemical plants. In this case, hot
14. What is the mechanism of entrapment? gases are allowed to escape through a vessel that is packed with a refractory material until the
In this method enzymes are physically entrapped inside a porous matrix. Bonds involved in
packing is hot. Air or other cool gas is then fed back to the plant through the hot bed, thereby
stabilizing the enzyme to the matrix may be covalent or non-covalent. The matrix used will be a
pre-heating the air or gas feed.
water soluble polymer. The form and nature of matrix varies with different enzymes. Pore size of
matrix is adjusted to prevent the loss of enzyme. Pore size of the matrix can be adjusted with the
18. Define fluidized bed reactor
concentration of the polymer used. Agar-agar and carrageenan have comparatively large pore A fluidized bed reactor (FBR) is a type of reactor device that can be used to carry out a variety of
sizes. The greatest disadvantage of this method is that there is a possibility of leakage of low multiphase chemical reactions.
molecular weight enzymes from the matrix.
19. Advantages of packed bed reactor 25.Define membrane reactor.
Membrane bioreactor (MBR) is the combination of amembrane process like microfiltration or
1. High conversion per unit mass of catalyst ultrafiltration with a suspended growth bioreactor, and is now widely used for municipal and
industrial wastewater treatment with plant sizes up to 80,000 population equivalent (i.e. 48
2. Low operating cost million liters per day).

3. Continuous operation 26. What is the significance of Thiele modulus in porous catalyst?
Thiele modulus

20. Advantages of fluidized bed reactor

1. Good mixing

2. Good uniformity of temperature

Effectiveness factor
3. Catalyst can be continuously regenerated with the use of an auxiliary loop

21. Disadvantages of packed bed reactor

 = 1 : the entire pellet volume is reacting at the same high rate because reactant is
1. Undesired thermal gradients may exist able to diffuse quickly through the pellet,
 = 0 : the pellet reacts at a slow rate, since the reactant is unable to penetrate into
2. Poor temperature control the pellet interior.

3. Channeling may occur

27. Name any four polymers used for enzyme immobilization.
4. Unit may be difficult to service and clean The polymers used for enzyme immobilization include vinylsulfone-hydroxymethyl
methacrylate, poly(vinylalcohol) beads, polyacrylamide gels, 2-fluoro-1-
22. Disadvantages of fluidized bed reactor methylpyridinium salt-activated Fractogel, hexamethylacrylate polymer (Sepharon Herma
E, an anion exchange polymer), chlormethylated polystyrene beads, and beaded cellulose.
1. Bed-fluid mechanics not well known Natural polymers, which are very hydrophilic, are popular support materials for enzyme
immobilization since the residues in these polymers contain hydroxyl groups, which are
2. Severe agitation can result in catalyst destruction and dust formation ideal functional groups for participating in covalent bonds.
3. Uncertain scale-up 28.Effectiveness factor
Overall productivity for a first order reaction in a spherical pellet
23. What are the reasons for non-ideality in a fluidized bed reactor? MAY/JUNE 2012
ψ, the “sphericity,” which is a measure of a particle’s nonideality in both shape and roughness. It
is calculated by visualizing a sphere whose volume is equal to the particle’s, and dividing the
surface area of this sphere by the actually measured surface area of the particle.
Effectiveness factor
24. Application of membrane filters
Membrane filtration is a technique that uses a physical barrier, a porousmembrane or filter, to
separate particles in a fluid. Particles are separated on the basis of their size and shape with the
use of pressure and specially designedmembranes with different pore sizes.
 = 1 : the entire pellet volume is reacting at the same high rate because reactant is 3. Explain in detail membrane reactor. NOV/DEC 2015
able to diffuse quickly through the pellet, Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
 = 0 : the pellet reacts at a slow rate, since the reactant is unable to penetrate into McGraw-Hill.(Page No-610)
the pellet interior.
4. How will you design the packed bed reactor for immobilized enzyme reaction?
29. DefineDamköhler number APR/MAY 2011
Damköhler number (Da)is used to determine the significant effect of external diffusion Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
resistance on the rate of enzyme catalytic reaction rate. McGraw-Hill.(Page No-609)

5. How will you design the fluidized bed reactor for immobilized enzyme reaction?
APR/MAY 2011
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-614)

 6. Derive the expression for effectiveness factors. MAY/JUNE 2014

 When Da >> 1, the external diffusion rate is limiting; Refer:Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
 Da << 1, the reaction rate is limiting; Inc.(Page No-122)
 Da ≈ 1, the external diffusion and reaction resistances are comparable.

