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Structure
Mechanism of Penicillin
The bacterial cell wall is made up of peptidoglycan that protects the cell
integrity, shape and prevents macromolecules from entering into the
cell.
It has cross-linking chains of NAG(N-acetylglucosamine) and NAM (N-
acetylmuramic acid) components covalently attached through small
peptides
Peptidoglycan is continuously synthesized and remodelled during growth
and division.
Therefore bacteria assemble them into a single macromolecule by
synthesizing the peptidoglycan components.
The peptide cross-linkages are formed by the activity of specific
enzymes called transpeptidases or Penicillin-Binding Proteins (PBPs).
Penicillin contains a four-membered beta-lactam ring inhibits the
transpeptidase.
By mimicking the last two D-alanine residues of the peptide, penicillin
is able to bind irreversibly the active site of the transpeptidase,
preventing the enzyme from cross-linking the peptidoglycan strands.
This leads to cell lysis as there won’t be any formation of peptidoglycan
and the cell is no more susceptible to osmotic stress.
Penicillin G
1. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5369031/
2. https://www.ncbi.nlm.nih.gov/books/NBK554560/
3. https://microdok.com/penicillin-mechanism-of-action/
Production of Penicillin
The penicillin nucleus contains two amino acids, L-cysteine and D-valine (Fig.
5.2) which are closely related in structure. Penicillins are N-acyl derivatives of
6-aminopenicillanic acid (6-APA). By adding side chains to the penicillin
nucleus, new penicillins are synthesized (Fig. 5.3). The semisynthetic
penicillins are prepared from 6-aminopenicillanic acid (6-APA). Acylation of
the 6-APA is easily and efficiently brought about by chemical reactions (Fig.
5.4) Different derivatives of semisynthetic penicillin are shown in Fig. 5.5.
Inoculum development:
Production media:
The exact composition of penicillin production media used in the industry for
production of penicillin is unknown. These media are considered to be trade
secrets of that particular fermentation industry. A typical medium described by
Jackson (1958) contains fermentable carbohydrates, such as corn steep liquor
solids (3.5%), lactose (3.5%) and glucose (1%) potassium dihydrogen
phosphate (0.4 %), calcium carbonate (1%), edible oil (04%), and penicillin
precursor. The pH of this medium after sterilization is 5.5 to 6.0. Inoculum
media are similar to production media except that lactose and precursors are
not added in the inoculum media. These media compositions may be slightly
changed to increase yields and meet economic changes.
Fermentation:
The media are placed in a fermentation vessel, sterilized, and inoculated with a
suspension of Penicillium chrysogenum (inoculum). A flowsheet for large-
scale production of penicillin is shown in Fig. 5.6. The fermentation vessel is
equipped with devices that allow continuous addition of nutrients, acids/bases
to maintain the pH (7 to 7.4), and cooling coils to maintain the temperature
(24°C).
Penicillin is not a single chemical compound, but a group of compounds of related structure
and activity. There are six penicillins – Penicillin G (benzyl penicillin), penicillin V
(phenoxy-methyl penicillin), penicillin F (A2-pentcnyl penicillin), penicillin X (p-
hydroxybenzyl penicillin), penicillin K (n-heptyl penicillin) and penicillin O (allyl-
mercaptomethyl penicillin). Penicillin is selective for gram-positive bacteria, some
spirochaetcs and the gram-negative diplococci (Neisseria).
Commercial Production of Penicillin:
Commercial production of penicillin is depicted in Fig. 40.7. The commercial production of
most of the antibiotics follows a similar plan. The major differences relate to the
microorganism, media composition, and extraction procedure. For penicillin production,
inoculum of a high yielding strain of P. chrysogenum is prepared as follows: inoculum is first
cultured in flasks in wheat bran nutrient solution for 1 week at 24°C.
The culture is then transferred to inoculum tanks, and grown for 1-2 days under aeration; this
supports heavy mycelial growth. The inoculum is now added to very large fermentors
containing the production medium, which consists of 10 per cent total glucose/molasses, 4-
5% corn-steep liquor solids, 0.5-0.8 per cent phenylacetic acid and 0.5% vegetable oil; pH is
adjusted to 6.0 and temperature is regulated at 25- 26°C.
Earlier media contained lactose, but it is no longer used. The fermentation is carried out
under aerobic conditions, and nutrient supply is maintained by regular feeding (fed-batch
culture). The fungus grows in a submerged culture mostly as mycelial balls.
The fermentation is carried out for 7 days. Initially, mycelial growth occurs, and
carbohydrates are used up. This reduces the carbohydrate level in the medium; this favours
penicillin production, which begins from the second day of fermentation. The pH of medium
rises to 8.0 by the 7th day, and penicillin production stops at this stage. Phenylacetic acid is
the precursor for the benzene ring side chain of Penicillin G.
ADVERTISEMENTS:
The side chain of 6-APA is variously modified to obtain a variety of derivatives of penicillin,
which are used in medicine. In addition, several other fungal metabolites are used as
antibiotics (Table 40.3).