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Industrial Production & Bioremediation

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Industrial Production & Bioremediation Microbes for industrial production Preservation of cultures Methods of industrial production Major products of industrial ... – PowerPoint PPT presentation

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Title: Industrial Production & Bioremediation


1
Industrial Production Bioremediation
  • Microbes for industrial production
  • Preservation of cultures
  • Methods of industrial production
  • Major products of industrial microbiology
  • Bioremediation
  • Biosensors microarrays

2
Microbes for industrial production
  • Finding microorganisms in nature
  • Only a small percentage of microbial species have
    been cultured
  • Bioprospecting Hunting for new microorganisms
    with potential for commercial exploitation
  • Great deal of interest in microbes from extreme
    environments
  • Challenge is to develop cost-effective techniques
    for their culture

3
Microbes for industrial production
  • Genetic manipulation
  • Altering the characteristics of existing known
    species to produce new and desirable
    characteristics
  • Mutations can be induced with mutagenic agents or
    UV irradiation
  • Example Development of high-yield cultures of
    Penicillium for penicillin production
  • Protoplast fusion can be used to fuse cells of
    eukaryotic microbes and microbes that are not
    phylogenetically related used especially for
    genetic manipulation in yeasts molds

4
Microbes for industrial production
  • Genetic manipulation
  • Altering the characteristics of existing known
    species to produce new and desirable
    characteristics
  • Mutations can be induced with mutagenic agents or
    UV irradiation
  • Example Development of high-yield cultures of
    Penicillium for penicillin production
  • Protoplast fusion can be used to fuse cells of
    eukaryotic microbes and microbes that are not
    phylogenetically related used especially for
    genetic manipulation in yeasts molds

5
Microbes for industrial production
  • Genetic manipulation
  • Altering the characteristics of existing known
    species to produce new and desirable
    characteristics
  • Mutations can be induced with mutagenic agents or
    UV irradiation
  • Example Development of high-yield cultures of
    Penicillium for penicillin production
  • Protoplast fusion can be used to fuse cells of
    eukaryotic microbes and microbes that are not
    phylogenetically related used especially for
    genetic manipulation in yeasts molds

6
Microbes for industrial production
  • Genetic manipulation
  • Altering the characteristics of existing known
    species to produce new and desirable
    characteristics
  • Mutations can be induced with mutagenic agents or
    UV irradiation
  • Example Development of high-yield cultures of
    Penicillium for penicillin production
  • Protoplast fusion can be used to fuse cells of
    eukaryotic microbes and microbes that are not
    phylogenetically related used especially for
    genetic manipulation in yeasts molds

7
Microbes for industrial production
  • Genetic manipulation
  • Altering the characteristics of existing known
    species to produce new and desirable
    characteristics
  • Mutations can be induced with mutagenic agents or
    UV irradiation
  • Example Development of high-yield cultures of
    Penicillium for penicillin production
  • Protoplast fusion can be used to fuse cells of
    eukaryotic microbes and microbes that are not
    phylogenetically related used especially for
    genetic manipulation in yeasts molds

8
Microbes for industrial production
  • Genetic manipulation
  • Altering the characteristics of existing known
    species to produce new and desirable
    characteristics
  • Mutations can be induced with mutagenic agents or
    UV irradiation
  • Example Development of high-yield cultures of
    Penicillium for penicillin production
  • Protoplast fusion can be used to fuse cells of
    eukaryotic microbes and microbes that are not
    phylogenetically related used especially for
    genetic manipulation in yeasts molds

9
Microbes for industrial production
  • Genetic manipulation
  • Site-directed mutagenesis is the insertion of
    short segments of DNA (using recombinant DNA
    technology) into a gene to lead to desired
    changes in its protein product
  • Recombinant DNA can be transferred between
    different organisms, creating combinations of
    genes with exhibit desired characteristics
  • Shuttle vectors Vectors (such as bacterial
    plasmids) that can replicate in more than one
    species
  • Expression vectors Vectors that have
    transcriptional promoters capable of mediating
    gene expression in the target species.

10
Microbes for industrial production
  • Genetic manipulation
  • Gene expression can be modified by altering
    transcriptional regulation, fusing proteins, and
    removing feedback regulation controls
  • This is used for pathway architecture, or
    metabolic pathway engineering, to increase or
    regulate production.
  • Natural genetic engineering
  • Growing cultures under marginal (stressful)
    growth conditions and selecting for new strains
    (spontaneous mutations) that have increased
    growth in those conditions

11
Preservation of cultures
  • Periodic transfer refrigeration
  • Mineral oil slant refrigeration
  • Washed culture refrigeration
  • Freezing
  • Freezing with 50 glycerol
  • Drying
  • Lyophilization (freeze drying)
  • Ultracold freezing

12
Methods of industrial production
  • Medium development
  • Lower-cost ingredients, such as crude plant or
    animal by-products, are used for
    cost-effectiveness
  • Manipulating the levels of a limiting nutrient
    may be critical to trigger or optimize the
    production of a desired product

