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CHE 172

COURSE OVERVIEW AND CLASS POLICIES

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Course Description Introduction to Biochemical Engineering

OBJECTIVES OF THE COURSE

• Provide concepts in microbiology and biochemistry

relevant to bioprocessing

• introduction to the mathematical aspects of cell growth

and application to bioreactor design

• introduction to chemical engineering aspects of

industrial sterilization and bioseparation (downstream

processing) technologies

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Course Requirements

• 4 Long Exams

• SARQ (Seatwork, Assignment, Recitation,

Quizzes)

• Oral/Written Report on the Industrial Production

of a Biotechnological Product (if there is still

time)

PREFINAL GRADE= Average of LE + SARQ + (Report)

FINAL GRADE= 70% PreF Grade + 30% Final Exam

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Course Requirements (contd)

Exemption from Final Exams:

A PreFinal Grade of 70% with

No Long Exam grade lower than 50%

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CHE 172 Lecture A

Introduction to Biochemical

Engineering and Industrial

Bioprocessing

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• the practical application of

biological agents

(living/dead cells or the sub-cellular components)

in technically useful operations ,

either in productive manufacture, services

operations or environmental management

BIOTECHNOLOGY

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Biological Agents

• Microorganisms (Bacteria, Yeasts,

Filamentous Fungi, etc)

• Plant/Animal Cells

• Sub-cellular components, Enzymes

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Technically Useful Operations

• Manufacture of an economically

useful/value added product

– Single cell protein, pharmaceuticals, industrial

chemicals, etc

• Waste

Treatment/Biodegradation/Bioremediation

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BIOTECHNOLOGY IS A BIG INDUSTRY

(variety of products)

Fermentation Product Typical organism used Approximate

World Market

(tons/yr)

Bulk organics

Ethanol

Acetone/butanol

Saccharomyces cerevisiae

Clostridium acetobutylicum

2 x 107

2 x 106

Biomass

Single-cell

protein

Candida utilis/ Pseudomonas,

methlotrophus, Baker’s yeast

0.5-1 x 106

Organic acids

Citric Acid

Lactic Acid

Aspergillus niger

Lactobacillus delbrueckii

2-3 x 106

2 x 105

Amino acids

L-glutamate

L-lysine

L-phenylalanine

Corynebacterium glutamicum

Brevibacterium flavum

Corynebacterium glutamicum

3 x 106

3 x 105

2 x 103

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BIOTECHNOLOGY IS A BIG INDUSTRY

(variety of products)

Fermentation Product Typical organism used Approximate World

Market (tons/yr)

Exocellular

polysaccharides

xanthan gum

dextran

Xanthomonas campestris

Leuconostoc mesenteroides

5 x 103

small

Enzymes

proteases

amylases

pectinases

Bacillus sp.

Bacillus amyloliquifaciens

Aspergillus niger

6 x 102

4 x 103

10

Vitamins

Vitamin B12

Propionicum shermanii

10

Pigments

shikonin

beta-carotene

Lithospermum erythrohizon (plant cell)

Blakeslea trispora

60 kg/yr

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BIOTECHNOLOGY IS A BIG INDUSTRY

(variety of products)

Fermentation Product Typical organism used Approximate

World Market

(tons/yr)

Vaccines

tetanus

hepatitis B

Clostridium tetani

Surface antigen in recombinant yeast

< 50 kg/yr

Therapeutic proteins

Insulin

Interferon

Recombinant E. coli

Recombinant E. coli

< 20 kg/yr

Monoclonal antibodies hybridoma cells < 20 kg/yr

Antibiotics

penicillins

cepaholosporins

tetracyclines

Penicillum chrysogenum

Cephalosporium acremonium

Streptomyces aureofaciens

3-4 x 105

1 x 105

1 x 105

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Biotechnology: An Interdisciplicary Endeavor

• The ability to harness capabilites of cells (development of new products and processes) usually starts in the laboratory:

Microbiology, Biochemistry , Cell physiology, Molecular biology/Genetics

• Bringing a “bioprocess” to industrial realization requires engineering skills and know-how:

Biochemical engineering or

Bioprocess Engineering

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What is the importance of biochemical

engineering in the biotechnology industry?

Biochemical engineering is one of the major areas of biotechnology important to its commercialization (Lee 1992).

Successful commercialization of

biotechnology requires the development of a

technologically viable and economically efficient process.

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Role of the biochemical engineer for

commercial realization of biotechnology

(1)Bioreactor: scale up, design, optimal

operation and control

(2)Downstream processing equipment:

design and operation

(3)Fermentation plant design

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ORIGINS AND EVOLUTION OF

BIOTECHNOLOGY

How did industrial fermentations

begin?

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EVOLUTION OF BIOTECHNOLOGY

The First Wave

• Microbial Production of

Food and Beverages

– 6000 BC Sumerians and

Babylonians were already

drinking beer (accidental

observation?)

– 4000 BC Egyptians were

already baking leavened

bread

– wine was known in the

Near East by the time of

the Book of Genesis

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EVOLUTION OF BIOTECHNOLOGY

The First Wave

• Microbial Production of Food and Beverages

– By 17th Century, the realization that

microorganisms had a role in wine, beer making

started

– Anton van Leeuwenhoek’s microscope

• Microbial Production of Food and Beverages

More of a “craft”;

involvement of microbes not yet known

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EVOLUTION OF BIOTECHNOLOGY

The First Wave

• Microbial Production of Food and Beverages

– Louis Pasteur gave definitive proof of the the

fermentative abilities of microorganisms

– Louis Pasteur can justifiably be considered as the

father of biotechnology

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EVOLUTION OF BIOTECHNOLOGY

The First Wave

• Microbial Production of Food and Beverages – Other Microbially bases processes: fermented milk,

yoghurt, cheeses, soy sauce, tempeh, etc.

