Antibiotics

Antibiotics are probably the most important group of compounds synthesized by

industrial microorganisms. Most antibiotics are secondary metabolites produced by
filamentous fungi and bacteria, particularly the actinomycetes. The best known and
probably the most medically important antibiotics are the β-lactams, penicillins
and cephalosporins; along with aminoglycosides, such as streptomycin, and the
broad-spectrum tetracyclines.

Over 100 β-lactams, mostly penicillins and cephalosporins, have been approved for
human use, and they account for over half of the antibiotics produced worldwide.
They specifically target the synthesis of peptidoglycan, a vital bacterial cell wall
component, which is not present in eukaryotic organisms, thus providing a high
level of selectivity. They primarily inhibit the cross-linking transpeptidation
reaction, resulting in the formation of incomplete peptidoglycan, severely
weakening the bacterial cell wall structure.

Penicillin

Penicillin was discovered by Fleming in 1928 following his famous observation of

an inhibitory zone surrounding a fungal contaminant, Penicillium notatum, on a
plate of Staphylococcus aureus. The discovery of penicillin and its later
characterization and purification ultimately led to major advancements in both
medicine and fermentation technology.

The basic structure of the penicillins is 6-aminopenicillanic acid (6-APA),

composed of a thiazolidine ring fused with a β-lactam ring whose 6-amino
position carries a variety of acyl substituents. In the absence of added side-chain
precursors to the fermentation medium of P. notatum or P. chrysogenum, a
mixture of natural penicillins is obtained from culture filtrates, notably penicillin G
and the more acid-resistant penicillin V.

Most penicillins are now semisynthetic, produced by the chemical modification of

natural penicillins. Modification is achieved by removing their natural acyl group,
leaving 6-APA, to which other acyl groups can be added to confer new properties.
Examples of semisynthetic penicillins include methicillin, carbenicillin and
ampicillin. The commercial production of penicillin is outlined in the figure below
Figure: Production of penicillin
Scale-up

Scale-up, in a broad sense, is a critical activity that enables a fermentation process

achieved in research and development to operate at a commercially viable scale for
manufacturing.

For a fermentation scale-up, the following parameters can be used to evaluate if

the scale-up is a success.

1. Product formation rate: First of all, the final product titer or yield at large scale
should equal that of small-scale studies.

2. Cell growth profile: A successful scale-up results in a comparable growth profile

between the large-scale fermentation and the small-scale fermentation

3. Operational aspects: The success of a scale-up is also judged by how efficiently

and effectively a new operation team at the receiving facility can execute the
scaled-up process, including media preparation, media transfer, and process
documents.

4. Product quality: Product quality should be examined to confirm a successful

scale-up.
Define metabolic engineering? What are the ways of enhancing production of
secondary metabolites? Give an example of metabolic engineering strategy.

Metabolic engineering is the practice of optimizing genetic and regulatory

processes within cells to increase the cells' production of a certain substance.

Many attempts have been made to enhance secondary metabolites production via
metabolic engineering. The many possible ways by which biosynthesis of secondary
metabolites may be enhanced include

 increasing the supply of precursors through primary metabolism,

 redirecting the flux of the needed cofactors to antibiotic biosynthesis,

 amplifying one or more of the enzymes involved in the biochemical

pathways and

 improving the kinetic characteristics of the enzyme.

For example, inactivation of cefR delays expression of the cefEF gene increases
penicillin N secretion and decreases cephalosporin production. Overexpression of
the cefR gene decreased up to 60% penicillin N secretion, saving precursors and
resulting in increased cephalosporin C production.