BCHE315L-BIOCHEMICAL ENGINEERING

LECTURE – 1

Dr. K. Sivagami - Associate Professor

Room#: SMV 108
Mobile: 9487628920
Email: sivagami.k@vit.ac.in
COURSE OBJECTIVES

1. Impart the basic knowledge and overview of biotechnology covering the

principles of cell and kinetics, bioreactor design, sterilization agitation and
aeration.

2. Understand the physical processes involved in bio-systems.

3. Apply the knowledge of chemical engineering principles to biological

processes.
COURSE OUTCOMES

1. Describe the significance and scope of biochemical processes with their

metabolic Pathways.
2. Understand basic principles of enzyme and microbial growth kinetics.
3. Apply basics of chemical engineering transport processes in designing
bioprocess systems.
4. Analyze bioreactor performance and their transient characteristic.
5. Distinguish downstream processing methods to purify and separate
biological products.
MODULE: 1 INTRODUCTION TO BIOCHEMICAL ENGINEERING

Number of hours: 3
An overview of industrial biochemical processes with typical examples -
comparing Chemical and Biochemical processes – Development and scope
of biochemical engineering as a discipline.

MODULE: 2 BASIC MICROBIOLOGY AND BIOCHEMISTRY

Number of hours: 5
Basics of Biology - overview of biotechnology - Diversity in microbial cells -
Cell constituents - Chemicals for life - Examples of microbial synthesis -
Major metabolic pathways – Bioenergetics - Glucose metabolism –
Biosynthesis.
MODULE: 3 ENZYMES & ENZYME KINETICS
Number of hours: 8
Enzymes - Classification of enzymes - Mechanism of enzymatic reactions –
Michaelis - Menten kinetics – Enzyme inhibition - Inhibition kinetics -
Enzyme denaturation and inactivation- Factors affecting the reaction rates
- Enzyme immobilization – kinetics of immobilized enzymes - Mass
transfer effects on immobilization.

MODULE: 4 KINETICS OF CELL GROWTH

Number of hours: 6
Typical growth characteristics of microbial cells - Factors affecting growth -
Unstructured models of microbial growth - Monod model - Modelling of
batch and continuous cell growth inhibition on cell growth - Immobilized
whole cells and their characteristics.
MODULE: 5 TRANSPORT IN MICROBIAL SYSTEMS
Number of hours: 7
Rheological behaviour of broth - Agitation and mixing - Power
consumption - gas/liquid transport in cells - Mass transfer coefficients and
its measurement – Oxygen transfer Factors affecting oxygen transfer rate -
Heat transport in microbial systems – Thermal death kinetics of
microorganism - batch and continuous sterilization - air and media
sterilization.
MODULE: 6 BIOREACTORS
Number of hours: 8
Classification of bioreactors - Batch and continuous types - Fed-batch
reactors - Free and immobilized whole-cell and enzyme reactors - Reactors
in series with and without recycle Transient behavior of bioreactors -
Design of reactors and scale up with examples.
MODULE: 7 DOWNSTREAM PROCESSES

Number of hours: 6
Different unit operations in down streaming with special reference to
filtration centrifugation - extraction - membrane separations -
crystallization - chromatographic techniques – drying – cell desruption
technologies. .

MODULE: 8 CONTEMPORARY ISSUES

Number of hours: 2
Classification of bioreactors - Batch and continuous types - Fed-batch
reactors - Free and immobilized whole-cell and enzyme reactors - Reactors
in series with and without recycle Transient behavior of bioreactors -
Design of reactors and scale up with examples.
BOOKS FOR SYLLABUS

Textbook
• Rao D.G., Introduction to Biochemical Engineering, 2012, 2nd ed., Tata
McGraw Hill, India.
• Harvey W.Blanch and Douglas S. Clark, Biochemical Engineering, 1997,
2nd ed., CRC Press, USA.

Reference Books
• Doran P.M., Bioprocess Engineering Principles, 2013, 3rd ed.,
Academic Press, United Kingdom
• Bailey J.B., Ollis D.F., Biochemical Engineering Fundamentals, 2010, 4th
ed., McGraw Hill, USA.
 COURSE EVALUATION
Type of Max. marks for which the Marks
Evaluation exam is conducted

CAT-I To be decided 50 Marks (15 marks)

CAT-II To be decided 50 Marks (15 marks)
2 Quizzes Each quiz will be conducted for a 20 marks
minimum of 10 marks (10 marks from each quiz)
Assignment In the form of a report, seminar, 10 marks
presentation, quiz, experiment, GD, etc.
as defined in the course syllabus/
course plan
FAT To be decided 40 marks

Total 100 marks

 BIOLOGY & BIOTECHNOLOGY

Biology –

• Branch of science which deals with the study of the living things.
• Activities which are performed by the living system or living cell
include locomotion, growth, respiration, reproduction, metabolism.

