FERMENTATION AND ITS TYPES
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The document provides an overview of fermentation processes, highlighting types of fermentation and notable products such as antibiotics (penicillin and streptomycin) and vitamins (B2 and B12). It details the production methods, including nutrient media composition, inoculum preparation, and recovery processes for these substances. Key differences between aerobic and anaerobic fermentation are also discussed, outlining the biochemical pathways and efficiency of energy production.
FERMENTATION AND ITS TYPES
- 1. FERMENTATION
- 2. CONTENTS Introduction Types of fermentation Production of – 1. Antibiotics : Penicillin & Streptomycin 2. Vitamins: B2 & B12 References 2
- 3. Introduction Pasteur definition: “life without air”, anaerobe red ox reactions in organisms. New definition: a form of metabolism in which the end products could be further oxidized. For example: a yeast cell obtains 2 molecules of ATP per molecule of glucose when it ferments it to ethanol 3
- 4. 4 Some important fermentation products PRODUCT ORGANISM USES Ethanol Saccharomyces cerevisiae Industrial solvents beverages Glycerol Saccharomyces cerevisiae Production of explosives Lactic acid Lactobacillus bulgaricus Food and pharmaceutical Acetone & Butanol Clostridium acetobutylicum solvents
- 5. Difference between FERMENTATION 5 AEROBIC FERMENTATION ANAEROBIC FERMENTATION Set of chemical reactions involved in production of energy by completely oxidizing food. Chemical breakdown of organic substrate into ethanol or lactic acid by micro-organism in absence of oxygen Occurs in both cytoplasm and mitochondria Occurs in cytoplasm Occurs in higher animals and plants Occurs in yeast ,parasites, and bacteria
- 6. 6 Uses molecular oxygen as final electron acceptor in ETC Does not use oxygen Produces 6 water molecules/glucose Does not produce water Glucose completely broken into CO2 & O2 Glucose incompletely oxidized either into ethanol & lactic acid NAD+ regeneration occurs in ETC NAD+ regeneration occurs during partial oxidation of Pyruvate 36 ATP is produced Only 2 ATP is produced Continued……
- 7. Penicillin Also known as Penicillium Notatum / Chrysogenum. It is common in temperate and subtropical region and can be found on salted food products, but it is mostly found in indoor enviroments, especially in damp or water damaged buildings. It is source of several β-lactum antibiotics, which inhibits biosynthesis of bacterial cell wall. 7
- 9. Production P. chyrosegenum is high yielding strain and therefore most widely used as production strain. Innoculum preparation- various sequential steps are necessary like: 1. A starter culture is needed for innoculation. 2. After getting growth on solid media, one or two growth stages should allowed in shaken flask cultures to create a suspension, which can be transfered to seed tanks for further growth. 9
- 10. 3. After about 24-28 hours, the content of seed tanks is transfered to primary fermentation tanks. 4. All the bio-parameters like temperature, pH, aeration, agitation,etc. should be properly maintained. A. BIO-PARAMETERS- pH- near 6.5 Temperature – 260- 280 C Aeration- Continous stream of sterilized air is pumped into it. Agitation- have baffles which allows constant agitation (200rpm). 10
- 11. B. NUTRIENT MEDIA Fermentation broth contains all necessary elements required for the proliferation of micro-organisms. Generally it contains- 1. Carbon source 2. Nitrogen source 3. Mineral source 4. Precorsors 5. Anti – foaming agents 11
- 12. 12 Composition of nutrient media for penicillin 1. Corn steep liquor 3.5% 2. Lactose 3.5% 3. Glucose 1% 4. Calcium carbonate 1% 5. Potassium hydrogen phosphate 0.4% 6. Edible oil 0.25% Phenyl acetic acid as preservative
- 13. C. RECOVERY The recovery of Penicillin is carried out in three successive stages: 1. Removal of mycelium 2. Counter current solvent extraction of penicillin 3. Treatment of crude extract 13
- 14. For increasing yield of penicillin Adding corn syrup Selection of strain Mutation and selection Sexual reproduction 14
- 15. streptomycin Streptomycin is an bactericidal antibiotic drug belonging to class of aminoglycosides. Derived from actinobacterium Streptomyces griseus. Used against gram negative bacteria especially. Chemical composition – It contains 3 sugars derived from glucose with C, N, O & H elements. Chemical formula- C21H39N7O12 15
- 16. INNOCULUM Streptomyces griseus maintained in soil stocks or lyophilized in carrrier are innoculated into sporulation medium which builds up mycelial innoculum. THE HOCKENHUL MEDIUM 1. Glucose 2.5% 2. Extracted soya meal 4% 3. Distillers dried soluble 0.5% 4. Sodium chloride 0.25% pH 7.3-7.5 16 Production
- 17. A. NUTRIENT MEDIA Medium is a nutritive substance in which cultures are grown for scientific purposes. The culture medium for streptomycin consists of: 1. Carbon source 2. Nitrogen source 3. Animal oils 4. Vegetable oils 5. Mineral oils 17
- 18. B. FERMENTATION PROCESS Spores of S. griseus are innoculated into medium to establish a culture with high mycelial biomass for introduction into innoculum tank, using innoculum to initiate the fermentation process. Yield in production vessel responds to high aeration & agitation condition Other condition are: Temperature 250-300C pH range 7-8 Time- 5-7 days 18
- 19. The fermentation process for production of streptomycin involves three phases : 1. Phase I (last up 24 hours) 2. Phase II (takeplace5-7 days) 3. Phase III (harvested before cell lysis) C. RECOVERY & PURIFICATION 1. Removal of mycelium 2. Recovery process 3. Purification 19
- 20. Cyanocobalamine ( B12) Cyanocobalamin is industrially produced stable cobalmin from which is not found in nature It is obtained exclusively by fermentation process. Merck began production of vitamin B12 by Pseudomonas dentrificans in 1952 & have improved the efficiency of culture more than 30 folds relative to the performance of original soil isolates by genetic manipulations and microbial screening. Mutagenic treatments have resulted in improved activity. 20
- 21. Mutagenic treatments have resulted in improved activity. Structure of cyanocobalamine : 21
- 22. 22 Downstream process of vitamin B12
- 23. Riboflavin (B2) Riboflavin has been produced commercially by chemical synthesis, by fermentation and by combination of fermentation and chemical synthesis. Fermentation route has been widely used as it produces the vitamin in single step, resulting in substantial cost savings. Most of the producers like BASF, Roche, ADM/ Aventis, Hubei Gaungii prefer fermentative production. 23
- 24. Although bacteria (Clostridium species) & yeasts (Candida species) are good producers, two closely related Ascomycete fungi, Eremothecium ashbyii & Ashbya gossypii, are considered the best riboflavin producers. 24
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