JEPPIAAR ENGINEERING COLLEGE

B.TECH – BIOTECHNOLOGY (R- 2013)

BT6502 BIOPROCESS ENGINEERING

III YEAR & V SEM

BATCH: 2016-2020

QUESTION BANK

PREPARED BY

Ms. R. SUGANYA, ASST. PROFESSOR

 VISION OF THE INSTITUTION

 To build Jeppiaar Engineering College as an institution of academic excellence in

technological and management education to become a world class University

MISSION OF THE INSTITUTION

 To excel in teaching and learning, research and innovation by promoting the principles
of scientific analysis and creative thinking.
 To participate in the production, development and dissemination of knowledge and
interact with national and international communities.
 To equip students with values, ethics and life skills needed to enrich their lives and
enable them to meaningfully contribute to the progress of society.
 To prepare students for higher studies and lifelong learning, enrich them with the
practical and entrepreneurial skills necessary to excel as future professionals and
contribute to Nation’s economy

PROGRAM OUTCOMES (PO)

Engineering knowledge: Apply the knowledge of mathematics, science,
PO 1 engineering fundamentals, and an engineering specialization to the solution of
complex engineering problems.
Problem analysis: Identify, formulate, review research literature, and analyze
PO 2 complex engineering problems reaching substantiated conclusions using first
principles of mathematics, natural sciences, and engineering sciences.
Design/development of solutions: Design solutions for complex engineering
problems and design system components or processes that meet the specified
PO 3
needs with appropriate consideration for the public health and safety, and the
cultural, societal, and environmental considerations
Conduct investigations of complex problems: Use research-based knowledge
and research methods including design of experiments, analysis and
PO 4
interpretation of data, and synthesis of the information to provide valid
conclusions.
Modern tool usage: Create, select, and apply appropriate techniques, resources,
PO 5 and modern engineering and IT tools including prediction and modeling to complex
engineering activities with an understanding of the limitations.
The engineer and society: Apply reasoning informed by the contextual
PO 6 knowledge to assess societal, health, safety, legal and cultural issues and the
consequent responsibilities relevant to the professional engineering practice.
Environment and sustainability: Understand the impact of the professional
PO 7 engineering solutions in societal and environmental contexts, and demonstrate
the knowledge of, and need for sustainable development.
Ethics: Apply ethical principles and commit to professional ethics and
PO 8
responsibilities and norms of the engineering practice.
 Individual and team work: Function effectively as an individual, and as a
PO 9
member or leader in diverse teams, and in multidisciplinary settings.
Communication: Communicate effectively on complex engineering activities
with the engineering community and with society at large, such as, being able to
PO 10
comprehend and write effective reports and design documentation, make effective
presentations, and give and receive clear instructions.
Project management and finance: Demonstrate knowledge and understanding
of the engineering and management principles and apply these to one’s own
PO 11
work, as a member and leader in a team, to manage projects and in
multidisciplinary environments.
Life-long learning: Recognize the need for, and have the preparation and ability
PO 12 to engage in independent and life-long learning in the broadest context of
technological change.

VISION OF THE DEPARTMENT

 To pursue excellence in producing bioengineers coupled with research attributes

MISSION OF THE DEPARTMENT

M1 To impart quality education and transform technical knowledge into career opportunities
M2 To establish a bridge between the program and society by fostering technical education
M3 To generate societal conscious technocrats towards community development
To facilitate higher studies and research in order to have an effective career /
M4
entrepreneurship

PROGRAM EDUCATIONAL OBJECTIVES (PEOS)

PEO - 1 To impart knowledge and produce competent graduates in the field of biotechnology
To inculcate professional attributes and ability to integrate engineering issues to broader
PEO - 2
social contexts.
PEO - 3 To connect the program and community by fostering technical education.

PEO - 4 To provide a wide technical exposure to work in an interdisciplinary environment

To prepare the students to have a professional career and motivation towards higher
PEO - 5
education.
 PROGRAM SPECIFIC OUTCOMES (PSOs)

Professional Skills: This programme will provide students with a solid foundation in the
field of Biological Sciences and Chemical engineering enabling them to work on engineering
PSO 1
platforms and applications in Biotechnology as per the requirement of Industries, and
facilitating the students to pursue higher studies.
Problem-solving skills: This programme will assist the students to acquire fundamental
PSO 2 and problem solving knowledge on subjects relevant to Biotechnology thereby encouraging
them to understand emerging and advanced concepts in modern biology.
Successful Career and Entrepreneurship: Graduates of the program will have a strong
successful career and entrepreneurial ability with the blend of inputs from basic science,
PSO 3
engineering and technology, thereby enabling them to translate the technology and tools in
various industries and/or institutes.
 BIO PROCESS ENGINEERING/BT6502
OBJECTIVES:
To provide the students with the basics of bioreactor engineering.
To develop bioengineering skills for the production of biochemical product using
integrated biochemical processes.

UNIT I OPERATIONAL MODES OF BIOREACTORS 9

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
reactor.

UNIT II BIOREACTOR SCALE – UP 8

Regime analysis of bioreactor processes, oxygen mass transfer in bioreactors –microbial oxygen
demands; methods for the determination of mass transfer coefficients;mass transfer correlations.
Scale up criteria for bioreactors based on oxygen transfer, power consumption and impeller tip
speed.

UNIT III BIOREACTOR CONSIDERATION IN ENZYME SYSTEMS 8

Analysis of film and pore diffusion effects on kinetics of immobilized enzyme

reactions;formulation of dimensionless groups and calculation of effectiveness factors. Design of
immobilized enzyme reactors – packed bed, fluidized bed and membrane reactors.

UNIT IV MODELLING AND SIMULATION OF BIOPROCESSES 11

Study of structured models for analysis of various bioprocess – compartmental models,models of

cellular energetics and metabolism, single cell models, plasmid replication and plasmid stability
model. Dynamic simulation of batch, fed batch, steady and transient culture metabolism.

UNIT V RECOMBINANT CELL CULTIVATION 9

Different host vector system for recombinant cell cultivation strategies and advantages. E.coli,
yeast Pichia pastoris/ Saccharomyces cereviseae, Animal cell cultivation, plant cell cultivation,
Insect cell cultivation. High cell density cultivation, process strategies, reactor considerations in
the above system.

TOTAL : 45 PERIODS

OUTCOMES:

Upon completion of Bioprocess Engineering course graduates will be able to

Select appropriate bioreactor configurations and operation modes based upon the nature of
bioproducts and cell lines and other process criteria.
 Apply modeling and simulation of bioprocesses so as to reduce costs and to enhance the
quality of products and systems. Plan a research career or to work in the biotechnology industry
with strong
foundation about bioreactor design and scale-up.
Integrate research lab and Industry; identify problems and seek practical solutions for
large scale implementation of Biotechnology.