11
 BIOLOGY & BIOTECHNOLOGY

Biotechnology –
It is the art of science of converting reactants into useful products by
the action of microorganisms and enzymes in the areas of agriculture,
food, medicine, pharmaceutical, environment .

Novel techniques in Biotechnology include

➢ recombinant DNA and cell fusion.

➢ Commercialization of these processes require need to design
effective bioreactor to cultivate the cells in the most optimum
conditions.
12
 APPLICATIONS OF BIOTECHNOLOGY
S.NO AREA PRODUCTS /APPLICATIONS
1 Pharmaceuticals Antibiotics, antigens, human growth hormones, insulin,
interferon, interleukins.
2 Animal agriculture Development of disease free, healthier high yielding food
animals.
3 Plant agriculture Development of plants with increased abilities of
photosynthesis and nitrogen fixation.
4 Specialty chemicals Amino acids, enzymes, vitamins, lipids, biopolymers.

5 Environmental applications Mineral leaching, metal concentration, pollution control,

toxic waste degradation, enhanced oil recovery.

6 Commodity chemicals Acetic acid, acetone, butanol, ethanol and other products
from biomass conversion processes.
7 Bioelectronics Biosensors, biochips. 13
 DEFINITIONS

Bioprocessing:
“Any process in which microbes or living organisms play a vital
role in getting transformation of the feed into useful products is
termed as bioprocessing”.

Biochemical Processes:
"A process that uses living cells or biomolecules to carry out a
chemical transformation leading to the production and ultimate
recovery of valuable products".
14
 SHORT HISTORY OF BIOCHEMICAL PROCESSES

Ancient Uses of Microorganisms (Before 1800 A.D.)

• Caveman to Earliest Recorded History --- aging of meats, cheeses, and
alcoholic beverages.
• Ancient Chinese and Japanese -- soy sauce from fermented beans.
• Ancient Greeks-Produced wine as early as 7000BC
• Ancient Egyptians (4000BC)-Baking Bread and during 2500 B.C. --
malting of barley and beer fermentation.
• Mesopotamian - Brewing of wine and beer are established in 2000 B.C.

15
 HISTORY OF BIOCHEMICAL PROCESSES

• Columbus lands in North America to find the native peoples drink

beer made from corn.

• Chinese use moldy soy bean curd to clear up skin infections

(1000B.C.)

• Central American native peoples use fungi to treat infected wounds.

• Middle Ages experimenters learn how to improve the taste of wine,

bread, beer, and cheese.

• Mankind did not know that these fermentation processes were being
carried out by microscopic forms of life. 16
 Old Science (1800-1940)

 From the discovery of the role of microscopic life in fermentations to

the use of non-sterile fermentations in organic molecule synthesis.

 1803 -- A French scientist, L.J. Thenard, announces that yeast used in

wine making were alive and that they were responsible for the
formation of alcohol. His findings were rejected by supporters of the
conventional notion that fermentations were chemical processes only.

 1838– Schleiden and Schwann proposed cell theory.

17
 BIRTH OF MODERN MICROBIOLOGY

• 1857 -- Louis Pasteur, another French Scientist, proves Thenard is

correct. Showed that certain diseases are caused by microorganisms.

Concludes that certain microorganisms are destroyed by other

microorganisms and suggests that human disease could be cured by
pitting microbe against microbe.

18
 OLD SCIENCE (1800-1940)

 1901 -- Rudolf Emmerich and Oscar Low, University of Munich,

isolate a primitive antibiotic, pyocyanase, from Pseudomonas
aeruginosa, a bacterium.

 Several hundred patients were successfully treated, but quality

control was poor and pyocyanase was abandoned as too hazardous

19
 OLD SCIENCE (1800-1940)

1900 – 1940:
 Production of bakers yeast in deep, aerated tanks.
 World War I -- Chaim Weismann solves a serious British ammunition
problem by converting corn maize mash into acetone, which is used
in the manufacture of the explosive cordite.
 1923 -- Pfizer opens the first commercial successful plant for citric
acid production from sugar.
 1928 -- Alexander Fleming discovers penicillin.
 Simple organic molecules such as glycerol, lactic acid, and butanol are
fermented on an industrial scale by fermentations
20
 NEW SCIENCE (1940-LATE 1970S)

• Fermentations of complex organic molecules requiring sterile

conditions which protect the non-robust, highly selected microbial
strains from competition by other microorganisms

• 1940 -- Dr. Howard Flory and Ernst Chain (England) and three
American pharmaceutical companies (Merck, Pfizer, and Squibb)
mass produce penicillin for WW-II effort.

