CH6705 Biochemical Engineering 2018-2019

St. JOSEPH’S COLLEGE OF ENGINEERING

DEPARTMENT OF CHEMICAL ENGINEERING
UNIT – I: INTRODUCTION
Part -A
1. List the substance that compose the cell wall. (Nov 2015)
Cellulose is one of the three major structural component of all plant cell walls with two other
components, hemicellulose and lignin.
2. Classify fungi, with a note on economic importance of fungi. (Nov 2015)
Fungi are plants devoid of chlorophylland are therefore unable to synthesize their own food. They
range in size and shape from single celled yeasts to multicellular mushrooms. Among them, yeasts
and molds are industrially important.
3. What are lipids? (Nov 2014)
Lipids are biological compounds which are soluble in polar solvents such as benzene, chloroform,
ether and are practically insoluble in water. Their relative insolubility lends to their presence
predominantly in the plasma and organelle membranes.
4. Why has the study of microorganisms become important suddenly? (May 2014)
To maintain balance on environment (microbial ecology)
(ii) Basis of food chains
(iii) Diseases causing microbes
(iv) Bioremediation
5. Give one example for different classes of Microorganisms. (Nov 2013)
Prokaryotes- bacteria – Escherichia coli
Eukaryotes – fungi – yeast – Sacchromycescerevisiae, mold – A. niger, algae – Euglena, protozoa –
Amoeba.
6. What are the various morphological forms of bacteria? (Nov 2014)
(i) Spirilla– rhodospirillum, (ii) Cocci– streptococcus and (iii) Bacilli – clostridium.
7. Classify bacteria based on gram reaction. (Nov 2011)
Gram reaction refers to response of bacteria to a relatively straight forward and rapid staining test.
Cells are first stained with a dye crystal violet then treated with iodine solution and washed with
alcohol. In response to the gram test bacteria are classified as gram positive and gram negative.
8. Compare bacterial cell and yeast cell. (Nov 2012)
Bacterial cell yeast cell
Show primitive cellular organization and do not show advanced cellular organization and possess
possess organized nucleus nucleus with a nuclear membrane and nucleolus
Bacterial cells are motile yeast cells are not motile
Size varies from 1-5μm size greater than 5μm
Cytoplasm does not contain well bound cytoplasm contains well bound Organelles
organelles
Cell wall contains peptodoglycan cell wall made up of cellulose
9. List out some of the enzymes used in medicinal industry. (May 2014)
i) Asparaginase (Leukemia)
(ii) Collagenase (Skin ulcer)
(iii) Lysozyme(Antibiotic)
(iv) Trypsin (inflammation)
10. Compare the role of enzymes and cells in the manufacture of biochemical product. (May
2014)
Enzymes are proteinaceous substances present in free state within cells which activate the reaction
taking place in the cell. Cell composes of various organelles, most important of which ribosomes
which serve as active sites for enzymatic reactions.
11.Defineheterogenous population. (May 2015)
Stratified sampling techniques are generally used when the population isheterogeneous, or
dissimilar, where certain homogeneous, or similar, sub-populations can be isolated (strata). Simple
random sampling is most appropriate when the entire population from which the sample is taken is
homogeneous.
12. What is cell fractionation? (May 2015)
Cell fractionation is the process used to separate cellularcomponents while preserving individual
functions of each component.
13. What are carbohydrates?

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Carbohydrates are organic compounds with general formula (CH2O) where n >3. These compounds
are found in all animal, plant and microbial cells. They serve both structural and storage functions.

14. What is cellulose?

Cellulose is a major structural component of all plant cells from algae to trees. Cellulose is the most
abundant organic compound occurring on earth. Each cellulose molecule is a long unbranched chain
of D – glucose subunits with a molecular weight ranging from 50,000 – 1,000,000.
15. What are proteins?
Proteins are most abundant organic molecules found in the cell. 30 – 70% of the cells dry weight is
protein. The prevalent components in protein are carbon, hydrogen, nitrogen and oxygen. In addition,
sulfur contributes to the three dimensional stabilization of proteins by the formation of disulfide bonds
between sulfur atoms at different locations along the polymer chain.
16. What is isoelectric point?
The acidic group (- COOH) and basic group (-NH2) of amino acid can ionize in aqueous solution.
The amino acid is positively charged at low pH and negatively charged at high pH. At an intermediate
pH value, the amino acid acts as a dipolar ion and zwittter ion which has no net charge. The pH at
which an amino acid has no net charge is termed as the isoelectric point.
17. What are actinomycetes?
Actinomycetes are a group of microorganisms with some properties of both fungi and bacteria. One
important characteristic is the susceptibity of actinomycetes to infection and disease by viruses which
can also attack bacteria.
18.What is mycelium?
Highly branched system of tubes which are vegetative structure of molds, higher class of fungi.
19. What are endospores? Explain their importance in bacterial cells. (Nov 2013)
Endospores are dormant parts of the cell which are capable of resisting heat, radiation and poisonous
chemicals. When the spores are returned to the surroundings through the cells membrane and the cell
wall suitable for cell function, endospores can germinate to normal functioning cells. This normal
biologically active cell state is often called vegetative form in order to distinguish it from the spores.
20. Give examples of bacteria capable of forming spores.
Bacillus, clostridium.
21. Mention one parameter that differentiates catalyst from a biocatalyst.
Biocatalyst is highly specific in comparison with catalyst.
22. What is the function of ribosomes?
They are grainy dark spots in the interior which are of great concern to the cells involved in
biochemical reactions. These are the sites where biochemical reactions take place.
23. What are the characteristic features of prokaryotic cells?
They do not contain a cell membrane bound nucleus. Prokaryotes are small and simple cells.
Prokaryotes exist in single without association with other cells. They may be spherical, rod like or
spiral and vary in size from 0.5 – 3.0μm.
24.What is the function of cell wall in prokaryotes and eukaryotes?
They are surrounded by a rigid cell wall. The thickness of the cell wall is approximately 200 A and
lends structural strength to the cell to preserve its integrity in a wide variety of external surroundings.
25. What are microbial strains? (Dec 2017)
A strain is "a population of organisms that descends from a single microorganism or pure culture
isolate. Strains within a species may differ slightly from one another in many ways."
26. What are vitamins? classify them.
Vitamins are organic substances which are required in trace amounts for normal cell function. The
vitamins which cannot be synthesized internally by an organism are termed as essential vitamins.
Classified into 2 heads – fat soluble vitamins and water soluble vitamins.
27. Compare chemical and biochemical processes. (Dec 2017)
Chemical Engineering concerns the translation of discoveries in Chemistry into
commercial processes for manufacture of new chemical entities.
Biochemical engineering concerns the engineering of discovery processes and the
translation of discoveries in Biochemistry and Medicine into commercial processes
for new biological entities such as medicines and therapeutics. It encompasses the
biology, engineering, mathematics and business behind this translation.
Chemical engineering largely involves the design and maintenance of chemical
processes for large-scale manufacture. Chemical engineers in this branch are usually
employed under the title of process engineers.