TEXT BOOKS:

1. Jens Nielson, John Villadsen and Gunnar Liden, “Bioreaction engineering principles”,2nd
Edition, Kulwer Academic, 2002
2. Harvey W. Blanch, Douglas S. Clark, Biochemical Engineering, Marcel Dekker, Inc

REFERENCES:

1. Anton Moser, “Bioprocess Technology: Kinetics and Reactors”, , Springer Verlag 2011.
2. Tapobrate Panda, “Bioreactors: Analysis and Design”, Tata McGraw Hill, 2011
3. Shijie Liu “Bioprocess Engineering” Elsevier, 2013
4. Atkinson, B, Mavituna, F, “Biochemical Engineering and Biotechnology Handbook”
Macmillan Publishers Ltd, New York, 1992.
5. James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”, McGraw Hill.
6. James M. Lee, “Biochemical Engineering”, PHI, USA 2002.
7. E. Heinzle, A. Biwer and C.Cooney “ Development of Sustainable Bioprocesses” John Wiley
& sons, 2006.

COURSE OUTCOMES (CO)

BT6502: BIOPROCESS ENGINEERING

The course graduates will be able to Select appropriate bioreactor configurations and operation
C302.1 modes based upon the nature of bioproducts and cell lines and other process criteria

The students will be able to Apply modeling and simulation of bioprocesses so as to reduce costs
C302.2
and to enhance the quality of products and systems

The students will be able to Plan a research career or to work in the biotechnology industry with
C302.3
strong foundation about bioreactor design and scale-up.

The students will be able to Integrate research lab and Industry; identify problems and seek
C302.4
practical solutions for large scale implementation of Biotechnology

The students will be able to understand the design of plant cultivation reactors, animal cell
C302.5 cultivation reactors and recombination techniques.
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
Engineering Fundamentals”, McGraw-Hill.
2. Cell recycle cultivation James E. Bailey & David F. Ollis, “Biochemical 602-603
Engineering Fundamentals”, McGraw-Hill.
3. Cell recycle cultivation in James E. Bailey & David F. Ollis, “Biochemical 603-605
waste water treatment Engineering Fundamentals”, McGraw-Hill.
4. Two stage cultivation. James E. Bailey & David F. Ollis, “Biochemical 605-609
Engineering Fundamentals”, McGraw-Hill.
5. Packed bed reactor James E. Bailey & David F. Ollis, “Biochemical 609-610
Engineering Fundamentals”, McGraw-Hill.
6. Airlift reactor James E. Bailey & David F. Ollis, “Biochemical 641-645
Engineering Fundamentals”, McGraw-Hill.
7. Fluidized bed reactor James E. Bailey & David F. Ollis, “Biochemical 614-617
Engineering Fundamentals”, McGraw-Hill.
8. Bubble column reactor. James E. Bailey & David F. Ollis, “Biochemical 610-614
Engineering Fundamentals”, McGraw-Hill.
UNIT II BIOREACTOR SCALE – UP (8)
9. Regime analysis of bioreactor James M. Lee, “Biochemical Engineering”, PHI, USA. 240-247
processes
10. Oxygen mass transfer in James M. Lee, “Biochemical Engineering”, PHI, USA. 241-242
bioreactors
11. Microbial oxygen demands; James M. Lee, “Biochemical Engineering”, PHI, USA. 261-262
12. Methods for the determination James M. Lee, “Biochemical Engineering”, PHI, USA. 264-265
of mass transfer coefficients;
13. Mass transfer correlations. James M. Lee, “Biochemical Engineering”, PHI, USA. 248-250
14. Scale up criteria for bioreactors James M. Lee, “Biochemical Engineering”, PHI, USA. 272-274
based on oxygen transfer,
power consumption and
impeller tip speed.
UNIT III BIOREACTOR CONSIDERATION IN ENZYME SYSTEMS (8)
15. Analysis of film and pore James M. Lee, “Biochemical Engineering”, PHI, USA. 202,
diffusion effects on kinetics of 216
immobilized enzyme 208
reactions;

16. Formulation of dimensionless Harvey W. Blanch, Douglas S. Clark, “Biochemical 128,

groups and calculation of 122
Engineering”, Marcel Decker Inc.
effectiveness factors.
17. Design of immobilized enzyme James E. Bailey & David F. Ollis, “Biochemical 609-611
reactors – packed bed
 Engineering Fundamentals”, McGraw-Hill.
18. Fluidized bed James E. Bailey & David F. Ollis, “Biochemical 614-617
Engineering Fundamentals”, McGraw-Hill.
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.
23. Plasmid replication Harvey W. Blanch, Douglas S. Clark, “Biochemical 251-257
Engineering”, Marcel Decker Inc.
24. Plasmid stability model. Harvey W. Blanch, Douglas S. Clark, “Biochemical 251-257
Engineering”, Marcel Decker Inc.
25. Dynamic simulation of batch, Harvey W. Blanch, Douglas S. Clark, “Biochemical 277-280
Engineering”, Marcel Decker Inc.
26. Fed batch, Harvey W. Blanch, Douglas S. Clark, “Biochemical 305-308
Engineering”, Marcel Decker Inc.
27. Steady and transient culture Harvey W. Blanch, Douglas S. Clark, “Biochemical 280-296
metabolism. 309-322
Engineering”, Marcel Decker Inc.
UNIT V RECOMBINANT CELL CULTIVATION (9)

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.
33. High cell density cultivation, James E. Bailey & David F. Ollis, “Biochemical 269-271
process strategies, reactor Engineering Fundamentals”, McGraw-Hill.
considerations in the above
system.

B.E./B.Tech. DEGREE EXAMINATION, NOVEMBER/DECEMBER 2017

FIFTH SEMESTER
BIOTECHNOLOGY
BT6502-BIOPROCESS ENGINEERING
(Regulations 2013)
Time: Three Hours Maximum: 100 Marks
Answer ALL questions
PART A (10X2=20)
1. Give the advantages of Fed Batch culture.
(i) Production of highest possible amount of PHA by balanced cell growth and PHA
accumulation.
(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.
(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
max specific growth rate in batch cultivation.

2. Draw and label the parts of bubble column reactor.

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
(iii) Surface Modification
(iv) Photochemical Immobilization
(v) Polymer Chemistry
6. What is effectiveness factor?
The effectiveness factor is defined as the ratio of the reaction rate actually observed to the
reaction rate calculated if the surface reactant concentration persisted throughout the
interior of the particle, Le., no reactant concentration gradient within the particle.
7. What is epigenetic system and metabolic system in a saturated model?
Epigenetics is the study of heritable changes in gene function that do not involve changes
in the DNA sequence. The Greek prefix epi- (ἐπι- "over, outside of, around")
in epigenetics implies features that are "on top of" or "in addition to" the
traditional genetic basis for inheritance. Epigenetics most often denotes changes in
a chromosome that affect gene activity and expression, but can also be used to describe
any heritable phenotypic change that does not derive from a modification of the genome,
such as prions.
Catabolism is the set of metabolic processes that break down large molecules. These
include breaking down and oxidizing food molecules. The purpose of the catabolic
reactions is to provide the energy and components needed by anabolic reactions which
build molecules.
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?