21
 NEW SCIENCE (1940-LATE 1970S)

• Pioria Illinois -- 1940's, government worker discovers a new strain

of Penicillium on a moldy cantaloupe which can produce 200
times more penicillin than Fleming's strain.
• Selman A. Waksman of Rutgers University discovers a new
antibiotic, streptomycin, for the treatment of tuberculosis.
• Fermentive syntheses of amino acids, vitamins, cortisone, nucleic
acids, polysaccharides, and enzymes.

22
ERA OF MOLECULAR BIOLOGY (LATE 1970S-PRESENT)

 1973 Herbert Boyer (University of California, San Francisco) and Stanley

Cohen (Stanford University) establish recombinant DNA technology

 The discovery of recombinant DNA technology and the birth of genetic

engineering allows for the efficient production of compounds not
indigenous to the host microorganism.

23
 Biochemical Engineering -Introduction

• Application of engineering principles to conceive, design, develop,

operate processes and products based on biochemical phenomena.

 Bio Chemical Processes:

"A process that uses living cells or biomolecules to carry out a

chemical transformation leading to the production and ultimate
recovery of valuable products".

24
 Biochemical Engineering -Introduction

• Biochemical Engineering:

“It is a branch of Chemical Engineering or Biological Engineering

that mainly deals with the design and construction of most
processes that involve biological organism.”

• Biomolecular engineering:

“Emphasis on the molecular basis biological phenomena influencing

broad range of industries”

25
 INTRODUCTION (CONTD…)

Biochemical Engineering includes:

• Basics of biology, overview of biotechnology, diversity of microbial
cells, cell constituents, chemicals for life.
• Kinetics of enzyme catalysis, immobilized enzymes, microbial growth
kinetics, design analysis and stability of bioreactors.
• Biological product synthesis and separation.
• Fusion of biology and chemical engineering , utilizes the techniques of
the later to enhance products from biological synthesis.

26
 INTRODUCTION (CONTD…)

Bioprocessing steps:
• Process involving microbial cells, raw materials usually biomass are
treated and mixed with nutrients for cell growth.
• Liquid mixture and medium are sterilized and introduced in to the
bioreactor/fermenter.
• The reactor is equipped with agitators, baffles, air spargers and
sensing devices.

27
 INTRODUCTION (CONTD…)

Bioprocessing steps:
• A pure strain of microorganism is introduced and the cells start to
multiply and reach maximum concentration.

• The medium is now depleted with and fermentation is stopped.

• The contents are then pumped out for product recovery and
purification in Batch or continuous process.

28
 ROLE OF BIOCHEMICAL ENGINEERS

“Perhaps the most notable contribution the engineer made was in the

advancement of sterile techniques, or the ‘contamination –free”

philosophy, in the design and operation of the fermentation vessel

and its associated maze of piping”

-Aiba et al.( 1965)

29
ROLE OF BIOCHEMICAL ENGINEERS (CONTD…)
PREPARATION
OF BIOMASS FOAM CONTROL pH CONTROL
Innoculum Stages Antifoam Addition Acid-Alkali Addition

Bioreactor
PRODUCT RECOVERY

CELL SEPARATION
BIOREACTOR Intracellular
product
Extracellular
1). CELL DISTRUPTION product
2). PRODUCT EXTRACTION

Free Cells,
Immoblized Cells PRODUCT
or CONCENTRATION
PROCESS
Enzyme Bioreactor

PRODUCT
SEPARATION

PURIFICATION

STERILIZATION
DRYING

RAW MATERIAS Air FINAL PRODUCT

Nutrients and Reactants
in Aqueous Solution 30
(may contain insoluble
organic and/or inorganic
materials)
ROLE OF BIOCHEMICAL ENGINEERS (CONTD…)

31

Industrial Bioreactor
 ROLE OF BIOCHEMICAL ENGINEERS (CONTD…)
Process design and development:
• Acquire knowledge of microbiology, biochemistry, molecular biology,
genetics etc to select a catalyst or to genetically modify for large scale
operations.
• Know about the kinetics of reactions , physical and chemical conditions
affecting the kinetics, operating conditions influencing the rate to design an
effective bioreactor.
• Develop reliable online sensing devices and ensure that the processes are
operated at the most economical points.
• Implement various separation techniques for downstream processing.
Achieve maximum purity with minimum cost.
• Develop novel techniques (at large scale) to separate biological materials.32
 ROLE OF BIOLOGICAL SCIENTISTS

Biochemical Engineers work with Biological Scientists

• To obtain best biological catalyst for a desired process.

