Презентация на тему "Microbial biotechnology"

Презентация: Microbial biotechnology
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Презентация на тему "Microbial biotechnology" по иностранным языкам. Состоит из 70 слайдов. Размер файла 0.76 Мб. Каталог презентаций в формате powerpoint. Можно бесплатно скачать материал к себе на компьютер или смотреть его онлайн.

  • Формат
    pptx (powerpoint)
  • Количество слайдов
    70
  • Слова
    английский язык
  • Конспект
    Отсутствует

Содержание

  • Презентация: Microbial biotechnology
    Слайд 1

    Microbial Biotechnology

  • Слайд 2

    What are microbes? Microbes are small single-celled organisms Either free-living or in colonies They can belong to any of the three domains

  • Слайд 3

    Eubacteria (Bacteria) Archaebacteria (Archaea) Eukaryota (Eukaryotes) Three Domains

  • Слайд 4

    Eubacteria Gram-negative and gram-positive prokaryotes Either autotrophs or heterotrophs Can be aerobic or anaerobic Mesophiles Examples: E. coli Lactobacillus Agrobacterium Staphylococcus

  • Слайд 5

    Archea Ancient domain, but only recently identified Through DNA analysis they were determined to differ significantly from eubacteria Found predominantly in extreme environments (Extremophiles) Thermophiles 50- 110°C Psychrophiles 0- 20°C Alkaliphiles pH>9 Halophiles 3- 20% salt Methanogens use H2 + CO2 to produce CH4

  • Слайд 6

    Eukaryotes Predominately yeasts/molds, protists, algae Sac shaped cells that form sexual spores Examples: Sacchromyces Penicillium Aspergillus Pichia

  • Слайд 7

    Commercial Uses of Microbes Products Bioconversion/Biocatalysis Agriculture Bioremediation Oil/Mineral Recovery

  • Слайд 8

    Fermentation is a process for the production of useful products through mass culture of single-cells The end products or the various intermediate products (metabolites) are siphoned off & purified for commercial use stirred tank reactor Fermenter or Bioreactor

  • Слайд 9

    http://www.wavebiotech.com/products/wave_bioreactor/system500/index.html http://www.pharmaceutical-technology.com/projects/lonza/lonza1.html 15 000L Fermenter 1000L Disposable Bag

  • Слайд 10

    Types of Products Produced in Microbes Amino Acids Vitamins Food Additives Enzymes Recombinant Protein Drugs Antibiotics Fuels Plastics

  • Слайд 11

    Enzyme: chymosin - the enzyme used to curdle milk products Hormone: bST - bovine somatotropin; used to increase milk production Examples of bacterially-expressed proteins:

  • Слайд 12

    1928: Alexander Fleming discovered the first antibiotic. He observed that Penicillium fungus made an antibiotic, penicillin, that killed S. aureus. 1940s: Penicillin was tested clinically and mass produced. Original Penicillium moulds produced less than 10 units of penicillin per ml of fermentation broth (1943) By 1955 Penicillium strains produced 8000 units/ml Mutation with UV, mustard gas, and X-Ray, strain selection / culture improvement Is this GMO?

  • Слайд 13

    How Are Microbes Modified? Artifical Selection Recombiant DNA Metabolic Engineering

  • Слайд 14

    Transgenic microbes are created when cDNAs for the protein product are cloned into expression vectors Human genes inserted into E. coli Genes from extremophiles are moved to mesophiles Due to the ease in culturing of mesophiles Mesophiles also have 5 to 10x higher growth rates Recombinant DNA Microbes

  • Слайд 15

    Enzyme/Drug

  • Слайд 16

    Metabolic Engineering, manipulation of pathways within an organism to optimize the production of a compound Done by turning off particular genes, either through mutation or deletion Products are also gained by altering the microbe’s environment Metabolically Engineered Cell Cheap Carbon Source Industrial Chemicals Chemical Precursors Biopolymers Vitamins Amino Acids

  • Слайд 17

    GlucoseNH4+ Pyruvate L-lactate D-Alanine L-Alanine 100% L-Alanine NAD+ NADH NADH NAD+ L-LDH AlaOH NADH NAD+ Racemase The microbe is forced to produce alanine at higher than normal amounts

  • Слайд 18

    Carotenoid production in E.coli cells

  • Слайд 19

    Fermentation Products Enzymes

  • Слайд 20

    Enzymes, the most common product produced by microbes Overall value of industrial enzymes is about $2.0 billion1 They are found in many household items that you would never think to have a biotechnology component Enzymes 1 2004 data