PART B (5X13=65)
11. (a) Explain in detail the design and operation of
(i) Packed Bed Reactor
(ii) Airlift Reactor
Ans: Refer Reference Book 5 – Pg. No: 609, 610, 641
(OR)
(b) Explain in detail the kinetics of cell cycle cultivation.
Ans: Refer Reference Book 5 – Pg. No: 603-605
12. (a) (i) Explain the static method for the determination of mass transfer coefficient in
aerated bioreactor.
Ans: Refer Reference Book 6 – Pg. No: 264-265
(ii) Calculate the Reynolds number and power required to agitate a 10,000 litre tank
filled with water. The diameter of the tank is 2.0 m and is agitated at 100 rpm by a
6-blade turbine type agitator. The agitator is half the tank diameter. (Np=4).
(OR)
(b) Explain in detail the regime analysis of a bioreactor process.
Ans: Refer Reference Book 5 – Pg. No: 240-244
13. (a) Explain the effect of inhibitors, temperature and pH on immobilized enzyme catalytic
activity.
Ans: Refer Reference Book 6 – Pg. No: 209-211
(OR)
 (b) Explain the characteristics of the following bioreactors
(i) Membrane Reactors
(ii) Fluidized Bed Reactor.
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
growth kinetics.
Ans: Refer Reference Book 5 – Pg. No: 174-176
(OR)
(b) Explain the kinetics of plasmid replication using a generalized model.
Ans: Refer Text Book 2 – Pg. No: 251-254
15. (a) Explain the strategies for High-cell density cultivation of E.coli.
Ans: Refer text Book 2 – Pg. No: 277-280
(OR)
(b) Explain in detail the guidelines for selecting a host vector system.
Ans: Refer Reference Book 6 – Pg. No: 257-259
PART C (1X15=15)
16. (a) Explain in detail with example, how insect cells are used for protein production.
Ans: Refer Reference Book 6 – Pg. No: 267-269
(OR)
(b) Write notes on the medical and analytical applications of immobilized enzymes.
Ans: Refer Reference Book 5 – Pg. No: 208,216

UNIT 1 OPERATIONAL MODES OF BIOREACTORS

TWO MARKS

1. Define residence time distribution.(APR/MAY 2017)

Residence time distribution (RTD) describes the distribution of times required for fluid elements
to pass through a continuous-flow system. RTD function and associated flow models have been
widely used to characterize hydrodynamic behavior of a variety of systems encountered in
agricultural, food, and biological engineering.

2. Define pasteurization. (APR/MAY 2017, APR/MAY 2016)

The treatment of foods or beverages with mild heat, irradiation, or chemical agents to improve
keeping quality or to inactivate disease-causing microorganisms. Originally, Louis Pasteur
observed that spoilage of wine and beer could be prevented by heating them a few minutes at
122–140°F (50–60°C).

3. Define sterilization(APR/MAY 2016, APR/MAY 2015)

A form of monetary action in which a central bank or federal reserve attempts to insulate itself
from the foreign exchange market to counteract the effects of a changing monetary base. The
sterilization process is used to manipulate the value of one domestic currency relative to another,
and is initiated in the forex market.
4. What is the role of draft tube? (APR/MAY 2015)
Gas hold-up and mass transfer were examined in a column with and without a draft tube. It was
found that the introduction of a draft tube increases the gas hold-up but decreases the volumetric
mass transfer coefficient in Newtonian fluid systems.

5. Write the applications of plug flow reactor.

Large-scale reactions
Fast reactions
Homogeneous or heterogeneous reactions
Continuous production
High-temperature reactions

6. Write the disadvantages of plug flow reactors. (APR/MAY 2014, APR/MAY 2013)
Disadvantages of plug flow reactors are that temperatures are hard to control and can result in
undesirable temperature gradients. PFR maintenance is also more expensive than CSTR
maintenance.

7. Write the advantages of plug flow reactors(APR/MAY 2014)

Plug flow reactors have a high volumetric unit conversion, run for long periods of time without
maintenance, and the heat transfer rate can be optimized by using more, thinner tubes or fewer,
thicker tubes in parallel.

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

The polymerase chain reaction (PCR), a common laboratory technique, employs artificial
synthesis in a cyclic manner to rapidly and specifically amplify a target DNA fragment from a
pool of DNA.

9. Define fluidization.
Fluidization is a process similar to liquefaction whereby a granular material is converted from a
static solid-like state to a dynamic fluid-like state. This process occurs when a fluid (liquid or
gas) is passed up through the granular material.

10. Define batch cultivation

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, the culture declines.
Cells, or products that the organisms have made, can then be harvested from the culture.

11. Define fed batch cultivation

Fed-batch culture is, in the broadest sense, defined as an operational technique in
biotechnological processes where one or more nutrients (substrates) are fed (supplied) to the
bioreactor during cultivation and in which the product(s) remain in the bioreactor until the end of
the run.
12. Define cell recycle cultivation
Continuous cultivation with cell recycling is used to obtain high cell concentration. Knowing that
secondary metabolite formation rate is proportional to cell concentration, continuous cultivations
with cell recycling is an alternative to increase the productivity

13. What are the two stages of cultivation?

Autotrophic
Mixotrophic

14. Define packed bed reactor

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
blockages and from poor oxygen transfer.

15. Define airlift reactor

Air-lift bioreactors are similar to bubble column reactors, but differ by the fact that they contain
a draft tube. The draft tube is always an inner tube (this kind of air-lift bioreactor is called "air-
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
equalizes shear forces in the reactor. The figure below illustrates the basic structure of an air-lift
bioreactor with an internal loop

16. Define fluidized bed reactor

A fluidized bed reactor (FBR) is a type of reactordevice that can be used to carry out a variety of
multiphase chemical reactions.

17. Define bubble column reactor

A bubble column reactor is an apparatus used to generate and control gas-liquid chemical
reactions. It consists of a vertically-arranged cylindrical column filled with liquid, at the bottom
of which gas is inserted

18. Advantages of packed bed reactor

1. High conversion per unit mass of catalyst
2. Low operating cost
3. Continuous operation

19. Advantages of airlift reactor

1. Good mixing
2. Good uniformity of temperature
3. Catalyst can be continuously regenerated with the use of an auxiliary loop

20. Advantages of fluidized bed reactor

 Uniform Particle Mixing: Due to the intrinsic fluid-like behavior of the solid material,
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
elimination of radial and axial concentration gradients also allows for better fluid-solid
contact, which is essential for reaction efficiency and quality.
 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
reactors allows for the ability to continuously withdraw product and introduce new reactants
into the reaction vessel. Operating at a continuous process state allows manufacturers to
produce their various products more efficiently due to the removal of startup conditions
in batch processes.