• To create best possible environment for the catalyst.
• To design and operate the bioreactors in the most efficient way.
• To separate desired products from the reaction mixtures in the most
economical way.

33
Typical Biochemical Engineering

Upstream processing

Downstream processing
Bioreactor
Chemical vs Biochemical processing
Chemical Vs Biochemical industry

36
 Biochemical Engineering – Salient Features

1. Mild reaction condition:

The reaction conditions for bioprocesses are mild. The typical condition is at
room temperature, atmospheric pressure, and neutral medium pH.
2. Specificity:
An enzyme catalyst is highly specific and catalyzes only one or a small number
of chemical reactions. A great variety of enzymes exist that can catalyze a very
wide range of reactions.
3. Effectiveness:
The rate of an enzyme-catalyzed reaction is usually much faster than that of
the same reaction when directed by nonbiological catalysts. A small
amount of enzyme is required to produce the desired effect.
 Biochemical Engineering – Salient Features

4. Renewable resources:
The major raw material for bioprocesses is biomass which provides both the
carbon skeletons and the energy required for synthesis for organic chemical
manufacture.

5. Recombinant DNA technology:

The development of the recombinant DNA technology promises enormous
possibilities to improve biological processes.
Biochemical Engineering – Disadvantages
Complex product mixtures:
➢ In cases of cell cultivation (microbial, animal, or plant), multiple
enzyme reactions are occurring in sequence or in parallel.
➢ The final product mixture contains cell mass, many metabolic by-
products, and a remnant of the original nutrients.
➢ The cell mass also contains various cell components.

2. Dilute aqueous environments:

➢ The components of commercial interests are only produced in
small amounts in an aqueous medium. Therefore, separation is
very expensive.
➢ Since products of bioprocesses are frequently heat sensitive,
traditional separation techniques cannot be employed.
➢ It requires novel separation techniques that have been developed
for analytical purposes, need to be scaled up.
Biochemical Engineering – Disadvantages
3. Contamination:
The fermenter system can be easily contaminated, since many
environmental bacteria and molds grow well in most media.

The problem becomes more difficult with the cultivation of plant or

animal cells because their growth rates are much slower than those
of environmental bacteria or molds.

4. Variability:
Cells tend to mutate due to the changing environment and may lose
some characteristics vital for the success of process. Enzymes are
comparatively ensitive or unstable molecules and require care in
their use.
 Role of Chemical Engineers
To carry out a bioprocess on a large scale, biochemical engineers need
to work together with biological scientists:
• To obtain the best biological catalyst (microorganism, animal cell,
plant cell, or enzyme) for a desired process.
• To create the best possible environment for the catalyst to perform by
designing the bioreactor and operating it in the most efficient way.
•To separate the desired products from the reaction mixture in the
most economical way.
Important Biochemical Processes
43
Classification of Biochemical Processes based on products

• Primary metabolite – Ethanol, lactic acid, some aminoacids

• Secondary metabolite - antibiotics, naphthalenes, nucleosides,
peptides and growth factors.
• Microbial cell – Yeast, single cell protein, vaccine
• Recombinant protein - Insulin
 Ethanol (Primary metabolite) – Batch process

Temp: 30°-35°C, pH: 4-6

Glucose < 100g/l,
ethanol < 50g/l
Glucose Ethanol

Saccharomyces cerevisiae (Baker’s Yeast)

WORK IN A TEAM ENVIRONMENT WITH CHEMISTS, BIOCHEMISTS,
MICROBIOLOGISTS, AND CHEMICAL ENGINEERS.

Steps in the development of a new biochemical process and role of

professionals.

1.Identify a desired reaction or product (chemist, biochemist).

2.Identify key enzyme(s) or microorganism (biochemist, microbiologist).

3.Process development (chemist, biochemist, microbiologist, chemical

engineer).

4.Design of bioreactor and recovery unit operations (chemical engineer).

5.Metabolic Engineering:

Application of engineering analysis to metabolic pathways within

microorganisms. 46