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Whereas biochemical engineering largely involves the design and maintenance of
biological processess for large-scale manufacturing. Chemical engineers in this
branch are usually employed under the title of biochemical or bioprocess engineers.
28. What are the difference between Eukaryotic and Prokaryotic cells? (May 2017)

29. Define diauxic growth. (May 2017)

Diauxic growth or diauxie is any cell growth characterized by cellular growth in two phases, and can
be illustrated with a diauxic growth curve. Diauxic growth, meaning double growth, is caused by the
presence of two sugars on a culture growth media, one of which is easier for the target bacterium to
metabolize.

PART – B
1. Explain the structures and features of prokaryotic and eukaryotic microbial species used in
industrial bioprocesses. (Nov 2013, Nov 2014)
2. What are the major classes of microorganisms? Cite the difference among these classes. (Nov
2015)
3.Classify the industrially important microbes, with an example, explain any one microbial synthesis
of product. (May 2014)
4. Compare chemical process over Biochemical process. (Nov 2014, May 2015)
5. Compare Prokaryotes with Eukaryotes based on various features. (Nov 2015)
6. Microbial fermentation processes can be classified according to the type of the fermentation
product. Give one example for each types of fermentation and compare them in terms of metabolism,
kinetics and product recovery. (Nov2015)
7. List the advantages and disadvantages of simple and complex media for microbial growth. (Nov
2015)
8. Explain briefly about the classification of microbial strains. (May 2015)
9. Write detailed note on the regulation of enzyme synthesis.
10. Write detailed note on the specific nutritional requirements of microorganisms in industrial
fermentation.
11. Explain in detail about cellular genetics. (Nov 2012)
12. Write short notes on the traditional and modern applications of biotechnology.
13. Discuss in detail about the role of a biochemical engineer.
14. Differentiate between chemical and biochemical reaction with suitable example and explain their
relative characteristics.

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15. Write about replication of nuclear material.
16. Write about the structure of DNA.
17.i) Explain in detail about the structure of cells. ii) Briefly discuss about cellular genetics. (May
2015)
18. What are the major classes of microorganisms? Cite the differences among these classes. (May
2017)
19. Microbial fermentation processes can be classified according to the type of fermentation product.
Give one example for each type of fermentation and compare them in terms of metabolism, kinetics
and product recovery. (May 2017)
20. Define simple and complex lipids. Describe enzymatic hydrolysis of lipids. (May 2017)
21. What are the four main biomolecules and their functions? (May 2017)
22. With a suitable example, elaborate the industrial biochemical process. (Dec 2017)
23. Classify microbial strains. Explain them with structure. (Dec 2017)
24. How microorganisms are classified? Support your answer with suitable examples for each type
and describe their characteristics and their utility in biochemical engineering. (May 2016)
25. List out industrially important microbes. Elaborate the production of any one microbial product.
(May 2016)

UNIT II: KINETICS OF ENZYME ACTION

Part – A
1. What is an Enzyme? (Nov 2013)
A substance produced by a living organism which acts as a catalyst to bring about a specific
biochemical reaction.
2. Give the nomenclature of enzymes. (Nov 2014)
Enzymes are named initially based on the substrate upon which they act. In the year 1961,
International Union of Biochemists proposed six major classes based on the general type of reaction.
(i) Oxidoreductase: They function in the oxidation-reduction reactions.
(ii) Transferases: They catalyze the transfer of the functional groups.
(iii) Hydrolases: They bring about the hydrolysis of compounds.
(iv) Lyases: They are specialized in the addition or removal of H2O, NH3, CO2, etc.
(v) Isomerases: They are used in the isomerization reactions.
(vi)Ligases: They catalyze the synthetic where two molecules are joined together.
The other way of classifying is: intracellular enzymes and extracellular enzymes. The intracellular
enzymes are functional within the cells where they are synthesized, whereas the extracellular enzymes
are active outside the cells, for example, digestive enzymes like trypsin, pepsin etc.
3. What are the methods available for immobilization of whole cells? (May 2014)
i) Entrapment techniques (ii) Adsorption Techniques
(iii) Selective binding of cells by immobilized macromolecules.
(iv) Covalent bonding of cell to support.
4. Mention the methods used in evaluating Michealis – Menten parameters. (Nov 2012)
Line weaver Burk plot
Langmuir plot method
Eadie – Hofstee plot method.
5. Mention the various approaches used in deriving the reaction rate for any enzyme catalysed
reaction (Nov 2013)
(i) Michealis – Menten approach. The equation is given by

(ii) Brigg’s – Haldane approach.