21. Advantages of bubble column reactor

Provide several advantages during operation and maintenance such as high heat and mass
transfer rates, compactness and low operating and maintenance costs.

22. Disadvantages of packed bed reactor

1. Undesired thermal gradients may exist
2. Poor temperature control
3. Channeling may occur
4. Unit may be difficult to service and clean

23. Disadvantages of airlift reactor

1. Bed-fluid mechanics not well known
2. Severe agitation can result in catalyst destruction and dust formation
3. Uncertain scale-up

24. Disadvantages of bubble column reactor

  considerable degree of back mixing in both the liquid and the gas phase
 short gas phase residence time
 higher pressure drop with respect to packed columns
 rapid decreasing of interfacial area above values of the aspect ratio greater than, say
12, due to the increased rate of coalescence

25. Disadvantages of fluidized bed reactor

 Increased Reactor Vessel Size: Because of the expansion of the bed materials in the
reactor, a larger vessel is often required than that for a packed bed reactor. This larger vessel
means that more must be spent on initial capital costs.
 Pumping Requirements and Pressure Drop: The requirement for the fluid to suspend
the solid material necessitates that a higher fluid velocity is attained in the reactor. In order to
achieve this, more pumping power and thus higher energy costs are needed. In addition,
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
in fine particles becoming entrained in the fluid. These captured particles are then carried out
of the reactor with the fluid, where they must be separated. This can be a very difficult and
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.
 Lack of Current Understanding: Current understanding of the actual behavior of the
materials in a fluidized bed is rather limited. It is very difficult to predict and calculate the
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
and may not reflect what was experienced in the pilot trial.
 Erosion of Internal Components: The fluid-like behavior of the fine solid particles
within the bed eventually results in the wear of the reactor vessel. This can require expensive
maintenance and upkeep for the reaction vessel and pipes.
 Pressure Loss Scenarios: If fluidization pressure is suddenly lost, the surface area of the
bed may be suddenly reduced. This can either be an inconvenience (e.g. making bed restart
difficult), or may have more serious implications, such as runaway reactions (e.g. for
exothermic reactions in which heat transfer is suddenly restricted).

26. What are the reasons for non-ideality in a fluidized bed reactor? MAY/JUNE 2012
Mass transfer,
Partitioning.
27. Define Recycle stream
A process stream that returns material from downstream of a process unit back to the process
unit.

28. Define bypass stream

It is a stream that skips one or more stages of the process and goes directly to another down
stream stage.

29. Define Purge Stream

Purge stream is a bled off to remove an accumulation of inerts or unwanted material that might
otherwise buildup in the recycle stream.

30. Explain external biomass feedback.

A cell separator such as a centrifuge or settling tank is used to concentrate biomass leaving the
reactor.Once biomass removed cell is recycled.

PART B

1. Explain packed bed reactor with applications and write down the design equations.
APR/MAY 2016,APR/MAY 2011,MAY/JUNE 2014
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-609-610)

2. Explain fluidized bed reactor with applications and write down the design
equations. APR/MAY 2011
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-614-617)

3. Explain in detail Airlift reactor, Bubble column reactor with their applications and
write down the design equations. APR/MAY 2016,APR/MAY 2011,MAY/JUNE
2013,2014
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-641)

4. Describe in detail Fed batch cultivation with the design equations. APR/MAY 2008
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-600)

5. Explain in detail cell recycle cultivation.APR/MAY 2008

Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-602)
 6. How the cell recycle cultivation is used in waste water treatment and explain the
stages of cell cultivation APR/MAY 2009
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-603)

UNIT 2 BIOREACTOR SCALE UP

TWO MARKS

1. Define mass transfer coefficient.

Determinations of volumetric mass transfer coefficient were conducted in a three-phase internal-
loop airlift reactor with an enlarged degassing zone. The effectof parameters such as the airflow
rate (riser superficial gas velocities between 0.01 and 0.5 m/s), solids loading (up to 30%, v/v),
solids density (1023 and 1048 kg/m3) and the liquid-phase properties on kLa was studied.

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
remains constant, and then the surface to volume ratio decreases during scale-up.

3. What is scale down?

Although scale up models and the use of characteristic time analysis are potentially attractive, a
more immediate approach to the rational scaling of reactors is scale down.

4. What are the effects of scale up?

(i) Decreases the relative contribution of surface aeration to oxygen supply.
(ii) Removal of dissolved carbon dioxide removal.

5. Write few mass transfer correlations. APR/MAY 2016

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
agreement between experimental data and the calculated values was obtained.

6. Write few bioreactors.

Ideal (stirred) Batch reactor Ideal tank reactor, also named Continuous Stirred Tank Reactor
(CSTR) Ideal Plug Flow Reactor (PFR)
7. Write the features of scale up.
Maintenance of constant power to volume ratios, constant kla, constant tip speed, a combination
of mixing time and renolds number and the maintenance of a constant substrate or product level
(usually dissolved oxygen concentration).

8. Write the difference between scale up and scale down.

Scale up
Generally, fermenters maintain a height to diameter ratio of 2 to 1. IfThe height to diameter ratio
remains constant, then the surface to volume ratio decreases during scale-up.
Scale down
Although scale up models and the use of characteristic time analysis are potentially attractive, a
more immediate approach to the rational scaling of reactors is scale down.

9. Write the advantages of fed batch mode of fermentation. APR/MAY 2015

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
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
provision of substrates solely required for product formation

10. What should be the value of impeller Reynolds number for laminar and turbulent
regime in a bioreactor? MAY/JUNE 2012
For ratios that are above 10,000 we know that the mixing regime is fully turbulent. For numbers
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.

11. List out the criteria to be followed for scaling up of a bioreactor. MAY/JUNE 2012

A. Constant power input per unit volume (P/V = constant).

B. Constant KLa

C. Constant mixing quality

D. Constant momentum factor (MF = ND. NWL (D – W)) = constant)

E. Similar drop size distribution (ds = constant)

F. Constant impeller tip speed (π NDi = constant)

G. Constant mixing rate number = (N/K)(Di/Dt)α constant

12. What is meant by flooding in bioreactor? APR/MAY 2011

When air flow dominates the flow pattern flooding is occur in the bioreactor. Flooding of
fermenter occurs due to inappropriate combination of air flow rate and speed of agitation.