30. Immobilization technique :Adsorption Explain UNIT 4 MODELLING AND SIMULATION OF BIOPROCESSES
TWO MARKS
Oldest method of enzyme immobilization 1. What is plasmid? APR/MAY 2016
Simplest method of enzyme immobilization A plasmid is an extra-chromosomal DNA molecule separate from the chromosomal DNA which
Nelson &Griffin used charcoal to adsorb invertase is capable of replicating independently of the chromosomal DNA.[1] In many cases, it is circular
Enzymes are adsorbed to external surface of support and double-stranded. Plasmids usually occur naturally in bacteria, but are sometimes found in
Support/carrier may be: eukaryotic organisms (e.g., the 2-micrometre-ring in Saccharomyces cerevisiae).
1. Mineral support (aluminium oxide, clay)
2. Organic support(starch) 2. What are the types of plasmids? APR/MAY 2016
3. Modified sepharose and ion exchange resins Fertility-F-plasmids, which contain tra-genes. They are capable of conjugation. Resistance-(R)
plasmids, which contain genes that can build a resistance against antibiotics or poisons.
Historically known as R-factors, before the nature of plasmids was understood. Col-plasmids,
which contain genes that code for (determine the production of) bacteriocins, proteins that can
PART B kill other bacteria. Degradative plasmids, which enable the digestion of unusual substances, e.g.,
toluene or salicylic acid. Virulence plasmids, which turn the bacterium into a pathogen
1. Describe in detail the analysis of film and pore diffusion effects in enzyme
3. Define Simulation of plasmids
immobilized in porous matrix. APR/MAY 2011,NOV/DEC 2015
The use of plasmids as a technique in molecular biology is supported by bioinformatics software.
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-208-216) These programmes record the DNA sequence of plasmid vectors; help to predict cut sites of
restriction enzymes, and to plan manipulations. Examples of software packages that handle
2. Explain in detail immobilized reactor. NOV/DEC 2015 plasmid maps are Lasergene, GeneConstructionKit, and Vector NTI.
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-606) 4. What is compartmental model? APR/MAY 2015
Compartmental models are, in theory, instantaneously mixed throughout, which means that the
outflow concentration for all solutes is equal to the concentration within the flowing fluid. This
allows ordinary differential equations (ODE's) to be used, for example, to calculate the change in It provides a flexible format for data exchange between different types ofdatabases.
concentration, C, within the unit and at its outflow: It can be helpful to view structured data as semi-structured (for browsingpurposes).
dC/dt = Flow*(Cin - C)/V, where Flow = volume of flow in and out, the volume, V, is a constant The schema can easily be changed
and Cin is the concentration of solute in the entering flow. . The data transfer format may be portable.

5. What is DNA replication? APR/MAY 2015, APR/MAY 2014 13. What are semi structured model?
DNA replication is the process of copying a double-stranded DNA molecule to form two double- The semi-structured model is a database model where there is no separation between the data and
stranded molecules. The process of DNA replication is a fundamental process used by all living the schema, and the amount of structure used depends on the purpose. The advantages of
organisms as it is the basis for biological inheritance. this model are the following: It can represent the information of some data sources that cannot be
constrained by schema.
6. What is dynamic simulation? APR/MAY 2014
A dynamic simulation can be used to estimate or illustrate the response, over time, to a change in 14. Write down the constituents of k and g compartments of the cell in Williams’s model.
the process. This primary concern of this site is dynamic models k compartments consists of RNA pools of small metabolites and g compartments consist of DNA
and proteins.
7. What types of simulations used in process engineering?
Static 15. What is meant by transient culture? MAY/JUNE 2012
Static simulations, typically used in process design, simulate the process at steady state Transientculture means it is short-lived culture (usually less than 24 hours). Ex: I have observed
conditions, usually at the design operating conditions. Time is not a variable. that in daily life people come together in groups for a short time and then disperse. Example, a
Dynamic bus queue or strangers on a train or in an aircraft. Individuals bring their own cultural attributes
Dynamic models consider time as a variable and simulate the process over a period of time. A
to this situation and build common ground with their fellow temporary group members. They
dynamic simulation can be used to estimate or illustrate the response, over time, to a change in
the process. This primary concern of this site is dynamic models form, in my terns, 'transient group culture'.

