Penicillin Production - The mould from which Fleming isolated penicillin, in was later

identified as Penicillium notatum. A variety of moulds belonging to other species and

genera were later found to yield greater amounts of the antibiotic and a series of closely
related penicillins.

The naturally occuring penicillins differ from each other in the side chain (R group).
Penicillin was produced by a surface culture method early in World War II. Sub-merged
culture methods were introduced by 1943 and are now almost exclusively employed.
Penicillin production needs strict asceptic conditions. Contamination by other
microorganisms reduces the yield of penicillin.

This is caused by the widespread occurrence of penicillinase producing bacteria which

inactivate the antibiotic. Secondly, penicillin production also needs tremendous around of
air. In all methods, deep tanks with a capacity of several thousand gallons arc filled with
a culture medium. The medium consists of cornsteep liquor, lactose, glucose, nutrient ,
salts, phenyl acetic acid or a derivative and calcium carbonate as buffer.

The medium is inoculated with a suspension of conidia of Penicillium chrysogenum. The

medium is constantly aerated and agitated, and the mould grows throughout as pellets.
After about seven days, growth is complete, the pH rises to 8.0 or above, and penicillin
production ceases. When the fermentation is complete, the masses of mould growth are
separated from the culture medium by centrifugation and filtration.

The complex process of extracting the penicillin from the clear fluid then begins. The
method involves various extractions with organic solvents and recrystallization. Penicillin
is assayed to determine its potency before being bottled and sold.

The potency of a batch of penicillin is determined by a biologic assay in which the

unknown is compared with a standard preparation of crystalline sodium penicillin G.
Cylender-plate method is used to determine the potency of penicillin. The test consists of
adding nutrient, .agar, previously inoculated, with a specified strain of Staphylococcus, to
a sterile Petri dish. Stainless steel cylender open at both the ends are placed on the agar.

The cylinders are filled with suitable dilutions of the working standards of penicillin and
of the unknown sample. The plates are incubated at 37°c for 16 to 18 hours. The
diameters of the zones of inhibition of the bacterial growth are measured. The antibiotic
activity of the unknown sample is determined by comparing its zones of inhibition with
those of the standard penicillin.