13. What are the different types of impeller used in bioreactors? APR/MAY 2011
1) High Head Closed Channel Impeller – high-efficiency design for pumping water and other
liquids at higher head pressures
2) Vortex Impeller – Used for pumping stringy solids and debris-laden liquids
3) Centrifugal Screw Impeller – Used for pumping oils and other viscous liquids
4) Propeller – Used for pumping high volumes of water at low heads
5) Shredder Impeller – Used for chopping solids to smaller pieces when they enter the pump
6) Closed Channel Impeller – Used for pumping sewage and wastewater
7) Mixed Flow Impeller – Used for high volume water pumping at low to medium heads
8) Semi-Open Impeller – Used for trash and debris laden liquids
9) Hardened Sand/Slurry Impeller – Used for pumping abrasive liquids
14. Write the equation for KLa. MAY/JUNE 2013
The increase in dissolved oxygen concentration is given by –

dCL / dt = KLa(C*-CL) (ii)

Taking logarithm after Integration of equation (ii) we have
ln(C*-CL) = - KLa.t

15. How will you calculate the power number?

NP = Power Number = P/(n3Da5r)

16. What are the factors affecting the value of KLa?

• A number of factors have been demonstrated to affect the KLa value. Such factors
include
– the air-flow rate employed in vessels,
– the degree of agitation inside vessels ,
– the rheological properties of the culture broth and
– the presence of antifoam agents.

17. Define oxygen mass transfer in bioreactors.

• The majority of fermentation processes are aerobic
• Therefore oxygen is an important requirement in aerobic processes like

• Therefore, 192 grams of oxygen are required for the complete oxidation of 180 grams of
glucose.

18. Define microbial oxygen demands.

Definition. Standard method for indirect measurement of the amount of organic pollution (that
can be oxidized biologically) in a sample of water. BOD test procedure is based on the activities
of bacteria and other aerobic microorganisms (microbes), which feed on organic matter in
presence of oxygen.

19. List out the methods for the determination of mass transfer coefficients.
 Static
 Dynamic
 Sulphite oxidation

20. Define mass transfer correlations.

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
transfer coefficient [mol/(s. · m2)/(mol/m3)], or m/s.

21. Scale up criteria for bioreactors based on oxygen transfer.

Q/VαNDi3/Di3=N
22. Scale up criteria for bioreactors based on power consumption
P/VαN3Di5/Di3=N3Di2

23. Scale up criteria for bioreactors based on impeller tip speed.

NDi

24. What are the effects on scale up?

The objective of a successful scale up is a homogeneous environment (temperature, pH and pO2,
nutrients, metabolites, cell density, etc.). This is achieved using optimisations based on:
 Agitation based parameters (mixing times, power input, tip speed)
 Gas based parameters (gassing rates)
 Heat transfer

25. write down the benefit of scale down.

 Production of data at much smaller effort and time
 Improved Safety ( Smaller is safer )
 Improve the knowledge of Process Characteristics

26. Define dilution rate?

At steady state the growth rate (µ) of the micro-organism is equal to the dilution rate
(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?
Each microorganism has a maximal growth rate (µMAX) (the rate of growth observed if
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.

28. What are the types of bioreactors?

A bioreactor may be classified as batch, fed batch or continuous (e.g. continuous stirredtank
reactor model). An example of a continuous bioreactor is the chemostat

29. Write the advantages of fed batch mode of fermentation.

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
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
provision of substrates solely required for product formation

30.Mension theReuse potential of MBR effluent

 The UMBR system can provide Good Quality effluent that is completely acceptable for
reuse.
 The reclaimed water can be directly reused for municipal watering, toilet flushing and car
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
quantities of water can be saved. As a result, the water industry would move towards a
more sustainable future.

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

2011,MAY/JUNE 2013
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-264-265)

3. Scale up criteria for bioreactors based on oxygen transfer, power consumption and
impeller tip speed. APR/MAY 2011
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-272-274)
4. Derive the expression for power requirement in an aerated bioreactor and in an
ungassed bioreactors. MAY/JUNE 2012
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-274)

5. What are the various factors which were followed while scaling up process in the
bioreactor? And comment on its significance. MAY/JUNE 2013
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-255)

6. Problem based on finding kLa in bioreactors. APR/MAY 2015, NOV/DEC 2015

Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-267)

UNIT 3 BIOREACTOR CONSIDERATION IN ENZYME SYSTEMS

TWO MARKS

1. Define metabolism. APR/MAY 2016

The term metabolism refers to all of the chemical reactions by which complex molecules taken
into an organism are broken down to produce energy and by which energy is used to build up
complex molecules.

2. What is PFR? APR/MAY 2016

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
variables, such as the dimensions of the reactor, can be estimated. PFRs are also sometimes
called as Continuous Tubular Reactors (CTRs).

3.Define diffusion.

Diffusion is the movement of molecules from a high concentration to a low concentration.

Many molecules diffuse across cell membranes.

4. Define kinetics. APR/MAY 2015

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.

5. Define enzyme.
Enzymes are biological catalysts, or chemicals that speed up the rate of reaction between
substances without themselves being consumed in the reaction.

6. What is dimensionless number?

A ratio of various physical properties (such as density or heat capacity) and conditions (such as
flow rate or weight) of such nature that the resulting number has no defining units of weight,
rate, and so on. Also known as nondimensional parameter.

7. What is Damkohler numbers? APR/MAY 2015

The Damköhler numbers (Da) are dimensionless numbers used in chemical engineering to relate
chemical reaction timescale to other phenomena occurring in a system.

8. What is Data acquisition?

Data acquisition is the sampling of the real world to generate data that can be manipulated by a
computer. Sometimes abbreviated DAQ or DAS, data acquisition typically involves acquisition
of signals and waveforms and processing the signals to obtain desired information.

9. Define thiele modulus. APR/MAY 2014, APR/MAY 2011

The Thiele modulus was developed by E.W. Thiele in his paper 'Relation between catalytic
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
of the catalyst particle.

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
external pellet surface and the interior of the pellet.

11. Write down the disadvantage of sulphite oxidation method?

Disadvantages i) slow,
ii) effected by surface active agents
iii) Rheology of soln not like media

12. Name any four polymers used in enzyme immobilization. MAY/JUNE 2012
(a). Alginate: A natural polymer derived from the cell wall of some algae. Calcium or
magnesium alginate is the most commonly used matrix. They are inert and have good water
holding capacity.
(b). Chitosan and chitin: They are structural polysaccharides occurring naturally in the cell wall
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.
(c). Collagen: It is the protenaceous support with good porosity and water holding capacity. The
side chains of the amino acids in the collagen and that of enzyme can form covalent bonds to
permanently hold the enzyme to the support.
(d). Carrageenan: It is a sulfated polysaccharide obtained from some red algae. Their good
gelling properties together with its high protein holding capacity makes it good support for
immobilizing enzymes.