6. Compare the role of enzymes and cells in the manufacture of a biochemical product.
(Nov 2014)
Each reaction in the cell is catalyzed by its own, specific enzyme to synthesis a biochemical product.
The enzyme catalysts regulate the structure and function of cells and organisms.
7.What is a cofactor? (May 2013)
It is a non – protein compound which combines withan inactive protein to give a catalytically active
complex.
8. Explain competitive inhibition. (May 2015)

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Competitive inhibition is a form of enzyme inhibitionwhere binding of the inhibitor to the active
site on the enzyme prevents binding of the substrate and vice versa. Most competitive
inhibitors function by binding reversibly to the active site of the enzyme.

9. Defineunits of enzyme activity. (May 2015)

The enzyme unit (U) is a unit for the amount of a particular enzyme. One U is defined as the amount
of the enzyme that produces a certain amount of enzymatic activity, that is, the amount that catalyzes
the conversion of 1 micro mole of substrate per minute.
10. Define specificity of the enzyme.
The enzyme catalyses only one specific reaction.
11. What do you understand by sterilization? (Nov 2011, May 2015)
An important aspect to be taken care of in bioprocesses unlike chemical reactions, because presence
of environmental bacteria and microorganisms consume the nutrients from the media, making the
process more difficult with the cultivation of plant or animal cells because their growth rates are much
slower than those of environmental bacteria or molds.
12. What is the unit of enzyme activity?
Unit of enzyme activity designate the amount of enzyme which gives a certain amount of catalytic
activity under a prescribed set of standard conditions for that particular enzyme.
13. State the assumptions to derive enzyme kinetics. (May 2010)
Total enzyme concentration remains constant.
Amount of substrate is very large in comparison with the enzyme added. Hence depletion of substrate
during enzyme substrate complex formation is assumed negligible.
14. Give an example of a strain used in confectionaries.
Sacchromycescerevisiae.
15. State the assumptions made in Brigg’s – Haldane approach for enzyme kinetics.
The enzymatic reaction is assumed to take place at pseudo – steady state wherein change of
intermediate substrate composition with time is negligible.
16. Mention the important kinetic measurements to be made over the samples removed from
isothermal vessel at different time intervals of the process.
Dry weight of microorganisms containing the enzyme, pH value, product concentration,
microscopical examination for identifying contamination.
17. What is the function of oxidoreductases.
Enzymes which bring about oxidation – reduction reactions, act on organic linkages to enhance
oxidation - reduction reactions.
18. What is a coenzyme? (May 2009)
An organic molecule of high complexity which is capable of converting an inactive protein to give a
catalytically active complex.
19. What is the function of transferases?
Enzymes which help in the transfer of functional groups are termed as transferases.
20. What is the function of hydrolases?
Hydrolases are enzymes which catalyse hydrolysis reactions.
21. Define turnover number. (May 2010)
The net number of substrate molecules reacting per catalyst site per unit time is turnover number.
22. What is substrate inhibition?
When a large amount of substrate is present, the enzyme catalyzed reaction diminishes by excess
substrate concentration. This phenomenon is called substrate inhibition. When S is larger than Max S
a decrease in substrate concentration causes an increase in reaction rate.
23.What are proteolytic enzymes?
Enzymes which have the tendency of attacking nitrogen carrying compounds. Wide application in dry
cleaning as detergents.
24. How are enzymes classified?

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Based on their applications, it is classified into three. They are
(i)Industrial enzymes
(ii) Analytical enzymes and
(iii) Medicinal enzymes.
25.Mention few commercial application of enzymes.
Amylase – glucose production – desizing textiles.
Pepsin – digestive aid, meat tenderizer.
26. State about the regulation of enzyme activity. (Dec 2017)
Enzyme activity may be turned "up" or "down" by activator and inhibitor molecules that bind
specifically to the enzyme.
27. Write short nots on enzyme immobilization? (Dec 2017)
Immobilization is defined as the imprisonment of cell or enzyme in a distinct support or matrix. The
support or matrix on which the enzymes are immobilized allows the exchange of medium containing
substrate or effector or inhibitor molecules.
28. What is meant by Turn-over number? (May 2017)
It is defined as the maximum number of chemical conversions of substrate molecules per second that
a single catalytic site will execute for a given enzyme concentration.
29. State the importance of Michaelis Constant (May 2017)
The Michaelis Constant, KM is very important in determining enzyme-substrate interaction. This value
of enzyme ranges widely and often dependent on environmental conditions such as pH, temperature,
and ionic strength. Secondly, it is, in some cases, able to detect the strength of the enzyme-substrate
complex (ES).