8. Define bioprocess engineering. 16. What is called promiscuous plasmids?

Bioprocess engineering include the production of biofuels, design and operation of fermentation Plasmids that have transfer systems that allow transfer of DNA to unrelated species are called
systems, development of food processing systems, application and testing of product separation promiscuous plasmids.
technologies, design of instrumentation to monitor and control biological processes.
17. How will you determine the KL value in growth associated model with inhibition.
9. Application of bioprocess engineering? MAY/JUNE 2012
Application areas commonly associated with bioprocess engineering include the production of The K parameter, defined as the ratio of the concentration of undissociated weak acid to the
biofuels, design and operation of fermentation systems, development of food processing systems, number of microorganisms in the medium, was determined. The K value is related to the degree
application and testing of product separation technologies,design of instrumentation to monitor of growth inhibition and provides new insight into the mode of action of weak organic acids
and control biological processes. against the studied yeasts.

10. What is semi-structured model? 18. Define single cell model.

The semi-structured model is a database model. In this model, there is no separation between the The cell, itself, is the biochemical reactor. The engineer often faces the challenge of designing
data and the schema, and the amount of structure used depends on the purpose. macroscopic bioreactors, operating polices and control strategies for the macroscopic reactor.
However, the engineer must recognize that the macroscopic reactor only ‘houses’ the cellular
11. What is fed batch reactor model? bioreactors; the reactions take place under the control of the cell’s own internal regulatory
The plug flow reactor (PFR) model is used to describe chemical reactions in continuous, flowing system. Thus, the design of the macroscopic reactor and its operating policy requires a
systems. The PFR model is used to predict the behavior of chemical reactors, so that key reactor quantitative and explicit linkage between the abiotic and biotic environment; between the
variables, such as the dimensions of the reactor, can be estimated. PFRs are also sometimes macroscopic control policy and the cellular regulatory system.
called as Continuous Tubular Reactors (CTRs).
19. What is plasmid replication?
12. Write the advantages of semi structured model. The replication strategy that a plasmid uses directly affects its copy number, host range, and
It can represent the information of some data sources that cannot be constrainedby schema. incompatibility group.
• Plasmid replication relies on the normal host DNA replication machinery.
• Plasmids will carry distinct origins of replication (oriV) and genes that enable and control the 28. Define bioprocess engineering.
frequency of their replication. Bioprocess engineering include the production of biofuels, design and operation of
fermentation systems, development of food processing systems, application and
20. Define plasmid stability. testing of product separation technologies, design of instrumentation to monitor and
Structural plasmid stability exists, when all generated plasmids have the correct base sequence. control biological processes.

21. Define metabolism. 29. Application of bioprocess engineering?

Metabolism is a term that is used to describe all chemical reactions involved in maintaining the Application areas commonly associated with bioprocess engineering include the
living state of the cells and the organism. Metabolism can be conveniently divided into two production of biofuels, design and operation of fermentation systems, development of
categories: Catabolism - the breakdown of molecules to obtain energy. food processing systems, application and testing of product separation technologies,
design of instrumentation to monitor and control biological processes.
22. Explain fed batch culture
Fed-batch culture is, in the broadest sense, defined as an operational technique in 30. Write the advantages of semi structured model.
biotechnological processes where one or more nutrients (substrates) are fed(supplied) to the It can represent the information of some data sources that cannot be constrained by schema.
bioreactor during cultivation and in which the product(s) remain in the bioreactor until the end of It provides a flexible format for data exchange between different types of databases.
the run. It can be helpful to view structured data as semi-structured (for browsing purposes).
The schema can easily be changed.
23. Explain batch culture The data transfer format may be portable.
batch culture A technique used to grow microorganisms or cells. A limited supply of nutrients
for growth is provided; when these are used up, or some other factor becomes limiting, PART B
the culturedeclines. Cells, or products that the organisms have made, can then be harvested from
the culture. 1. Describe the design consideration to be taken in account during the bio processing
of animal cell culture. NOV/DEC 2015
24. Differentiate steady state and unsteady state culture.what is the difference between Refer: Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
transient and periodic flow? Inc.(Page No-203)
Steady state flow implies a uniform flow field that has no change in properties, e.g.
velocity,thermal properties (such as temperature), etc.With unsteady flow, properties of the flow
arechanging.Transient flow implies an unsteady state flow that is changing or developing as Explain in detail plasmid replication. APR/MAY 2016, APR/MAY 2015,MAY/JUNE 2012
afunction of time – often observed in flow start up. With periodic flows, the flow field is Refer:Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
unsteadybut oscillates around a mean. Inc.(Page No-251-257)

