1. BCM302 Food and Beverage Biotechnology
Topic 1: Biotechnology: old and new

2. Today’s Lecture What is biotechnology?
How did molecular biotechnology emerge?
Determination of DNA structure.
First recombinant DNA and cloning experiments.

3. Learning objectives Define “biotechnology”.
Describe emergence of molecular biotech.
Classical biotech vs ancient and modern biotech.
List events that led to determination of DNA structure.
List events that built upon DNA structure and formed foundation of molecular biology.
Be familiar with first recombinant DNA, electrophoresis, and DNA cloning experiments.

4. What is biotechnology?
“any technique that uses living organisms or substances from those organisms to make or modify a product, to improve plants or animals, or to develop microorganisms for specific uses”

5. Biotechnology is multidisciplinary
Molecular Biology
Microbiology
Biochemistry
Genetics
Chemical engineering
Cell biology
Molecular Biotechnology
Crops
Drugs
Vaccines
Diagnostics
Livestock

6. Applications of Biotechnology
Virus-resistant crop plants and livestock.
Diagnostics for detecting genetic and acquired diseases.
Therapies that use genes to cure diseases.
Recombinant vaccines to prevent diseases.
Biotechnology aiding the environment eg. bioremediation.

7. Ancient Biotechnology
Paleolithic people settled and developed agrarian societies ~ 10,000 years ago.
Early farmers in Near East & Egypt had wheat, barley, chick-pea, cattle, sheep, goats.
Archaeologists have found ancient farming sites in Americas, Far East, Europe.

8. Ancient Biotechnology
People collected seeds of wild plants for cultivation, domesticated some species of wild animals, performed selective breeding.
Farmers saved seeds and tubers from season to season for thousand of years.

10. Ancient Plant Germplasm
Vavilov, Russian plant geneticist, developed 1st organised plan for crop genetic resource management.
Worldwide centres for germplasm storage.
Germplasm in danger due to agricultural expansion and use of herbicides.
Global effort to salvage germplasm for gene banks.

11. History of Fermented Foods
Fermentation - microbial process, enzymes control transformations of organic compounds.
Production of foods such as bread, yoghurt, cheese, wine, beer.
Fermented dough discovered by accident - old, uncooked dough and yeast such as Saccharomyces winlocki underwent fermentation.

12. History of Fermented Foods
Pasteur discovered yeast’s role in baking and production of baker’s yeast.
~4000 BC Chinese used fermentation to produce yogurt, cheese, fermented rice, soy sauces.
Milk to produce cheese, cream, yogurt, sour cream, butter.

13. Modern cheese manufacturing
Inoculation of milk with lactic acid bacteria
Addition of enzymes eg. rennet to curdle casein (a milk protein)
Heating
Separation of curd from whey
Draining whey
Salting
Pressing the curd
Ripening

14. History of Fermented Beverages
BC, beer making using cereal grains (sorghum, corn, rice, millet, wheat).
1680, Anton van Leeuwenhoek looked at samples of fermenting yeast under a microscope.
, Pasteur established yeast and other microbes responsible for fermentation.
Wine probably made by accident, when grape juices were contaminated with yeast and other microbes.

15. Classical Biotechnology
Improved fermentation for production of many new compounds.
Brewers began producing alcohol on large scale in early 1700s.
Top fermentation - English, Dutch, Belgian, red beers; yeast rises to top of liquid.
Bottom fermentation - US, Europe, pale ales; yeast remains at bottom.
1911, brewers developed method for measuring acid produced to better control beer quality.

16. Classical Biotechnology
, glycerol, acetone, butanol, lactic acid, citric acid, and yeast biomass for baker’s yeast developed.
Industrial fermentation established during WW I - large amounts of glycerol needed for explosives.
Modern fermenter or bioreactor for mass-production of antibiotics eg. penicillin.

17. Classical Biotechnology
Products with therapeutic value:
- amino acids
- vitamins
- enzymes
- microbes as sources of protein secondary metabolites.

18. Classical Biotechnology

19. Enzymes (Table 1.5). - insert

20. Classical Biotechnology
Pharmaceutical compounds eg. antibiotics.
Many chemicals, hormones, and pigments.
Biomass for commercial and animal consumption (eg. single-cell protein).

21. Biotech Revolution: Old meets new
Fermentation and genetic engineering used in food production since 1980s.
Genetically engineered organisms cultured in fermenters, modified to produce large quantities of desirable enzymes
Enzymes are extracted and purified.

22. Biotech Revolution: Old meets new
Enzymes used in production of milk, cheese, beer, wine, lollies, vitamins, mineral supplements.
Genetic engineering used to increase amount and purity of enzymes, to improve enzyme’s function, to provide more cost-efficient method for production.
One of the first produced was chymosin, used in cheese production.

23. Foundations of Modern Biotechnology
Compound microscope to examine cell structure and living organisms in pond water and bacteria.
1838 – 1858 development of plant and animal cell theory, “all cells arise from cells” (Rudolf Virchow).
Microscopes, tissue preservation technology, and stains allowed better understanding of cell structure and function.

24. Biochemistry & Genetics
Elucidation of cell function.
Organic compounds made by living organisms can be made from inorganic compounds in the lab.
Pasteur, pasteurization as means of preserving wine by heating before production of lactic acid (wine spoilage).

25. Biochemistry & Genetics
1896, Buchner converted sugar to ethyl alcohol using yeast extracts, biochemical transformations can occur without cells.
1920s and 1930s, biochemical reactions of many important metabolic pathways established.
By 1935 all twenty amino acids were isolated.