PART - B
1. How the enzyme commission developed a system of classification and its recommendations on
nomenclature. (Nov 2014, May 2014, Nov 2015)
2. The reaction data (moles/liter.min) for asparaginase in presence of an inhibitor (I) is given
Substrate 0.2 M 0.02 M
concentration I/V I(M) 1/V I(M)
0.22 0 0.68 0
0.33 0.0012 1.02 0.0012
0.88 0.0060 3.46 0.0060
i) Determine the type of inhibition ii) Calculate the K1(Nov2015)
3. Describe the different mechanism for enzyme immobilization with suitable examples. List its
advantages and disadvantages. (Nov 2015)
4. Discuss the various methods of determining kinetic parameters for Michaelis – Menten type
Kinetics. (Nov 2014)
5. Explain Oxidoreductases, Transferases, and Hydrolases in detail with examples and reactions
involved.
6. i) Explain briefly about factors affecting cell growth. ii) using a neat flow sheet explain in detail
about the high fructose corn syrup production using immobilized glucose isomerase. (May 2015)
7. Write detail notes on i) Active site in enzymes ii) Role of enthalpy on enzyme catalysis
8. Describe the various factors that affect enzyme activity. (Nov 2014)
9. Derive an expression for Competitive inhibition.
10. Derive the Michaelis-Menten equation for single substrate enzyme catalysed reactions. Write the
significance Michaelis-Menten parameters. (Nov 2014, May 2015)
11. Describe elaborately the different types of enzyme inhibition. Compare the competitive inhibition
with uncompetitive and noncompetitive inhibition.
12. Mathematically explain in detail about the transition and collision theory and Compare.
13. Explain any three reactions involving oxidoreductase.
14. Explain with neat diagram the various model of enzyme action on substrate.
15.Write a detailed account on the classification and nomenclature of enzymes by enzyme
commission. (May 2017)
16. Discuss and compare non-competitive and un-competitive inhibition based on their kinetics. (May
2017)
17. Describe the different mechanism for enzyme immobilization with suitable examples. List their
advantages and disadvantages. (May 2017)
18. Discuss why the lock and key model could lead to an inefficient enzyme mechanism and induced
fit model to an efficient enzyme mechanism. (May 2017)
19. Give a detailed description about the immobilized enzyme technology. (Dec 2017)

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20. Give a detailed account on the effect of external mass transfer resistance. (Dec 2017)
21. Explain the different methods of evaluation of parameters in Michaelis-Menton equation. Derive
the equation. (May 2016)
22. What do you understand by immobilized enzymes? Describe any one method in detail for
immobilization of enzymes. What are its limitations? (May 2016)

UNIT III: KINETICS OF MICROBIAL GROWTH

Part-A
1. What are the phases involved in the growth cycle of a batch cultivation. (Nov 2013)
On suspending a microbial strain into a medium containing nutrients 4 phases are involved as a part of
microbial strain growth. Lag phase, growth phase, stationary phase, death phase.
2.What are the advantages of immobilization? (May 2014)
With enzymes in solution, there are chances of enzymes to leave the reactor along with the product.
This will not only need the introduction of fresh enzymes to maintain uniform concentration of
enzymes until the completion of the reaction but will also contaminate the product.
Immobilisation increases the catalytic activity efficiency of the enzymes by allowing the enzymes to
remain inside the reactor till the completion of reaction.
3. Explain a structured model and an unsegregated model with suitable examples. (May 2015,
Nov 2015)
Population Cell components
Unstructured Structured
Distributed Cells are represented by a single component, Multiple Cell components,
which is uniformly distributed throughout the uniformly distributed
culture throughout the culture inter at
with each others.
Segregated Cells are represented by a single component, Cells are composed of multiple
but they form a heterogeneous mixture components and form a
heterogeneous mixture.
4. What is doubling time? (Nov 2014)
The time required to double cell population in any inoculum.

5. Draw the growth curve for batch cell cultivation. (May 2014)

6.What is enzyme inhibition? (Nov 2011)

The presence of a modulator would activate the enzyme activity is termed as enzyme inhibition.
7. What are the methods available for cell immobilization? (Nov 2011, May 2014)
(i) Attachment to micro carriers;
(ii) Entrapment within a porous matrix; and
(iii) Containment behind a barrier.
8. Define yield coefficient. (May2015)
The maximum specific growth rate is given by: mm =Y.k. where, Y is the maximumyield
coefficient and is defined as the ratio of maximum mass of cells formed to the mass of substrate
utilized. The coefficients Y, kd, k and Ks are designated as kineticcoefficients.
9. What is immobilization? (Nov 2013)
Refers to confinement or localization of enzymes so that it can be reused continuously.
10. What is the unit of specific growth rate?
1/ time.
11. What are the assumptions made in deriving growth kinetics?

