2003 LECTURES , Biotechnology

Introduction to chemical engineeringIntroduction to chemical engineering

Biotechnology and the Biotechnology and the role of chemical role of chemical

engineersengineers

Lila Gierasch How do chains of amino acids fold in 3D to make active proteins inside the cell?

Igor Kaltashov How is the ability of transport proteins to bind and release ligands when necessary controlled by their environment?

In the CBI Program we study …

How proteins work: structure is related to function.

Chemical

Glucophage® 166

Vioxx® 314

Prozac® 346

Zantac® 351

Paxil® 375

Claritin® 383

Zocor® 419

Augmentin® 420

Crixivan® 712

Taxol® 854

Complexity – Difference in Weight

Biotechnology products

Neupogen® 18,800

Intron-A® 19,625

Humatrope® 22,125

Avonex® 22,500

Epogen® 30,400

Pulmozyme® 37,000

ReoPro® 47,615

Enbrel® 75,000

Zenapax® 144,000

Rituxan® 145,000

The molecular weights (in daltons, the standard unit of molecular mass) of some popular drugs:Source: Lisa Raines, Genzyme 2001

What is biotechnology? What is biotechnology? Biotechnology = bios (life) + logos (study of or essence)Biotechnology = bios (life) + logos (study of or essence)

Literally ‘the study of tools from living things’Literally ‘the study of tools from living things’

CLASSIC: The word "biotechnology" was first used in 1917 CLASSIC: The word "biotechnology" was first used in 1917 to describe processes using living organisms to make a to describe processes using living organisms to make a product or run a process, such as industrial fermentations. product or run a process, such as industrial fermentations. ((Robert BudRobert Bud,, The The Uses Uses of Life of Life: A: A History of Biotechnology History of Biotechnology))

LAYMAN: Biotechnology began when humans began to LAYMAN: Biotechnology began when humans began to plant their own crops, domesticate animals, ferment juice plant their own crops, domesticate animals, ferment juice into wine, make cheese, and leaven bread (into wine, make cheese, and leaven bread (AccesExcellenceAccesExcellence) )

What is biotechnology? • GENENTECH: Biotechnology is the process of

harnessing 'nature's own' biochemical tools to make possible new products and processes and provide solutions to society's ills (G. Kirk Raab, Former President and CEO of Genentech)

• WEBSTER’S: The aspect of technology concerned with the application of living organisms to meet the needs and ends of man.

• WALL STREET: Biotechnology is the application of genetic engineering and DNA technology to produce therapeutic and medical diagnostic products and processes. Biotech companies have one thing in common - the use of genetic engineering and manipulation of organisms at a molecular level.

What is biotechnology?

• Using scientific methods with organisms to produce new products or new forms of organisms

• Any technique that uses living organisms or substances from those organisms or substances from those organisms to make or modify a product, to improve plants or animals, or to develop microorganisms for specific uses


• GMO - genetically modified organisms• GEO - genetically enhanced organisms

• In both cases, the natural genetic material of the organism has been altered.

• Roots in bread-making, wine brewing, cheese and yogurt fermentation, and classical plant and animal breeding


• Biotechnology is a multidisciplinarian in nature, involving input from

• Engineering• Computer Science• Cell and Molecular Biology• Microbiology• Genetics• Physiology• Biochemistry• Immunology• Virology• Recombinant DNA Technology Genetic manipulation of

bacteria, viruses, fungi, plants and animals, often for the development of specific products

What are the stages of biotechnology?

• Ancient Biotechnology• early history as related to food and shelter,

including domestication

• Classical Biotechnology• built on ancient biotechnology• fermentation promoted food production• medicine

• Modern Biotechnology• manipulates genetic information in organism• genetic engineering

•Long history of fermented foods since people began to settle (9000 BC) (fervere –to boil)

•Often discovered by accident!

•Improved flavor and texture

•Deliberate contamination with bacteria or fungi (molds)

•Examples:•Bread•Yogurt•Sour cream•Cheese•Wine•Beer

Ancient biotechnology

Fermented foods and beverages

•Dough not baked immediately would undergo spontaneous fermentation would rise Eureka!!