13
Methods of industrial production
  • Scaleup
  • Successive optimization of growth product yield
    from a small scale (such as a shaking flask or
    small fermenter) to a large scale (such as
    industrial scale fermenters)
  • Mixing, aeration, pH control, foaming,
    formation of filamentous growth or biofilms are
    significant issues in scaleup

14
Methods of industrial production
  • Methods for mass culture
  • Batch fermentation
  • Continuous culture (chemostat)
  • Lift-tube fermentation
  • Solid-state fermentation
  • Fixed-bed reactors
  • Fluidized-bed reactor
  • Dialysis culture unit

15
Methods of industrial production
  • Primary secondary metabolites
  • Primary metabolites are produced during the
    growth phase of the microbe. Examples amino
    acids, nucleotides, fermentation end products,
    and many types of enzymes
  • Secondary metabolites accumulate during periods
    of nutrient limitation and waste buildup.
    Examples many antibiotics and mycotoxins

16
Major products
  • Antibiotics
  • Examples penicillin streptomycin
  • The yield of both of these antibiotics are
    optimized by nutrient limitation (carbon
    nitrogen)
  • Recombinant DNA products
  • Proteins produced from genes introduced into
    microbes via recombinant DNA techniques, such as
    enzymes, peptide hormones, recombinant vaccines

17
Major products
  • Amino acids
  • Glutamic acid (monosodium glutamate) is produced
    by regulatory mutants of Corynebacterium
    glutamicum that have a modified Krebs cycle that
    can be manipulated to shift ?-ketoglutarate to
    glutamate production
  • Lysine is produced by a Corynebacterium
    glutamicum strain in which homoserine lactone
    synthesis is blocked

18
Major products
  • Other organic acids
  • Acetic acid, citric acid, fumaric acid, gluconic
    acid, itaconic acid, kojic acid, lactic acid
  • Speciality compounds
  • A variety of drugs (cholesterol drugs,
    immunosuppressants, antitumor drugs), ionophores,
    enzyme inhibitors, pesticides
  • Biopolymers
  • Microbial-produced polymers, mostly
    polysaccharides, useful as thickening or gelling
    agents in foods, pharmaceuticals, paints, etc.

19
Major products
  • Biosurfactants
  • Microbial-produced detergents, such as
    glycolipids used in bioremediation applications
    such as oil spill cleanups
  • Bioconversions
  • Using a microbe as a biocatalyst to convert a
    substrate into a desired product for example, in
    the modification of steroid hormones

20
Bioremediation
  • Biodegradion in natural communities
  • Includes
  • minor changes in organic molecules, leaving the
    main structure still intact
  • fragmentation of an organic molecule into smaller
    organic molecules, still resembling the original
    structure
  • complete mineralization of an organic molecule to
    CO2
  • Recalcitrant compounds are organic compounds that
    are resistant to biodegradation

21
Bioremediation
  • Biodegradion in natural communities
  • Halogenated compounds, especially halogenated
    aromatic compounds (such as polychlorinated
    biphenyls) are often recalcitrant
  • The presence of halogens in a meta position makes
    the compound more recalcitrant
  • Often one stereoisomer of an organic compound
    will be biodegradable, while another isomer will
    be recalcitrant
  • Specific organisms in an environment may be able
    to degrade recalcitrant compounds, at varying
    rates depending on the conditions

22
Bioremediation
  • Biodegradation in natural communities
  • Sometimes partial degradation of a compound may
    yield compounds that are worse for example,
    trichloroethylene can be degraded to form highly
    carcinogenic vinyl chloride
  • Another example of detrimental biodegradation is
    microbial corrosion of metal pipes

23
Bioremediation
  • Stimulating biodegradation
  • Biodegradation by naturally-occurring organisms
    may be stimulated by
  • Adding essential nutrients to the contaminated
    area
  • Providing aeration or limiting aeration,
    depending on whether the contamination is better
    degraded under aerobic or anaerobic conditions
  • Using plants and the microbial communities of
    their rhizospheres (phytoremediation)
  • Using microbes for metal bioleaching from
    minerals

24
Bioremediation
  • Bioaugmentation
  • Adding microbes not normally found in an
    environment to try to alter or accelerate the
    biodegradation process
  • When the microbes are added without consideration
    of their normal habitat (e.g., just adding a
    pure culture), there may be short-term
    improvement but the added microbe usually fails
    to establish a stable population
  • Better results are may be seen when the added
    organisms microenvironment (nutrients, oxygen,
    aeration, etc.) are included in the
    bioaugmentation strategy

25
Biosensors microarrays
  • Biosensors
  • Devices in which a biospecific molecule (e.g., a
    monoclonal antibody or a hormone receptor
    protein) is attached to a transducer (often a
    piezoelectrically-active quartz chip)
  • When the biosensor binds to its target, it
    slighty twists the transducer, creating a small
    electrical current that can be amplified,
    detected, and measured

26
Biosensors microarrays
  • Microarrays
  • A series of microscopic DNA spots on a glass,
    plastic, or silicon backing used to monitor
    levels of gene expression for thousands of genes
    simultaneously, or to determine differences in
    genotype
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