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Cells can be represented by a single component such as cell mass, cell number, concentration of
protein, DNA, RNA. The population of cell mass is distributed uniformly throughout the culture.
12. What is growth rate and what is meant by limiting nutrient?
Growth rate is defined as rate of change of cell number or cell mass with respect to time. A nutrient
whose concentration change controls the growth rate of microorganism is termed as limiting nutrient.
13. What marks the beginning of death phase?
The toxic product buildup as a result of cell metabolism with complete nutrient depletion.
14. What are the functions of a semipermeable membrane?
Selectively permeates the essential nutrients required for cell metabolisation. Also helps in the
diffusion of fermentation products from within the cell to the inoculum.
15. What factors control the length of lag phase in the growth cycle?
Type and age of the microorganism, size of the inoculum, culture conditions.
16. What are the different types of inhibition?
Enzyme, substrate, product.
17. What are the factors affecting enzyme action?
Presence of modulators, pH, temperature, mechanical forces, chemical agents, irradiation.
18. What is the optimum pH for enzymatic reactions?
All enzymes do not have optimum pH value. The optimum pH of some enzymes lies in the acidic
range, some basic range. Catalytic activity of enzyme increases with increase in pH of the enzyme in
solution and passes through a maximum at the optimum pH. The catalytic activity declines smoothly
and symmetrically as pH is varied away from the optimum value.
19. Explain apoenzyme, holoenzyme and coenzyme.
A cofactor is a non protein compound which combines with an inactive protein called apoenzyme to
give a ctalytically active complex called holoenzyme. A complex organic molecule called as
coenzyme may serve as a cofactor.
20. What are modulators?
Chemical species other than substrate which combine with enzyme to alter their catalytic activity are
called modulators.
21. What are the methods available for enzyme immobilization?
Chemical – covalent bond attachment of enzymes, cross linking
Physical – entrapment of enzymes, microencapsulation of enzymes.
22. Define division rate.
Rate of cell division per unit time.

23. What is silanisation?

A method for modification of support surface by coating the support with organic functional groups
by using organofunctionalsilane reagent. This modification enhances better attachment of the enzyme
onto the support.
24. What is microencapsulation?
An artificial method of enzyme immobilization within porous semipermeable polymeric membrane.
25. Mention some materials used in constructing microcarriers.
Ion exchange resins, dextran based beadscoated with gelatin, plyacrlamide beads, polystyrene beads,
cellulose beads.
26. Mention the disadvantages exhibited by entrapment of cells formed insitu.
Cell leakage due to cell division occurring within individual beads thereby preventing the enzymes to
remain inside the reactor to enhance further enzymatic reactions.
27. What do you understand by cross linking of enzymes?
Most widely used multifunctional reagent is glutaraldehyde which establishes intermolecular cross
links of amino groups.
28. Mention some materials used in constructing synthetic water – soluble supports for enzyme
immobilization.
Acrylamide based polymers, styrene based polymers, methacrylic acid based polymers, etc.
29. Give some examples of non – essential amino acids residues suitable for enzyme
immobilization.
Alpha, beta, gamma – carboxyl groups, alpha, beta amino groups, phenolic, hydroxyl groups.
30.What are the various modes of expressing cell concentration?
Number of cells per unit volume of inoculum,
Wet cell weight per unit weight of inoculum
Dry cell weight per unit weight of inoculum.
31. Is the growth rate based on cell mass and cell number identical?

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The growth rate based on the number of cells and that based on cell weight are not the same. When
the mass of an individual cell increases without division, growth weight based on cell weight
increases, while growth rate based on number of cells remains the same.
32. What are the factors affecting cell growth?
Nutrient and toxin concentration.
33. Write about the cell growth in continuous culture. (Dec 2017)
When cells are cultured in a container of fixed volume, both the population density (i.e., the number
of cells per unit volume of culture) and the population size (i.e., the total number of cells in the
culture) increase at the same rate during the exponential phase of growth.
PART – B
1. Write about the microbial growth pattern and kinetics in microbial batch culture. (Nov 2013)
2. State and explain the factors that affect enzymatic reaction rates. (May 2014)
3. Write about Monod’s Growth Kinetics. (Nov 2013, May 2015)
4. Compare and contrast between free cell and immobilized cell reactors with an example. (May
2014)
5. Lactate is produced in a growth associated manner and yield coefficient of lactate formation Yp/s=
0.8 g/l. The feed contains 1 g/l of glucose. The culture is growing in a 5 litre chernostat being a fed at
2.75 liter/hr. Calculate 1) Dilution rate 2) Residence time 3) Biomass concentration 4) Product
concentration. (Nov 2015)
6. Aerobic degradation of benzoic acid by a mixed culture of microorganisms can be represented by
the following reaction.
C6H5COOH+ aO2+ bNH3→cC5H7NO2+dH2O+eCO2
i) Determine the yield coefficients Y x/s and Y x/ O2 (ii) Determine degree of reduction for the
substrate and bacteria. (Nov 2015)
7. Write about cell immobilization and its methods. (Nov 2014, May 2015)
8. i) A simple batch fermentation of a n aerobic bacterium growing on methanol gave the results
shown in the table. Calculate 1) Maximum growth rate 2) Yield on substrate (Y x/s) 3) Mass doubling
time 4) Saturation constant 5) Specific growth rate at t=14h. (Nov 2015)
Time (h) 0 2 4 8 10 12 14 16 18
X(g/I) 0.2 0.211 0.31 0.98 1.77 3.2 5.6 6.15 6.2
S(g/I) 9.2 9.21 9.07 8.03 6.8 4.6 0.92 0.077 0
ii) Define Decimal reduction time. What is its significance? (Nov 2015)
9. Write about reactor configurations for immobilized cells.
10. Write and compare about the fixed and fluidized bed reactors for immobilized enzymes and
microorganisms. (Nov 2012)
11. Briefly explain about modeling of batch and continuous cell growth. (May 2014, Nov 2014, May
2015)
12. i) Explain in detail about typical growth curve for batch cultivation. ii) Discuss about free cell
reactors. (May 2015)
13. Derive Monod equation from mechanistic considerations.
14. Discuss the ways to determination of growth parameters Ks and μm.
15. Write about Monod’s Model.
16.i) Write about transport across cell membranes. ii)Write about the Environmental factors affecting
growth.
17. Explain the models suitable for the cellular growth of structured and cybernetic models. (Dec
2017)
18. With example, give the design and explain the analysis of biological reactors. (Dec 2017)
19. Explain in detail about the modelling of batch and continuous cell growth. (May 2016)
20. Give a brief account of immobilized cell reactors with suitable diagrams. (May 2016)