13. What are the distinct advantages of membrane reactors?

Making a gaseous product in a membrane reactor generally affects the way that pressure affects
the extent of reaction at thermodynamic pseudo-equilibrium. In an ordinary flow reactor, 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
the feed composition and the exhaust equilibrium constant, the latter being determined by the
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
temperature, and feed composition. There is also a significant (and beneficial) effect that derives
from the controlled removal of a product or addition of reactant.

14. What is the mechanism of entrapment?

In this method enzymes are physically entrapped inside a porous matrix. Bonds involved in
stabilizing the enzyme to the matrix may be covalent or non-covalent. The matrix used will be a
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
concentration of the polymer used. Agar-agar and carrageenan have comparatively large pore
sizes. The greatest disadvantage of this method is that there is a possibility of leakage of low
molecular weight enzymes from the matrix.
Examples of commonly used matrixes for entrapment are:
(1). Polyacrylamide gels
(2). Cellulose triacetate
(3). Agar
(4). Gelatin
(5). Carrageenan
(6). Alginate

15. Write down the factors which influence the pore diffusion of immobilized enzyme.
Analysis of the effect of internal diffusion is complicated by such factors as the shape of the
particles, the distribution in size and shape of the pores, the total volume of the pores with
respect to the particle volume (porosity, h), the depth to which the pores penetrate the particles
(e.g. pellicular particles)

16. Compare the role of enzymes and cells in the manufacture of a biochemical product.
The basic function of an enzyme is to increase the rate of a reaction. Most cellular reactions
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.

Enzymes are what make all the chemical reactions in the cell possible. The human body is made
up of trillions of cells, and there are different cells for differentfunctions. Cells are little bundles
of chemical reactions. They reproduce, they create energy, and they break molecules down and
build them up.

17. Define packed bed reactor

A packed bed is a hollow tube, pipe, or other vessel that is filled with a packing material. The
packing can be randomly filled with small objects like Raschig rings or else it can be a
specifically designed structured packing. Packed beds may also contain catalyst particles or
adsorbents such as zeolite pellets, granular activated carbon, etc.

The purpose of a packed bed is typically to improve contact between two phases in a chemical or
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
gases are allowed to escape through a vessel that is packed with a refractory material until the
packing is hot. Air or other cool gas is then fed back to the plant through the hot bed, thereby
pre-heating the air or gas feed.

18. Define fluidized bed reactor

A fluidized bed reactor (FBR) is a type of reactor device that can be used to carry out a variety of
multiphase chemical reactions.
19. Advantages of packed bed reactor

1. High conversion per unit mass of catalyst

2. Low operating cost

3. Continuous operation

20. Advantages of fluidized bed reactor

1. Good mixing

2. Good uniformity of temperature

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

21. Disadvantages of packed bed reactor

1. Undesired thermal gradients may exist

2. Poor temperature control

3. Channeling may occur

4. Unit may be difficult to service and clean

22. Disadvantages of fluidized bed reactor

1. Bed-fluid mechanics not well known

2. Severe agitation can result in catalyst destruction and dust formation

3. Uncertain scale-up

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.

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.
 25.Define membrane reactor.
Membrane bioreactor (MBR) is the combination of amembrane process like microfiltration or
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
million liters per day).

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

Thiele modulus

Effectiveness factor

 = 1 : the entire pellet volume is reacting at the same high rate because reactant is
able to diffuse quickly through the pellet,
 = 0 : the pellet reacts at a slow rate, since the reactant is unable to penetrate into
the pellet interior.

27. Name any four polymers used for enzyme immobilization.

The polymers used for enzyme immobilization include vinylsulfone-hydroxymethyl

methacrylate, poly(vinylalcohol) beads, polyacrylamide gels, 2-fluoro-1-
methylpyridinium salt-activated Fractogel, hexamethylacrylate polymer (Sepharon Herma
E, an anion exchange polymer), chlormethylated polystyrene beads, and beaded cellulose.
Natural polymers, which are very hydrophilic, are popular support materials for enzyme
immobilization since the residues in these polymers contain hydroxyl groups, which are
ideal functional groups for participating in covalent bonds.

28.Effectiveness factor
Overall productivity for a first order reaction in a spherical pellet

Effectiveness factor
 = 1 : the entire pellet volume is reacting at the same high rate because reactant is
able to diffuse quickly through the pellet,
 = 0 : the pellet reacts at a slow rate, since the reactant is unable to penetrate into
the pellet interior.

29. DefineDamköhler number

Damköhler number (Da)is used to determine the significant effect of external diffusion
resistance on the rate of enzyme catalytic reaction rate.


 When Da >> 1, the external diffusion rate is limiting;
 Da << 1, the reaction rate is limiting;
 Da ≈ 1, the external diffusion and reaction resistances are comparable.

30. Immobilization technique :Adsorption Explain

Oldest method of enzyme immobilization

Simplest method of enzyme immobilization
Nelson &Griffin used charcoal to adsorb invertase
Enzymes are adsorbed to external surface of support
Support/carrier may be:
1. Mineral support (aluminium oxide, clay)
2. Organic support(starch)
3. Modified sepharose and ion exchange resins

PART B

1. Describe in detail the analysis of film and pore diffusion effects in enzyme
immobilized in porous matrix. APR/MAY 2011,NOV/DEC 2015
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-208-216)

2. Explain in detail immobilized reactor. NOV/DEC 2015

Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-606)
 3. Explain in detail membrane reactor. NOV/DEC 2015
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-610)

4. How will you design the packed bed reactor for immobilized enzyme reaction?
APR/MAY 2011
Refer:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
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

Refer:Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
Inc.(Page No-122)

UNIT 4 MODELLING AND SIMULATION OF BIOPROCESSES

TWO MARKS
1. What is plasmid? APR/MAY 2016
A plasmid is an extra-chromosomal DNA molecule separate from the chromosomal DNA which
is capable of replicating independently of the chromosomal DNA.[1] In many cases, it is circular
and double-stranded. Plasmids usually occur naturally in bacteria, but are sometimes found in
eukaryotic organisms (e.g., the 2-micrometre-ring in Saccharomyces cerevisiae).

2. What are the types of plasmids? APR/MAY 2016

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
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

3. Define Simulation of plasmids

The use of plasmids as a technique in molecular biology is supported by bioinformatics software.
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
plasmid maps are Lasergene, GeneConstructionKit, and Vector NTI.

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
concentration, C, within the unit and at its outflow:
dC/dt = Flow*(Cin - C)/V, where Flow = volume of flow in and out, the volume, V, is a constant
and Cin is the concentration of solute in the entering flow.

5. What is DNA replication? APR/MAY 2015, APR/MAY 2014

DNA replication is the process of copying a double-stranded DNA molecule to form two double-
stranded molecules. The process of DNA replication is a fundamental process used by all living
organisms as it is the basis for biological inheritance.

