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Microbial Biotechnology

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Chapter 5 Microbial Biotechnology The rich abundance of bacteria and other microbes provides a wealth of potential biotech applications. * Before you can consider the ... – PowerPoint PPT presentation

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Title: Microbial Biotechnology


1
Microbial Biotechnology
  • Chapter 5

2
The Structure of Microbes
  • Prokaryotes
  • Archaebacteria
  • Includes halophiles, thermophiles,
    extremophiles
  • Eubacteria
  • On skin, soil, water, can be pathogenic

3
The Structure of Microbes
  • Characteristics of Prokaryotes
  • Generally smaller than Eukaryotes
  • No nucleus
  • Cell wall composed of peptidoglycan
  • Conjugation (transfer of DNA by cytoplasmic
    bridge)
  • Transduction (DNA is packaged in a virus and
    infects recipient bacterial cells)
  • 20 minute growth rate (binary fission)

4
Yeast are Important Too!
  • Single celled eukaryote
  • Kingdom Fungi
  • Over 1.5 million species
  • Source of antibiotics, blood cholesterol lowering
    drugs
  • Able to do post translational modifications
  • Grow anaerobic or aerobic
  • Examples Pichia pastoris (grows to a higher
    density than most laboratory strains), has a no.
    of strong promoters, can be used in batch
    processes

5
Microorganisms as Tools
  • Microbial Enzymes
  • Taq (DNA polymerase), cellulases, proteases,
    amylases

6
Microorganisms as Tools
  • Bacterial Transformation
  • The ability of bacteria to take in DNA from their
    surrounding environment
  • Bacteria must be made competent to take up DNA

7
Microorganisms as Tools
  • Cloning and Expression Techniques
  • Fusion Proteins

8
Microorganisms as Tools
  • Microbial Proteins as Reporters
  • Examples the lux gene which produces luciferase
  • Used to develop a fluorescent bioassay to test
    for TB

9
Microorganisms as Tools
  • Yeast Two-Hybrid System
  • Used to study protein interactions

10
Using Microbes for a Variety of Everyday
Applications
  • Food Products
  • Rennin used to make curds (solid) and whey in
    production of cheese
  • Recombinant rennin is known as chymosin (first
    recombinant food ingredient approved by FDA)

11
Using Microbes for a Variety of Everyday
Applications
  • Food Products
  • Energy production in bacteria
  • Aerobic or anaerobic

12
Using Microbes for a Variety of Everyday
Applications
  • Food Products
  • Fermentation
  • (anaerobic respiration)
  • Lactic acid fermentation
  • Used to make cheese, yogurt, etc.
  • Ethanol fermentation
  • Used to make beer and wine

13
Using Microbes for a Variety of Everyday
Applications
  • Field Applications of Recombinant Microorganisms
  • Ice-minus bacteria (remove ice protein producing
    genes from P. syringae)
  • P. fluorescens containing the gene that codes for
    the bacterial toxin from Bacillus thuringiensis
    (kills insects) Bt toxin!

14
Using Microbes for a Variety of Everyday
Applications
  • Therapeutic proteins
  • Recombinant insulin in bacteria

15
Using Microbes for a Variety of Everyday
Applications
  • Using Microbes Against Other Microbes
  • Antibiotics
  • Act in a few key ways
  • Prevent replication
  • Kill directly
  • Damage cell wall or prevent its synthesis

16
Vaccines
  • First was a vaccine against smallpox (cowpox
    provides immunity)
  • DPT-diphtheria, pertussis, and tetanus
  • MMR measles, mumps, and rubella
  • OPV- oral polio vaccine (Sabin)

17
Vaccines
  • A Primer on Antibodies
  • Antigen- foreign substances that stimulate an
    immune response
  • Types of leukocytes or white blood cells
  • B-lymphocytes antibody-mediated immunity
  • T-lymphocytes cellular immunity
  • Macrophages cell eating (phagocytosis)

18
Vaccines
  • Antigens stimulate antibody production in the
    immune system

Light chain
Heavy chain
IgA first line of defense IgG and IgM
activates macrophages
19
Vaccines
  • Mechanism of Antibody Action

20
Vaccines
  • How are vaccines made?
  • They can be part of a pathogen (e.g. a toxin) or
    whole organism that is dead or alive but
    attenuated (doesnt cause disease)
  • Subunit (toxin) or another part of the pathogen
  • Attenuated (doesnt cause disease)
  • Inactivated (killed)
  • What about flu vaccines (why do we have to get a
    shot every year?)

21
Recombinant Vaccines
  • Vaccines provide immunity to infectious
    microorganisms

Attenuated Vaccine
Inactivated Vaccine
Subunit Vaccine
22
Recombinant Vaccines
  • Recombinant Vaccines
  • A vaccine produced from a cloned gene

Video Constructing Vaccines
23
Recombinant Vaccines
  • DNA vaccines
  • Direct injection of plasmid DNA containing genes
    encoding specific antigenic proteins

24
Bacterial and Viral Targets for Vaccines
  • HIV

25
Microbial Genomes
  • Microbial Genome Program (MGP) the goal is to
    sequence the entire genomes of microorganisms
    that have potential applications in environmental
    biology, research, industry, and health
  • Sequencing Strategies?

26
Microbial Genomes
  • Why study viral genomes?
  • Decipher genes and their products so that agents
    that block attachment, block replication can be
    made

27
Microbial Diagnostics
  • Using Molecular Techniques to Identify Bacteria
  • RFLP
  • PCR and Real time PCR
  • Sequencing

28
Molecular Diagnostics
  • Application Molecular Epidemiology
  • Pulse Net monitors disease outbreaks related to
    different strains of food-borne pathogens

29
Microbial Diagnostics
  • Microarrays for tracking contagious disease
  • PulseNet used to identify outbreaks

30
Microbial Diagnostics
  • Combating Bioterrorism
  • The use of biological materials as weapons to
    harm humans or animals and plants we depend on
    for food
  • Examples in History
  • Throwing plague infected dead bodies over the
    walls of their enemies

31
Microbial Diagnostics
  • Using Biotech Against Bioweapons
  • Postal service x-raying packages
  • Antibody tests in the field
  • PCR tests in the field
  • Protein Microarrays for detecting bioweapon
    pathogens
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