UNIT IV: TRANSPORT PHENOMENA

PART – A
1. Give the transport resistance in gas – liquid mass transfer. (Nov 2015)
Diffusion from bulk gas to gas – liquid interface
Movement through gas – liquid interface
Diffusion of solute through the relatively unmixed region adjacent to the bubble into the well mixed
bulk liquid. Transport of solute through the bulk liquid to a second relatively unmixed region
surrounding the cells. Transport through second unmixed liquid region associated with the cells.
Diffusive transport through cellular floc, mycelia or soil particle.
Transport across cell envelope and into intracellular reaction site.

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2. What are the factors affecting transport resistances in gas – liquid mass transfer. (Nov 2015)
Air bubble hydrodynamics, temperature, cellular activity, density of fermentation broth, fermentation
broth composition, interfacial area.
3. Give the names of some gas – liquid contacting molecules.
Freely rising air bubbles in contact with falling fluid as in sparged liquid fermenter. Mechanical
agitation as in stirred tank bioreactor operated in continuous or batch mode.
4. Define Power Number. (Nov 2014, May 2014)
It is defined as the drag force to inertia force used for power consumption calculations.

5. What is volumetric mass transfer coefficient? (May 2014)

In case of packed tower fermenters interfacial area between gas and fermentation broth is not directly
measured wherein the flux of mass transfer cannot be determined but only the rate of mass transfer as
the product of flux and interfacial area may be determined. Volumetric mass transfer is obtained by
dividing the rate of mass transfer by the volume of the packing.
6. What are the factors affecting KLa values. (Nov 2014)
Liquid phase solute diffusivity, continuous phase viscosity, gas liquid interfacial resistance.
7. Define Newtonian fluid. (Nov 2013)
The fluid which follows Newton’s law of viscosity ie., rate of shear stress is directly proportional to
shear strain are called Newtonian Fluids.
8. Define Non – Newtonian fluid.
Fluids for which rate of shear stress is not proportional to shear strain.
9. Define Yield Factor. (Nov 2014)

10. Define sterilization. (Nov 2012, May 2015)

The method of destroying foreign organisms in fermentation medium or fermentation equipment is
called sterilization.
11. What are the advantages of moist heat over dry heat in sterilization.
The intrinsic heat resistance of vegetative bacterial cells is greatly increased in completely dry state.
Death rate is much lower for dry cells than for moist cells. Heat conduction in dry air is less rapid than
in steam.
12. What is disinfection?
Disinfection is a method of sterilization to remove or reduce the risk from pathogenic organisms using
chemicals like quaternary ammonium compounds, acids and alkalies.
13. Define Del factor.
Defined as the logarithm of ratio of cell concentration in terms of cell number before sterilization and
after sterilization.
14. Give the relation illustration the dependence of death rate constant on temperature.
Dependence of death rate constant on temperature is given by Arrhenius equation.
15. Mention the important characteristics that a support should possess.
Supports should be highly susceptible to mechanical attrition; supports should be porous.
16. What are the mechanisms of transport across the cell?
Passive and facilitated diffusion, active transport.
17. Enumerate the purpose of aeration and agitation in activated sludge reactors.
To provide oxygen for microbial respiration
To suspend and mix the sludge and other particulates
To strip out volatile metabolic products such as carbon dioxide.
18. What is a fed batch reactor?
Batch reactors to which desirable liquid stream is added as the reaction process occurs unlike the
batch reactors in which feed and all required material for fermentation to take place are fed at once.
19. Define aspect ratio.
The height to diameter ratio of a cylindrical bioreactor.
20.What is a sparger?
Device used for introducing a stream of gas in the form of bubbles into a liquid.
21. What is the unit of mass transfer coefficient?
The unit of mass transfer coefficient is mol/m2. s.
22. What is the use of sparger in a fermentor. (May 2017)
In aerobic cultivation process the purpose of the sparger is to supply oxygen to the growing cells.