•Uncooked fermented dough could be used to ferment a new batch no longer reliant on “chance fermentation”

•1866 – Louis Pasteur published his findings on the direct link between yeast and sugars CO2 + ethanol (anaerobic process)

•1915 – Production of baker’s yeast – Saccharomyces cerevisiae

Ancient biotechnology

Fermented foods and beverages

•Different types of beer•Vinegar•Glycerol•Acetone•Butanol•Lactic acid•Citric acid•Antibiotics – WWII (Bioreactor developed for large scale production, e.g. penicilin made by fermentation of penicillium)

•Today many different antibiotics are produced by microorganisms•Cephalosporins, bacitracin, neomycin, tetracycline……..)

Classical biotechnology

Industry today exploits early discoveries of the fermentation process for production of huge numbers of products

• Substrate + Microbial Enzyme Product

• Examples:• Cholesterol Steroids (cortisone, estrogen, progesterone) (hydroxylation reaction -OH group added to cholesterol ring)


Chemical transformations to produce therapeutic products

• Amino acids to improve food taste, quality or preservation

• Enzymes (cellulase, collagenase, diastase, glucose isomerase, invertase, lipase, pectinase, protease)

• Vitamins

• Pigments


Microbial synthesis of other commercially valuable products

• Cell biology• Structure, organization and reproduction

• Biochemistry• Synthesis of organic compounds• Cell extracts for fermentation (enzymes versus whole cells)

• Genetics• Resurrection of Gregor Mendel’s findings 1866 1900s

• Theory of Inheritance (ratios dependent on traits of parents)• Theory of Transmission factors

• W.H. Sutton – 1902• Chromosomes = inheritance factors

• T.H. Morgan – Drosophila melanogaster

Modern biotechnology

• Molecular Biology

• Beadle and Tatum (Neurospora crassa)• One gene, one enzyme hypothesis

• Charles Yanofsky colinearity between mutations in genes and amino acid sequence (E. coli)• Genes determine structure of proteins

• Griffith’s 1928 experiment• Bacterial transformation of S. pneumoniae from rough to smooth• 1944 – Avery, MacLeod and McCarty identify transforming principle as DNA


• Hershey and Chase – 1952 • T2 bacteriophage – 32P DNA, not 35S protein is the material that encodes genetic information

• Watson, Crick, Franklin and Wilkins (1953)• X-ray crystallography • 1962 – Nobel Prize awarded to three men• Chargaff – DNA base ratios• Structural model of DNA developed

• Messelson and Stahl• 14N/15N semi-conservative replication confirmed

• DNA Revolution – Promise and Controversy!!!• Scientific foundation of modern biotechnology based on knowledge of DNA, its replication, repair and use of enzymes to carry out in vitro splicing DNA fragments


• Breaking the Genetic Code – Finding the Central Dogma

• An “RNA Club” organized by George Gamow (1954) assembled to determine the role of RNA in protein synthesis

• Vernon Ingram’s research on sickle cell anemia (1956) tied together inheritable diseases with protein structure

• Link made between amino acids and DNA

• Radioactive tagging experiments demonstrate intermediate between DNA and protein = RNA

• RNA movement tracked from nucleus to cytoplasm site of protein synthesis


• DNA RNA Protein Transcription Translation

Genetic code determined for all 20 amino acids by Marshal Nirenberg and Heinrich Matthaei and Gobind Khorana – Nobel Prize – 1968

• 3 base sequence = codon


What are the areas of biotechnology?

• Organismic biotechnology• uses intact organisms and does not alter genetic

material

• Molecular Biotechnology• alters genetic makeup to achieve specific goals

Transgenic organism: an organism with artificially altered genetic material

What are the benefits of biotechnology?

• Medicine• human• veterinary• biopharming

• Environment• Agriculture• Food products• Industry and manufacturing

What are the applications of biotechnology?

• Production of new and improved crops/foods, industrial chemicals, pharmaceuticals and livestock

• Diagnostics for detecting genetic diseases• Gene therapy (e.g. ADA, CF)• Vaccine development (recombinant vaccines)• Environmental restoration• Protection of endangered species• Conservation biology• Bioremediation• Forensic applications• Food processing (cheese, beer)

Documents

2003 LECTURES , Biotechnology