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Penicillin prodn
- 1. Y5E02 BIOPROCESS TECHNOLOGY – Unit IV – Penicillin Production Dr. S. SIVASANKARA NARAYANI ASSISTANT PROFESSOR DEPARTMENT OF MICROBIOLOGY AYYA NADAR JANAKI AMMAL COLLEGE SIVAKASI
- 2. UniT -IV • Microbial production technology: antibiotics (penicillin) - organic acids (citric acid and acetic acid) - vitamins (b12 and riboflavin) - *enzymes (amylase) - alcoholic beverages (beer and wine). Single cell protein. 08-08-2020Dr.SS
- 3. Antibiotics • The term antibiotic has been defined by selman waksman as being an organic compound produced by one microorganism that, at great dilutions, inhibits the growth of or kills another or even group of other harmful microorganisms. • Antibiotics are available in various forms. Ointment or cream form - treat bacterial infection on the surface. Treat internal infection it can be directly injected into the bloodstream which finally Distributed throughout the body. Antibiotics are produced primarily by bacteria, streptomycetes, nocardia and fungi. 08-08-2020Dr.SS
- 4. 1 2 3 Damaging cell membranes Interfere with Protein synthesis Affects cell wall Mode of Action 08-08-2020Dr.SS
- 5. 1 2 * Blocking cell metabolism Penicillin – cell wall synthesis, inhibits the cross linking steps of peptidoglycan synthesis Inhibits nucleic acid synthesis Mode of Action 08-08-2020Dr.SS
- 6. Penicillin • First discovered natural antibiotic • 1928, Alexander fleming • Penicillium molds – Penicillium notatum • 6 – aminopencillinic acid (6 APA) – B –lactam ring • Two types – Natural and Semisynthetic 08-08-2020Dr.SS
- 7. 08-08-2020Dr.SS Process Microorganism Inoculum preparation Bio parameters Raw materials – C, N, &Mineral Precursors Antifoam agents Production medium Recovery Marketing
- 8. Microorganism • P. Notatum & P. Chrysogenum. • P. Chrysogenum is high yielding strain • The production strain is improved by mutation with the help of x-rays or any other agents to give high yield. • Strain should be carefully maintained because P. Chrysogenum is genetically unstable. 08-08-2020Dr.SS
- 10. 08-08-2020Dr.SS Inoculum Preparation Cold store – agar plate Shaken flask culture Seed tank (stainless steel – all the nutrients, equipped with agitators, sterilized air) 24 – 48 hours of incubation Primary fermentation tank Larger version of seed tank ( favourble condition for organism
- 11. Bioparameters • PH: near 6.5 temperature: 26°c to 28°c aeration: a continuous stream of sterilized air is pumped into it. Agitation: have baffles which allow constant agitation. 08-08-2020Dr.SS
- 12. Raw Materials Carbon source • Lactose – 6% , glucose and sucrose • Molasses, soy meal Nitrogen source • ammonium sulfate, ammonium acetate, ammonium lactate or ammonia gas • corn steep liquor Mineral source • Phosphorus, sulfur, magnesium, zinc, iron, and copper • water soluble salts. Precursors • phenyl acetic acid - penicillin-G ,hydroxy phenyl acetic acid - penicillin-X. Phenoxy acetic acid - for penicillin-V production. • When corn steep liquor is provided as nitrogen source, it also provides phenyl acetic acid derivatives; therefore it is widely used in the production of penicillin-G. Antifoam agents • lard oil, octadecanol and silicones 08-08-2020Dr.SS
- 13. 08-08-2020Dr.SS Imporant things have to keep in mind before choosing raw materials 1.Ease of extraction and purification of antibiotics. 1.An abundant growth of mycelium 1.Maximum accumulation of penicillin
- 14. Production Medium • Lactose: 3.5 to 6 • Calcium carbonate: 1.0 • Cornsteep liquor: 3.5 • Glucose: 1.5 • Phenyl acetic acid: 0.5 • Sodium hydrogen phosphate: 0.4 • Antifoaming agent: edible oil: 0.25 note : concentrations are given in % 08-08-2020Dr.SS
- 15. Recovery • Removal of mycelium • Counter current solvent extraction of penicillin • Treatment of crude extracts 08-08-2020Dr.SS
- 16. 08-08-2020Dr.SS Recovery Fermentation broth – filtered – rotary vaccum evaporator Remove mycelium and soilds Phosporic acid or sulfuric acid added to lower pH (2 to 2.5) Broth – extracted - Podbielniak Counter Current Solvent Extractor -methyl isobutyl ketone, amyl acetate or butyl acetate. filteration •Extracted into water from the organic solvent by potassium or sodium hydroxide to form a salt of the penicillin. again acidified & re-extracted with methyl isobutyl ketone •shifts between water and solvent help in purification of the penicillin. •penicillin to crystalize as sodium or potassium penicillinate •crystalline penicillin salts are then washed and dried. •government standards.
- 17. 08-08-2020Dr.SS Cont…. •Spent solvents resulting from the above procedure are recovered for re use •crude extract of penicillin is passed out - charcoal treatment •Sterile vials are used for packaging of an antibiotic either as a powder or suspension. •For oral use it is tableted - film coating. •government standards before being marketed.
- 19. Questions to think • What are the main things have to keep in mind before the penicillin production? • What is the role of precursors in the medium? • Steps involved in the recovery process • Role of charcoal in the penicillin extraction • Pointout the use of cornsteep liquor • Lactose and production medium relationship - specify 08-08-2020Dr.SS
- 20. 08-08-2020Dr.SS