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PART – B
1. Discuss the different resistances to gaseous transfer through liquid for use in intercellular reactions.
(Nov 2014)
2. Explain the mass transfer coefficient and its role in scale-up. (Nov 2014)
3. i) Discuss the aeration and agitation in fermenters.
ii) Discuss the mass transfer theories applied to oxygen transfer. (May 2014)
4. Write about sterilization of fermentation media and system. (Nov 2014)
5. Give a brief account on i) sterilization cycle (May 2015). ii) Newtonian and non-Newtonian fluids.
(May 2014)
6. i) Outline the various heat transfer configurations for bioreactors used to improve the heat transfer
rate? (May 2015)
ii) Results from a static gassing out method is given,
Time (min) 0 1 2 3 4 5 6
%00 0 21 43 60 74 85 90
Calculate volumetric mass transfer coefficient for the reactor. (Nov 2015)
7. A fermenter used for antibiotic production must be kept at 27 °C. After considering the oxygen
demand of the organism and the heat dissipation from the stirrer, the maximum heat transfer rate
required is estimated as 600 Kw. Cooling water is available at 10 °C; the exit temperature of the
cooling water is calculated as 25°C. The heat transfer coefficient for the fermentation broth and
cooling water are 2050 and 14000 Wm-2°C-1. It is proposed to install a helical cooling coil inside the
fermenter, the outer diameter of the coil pipe is 8 cm, the pipe thickness is 5 mm and the thermal
conductivity of the steel is 65 Wm-2 ° C-1. An average internal fouling factor of 800 Wm-2 ° C-1 is
expected; the fermenter side surface of the coil is kept relatively clean. What length of cooling coil is
required? (Nov 2015)
8. i) Explain briefly about gas liquid contacting modes. ii) Explain in detail about enhancement of
oxygen transfer. (May 2015)
9. Write about power consumption in an agitator.
10. Explain the steps involved in the determination of KLa by Sulphite Oxidation, static and dynamic
method. (Nov 2013)
11. Explain the standard tank design concept.
12. Explain the various criteria and problem associated with the scale-up of bioreactors.
13. Write about scale-up method in fermentation system.
14. Derive an expression for thermal death kinetics. ii) Write about batch and continuous sterilization.
15. Describe the various steps encountered in the transfer of oxygen from gas phase to microbial cell.
(May 2017)
16. Outline the various heat transfer configurations for bioreactors used to improve the heat transfer
rate? (May 2017)
17. Why do foams form in fermentation systems and what problems to they cause? How can foam
formation be controlled? (May 2017)
18. Give a detailed description about the transport phenomena in bioprocess systems. (Dec 2017)
19. Elaborate the scaling of mass transfer equipment. (Dec 2017)
20. How do agitation and aeration affect microbial growth? How does aeration help in agitation and
mixing? Explain (May 2016)
21. Discuss the mass transfer theories applied to oxygen transfer. (May 2016)
22. Discuss in detail about the sterilization cycles. Enumerate various methods of sterilization that are
used for industrial fermentation. (May 2016)

UNIT V: DOWN STREAM PROCESSING

Part - A
1. What is Scale up? (May 2015, Nov 2014)
Scaling-up is a procedure where the results of small scale experiments are used as the basis for the
design, testing and implementation of a larger scale system.
2. Define a pilot scale. (Nov 2013)
The size of a system between the small laboratory model size (bench scale) and a full-size system.
3. What are the different methods of sterilization? (May 2014)
The various methods of sterilization are:
1. Physical Method (a) Thermal (Heat) methods (b) Radiation method (c) Filtration method
2. Chemical Method

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3. Gaseous method
4. Compare between batch and continuous bio reactors. (May 2014/2016)
Batch Reactors:
Simplest type of mode of reactor operation. In this mode, the reactor is filled with medium and the
fermentation is allowed to proceed. When the fermentation has finished the contents are emptied for
downstream processing. The reactor is then cleaned, re-filled, re-inoculated and the fermentation
process starts again.
Continuous reactors:
Fresh media is continuously added and bioreactor fluid is continuously removed. As a result, cells
continuously receive fresh medium and products and waste products and cells are continuously
removed for processing. The reactor can thus be operated for long periods of time without having to
be shut down.
5. What is Residence Time Distribution? (May 2015)
The residence time distribution (RTD) of a chemical reactor is a probability distribution function that
describes the amount of time a fluid element could spend inside the reactor
6. Define micro mixing. (May 2015)
In pharmaceutics, micro mixing is a process in which ingredient particles rearrange to form a blend.
Development of pharmaceutical formulations requires understanding how the ingredients blend with
each other and how the blending progresses through different stages.
7. What are the steps involved in the construction of microbial biosensors.
Culture of microorganisms, preparation of microbe immobilized membrane, construction of microbial
biosensor.
8. What is PTFE?
Poly tetra fluoro ethylene membrane used in gas sensors.
9. Give the classification of microbial biosensor.
Classified either as respiration activity measurement or electrochemically active metabolite
measurement.
10. What are the important points while immobilizing microorganisms for biosensors?
In the case of oxygen electrode based sensors gas permeability through the microorganism
immobilized enzyme is important. If it is based on the function of living cells a very gentle method for
microbe immobilization must be selected.
11. What is a thermistor?
A device used in the measurement of metabolite heat evolved from the immobilized microorganism
used in microbial biosensors.
12. Mention the steps involved in isolation of products from bioreactor broth.
(i) Removal of particulates; (ii) primary isolation; and (iii) purification.
13. What is ultrafiltration?
A membrane separation process used in the retention of molecules of size 10 to 1000 in diameter.
Useful in both product concentration and purification.
14. What is salting – out.
A purification technique used to precipitate proteins in solution using salts.
15. What is the role of spacer arm in affinity chromatographic media?
Provides flexibility of ligand within the matrix. Thus proteins are easily isolated.
16. Mention the advantages of using ammonium sulphate.
Highly soluble at low temperatures, less inexpensive.
17. Mention some membrane separation techniques used in protein isolation.
Microfiltration, ultrafiltration, reverse osmosis, electrophoresis.
18. What is electrophoresis?
Separation of charged species based on their specific migration rates in an electric field .
19. What factors control the separation of one protein from another in gel filtration
chromatography?
(i) Chromatography column length; and
(ii) Rate of diffusion of protein molecules from the feed.
20. Give the advantage of airlift in comparison with other bioreactors. (Nov 2015)
No moving parts, simple in construction, high gas absorbance efficiency, and good heat transfer.
21. What is minimum fluidization velocity?
The superficial liquid velocity needed to just suspend the solids from a settled state.
22. State the advantages of membrane reactor.
A considerable advantage of membrane reactor lies in the fact that the biocatalyst can be used in their
native form, thus avoiding exposure to the usual inactivating steps.

