Production of penicillin and citric acid

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Production of penicillin and citric acid

1. PRODUCTION OF PENICILLIN & Citric acid By Dr. Vijay Sharma Pharma Knowledge Subscribe and share 1

2. What is an antibiotic ? The term antibiotic is refers to an organic compound that is 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. For example, A s ointment or cream to treat bacterial infection on the surface. As tablet/capsules/injection to treat internal infection.

3. Antibiotics are produced primarily by bacteria, Streptomycetes , Nocardia and fungi. However, antibiotics produced by Streptomycetes spp. Having greatest commercial applications. On the basis of mode of action antibiotics can be divided as: Antibiotics affecting cell wall Antibiotics damaging cell membrane Antibiotics interfering with protein synthesis Antibiotics inhibiting nucleic acid synthesis Antibiotics blocking cell metabolism What is antibiotic

4. Penicillin The first discovered natural antibiotic was Penicillin which was obtained from multicellular fungi, “ Penicillium molds ”. Penicillin is a group of compounds having 6-amino penicillinic acid , contains ring like structure termed as a β- lactam ring . Penicillin are of two different types, Natural Penicillin (directly harvested from the penicillium mold). Semi-synthetic Penicillin consists of the basic Penicillin nucleus (6-APA), but with new side chain. It includes Ampicillin , Methicillin , Penicillin V, etc.

5. Penicillin The first discovered natural antibiotic was Penicillin by Dr Flemming ; which was obtained from multicellular fungi, “ Penicillium molds ”. Penicillin is a group of compounds having 6-amino penicillinic acid , contains ring like structure termed as a β- lactam ring . Penicillin are of two different types, Natural Penicillin (directly harvested from the penicillium mold). Semi-synthetic Penicillin consists of the basic Penicillin nucleus (6-APA), but with new side chain. It includes Ampicillin , Methicillin , Penicillin V, etc.

6. Production of Penicillin Inoculum P. notatum & P. chrysogenum are two species used. P. chrysogenum is high yielding strain and therefore most widely used as production strain. A starter culture transferred to an agar containing plate to allow growth. After getting growth on solid media, one or two growth stages should allowed in shaken flask cultures to create a suspension, which can be transferred to seed tanks for further growth. Feed tank contain all the nutrients including Growth factors like vitamins & amino acid as well as it may maintain temperature, pH, agitation, aeration etc.

7. Media Components Percent(%) Use Lactose 3.5 to 6 C- sourse Calcium Carbonate 1.0 buffer Corn steep Liquor 3.5 N-source Glucose 1.0 C- sourse ) Phenyl acetic acid 0.5 Precursor Sodium hydrogen phosphate 0.4 catalyst Edible oil 0.25 Bio parameters pH : near 6.5 Temperature: 26°C to 28°C Aeration: a continuous stream of sterilized air is pumped into it. Agitation: achieved by turbine type impellers for constant agitation. Time period 5-6 days Production of Penicillin

8. Recovery and purification Vacuum filter is used for separation of mycelium from the broth on a rotary. The lowering of pᴴ is done by adding phosphoric acid or sulphuric acid for conversion of penicillin to the anionic form occurs at low pᴴ (2.0 to 2.5). For removal of pigment and other impurities from solvent containing penicillin it is treated with active charcoal. The product of penicillin is then crystallized into sodium or potassium penicillin. Production of Penicillin

9. Citric Acid Citric acid is the most important commercial product, which is found in almost all plant & animal tissues. Citric acid is the most important organic acid produced in tonnage and is extensively used in food and pharmaceutical industries. Citric acid is a weak organic acid found in citrus fruits(lemon). It is natural preservative and is also used to add an acidic taste to food and soft drinks. Million tonnes of citric acid are produced every year by fermentation.

10. Inoculum The fungus Aspergillus niger is used. Mutant strain of Aspergillus niger is used for improved production. It can tolerate high sugar concentration and low pH with reduced synthesis of undesired byproducts. Industrial fermentation process is carried out in three different ways as: Surface fermentation Submerged fermentation Solid state fermentation Production of Citric acid

11. Surface Fermentation a. Solid Surface Fermentation Carried out in some Asian countries like Japan using A. niger . Solid Substrate such as wheat bran or pulp from sweet potato starch are used, as culture media. Fermentation is done at pH is kept 4-5 at incubation temp. of 28 C for 80-100 h. Recovery and isolation is done using hot water . Production of Citric acid

12. Surface Fermentation b . Liquid Surface Fermentation This is oldest method & carried out using liquid nutrient such as beet carried out in aluminium trays filled with nutrient media. Inoculum in the form of spores sprayed. Temperature spread 30 C The pH of medium fall below 2 , as mycellium grows. After 7-15 days , the mycellium is separated, pressed mechanically and washed to get maximum amout of citric acid. Production of Citric acid

13. SUBMERGED FERMENTATION : In this case, the strains are inoculated of about 15cm depth in fermentation tank. The culture is enhanced by giving aeration using air bubbles. And its allowed to grow for about 5 to 14 days at 27 to 33 degree Celsius. The citric acid produced in the fermentation tank and it is purified. Production of Citric acid

14. Solid state fermentation: It is simplest method for citric acid production. Solid state fermentation is also known as Koji process , was first developed in Japan. Citric acid production reached a maximum(88g/kg dry matter) when fermentation as carried out with cassava having initial moisture of 62% at 26degree Celsius for 120 hours. Production of Citric acid

15. Recovery The mycellium is separated by filtration of culture broth and washing. Oxalic acid as unwanted byproduct is separated by precipitation by adding lime. The liquid is transferred to recovery process Separation of citric acidis done at pH 7.2 and temp. 70-90 degree by precipitation. Further purification is done by dissolving citric acid in sulfuric acid. Finally treated with activated charcoal, cation , anion exchanger and crystallize Production of Citric acid

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