  • Слайд 21

    Enzyme NameGE OrganismUse (examples) -acetolactate bacteria Removes bitter substances decarboxylase from beer  -amylase bacteria Converts starch to simple sugar Catalase fungi Reduces food deterioration Chymosin bacteria or fungi Clots casein to make cheese -glucanase bacteria Improves beer filtration Glucose isomerase bacteria Converts glucose to fructose Glucose oxidase fungi Reduces food deterioration Lipase fungi Oil and fat modification Maltogenic amylase bacteria Slows staling of breads Pectinesterase fungi Improves fruit juice clarity Protease bacteria Improves bread dough structure xylanase (hemicellulase) bacteria or fungi Enhances rising of bread dough http://www.geo-pie.cornell.edu/crops/enzymes.html

  • Слайд 22

    Detergent Enzymes

  • Слайд 23

    Detergents are the largest application of industrial enzymes Traditionally these are lipolases,proteases & amylases A recent innovation is the addition of mannanase This enzyme aids in removing stains containing guar gum These enzymes are engineered to improve stability in the presence of detergent, alkaline pH, and cold water

  • Слайд 24

    Subtilisin, a protease used in laundry detergents The recombinant protein was engineered to remain active in the presence of bleach Bleach caused the oxidation of one amino acid (methionine) and the enzyme lost 90% of its activity By replacing this amino acid with alanine, the engineered enzyme was no longer sensitive to oxidation

  • Слайд 25

    Directed evolution is the most recent tool utilized in the creation of new and better enzymes (& other proteins) http://www.rsc.org/chemistryworld/Issues/2004/July/rational.asp

  • Слайд 26

    Subtilisin normally functions in aqueous solution Mutations were introduced randomly throughout the structure of the enzyme Only 0.1–1% of the mutations were beneficial, but… Activity in 60% dimethylformamide was improved 256-fold doi:10.1016/S1367-5931(02)00396-4  

  • Слайд 27

    Enzymes for Feed

  • Слайд 28

    Enzymes are used in animal feed to breakdown cellulose (cellulase) New use of enzymes (phytases) which breakdown phytic acid This allows better utilization of plant phosphorus stores Allowing bone-meal to be removed from feeds The latest generation of phytases are from fungus and have been engineered to survive high temperatures used during food processing 65% of poultry and 10% of swine feeds contain enzymes

  • Слайд 29

    Where do the genes for these enzymes come from? Nature is still an important source (Gene Prospecting) ~

  • Слайд 30

    Recombinant Drugs Besides antibiotics which are derived from microorganisms Protein medicines are produced by inserting human genes into microbes

  • Слайд 31

    1982, FDA approves the first recombinant protein drug, human insulin produced by E. coli developed by Genentech Today there are >75 recombinant protein drugs approved by the FDA with 100s more being studied Currently the global market for recombinant protein drugs is $47.4 billion1 (2006)

  • Слайд 32

    Product Microbe Purpose Insulin E. coli Diabetes treatment Interleukin-2 E. coli Cancer/immune system stimulant EGF E. coli wound healing Interferons E. coli/yeast Cancer/virus treatments Prourokinase E.coli/yeast Anticoagulant/heart attacks CSF E. coli/yeast Immune stimulant Taxol E. coli ovarian cancer

  • Слайд 33

    Other Products From Microbes Fuels, Plastics, Medications

  • Слайд 34

    Ethanol Production Produced via anaerobic fermentation by yeast Corn starch is hydrolyzed to glucose monomers

  • Слайд 35

    Problem with Corn Ethanol Ethanol contains 76000BTU/gal Takes ~98000BTU/gal to produce from corn sugar Gasoline contains 112000BTU/gal Costs 22000BTU/gal to extract and refine A BTU (British thermal unit) is defined as the amount of heat required to raise the temperature of one pound of water by one degree Fahrenheit 2004 3.4 billion gallons of ethanol were produced U.S. consumes 140 billion gallons of gasoline/yr

  • Слайд 36

    Plastics Polyhydroxyalkanoate (PHA) is a polymer made by some microbes as a way of storing carbon Up to 80% of the microbe’s biomass is plastic PHA is sold to make shampoo bottles in Germany, and disposable razors in Japan The microbe Pseudomonas putida converts styrene to PHA http://www2.unil.ch/ibpv/WWWPoirier/images/Fig1.jpg

  • Слайд 37

    http://www2.unil.ch/ibpv/WWWPoirier/images/Fig2.jpg

  • Слайд 38

    Bioconversion Utilization of microbes to modify a compound Useful when multi-step chemical synthesis is expensive or inefficient Often microbial conversion is combined with traditional chemistry to reduce the steps necessary The most common use of bioconversion is in the synthesis of steroids such as hormones & corticosteroids

  • Слайд 39

    starting product End products

  • Слайд 40

    Microbes and Agriculture

  • Слайд 41

    Frost damages many crops such as citrus trees & strawberries When fruit freeze the ice crystals form As the plants thaws they are effectively turned to mush Frost damage to an orange leaf and fruit Frost Damage

  • Слайд 42

    Some ice crystal nucleation is due to bacterial activity Pseudomonas syringae promotes the development of ice at 0 to 2°C If the bacteria are not present ice does not form until between –6 and –8°C

