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FE 462 BIOCHEMICAL ENGINEERING

FE 462 BIOCHEMICAL ENGINEERING. INOCULUM. DEFINITION OF INOCULUM Living organisms or an amount of material containing living organisms (such as bacteria or other microorganisms) that is added to initiate or accelerate a biological process, i.e., biological seeding. CRITERIA;

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FE 462 BIOCHEMICAL ENGINEERING

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  1. FE 462 BIOCHEMICAL ENGINEERING INOCULUM

  2. DEFINITION OF INOCULUM • Living organisms or an amount of material containing living organisms (such as bacteria or other microorganisms) that is added to initiate or accelerate a biological process, i.e., biological seeding. • CRITERIA; • Healthy, active state - minimize lag period • Available in sufficient quantities • Suitable morphological form • Free of contamination • Stable - retain its product forming properties

  3. CHOICE OF MICROORGANISM; • Nutritional characteristics - cheap medium • Optimum environmental conditions • Productivity - substrate conversion, product yield, rates. • Amenability to genetic manipulation • Ease of handling and safety (suitability)

  4. Inocula • Pure Monocultures • Advantages and disadvantages of pure cultures • Advantages: easy to obtain (isolate, genetically modify, or purchase; better control of products) • Disadvantages: subject to contamination and genetic change

  5. Processes requiring monocultures i.e PURE CULTURE FERMENTATIONS - industrial ethanol - alcoholic beverages - fermented foods - pharmaceuticals - acetone-butanol - acetic acid - single cell protein - industrial enzymes - biotech products (insulin, growth hormone)

  6. Culture collections Culture collections:for academia, industry, agriculture, medicine and research. specialized in the long-term preservation and storage of microorganisms (documentation associated with these materials.) may be private and profit-oriented, private and non-profit, or public (academic or governmental); cultures may be distributed for a fee or, more rarely, free of cost.

  7. Culture collections supply of industrial microorganisms • AbbreviationNameLocation • ATCC American Type Culture Collection Rockville, MD, U.S. • CBS Centraalbureau voor Schimmenlculturen Baarn, The Netherlands • CDDA Canadian Department of Agriculture Ottawa, Canada • CMI Commonwealth Mycological Institute Kew, United Kingdom • FAT Faculty of Agriculture, Tokyo University Tokyo, Japan • IAM Institute of Applied Microbiology University of Tokyo, Japan • NCIB National Collection of Industrial Bacteria Aberdeen, Scotland • NCTC National Collection of Type Cultures London, United Kingdom • NRRL Northern Regional Research Laboratory Peoria, IL, United States • PCC Pasteur Culture Collection Paris, France

  8. Preservation of pure cultures • 1. Culture Transfer • contamination • genetic change • 2. Refrigeration from 0o to 5oC • short term storage • 3. Low Temperature Freezing • ultra low temp. freezer (-80oC) • liquid nitrogen (-196oC) • 4. Lyophilization • freeze with dry ice and acetone • sublime off water (dries cells without disruption) • use of skim milk, glycerol, or sucrose to protect cells • 5. Mineral Oil • 6. Dry Spores

  9. Mixed Cultures • Advantages and disadvantages of mixed cultures • - Advantages: obtained by enrichment or purchased; can't be patented; contamination not as much of problem • - Disadvantages: control of culture and products is less definite;

  10. Mixed culture fermentations - breads: sour dough, soda cracker - wines - vegetables: pickles, sauerkraut - dairy products: yogurt, sour cream - ensiling - composting - anaerobic digestion - soil and groundwater remediation - bioleaching - microbial enhanced oil recovery - microbial metals recovery - waste treatment

  11. STORAGE AND PRESERVATION; • Essential that isolates / cultures retain desirable characteristics over long periods of time. • METHODS; •  Storage at reduced temperatures; • 1. Slopes - refrigerator (4 oC), freezer (-20 oC), • protec beads (-80 oC), • 2. Fungal spores in water (5 oC) • 3. Liquid nitrogen (-150 to -196 oC) •  Storage in dehydrated form; • 1. Soil + culture dried. Used for fungi • 2. Lyophilization \ freeze drying. Freezing of culture followed by drying under vacuum which results in sublimination of cell water

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