25. What are the origin of plasmid replication? Discuss in detail compartmental model. APR/MAY 2016, MAY/JUNE 2013,2014
The origin of replication (also called the replication origin) is a particular sequence in a genome Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-174)
at which replication is initiated. This can either involve thereplication of DNA in living
organisms such as prokaryotes and eukaryotes, or that of DNA or RNA in viruses, such as
2. Explain in detail the structured model for analysis of various bioprocess.APR/MAY
double-stranded RNA viruses.
2011
26.Define static method Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-174)
Static simulations, typically used in process design, simulate the process at steady state
conditions, usually at the design operating conditions. Time is not a variable. 3. Discuss in detail Single cell model. NOV/DEC 2015
Refer:Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
27. Define Dynamic method Inc.(Page No-244-246)
Dynamic models consider time as a variable and simulate the process over a period of time. A
dynamic simulation can be used to estimate or illustrate the response, over time, to a change in
the process. This primary concern of this site is dynamic models 4. Explain in detail dynamic simulation of batch, fed batch, steady and transient
culture metabolism MAY/JUNE 2013,2014 APR/MAY 2011
 Refer:Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker many simpler proteins can be produced using rDNA in bacterial cultures, more complex proteins
Inc.(Page No-277-280) that are glycosylated(carbohydrate-modified) currently must be made in animal cells

UNIT 5 RECOMBINANT CELL CULTIVATION 9. What is meant by plant cell cultivation?