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EVOLUTION OF BIOTECHNOLOGY

The Second Wave

• Biotechnological processes initially

developed under non-sterile conditions many industrial compounds such as ethanol, acetic acid, organic

acids, butanol and acetone were produced by the end of the 19th

century by microbial fermentation procedures that were open to the

environment

the control of contaminating microorganisms were achieved by

careful manipulation of the ecological environment and not by

complicated engineering practices.

municipal composting of solid wastes and wastewater treatment are

outstanding examples of non-sterile biotechnology

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EVOLUTION OF BIOTECHNOLOGY

The Third Wave • The introduction of sterility to

biotechnological processes

– A 1940’s: a new direction in biotechnology with the introduction of complicated engineering techniques to the mass cultivation of microorganisms to ensure that the particular biological process could proceed at higher yields with the exclusion of contaminating microorganisms. (birth of biochemical engineering thought to be here)

– period of increasing volumes of biotechnological activies: antibiotics, amino acids, organic acids, enzymes, steroids, polysaccharides, vaccines Modern Bioreactor

http://www.biotopics.co.uk/edexcel/fermtr.gif

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EVOLUTION OF BIOTECHNOLOGY

The Third Wave

• The introduction of sterility to biotechnological processes

– A 1940’s: a new direction in biotechnology with the introduction of complicated engineering techniques to the mass cultivation of microorganisms to ensure that the particular biological process could proceed at higher yields with the exclusion of contaminating microorganisms. (birth of biochemical engineering thought to be here)

– period of increasing volumes of biotechnological activies: antibiotics, amino acids, organic acids, enzymes, steroids, polysaccharides, vaccines

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EVOLUTION OF BIOTECHNOLOGY

The Fourth Wave

• Explosive developments in molecular biology and process control have created new and exciting opportunities to create new frontiers and to improve greatly the efficiency and economics of the established biotechnological industries

• production of human insulin from E. coli,

• monoclonal antibodies for detection and treatment of diseases

• plant tissue/animal cell culture

• protoplast fusion

• artificial intelligence for control of bioreactors

• artificial organs, stem cells

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Stem cells are the source, or “stem,” for all of the

specialized cells that form our organs and tissues.

That’s why stem cells are able to change into other types

of cells, something no other kind of cell can do.

Each time a stem cell divides, it can remain a stem cell or

change into a heart, blood, brain, or other type of cell.

Theoretically, stem cells can even divide without limit to

replenish themselves and other cells.

STEM CELL TECHNOLOGY

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Stem cells are found in very early embryos and a few adult

organs. In embryos, stem cells produce the first cells of the

heart, brain, and other organs.

In adults, stem cells can be found in a few kinds of tissues

that need constant replenishing. For example, stem cells in

bone marrow produce new blood cells to replace those lost

through normal wear and tear or injury.

An important difference between embryonic and adult

stem cells involves how many different types of cells they

can develop into.

Embryonic stem cells have the potential to form just about

any kind of cell in the body, but

Adult stem cells are only able to form a few new cell types.

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THE BIRTH OF BIOCHEMICAL

ENGINEERING

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The Birth of Biochemical

Engineering

• Necessity to produce large quantities of more effective antimicrobials for the war effort

(WW II)

• Many soldiers dying from infectious wounds

• Sulfa drugs losing effectivity due to resistance

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The Birth of Biochemical

Engineering

• Scientists at Oxford University rediscover’s Alexander

Flemming’s earlier publication on the germicidal

properties of “mold juice” which was largely unnoticed

• Germicidal component named “Penicillin”, after the

genus of the mold

• Oxford University scientists proved that penicillin

could effectively treat wound infections

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The Birth of Biochemical Engineering

• Penicillin was heralded as a “wonder drug” at the

time

• Intial attempts for mass production was space

consuming with very low product yields

• (as low as 0.001 g/L)

• Pfizer was the first pharmaceutical company to take the challenge of mass producing penicillin

• After 3 years of difficulty (low yield and stability of product), and new approach using deep tank fermentation (“submerged fermentation”) was attempted.

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The Birth of Biochemical Engineering

• The chemical engineering techiques learned for high penicillin production by fermentation in a stirred tank reactor became the foundation for the biochemical engineering field

• The penicillin yield increased 50-fold

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• The penicillin process also established a

paradigm for bioprocess development and

biochemical engineering. This paradigm

still guides much of the profession’s

thinking.

• The mind set of bioprocess engineers was

cast with the penicillin experience.

The Birth of Biochemical Engineering

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Commercialization of Biotechnology

Involves Scale Up from Laboratory

Scale (small) to Production Scale

(large)

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Small Scale vs Large Scale How Does the Story Change?

SMALL

SCALE

LARGE

SCALE

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Small Scale vs Large Scale

How does the story change when you implement the following stages of fermentation from small to large scale?

(1) Medium preparation (2) Sterilization (3) Inoculation (4) Main Fermentation (5) Product Separation and Purification (downstream processing)