  • Слайд 43

    A strain of P. syringae called “ice minus” was developed Plants were to be sprayed with the ice minus strain This inhibits colonization by the “ice plus” (wild) strain The EPA declared the new strain to be a pesticide This made the review process lengthy and burdensome The company thought it too expensive to pursue However the “ice plus” strain has found a purpose…

  • Слайд 44

    Bacillus thuringiensis (Bt) is an aerobic spore-forming bacterium During sporulation producesinsecticidal crystal protein (ICP), a toxin (Cry) The toxin brakes down quickly in the environment They have no toxicity to humans & there is no withholding period on produce sprayed with Bt Cry toxins vary in their toxicity and specificity Microbial Pesticides

  • Слайд 45

    http://www.bioc.cam.ac.uk/~dje1/

  • Слайд 46
  • Слайд 47

    Bioremediation

  • Слайд 48

    Bioremediation is reclaiming or cleaning of contaminated sites using microbes or other organisms This entails the removal, degradation, or sequestering of pollutants &/or toxic wastes http://www.cleanearthltd.com/en/contamination_cleanup/index.php

  • Слайд 49

    Bacteria are isolated based on their efficiency at digesting & converting the waste The bacteria are tested for performance and safety Bacteria are placed back in the waste environment in high concentrations The bacteria grow & in the process digest & convert the waste into CO2 and H20

  • Слайд 50

    What can be cleaned up using bioremediation? Oil spills Waste water Plastics Chemicals (PCBs) Toxic Metals

  • Слайд 51

    Oil/Wastewater Cleanup

  • Слайд 52

    Bioremediation Bacteria degrade organic matter in sewage. Bacteria degrade or detoxify pollutants such as oil and mercury

  • Слайд 53

    Microbes that digest hydrocarbons found throughout the environment These naturally occurring microbes are utilized during a spill to clean shore lines Fertilizer is added to supply the nutrients phosphorus and nitrogen This was approach was used after the Exxon Valdez Stimulated the natural rate of biodegradation by 2 to 5x There have yet to be any other instances of this being used on a large-scale

  • Слайд 54

    Exxon Valdez off the Coast of Alaska http://blogs.abcnews.com/photos/uncategorized/exxon_valdez.jpg

  • Слайд 55

    Before After Smaller scale cleanup is feasible For 3 months nutrients and microbes were sprayed on this field After 11 months the site was deemed clean 6000yards3 petroleum conc. Before 4000ppm After 100ppm

  • Слайд 56

    Before After Treatment of domestic sewage or industrial waste Utilizes aeration to oxygenate allowing aerobic microbes to digest solid waste Wastewater

  • Слайд 57

    Plastic Degradation

  • Слайд 58

    140 million tons of plastics are produced each year Traditional plastics are very stable and do not degrade Some plastics have been shown to be biodegradable Strains of bacteria have been isolated that breakdown: Polyurethane Polyvinyl alcohol Nylon-66 The degradation pathways are currently under study

  • Слайд 59

    Chemicals Polychlorinated biphenyls (PCBs)

  • Слайд 60

    PCBs have low water solubility, good insulating properties, high boiling points and resistance to chemicals The largest uses for PCBs was in capacitors, transformers, & as plasticizers

  • Слайд 61

    1977, Monsanto (main producer) stops all PCB production Millions of lbs of PCBs are still in place around the world The stability properties that made PCBs so useful have allowed them to persist in the environment Most people in industrialized countries have PCBs in their tissue

  • Слайд 62

    Microbes that dehalogenate PCBs have been isolated This process is referred to as halorespiration Involves the replacement of the Cl with an –OH This process is multi-step with four enzymes required These enzymes are now the target of protein engineering to optimize their performance

  • Слайд 63

    Heavy Metal Clean up

  • Слайд 64

    Uranium processing has left contaminated groundwater sites across the United States and the world Traditional “pump-and-treat” methods take decades and expose workers to toxic levels of uranium Geobacter to convert soluble uranium to insoluble uraninite Uraninite stays put instead of mixing with water used for drinking or irrigation The microbes are encouraged to multiply by injecting acetate In ~50 days, 70% of the uranium is converted into uraninite

  • Слайд 65

    Biomining

  • Слайд 66

    Microbe assisted mining has gone on for millennia Early copper miners used microbes to leach copper from ore without even knowing it Low-grade ore and mine tailings are exploited biologically Sulfides of metals like zinc, copper, nickel, cobalt, iron, tungsten, lead are insoluble in water These sulfides are converted to sulfate which are soluble The sulfates leach out of the ore and are then extracted

  • Слайд 67

    Cu2S not soluble CuSO4 is soluble

  • Слайд 68

    Commercial Bioleaching Tanks

  • Слайд 69
  • Слайд 70
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