TWO MARKS Plant cell cultures are typically grown as cell suspension cultures in a liquid medium or as callus
cultures on a solid medium. The culturing of undifferentiated plant cells and calli requires the
1. Define Animal cell culture? APR/MAY 2016, APR/MAY 2015 proper balance of the plant growth hormones auxin and cytokinin.
Animal cell culture (ACC) is the process of culture of animal cells outside the tissue (ex vivo)
from which they were obtained. The process of ACC is carried out under strict laboratory 10. Write down the Application of high cell density cultivation.
In this system the amount of an enzyme determines the glucose release rate and all optimisation
conditions of asepsis, sterility and controlled environment involving temperature, gases and
pressure. can be done at the microwell or deepwell plate stage. Here we show at the example of ADH that
such preoptimised processes simply can be transferred to a rocking-motion-system, without the
need for setting up any extra control. This would provide a simple scale up to the 100 scale. On
2. Define cell culture?
top of this, further modifications can be done for improving the volumetric yield further
Cell culture is the process by which prokaryotic, or eukaryotic cells are grown under controlled
conditions. In practice the term "cell culture" has come to refer to the culturing of cells derived
from multicellular eukaryotes, especially animal cells. 11. Explain the reactor considerations in animal cell cultivation.
 It is critical to find a ‘HAPPY’ environment for cell cultures.
 Happy environment = allows cells to increase in number by undergoing cell division
3. What are the applications of cell culture?
(mitosis).
Mass culture of animal cell lines is fundamental to the manufacture of viral vaccines and many
 Provide the cells with appropriate temp, good substrate for attachment and proper culture
products of biotechnology. Biological products produced by recombinant DNA (rDNA)
technology in animal cell cultures include enzymes, synthetic hormones, immunobiologicals medium.
 Temperature:Cold-blooded vertebrates – 18-25°C,Mammalian cells – 36-37°C
(monoclonal antibodies, interleukins, lymphokines), and anticancer agents.
12. Explain the common features of animal cell bioreactor?
4. Explain high cell density cultivation? APR/MAY 2015 1) Reactor should be gently agitated and aerated. Agitation speed ≈20rpm.
In particular, microbial high cell density cultures have a high metabolic oxygen demand. In these
Bubble-column & airlift reactor operating at high aeration may cause damage of cells
cultures the oxygen transfer rate of the bioreactor determines the maximum biomass
2) Supply of CO2-enriched air
concentration. Unfortunately, the solubility of oxygen is even decreasing with increasing cell
3) Removal of toxic products from metabolism eg lactic acid, ammonium
densities due to a higher viscosity of the cell suspension. Therefore, in order to enhance the
existing oxygen transfer limitation for aerobic high cell density cultivation in disposable
13. List out the advantages of animal cell bioreactor?
bioreactors, an aeration membrane or disposable spargers and baffles have been inserted or the
 Ability to perform post-translational modifications and achieve a product close to that
medium was vibrated.
produced in vivo
 The used of other expression systems (yeast, plant or insect cell) more limited because
5. What is meant by host?
despite their potential economic advantages for culture and higher yields of these
An animal or plant on or in which a parasite or commensal organism lives.
systems, their glycosylation capacities do not resemble those of the mammalian cells.
 Prokaryotes cells can be easily manipulated and grown in large scale but they lack the
6. Define recombinant cell cultivation? APR/MAY 2014 necessary glycosylation machinery and so the proteins produced are not glycosylated.
Relating to or denoting an organism, cell, or genetic material formed by recombination.
 Lower eukaryotes cells produced differ significantly from those present in human
glycoproteins.
7. Advantages of recombinant cell cultivation?
 So mamalian cells are the chosen host for the production of human glycoproteins because
Precise control over growth conditions, batch-to-batch product consistency, a high level of
it has been recognized that they meet the criteria for an appropriate glycosylation of
containment and the ability to produce recombinant proteins in compliance with good
recombinant human glycoprotein.
manufacturing practice.
 The most commonly used cell lines are hamster-derived Chinese hamster ovari (CHO and
baby hamster kidney (BHK) cells
8. Write down the Application of animal cell cultivation. APR/MAY 2014  These cells are chosen because of their favorable growth characteristics in culture both as
Mass culture of animal cell lines is fundamental to the manufacture of viral vaccines and other anchorage-dependent or suspension cells.
products of biotechnology.
14. Define E.coli? 3. Infect the culture at your desired MOI Volume of inoculum needed = MOI ( iu/cell ) X
A bacterium commonly found in the intestines of humans and other animals, some strains of number of cells Titer of virus ( iu/ml )
which can cause severe food poisoning.
21. Process strategies of high cell density cultivation.
15. What is yeast? the application of more advanced control strategies, such as exponential feeding procedures and
A microscopic fungus consisting of single oval cells that reproduce by budding, and capable of feed-back control of the pO2. Pulsing of extra oxygen provides some advantage in overcoming
converting sugar into alcohol and carbon dioxide.A greyish-yellow preparation of the yeast the low oxygen transfer rates.
fungus obtained chiefly from fermented beer, used as a fermenting agent, to raise bread dough,
and as a food supplement. 22. List out the host vector system.
Prokaryotes
16. What is Pichia pastoris? 1) Gram Negative
Pichia pastoris is a species of methylotrophic yeast. Pichia is widely used for protein production 2) Gram Positive
using recombinant DNA techniques. Hence it is used in biochemical and genetic research in Eukaryotes
academia and the biotechnical industry. 1) Yeast
2) Fungi
17. What is Saccharomyces cereviseae? 3) Insect Cells
Saccharomyces cerevisiae is a species of yeast. It has been instrumental to winemaking, baking, 4) Plant Cell Lines
and brewing since ancient times. 5) Animal Cell lines
6) Transgenic Animals
18. Process strategies of animal cell cultivation. 7) Transgenic Plants
(1) cell lines capable of synthesizing the required molecules at high productivities that ensure
low operating cost; 23. What is a recombinant cell?
(2) culture media and bioreactor culture conditions that achieve both the requisite productivity Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of
and meet product quality specifications; genetic recombination (such as molecular cloning) to bring together genetic material from
(3) appropriate on-line and off-line sensors capable of providing information that enhances multiple sources, creating sequences that would not otherwise be found in biological organisms.
process control; and
(4) good understanding of culture performance at different scales to ensure smooth scale-up. 24. What is a primary culture?
Successful implementation also requires appropriate strategies for process development, scale-up Primary culture refers to the stage of the culture after the cells are isolated from the tissue and
and process characterization and validation that enable robust operation and ensure compliance proliferated under the appropriate conditions until they occupy all of the available substrate (i.e.,
with current regulations reach confluence).