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
the process. This primary concern of this site is dynamic models

7. What types of simulations used in process engineering?

Static
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.
Dynamic
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

8. Define bioprocess engineering.

Bioprocess engineering include the production of biofuels, design and operation of fermentation
systems, development of food processing systems, application and testing of product separation
technologies, design of instrumentation to monitor and control biological processes.

9. Application of bioprocess engineering?

Application areas commonly associated with bioprocess engineering include the production of
biofuels, design and operation of fermentation systems, development of food processing systems,
application and testing of product separation technologies,design of instrumentation to monitor
and control biological processes.

10. What is semi-structured model?

The semi-structured model is a database model. In this model, there is no separation between the
data and the schema, and the amount of structure used depends on the purpose.

11. What is fed batch reactor model?

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
variables, such as the dimensions of the reactor, can be estimated. PFRs are also sometimes
called as Continuous Tubular Reactors (CTRs).

12. Write the advantages of semi structured model.

It can represent the information of some data sources that cannot be constrainedby schema.
 It provides a flexible format for data exchange between different types ofdatabases.
It can be helpful to view structured data as semi-structured (for browsingpurposes).
The schema can easily be changed
. The data transfer format may be portable.

13. What are semi structured model?

The semi-structured model is a database model where there is no separation between the data and
the schema, and the amount of structure used depends on the purpose. The advantages of
this model are the following: It can represent the information of some data sources that cannot be
constrained by schema.

14. Write down the constituents of k and g compartments of the cell in Williams’s model.
k compartments consists of RNA pools of small metabolites and g compartments consist of DNA
and proteins.

15. What is meant by transient culture? MAY/JUNE 2012

Transientculture means it is short-lived culture (usually less than 24 hours). Ex: I have observed
that in daily life people come together in groups for a short time and then disperse. Example, a
bus queue or strangers on a train or in an aircraft. Individuals bring their own cultural attributes
to this situation and build common ground with their fellow temporary group members. They
form, in my terns, 'transient group culture'.

16. What is called promiscuous plasmids?

Plasmids that have transfer systems that allow transfer of DNA to unrelated species are called
promiscuous plasmids.

17. How will you determine the KL value in growth associated model with inhibition.
MAY/JUNE 2012
The K parameter, defined as the ratio of the concentration of undissociated weak acid to the
number of microorganisms in the medium, was determined. The K value is related to the degree
of growth inhibition and provides new insight into the mode of action of weak organic acids
against the studied yeasts.

18. Define single cell model.

The cell, itself, is the biochemical reactor. The engineer often faces the challenge of designing
macroscopic bioreactors, operating polices and control strategies for the macroscopic reactor.
However, the engineer must recognize that the macroscopic reactor only ‘houses’ the cellular
bioreactors; the reactions take place under the control of the cell’s own internal regulatory
system. Thus, the design of the macroscopic reactor and its operating policy requires a
quantitative and explicit linkage between the abiotic and biotic environment; between the
macroscopic control policy and the cellular regulatory system.

19. What is plasmid replication?

The replication strategy that a plasmid uses directly affects its copy number, host range, and
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
frequency of their replication.

20. Define plasmid stability.

Structural plasmid stability exists, when all generated plasmids have the correct base sequence.

21. Define metabolism.

Metabolism is a term that is used to describe all chemical reactions involved in maintaining the
living state of the cells and the organism. Metabolism can be conveniently divided into two
categories: Catabolism - the breakdown of molecules to obtain energy.

22. Explain fed batch culture

Fed-batch culture is, in the broadest sense, defined as an operational technique in
biotechnological processes where one or more nutrients (substrates) are fed(supplied) to the
bioreactor during cultivation and in which the product(s) remain in the bioreactor until the end of
the run.

23. Explain batch culture

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,
the culturedeclines. Cells, or products that the organisms have made, can then be harvested from
the culture.

24. Differentiate steady state and unsteady state culture.what is the difference between
transient and periodic flow?
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
afunction of time – often observed in flow start up. With periodic flows, the flow field is
unsteadybut oscillates around a mean.

25. What are the origin of plasmid replication?

The origin of replication (also called the replication origin) is a particular sequence in a genome
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
double-stranded RNA viruses.

26.Define static method

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.

27. Define Dynamic method

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
28. Define bioprocess engineering.
Bioprocess engineering include the production of biofuels, design and operation of
fermentation systems, development of food processing systems, application and
testing of product separation technologies, design of instrumentation to monitor and
control biological processes.

29. Application of bioprocess engineering?

Application areas commonly associated with bioprocess engineering include the
production of biofuels, design and operation of fermentation systems, development of
food processing systems, application and testing of product separation technologies,
design of instrumentation to monitor and control biological processes.

30. Write the advantages of semi structured model.

It can represent the information of some data sources that cannot be constrained by schema.
It provides a flexible format for data exchange between different types of databases.
It can be helpful to view structured data as semi-structured (for browsing purposes).
The schema can easily be changed.
The data transfer format may be portable.

PART B

1. Describe the design consideration to be taken in account during the bio processing
of animal cell culture. NOV/DEC 2015
Refer: Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
Inc.(Page No-203)

Explain in detail plasmid replication. APR/MAY 2016, APR/MAY 2015,MAY/JUNE 2012

Refer:Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
Inc.(Page No-251-257)

Discuss in detail compartmental model. APR/MAY 2016, MAY/JUNE 2013,2014

Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-174)

2. Explain in detail the structured model for analysis of various bioprocess.APR/MAY

2011
Refer:James M. Lee, “Biochemical Engineering”, PHI, USA.(Page No-174)

3. Discuss in detail Single cell model. NOV/DEC 2015

Refer:Harvey W. Blanch, Douglas S. Clark, “Biochemical Engineering”, Marcel Decker
Inc.(Page No-244-246)

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
Inc.(Page No-277-280)

UNIT 5 RECOMBINANT CELL CULTIVATION

TWO MARKS

1. Define Animal cell culture? APR/MAY 2016, APR/MAY 2015

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
conditions of asepsis, sterility and controlled environment involving temperature, gases and
pressure.

2. Define cell culture?

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.

3. What are the applications of cell culture?

Mass culture of animal cell lines is fundamental to the manufacture of viral vaccines and many
products of biotechnology. Biological products produced by recombinant DNA (rDNA)
technology in animal cell cultures include enzymes, synthetic hormones, immunobiologicals
(monoclonal antibodies, interleukins, lymphokines), and anticancer agents.

4. Explain high cell density cultivation? APR/MAY 2015

In particular, microbial high cell density cultures have a high metabolic oxygen demand. In these
cultures the oxygen transfer rate of the bioreactor determines the maximum biomass
concentration. Unfortunately, the solubility of oxygen is even decreasing with increasing cell
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
bioreactors, an aeration membrane or disposable spargers and baffles have been inserted or the
medium was vibrated.