26. Biochemistry & Genetics
1920 – 1940, ultracentrifuge developed.
First electron microscope.
1857, Mendel studied genetic nature of organisms by cross-pollinating pea plants to examine traits such as petal color, seed color, and seed texture.
1869, Miescher isolated nuclein from nuclei of white blood cells. The substance contained nucleic acids.

27. Biochemistry & Genetics
1882, Flemming described threadlike bodies visible during cell division, and equal distribution of this material to daughter cells. He was actually viewing chromosomes during mitosis (cell division).
1903, Sutton, chromosomes were carriers of Mendel’s units of heredity by studying meiosis, cell division that produces reproductive cells.
1909, Johannsen named Mendel’s units of inheritance genes in 1909.

28. Nature of the Gene Experiments linked genes with proteins.
Links between genes and enzymes determined through experiments with fruit fly Drosophila and fungus Neurospora.
Experiments with E. coli showed that genes ultimately determine structure of proteins.

29. Nature of the Gene Experiments linked genes with proteins.
Links between genes and enzymes determined through experiments with the fruit fly Drosophila and the fungus Neurospora.
Experiments with E. coli showed that genes ultimately determine structure of proteins.
(Insert Figure 1.11):

30. Nature of the Gene

31. Nature of the Gene
In 1952, Alfred Hershey and Martha Chase performed an experiment that determined once and for all that DNA is the genetic material (Figure 1.13):

32. Nature of the Gene 1953, Watson and Crick determined structure of DNA.
Franklin and Wilkins provided X-ray diffraction data.
Chargaff determined ratios of nitrogen bases in DNA.

33. Nature of the Gene
Experiments also determined how information in gene is used, such as manipulation of enzymes involved in DNA replication and repair.
Recombinant DNA technology allowed cutting and linking of different pieces of DNA
The new piece of DNA could be placed into a new host.

34. Nature of the Gene
1950s and 60s research focused on two main questions:
1. How does DNA sequence of gene relate to sequence of amino acids that make up protein?
2. What is cell decoding process that produces a protein from information encoded by gene?

35. Nature of the Gene
1956, sequence of deoxyribonucleotides determined information, or message, of DNA.
1957, process of DNA replication demonstrated.
1957, Watson & Crick - DNA bases determine amino acid sequence of protein.
In 1960, RNA discovered, and noted as a messenger between nucleus and ribosome.

36. 1966, complete 64-triplet genetic code determined.
The genetic code

37. Recombinant DNA technology
1971, Berg et al., performed first recombinant DNA experiments, manipulating DNA and placing them into bacteria.
Two DNA molecules joined from different sources.
Mertz and Davis used EcoRI and DNA ligase to combine pieces of DNA.

39. Recombinant DNA technology
Plasmid DNA - EcoRI used to generate DNA fragments for insertion into Cohen’s plasmids.
Gel electrophoresis to separate DNA fragments.
Biotech Revolution: Breaking the Genetic Code.

40. Recombinant DNA technology
1961, first attempt to break genetic code using synthetic messenger RNA (mRNA) such as UUU, AAA, and CCC.
Binding assay developed that allowed determination of which triplet codons specified which amino acids by using RNA sequences that were made of specific codons.

41. First DNA cloning experiment
Joined specific DNA fragments in a vector and transformed an E. coli cell, using EcoRI - technique was patented.
Bacterial DNA placed into an unrelated bacterial species, using Salmonella and Staphylococcus in E. coli.

42. First DNA cloning experiment
RNA genes from frog transferred into E. coli.
Genes from other species could be transferred to bacteria.
1980, patent for basic methods of DNA cloning and transformation awarded to Boyer and Cohen
A second patent granted rights to any organism that was engineered using patented methods.

43. Potential benefits of biotechnology
Diagnosis and prevention or cure wide range of infectious and genetic disorders.
Increase crop yields by creating plants which are resistant to pests and environmental stresses.
Develop animals with enhanced genetically determined attributes.
Facilitate removal of pollutants from the environment.

44. Cause for concern? Public reactions to Recombinant DNA Technology.
DNA cloning methods sparked debate among scientists, ethicists, media, venture capitalists, lawyers, and others.
1980s, concluded that no disasters had occurred through use of recombinant DNA technology, the technology does not pose a threat to human health or environment.
Concerns have focused on both applications and ethical implications.

45. Cause for concern?
Gene therapy experiments have raised question of eugenics (artificial human selection) as well as testing for diseases currently without a cure.
Animal clones developed, fears this may lead to human clones.
Genes from genetically modified crop plants may cause herbicide-resistant weeds.

46. Cause for concern? Fears are focusing on genetically engineered foods.
Hundreds of genetically modified disease, pest, and herbicide-resistant plants awaiting approval for commercialisation.
Genes involved in disease are being identified.
New medical treatments are being developed.
Molecular “pharming,” where plants are being used to produce pharmaceuticals, is being developed.

47. Concerns Will GMOs be harmful to the environment?
Will use of GMOs reduce natural diversity?
Will diagnostics undermine privacy?
Should genetically engineered animals be patentable?
Will financial support for molecular biotechnology constrain the development of other technologies?

48. Concerns
Will emphasis on commercial success mean that benefits of molecular biotechnology will be available only to wealthy?
Will agricultural biotechnology undermine traditional farming practices?
Will medical therapies based on molecular biotechnology supersede equally effective traditional treatments?
Will quest for patents inhibit free exchange of ideas among research scientist?

49. Commercialisation of molecular biotechnology
Ultimate objective of most biotech. companies is to develop commercial products.
New companies are often set up to develop new product from new scientific discovery.
New discovery is generally covered by patent.