19. Process strategies of plant cell cultivation. 25. What does meant to culture cells?
In order to successfully cultivate the plant cells at large scale, several engineering parameters Cell culture is the complex process by which cells are grown under controlled conditions,
such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for generally outside of their natural environment. In practice, the term "cell culture" now refers to
selection of a suitable bioreactor. The media ingredients, their concentrations and the the culturing of cells derived from multi-cellular eukaryotes, especially animal cells.
environmental factors are optimized for maximal synthesis of a desired metabolite. Increased
productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, 26. Define Cell strain?
fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular A cell strain is derived either from a primary culture or a cell line by the selection or cloning of
metabolites. Proper understanding and rigorous analysis of these parameters would pave the way cells having specific properties or characteristics which must be defined. Cell strains are cells
towards the successful commercialization of plant cell bioprocesses.
that have been adapted to culture but, unlike cell lines, have a finite division potential. Non-
immortalized cells stop dividing after 40 to 60 population doublings and, after this, they lose
20. Process strategies of insect cell cultivation. their ability to proliferate (a genetically determined event known as senescence).
1. Seed the experimental culture at between 0.5-1 x 106 cells per ml and allow to grow
overnight. 27.How will you Determine the Cell Viability (for all cell types)
2. Confirm log growth of cells by observation of increase cell counts. If the culture did not grow
After the cells have been suspended into the medium, 0.2 ml of the cell suspension can be mixed
overnight, discard and start again.
with 0.3 ml PBS and 0.5 ml trypan blue (final concentration, 0.2% w/v) in a small test tube. An
aliquot is then placed on a hemacytometer and counted with the compound microscope. The 3. Explain in detail insect cell cultivation APR/MAY 2015
number of viable cells (those not taking up the stain) can be determined and used for initiating Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
the new culture with a precise number of viable cells. I personally find this is a time consuming McGraw-Hill.(Page No-267)
step that does not greatly improve the probability of maintaining healthy cultures but this is
largely because I feel confident in recognizing healthy cells just by examining them in the flask
4. How the high cell density cultivation was processed explain in detail?
with the inverted microscope. Beginners may want to include this step until they gain confidence
in their visual inspection of cells. Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-269)
28.Explain Micro Propagation
It is in vitro asexual propagation of crop plants. This technique is advantages over the
5. In what way recombinant cell cultivated in E.coli explain clearly. APR/MAY 2014
conventional practice of asexual propagation as only a small amount of plant is needed, species
Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
highly resistant to conventional bulk propagation can be propagated by this method and it is non
season dependent. Micro propagation is used for rapid multiplication of stocks, elimination of McGraw-Hill.(Page No-259)
diseases, germ plasm preservation, and induction of mutation.

29.Define Embryo Culture

The embryos are isolated from young seeds and placed on a solid medium containing nutrients
and vitamins. Embryos are cultured at 25°c,first in dark until seedlings are about 2 cm long and
root formation has started, and than in light until the seedlings can be planted in soil.

30.Short notes on Protoplast Culture

It is one of the most significant and recent developments in the field of plant tissue culture. the
protoplast are usually isolated from cultured cell or leaf mesophyll cell by treating them with
enzyme solutions. the isolated protoplast may be used to regenerate the plants directly, or for the
production of somatic hybrids through fusion.

PART B

Discuss animal cell cultivation and explain the products of animal cell culture. APR/MAY
2016, NOV/DEC 2015
Ref: James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-261)

1. Discuss plant cell cultivation.

Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-265)

2. Explain in detail Different host vector system for recombinant cell cultivation.
NOV/DEC 2015,MAY/JUNE 2016, 2014
Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-257)