5. What is meant by host?

An animal or plant on or in which a parasite or commensal organism lives.

6. Define recombinant cell cultivation? APR/MAY 2014

Relating to or denoting an organism, cell, or genetic material formed by recombination.

7. Advantages of recombinant cell cultivation?

Precise control over growth conditions, batch-to-batch product consistency, a high level of
containment and the ability to produce recombinant proteins in compliance with good
manufacturing practice.

8. Write down the Application of animal cell cultivation. APR/MAY 2014

Mass culture of animal cell lines is fundamental to the manufacture of viral vaccines and other
products of biotechnology.
many simpler proteins can be produced using rDNA in bacterial cultures, more complex proteins
that are glycosylated(carbohydrate-modified) currently must be made in animal cells

9. What is meant by plant cell cultivation?

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
proper balance of the plant growth hormones auxin and cytokinin.
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
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
top of this, further modifications can be done for improving the volumetric yield further

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
(mitosis).
 Provide the cells with appropriate temp, good substrate for attachment and proper culture
medium.
 Temperature:Cold-blooded vertebrates – 18-25°C,Mammalian cells – 36-37°C

12. Explain the common features of animal cell bioreactor?

1) Reactor should be gently agitated and aerated. Agitation speed ≈20rpm.
Bubble-column & airlift reactor operating at high aeration may cause damage of cells
2) Supply of CO2-enriched air
3) Removal of toxic products from metabolism eg lactic acid, ammonium

13. List out the advantages of animal cell bioreactor?

 Ability to perform post-translational modifications and achieve a product close to that
produced in vivo
 The used of other expression systems (yeast, plant or insect cell) more limited because
despite their potential economic advantages for culture and higher yields of these
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
necessary glycosylation machinery and so the proteins produced are not glycosylated.
 Lower eukaryotes cells produced differ significantly from those present in human
glycoproteins.
 So mamalian cells are the chosen host for the production of human glycoproteins because
it has been recognized that they meet the criteria for an appropriate glycosylation of
recombinant human glycoprotein.
 The most commonly used cell lines are hamster-derived Chinese hamster ovari (CHO and
baby hamster kidney (BHK) cells
 These cells are chosen because of their favorable growth characteristics in culture both as
anchorage-dependent or suspension cells.
14. Define E.coli?
A bacterium commonly found in the intestines of humans and other animals, some strains of
which can cause severe food poisoning.

15. What is yeast?

A microscopic fungus consisting of single oval cells that reproduce by budding, and capable of
converting sugar into alcohol and carbon dioxide.A greyish-yellow preparation of the yeast
fungus obtained chiefly from fermented beer, used as a fermenting agent, to raise bread dough,
and as a food supplement.

16. What is Pichia pastoris?

Pichia pastoris is a species of methylotrophic yeast. Pichia is widely used for protein production
using recombinant DNA techniques. Hence it is used in biochemical and genetic research in
academia and the biotechnical industry.

17. What is Saccharomyces cereviseae?

Saccharomyces cerevisiae is a species of yeast. It has been instrumental to winemaking, baking,
and brewing since ancient times.

18. Process strategies of animal cell cultivation.

(1) cell lines capable of synthesizing the required molecules at high productivities that ensure
low operating cost;
(2) culture media and bioreactor culture conditions that achieve both the requisite productivity
and meet product quality specifications;
(3) appropriate on-line and off-line sensors capable of providing information that enhances
process control; and
(4) good understanding of culture performance at different scales to ensure smooth scale-up.
Successful implementation also requires appropriate strategies for process development, scale-up
and process characterization and validation that enable robust operation and ensure compliance
with current regulations

19. Process strategies of plant cell cultivation.

In order to successfully cultivate the plant cells at large scale, several engineering parameters
such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for
selection of a suitable bioreactor. The media ingredients, their concentrations and the
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,
fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular
metabolites. Proper understanding and rigorous analysis of these parameters would pave the way
towards the successful commercialization of plant cell bioprocesses.

20. Process strategies of insect cell cultivation.

1. Seed the experimental culture at between 0.5-1 x 106 cells per ml and allow to grow
overnight.
2. Confirm log growth of cells by observation of increase cell counts. If the culture did not grow
overnight, discard and start again.
3. Infect the culture at your desired MOI Volume of inoculum needed = MOI ( iu/cell ) X
number of cells Titer of virus ( iu/ml )

21. Process strategies of high cell density cultivation.

the application of more advanced control strategies, such as exponential feeding procedures and
feed-back control of the pO2. Pulsing of extra oxygen provides some advantage in overcoming
the low oxygen transfer rates.

22. List out the host vector system.

Prokaryotes
1) Gram Negative
2) Gram Positive
Eukaryotes
1) Yeast
2) Fungi
3) Insect Cells
4) Plant Cell Lines
5) Animal Cell lines
6) Transgenic Animals
7) Transgenic Plants

23. What is a recombinant cell?

Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of
genetic recombination (such as molecular cloning) to bring together genetic material from
multiple sources, creating sequences that would not otherwise be found in biological organisms.

24. What is a primary culture?

Primary culture refers to the stage of the culture after the cells are isolated from the tissue and
proliferated under the appropriate conditions until they occupy all of the available substrate (i.e.,
reach confluence).

25. What does meant to culture cells?

Cell culture is the complex process by which cells are grown under controlled conditions,
generally outside of their natural environment. In practice, the term "cell culture" now refers to
the culturing of cells derived from multi-cellular eukaryotes, especially animal cells.

26. Define Cell strain?

A cell strain is derived either from a primary culture or a cell line by the selection or cloning of
cells having specific properties or characteristics which must be defined. Cell strains are cells
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
their ability to proliferate (a genetically determined event known as senescence).
27.How will you Determine the Cell Viability (for all cell types)
After the cells have been suspended into the medium, 0.2 ml of the cell suspension can be mixed
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
number of viable cells (those not taking up the stain) can be determined and used for initiating
the new culture with a precise number of viable cells. I personally find this is a time consuming
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
with the inverted microscope. Beginners may want to include this step until they gain confidence
in their visual inspection of cells.
28.Explain Micro Propagation
It is in vitro asexual propagation of crop plants. This technique is advantages over the
conventional practice of asexual propagation as only a small amount of plant is needed, species
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
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)
3. Explain in detail insect cell cultivation APR/MAY 2015
Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-267)

4. How the high cell density cultivation was processed explain in detail?
Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-269)

5. In what way recombinant cell cultivated in E.coli explain clearly. APR/MAY 2014
Ref:James E. Bailey & David F. Ollis, “Biochemical Engineering Fundamentals”,
McGraw-Hill.(Page No-259)