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23. Discuss on trickle bed reactors. (Nov 2011)

TBR are 3 phase systems containing a packed bed of heterogeneous biocatalyst and flowing gas and
liquid phases. Feed consists of both liquid and gas phase. Biochemical reaction takes place by contact
of reactant in the liquid phase and the reactant from the gas phase at the catalyst surface.
24. Write down the performance equation of continuous reactor. (Nov 2013)

25. What are microbial biosensors?

They are composed of immobilized microorganisms and an electrochemical device. These sensors are
suitable for online control of biochemical processes.

26. What is the purpose of recycling in a bioreactor? (Nov 2014)

For industrial processes, cells, broth, or both, may be reused. Recycle of spent broth has the added
advantage of saving expensive substrate that would otherwise be needed for new cell growth. Usually
the carbohydrates in a bioprocess are pretty well exhausted, but much of the expensive nitrogenous
ingredients remains after the process.
27.Name some electrodes used in construction of biosensors. How does contamination affect
biosensors?
Oxygen, ammonia, pH electrodes.
contamination affect biosensors: Contamination will affect selectivity and sensitivity of microbial
sensors.
28. What are the various methods of immobilizing enzymes in microbial biosensors?
Membrane entrapment, gel entrapment.

29. How is crystallization process carried out? (Dec 2017)

A solid product that has been separated from a liquid reaction mixture by filtration or
crystallization will still contain some of the solvent. It can be dried by placing it in an oven or into a
desiccator. This technique is used to separate a solid from a liquid.

PART – B
1. When using a Ruston turbine for mixing an ungassed Newtonian fluid, if you were to increase the
impeller rotational speed by 2, by how much would the power requirements increase? Explain when
gas is sparged into the fermentation broth do the power requirements increase or decrease? Why?
(Nov 2015)

2. Discuss the design of bioreactors and scale up with an example. (Nov 2013, May 2014)
3. Medium at a flow rate of 2 m3 h-1 is to be sterilized by heat exchange with steam in a continuous
sterilizer. The liquid contains bacterial spores at a concentration of 6 X 10 12 m-3; the activation energy
and Arrhenius constant for thermal destruction of these contaminants are 283 kJ gmol -1 and 5.7
X10.39 h-1, respectively. A contamination risk of one organism surviving every 60 days’ operation is
considered acceptable. The sterilizer pipe has as inner diameter of 0.1m; the length of the holding
section is 24m. The density of the medium is 1000 kgm-3 and the viscosity is 3.6 kgm-1h-1. What
sterilizing temperature is required? (Nov 2015)
4. What are the main parameters to be monitored in the fermenter? What are the problems do they
cause if it is not controlled? (Nov 2015)
5. Discuss in detail about high performance bioreactors. (May 2014, Nov 2014)
6. Compare and contrast between batch and continuous reactor with respect to bioprocess.
(Nov 2014)
7. Explain the reactors in series with recycle. (Nov 2013)
8. A stirred tank reactor is to be scaled down from 10 m3 to 0.1 m3. The dimensions of the large tank
are Dt= 2m; Dim = 0.5m; N=100 rpm; H=3Dt
i) Determine the dimensions of the mall tank (Dt, Dim, H) by using geometric similarity.
ii) What would be the required rotational speed of the impeller in the tank if the following criteria
were used? a) Constant tip speed b) constant impeller Re number? (Nov 2015)
9. i) write brief notes on immobilized whole cell and enzyme reactors. ii) Discuss about sterile and
non-sterile operations. (May 2015)

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10. i) Briefly explain the design of ideal plug flow tubular reactor. ii) explain in detail about reactors
in series with recycle. (May 2015)
11. Explain the basic steps and unit operations involved in the downstream processing operations
12. Discuss about various instruments and control apparatus used in bioreactors.
13. Describe the working of Fluidized bed reactor. ii) Explain the working of bubble column reactor.

14.Write about conventional packed bed reactors.

15. Write about the classification of bioreactors and also enlist the advantages and disadvantages of
any three basic fermenters.
16. Describe the working and classification of loop fermenter.
17. Write about the reactor configurations for conducting enzyme catalyzed process.
18.Describe the role and importance of downstream processing in bioprocess.
19. Give an account of the action of enzymes and chemical agents on cell disruption? (May 2017)
20. Write short notes on Gel Filtration Chromatography, Electrophoresis and Ultra Filtration. (May
2017)
21. What are aqueous biphasic systems? Explain the steps involved in the aqueous two-phase
extraction? (May 2017)
22. With a relevant example, explain the strategy of recovering and purifying a product. (Dec 2017)
23. Give a detailed account on the separation of soluble products using Dialysis and Ultra-Filtration
techniques. (Dec 2017)
24. Write short notes on (May 2016)
i. Enzyme Reactors
ii. High performance bioreactors.
25. Write in detail about (May 2016)
i. Sterile and Non Sterile operations
ii. Reactors in series with recycle and methods of scale up

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