Fermentation process of antibiotics

BACKGROUND

Antibiotics are a type of drugs or medicines that hinder the growth of bacteria.¹ They generally kill the microorganisms and also cure bacterial infections. Any drug that kills any microorganisms or germs is known as antibiotics.² The first ever antibiotic discovered was Penicillin.

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The antibiotic penicillin which is β lactamase antibiotic is Penicillin.³ Penicillin was among the first antibiotics used to kill the microorganisms and cure bacterial infections. In 1928 Alexander Fleming discovered it.⁴ For the production of Penicillin, fermentation process was carried out. The fermentation is thus a chemical process by which the molecules broke down anaerobically.⁵

So the basic objective of the test is to show the production of antibiotic-penicillin by process of industrial fermentation [Fed-Batch Fermenter].

REQUIREMENTS

Chemicals:     

    Corn steep 8.5%- 21.88 gm/ltr

    Glucose: 1%

    Butyl acetate

    Phosphate buffer

    Chloroform

    Ether solution

    Sodium bicarbonate

Apparatus:    

    Fermenter

    Centrifuges

    Rotary vacuum filter

    Fluid bed dryer

PROCEDURE

Media preparation

Media preparation is necessary as it involves the utilisation of microorganism to obtaine the products. In Penicillium fungus, the medium contains its carbon source found in corn steep and glucose. Medium consists of salts such as Sodium Nitrate, Magnesium sulphate and Potassium phosphate.

Heat sterilisation

Medium is sterilised at high heat and high pressure or directly sterilised together by fermenter. The medium is heated at 121°C. High temperature is used to minimise the degradation of components of media.

Fermentation

Basically penicillin fermentation is done in fed-batch mode as in this process glucose is not added in high amount from the beginning of growth which further results in low yield of penicillin production as excess glucose inhibits penicillin production.⁶ Penicillin is a secondary metabolite of the fungus, so the fed batch mode is ideal for such products as it allows high production of penicillin. The temperature is generally maintained at 20-24°C and pH maintained at 6.0-6.5. The air bubbles are sprinkled to provide enough oxygen. The rotor is required as it mixes the culture evenly throughout the culture medium.

Culture

The culture is developed by Penicillium spores addition into a liquid medium. When it grows up to an acceptable amount, it is inoculated into the Fermenter.

Inoculation: Medias used in the manufacture of penicillin is inoculated by several methods, such as

1. Surface culture: the medium surface is inoculated with dry spores. The spore material is applied to cover the surface as uniformly.
2. Submerged culture: production medium inoculated with dry spores, by pellet inocula prepared in sterile soap solutions in sterile water containing 100 ppm sodium lauryl sulphonate. Under submerged conditions by growing mycelium from mold spores Pellet inoculation is prepared.

Biomass removal

Biomass such impurities are separated from the culture medium containing the penicillin product. For removal of these impurities Rotary vacuum filter is commonly used. As the pH will be high upto 8.5 so to maintain the pH between 6.0-6.5 phosphoric acid is added.

Solvent addition

Organic solvent butyl acetate is added to dissolve the penicillin in the filtrate. After addition, penicillin is then present in solution mode & other solids present are waste.

Centrifugal extraction

This is done for separation of the solid waste from the liquid solution containing Penicillin. Generally a tubular bowl centrifuge or disk centrifuge or a chamber bowl centrifuge is used for the separation. Penicillin dissolved in the solvent undergoes continuous extractions to obtain purified penicillin product. After mixing of the butyl acetate, phosphate buffer is added then mixed with chloroform solution and then again mixed with phosphate buffer, finally adding ether solution. After addition of ether solution Penicillin will be present in higher concentration. To obtain penicillin-sodium salt sodium bicarbonate is added. It is thus obtained by continued centrifugation which completely removes the solid wastes from the penicillin-sodium salt.

Drying

Drying completely removes the moisture content from the penicillin salt. This is done through drying method-fluid bed where gas(hot) is pumped into chamber base (where penicillin is present) passing to a vacuum chamber resulting in obtaining a dryer form of Penicillin.

Isolation, Purification, Refining, Packaging And Shipping (Source: www.madehow.com)

        PENICILLIN STRUCTURE (SOURCE: WIKIPEDIA)

CONCLUSION

Penicillin is thus stored in dry containers & in dry environment. Further it is packaged as liquid for or as pills

REFERENCES

1. NHS.”Antibiotics” 2015
2. Europian Centre for disease prevention and control; “Factsheet for experts” 2014
3. Holten KB, Onusko EM; “Appropriate prescribing of oral beta-lactam antibiotics”. 2000: 62(3): 611-20
4. American Chemical Society “Discovery and Development of Penicillin”;(2015)
5. Klein, Donald W, Lansing M, Harley, “Microbiology 6^th Edition; New York; McGraw-hill; ISBN 978-0-07
6. Journal of Chemical Technology and Biotechnology.”The theory of fed batch culture with reference to penicillin fermentation”. 1974. Vol 24, issue 7:415-424.

Also read:

• Isolation, screening and purification of an unknown organism from soil
• Cell Immobilization Techniques
• Fermentation process of alcohol production
• Analgesic activity study of drugs by hot plate (Eddy’s hot plate) method
• Study of local anesthetics by Infiltration anaesthesia in guinea pig
• Study of local anesthetics by Surface anaesthesia on the cornea of rabbits
• Study of local anesthetics by Spinal anaesthesia in rats

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