Lecture 1 fermentation biotechnology


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Lecture 1 fermentation biotechnology

  1. 1. Topics Fermentation Biotechnology Introduction Microbiology Metabolism (Metabolic pathways) Prof. S.T. Yang Medium formulation; sterilization Dept. Chemical & Biomolecular Eng. The Ohio State University Growth and fermentation kinetics Industrial Biotechnology Industrial Biotechnology Today Tomorrow Baby whole carrots; Fresh-for-two-week Seedless mini melon; Sweeter peas and tomatoes; Insect-protected crops peppers; Colored cotton High-laurate oil, for soaps and detergents Healthy low-saturated oil; Bst-stimulated milk; Vaccinate eggs Faster growing salmon; Non-allergenic cats Antibiotics, Vitamins, … AIDS vaccine, cancer drugs/vaccines Citric acid, glutamic acid, lysine, … Artificial tissues and organs Insulin; Hepatitis B Vaccine; tPA, EPO, .. Bioplastics; Biochemicals; Biomaterials Ethanol, Methane gas Hydrogen, biofuels 1
  2. 2. Four Types of Fermentation Products Industrial fermentation products Production Microorganism Applications Cells (biomass) (metric tons) Proteins, enzymes (cell components) Citric acid 1,200,000 A. niger Food Ethanol 26,000,000 S. cerevisiae Fuel Metabolites Glutamate 1,000,000 C. glutamicum Flavoring Lactic acid 400,000 Lactobacillus sp. Food, Plastics • Primary metabolites Lysine 800,000 C. glutamicum Feed Penicillin 60,000 P. chrysogenum Drug • Secondary metabolites Xanthan gum 100,000 X. campestris Food, Oil drilling Biotransformation (steroids) Changing the history Changing the history - naturally-occurring organisms - genetically modified organisms Product Application Organism Product Application Organism Bacitracin Antiobiotics Bacillus strain Bovine growth hormone Milk production Escherichia coli Citric acid Food flavoring Aspergillus niger Cellulase Cellulose hydrolysis Escherichia coli Invertase Candy Saccharomyces cerevisiae Human growth hormone Growth deficiencies Escherichia coli Lactase Digestive aid Escherichia coli Human insulin Diabetics Escherichia coli Pectinase Fruit juice Aspergillus niger Monoclonal antibodies Therapeutics Mammalian cell culture Penicillin Antibiotics Penicillium notatum Ice-minus Prevent ice from plants Pseudomonas syringae Riboflavin Vitamin Ashbya gossypii Sno-max Make snow Pseudomonas syringae Subtilisin Laundry detergent Bacillus subtilis tPA Blood clots Mammalian cell culture Tetracycline Antibiotics Streptomyces aureofaciens Tumor necrosis factor Kill/inhibit tumor cells Escherichia coli Xanthan gum Rheology modifier Xanthomonas campestris 2
  3. 3. Classification of microorganisms Prokaryotes Prokaryotes – Bacteria, Blue-green algae Unicellular: bacteria Multicellular: cyanobacteria Eukaryotes – Fungi (molds, yeasts), Do not contained membrane-contained algae, Protozoe nucleus Archaebacteria Can accept a wide variety of nutrients Rapid growth Viruses Versatile biochemical metabolism Structure of bacterial cell Essential structure Essential structures Cell wall: • 20 nm thick Non-essential • Consists of peptidoglycan structures • Structural strength and shape Cell membrane: • 7-9 nm thick • Lipid bilayer • Semi-permeable - controls the transfer of chemicals and nutrients 3
  4. 4. Essential structure (cont’d) Essential structure (cont’d) Nuclear body: Cytoplasm: • DNA • Fluid material • Control center for all operation • No nuclear membrane Mesosomes: • No mitotic apparatus during replication • Bacteria do not have mitochondria, but have Ribosomes: mesosomes, which are extensions of the • Sites of important biochemical reactions cytoplasmic membrane • Protein synthesis • ATP Non-essential structure Non-essential structure (cont’d) Pili (fimbria): Capsule (slime layer): • Gram(-) rods • Secreted by cells to increase viscosity and • Sexual conjugation impede diffusion • Adhesive to animal and plant cells • Coating to cell wall • Insert surface Volutin (mitochromatic) granules: Flagella: • Highly refractile globules • Motility of bacteria • Sourse of stored food, e.g. PHB • Can be polar or peritichous • Appearance influenced by age 4
  5. 5. Non-essential structure (cont’d) Actinomyces spp. Chromatophores: Beaded appearance of branched filamentous • photosynthetic rod shaped bacteria • Counterpart of chloroplasts for plant cells Cells are smaller Endospores: No nuclear membrane • Highly resistant to destructive effects of chemical and physical agents No lysozyme • Contain large amounts of dipicolinic acid Extremely important as a source of powerful (DPA) antibiotics Cyanobacteria Classification of bacteria Gas vesicles Grain stain Gas vesicles are aggregates of hollow cylindrical structures Morphological type – cocci, rod, spiral composed of rigid proteins. They are impermeable to water, but permeable to gas. The amount of gas in the vacuole is Spore forming or not under the control of the microorganism. Metabolism of sugar (carbon) substrates Gas vesicles are found in Cyanobacteria, which are Growth requirements: photosynthetic and live in aquatic systems. In these lakes • Oxygen – aerobic, anaerobic or facultative and oceans, the Cyanonbacteria want to control their • Nutrients position in the water column to obtain the optimum amount of light and nutrients. Genetic composition (GC content) 5
  6. 6. Gram stain Eukaryotes Unicellular: yeasts, algae Multicellular: molds, algae Posses a membrane-contained organelles Larger than prokaryotes Complex cell structure Spatial organization and differentiation Yeast Yeast Elliptical or spherical cells Size: 5 -10 microns Form spores Vegetative growth is by budding Aerobic and anaerobic growth Colony on agar plate similar to bacteria’s Important in beverage alcohol industry, ethanol, baker yeast and single cell proteins (SCP) 6
  7. 7. Mold (Filamentous Fungus) Mold Filamentous Hyphae Complicated life cycles Sexual and asexual spores Aerobic Mostly pathogens of plants Important in industrial fermentations: • Organic acids (citric, gluconic, gibberellic acid) • Antibiotics (penicillin, griseofulvin) • Enzymes (cellulase, protease, amylase) Cause spoilage in paper, fabrics and food Eukaryotes Animal and plant cells Plant cells Either as callus (undifferentiated plant tissue) Or as aggregated cells in suspension Can produce many commercially important compounds (perfumes, dyes, medicines and opiates) Can catalyze highly specific useful transformations 7
  8. 8. Eukaryotes Animal cells Eukaryote v.s. Prokaryote Tissue derived cells Primary cell lines secondary cell lines established, permanent cell lines Anchorage dependent cells Microcarrier culture techniques Flagella Important in large scale production of vaccines and other useful biochemicals and therapeutics Prokaryotes v.s. Eukaryotes Prokaryotes v.s. Eukaryotes Genome Organelles Characteristics Prokaryotes Eukaryotes Characteristics Prokaryotes Eukaryotes Mitochondria No More than one No. of DNA molecules One More than one Endoplasmic reticulum No Yes DNA in organelles No Yes Golgi apparatus No Yes DNA observed as chromosomes No Yes Photosynthetic apparatus Single protein Complex structure, Nuclear membrane No Yes Simple structure with microtubulus Mitotic/meiotic division of nucleus No Yes Ribosome Smaller, 70s Larger, 80s Formation of partial diploid Yes No Spore Endospores - High Endo/Exospores heat resistance low heat resistance 8
  9. 9. Archaebacteria Phylogenetic Tree of Life A third ancestral type of cells Absence of peptidoglycan from the cell wall Possess unusual lipids Methanogens, extreme halophiles, thermoacidophiles Viruses Bacteriophages Not cellular Possible contaminants of bacterial Informational parasites fermentation “Alive” when inside the host Vaccine production from animal viruses Characteristics of living things Smallest microbes Recombinant DNA techniques < 0.2 micron in size Cloning vehicle in genetic engineering Genetic materials: DNA or RNA 9
  10. 10. Protozoa Algae Small, single- Photosynthetic celled animal Some are Prokaryotes and some are Most live in water Eukaryotes (oceans, lakes, rivers, ponds) Eat bacteria Some are parasites – e.g., Malaria Metabolism Growth Requirement for Microbe Catabolism Nutrients - Carbon, nitrogen, and energy • Glycolysis sources; P, S, minerals, vitamins, etc. • Aerobic (Respiration) Temperature - mesophilic or thermophilic • TCA (Krebs) cycle pH - neutral, acidic, or basic • Electron transport chain • Anaerobic (Fermentation) Water activity - halophilic Anabolism (Biosynthesis) Oxygen - aerobic or anaerobic 10