Lecture-10

SEMISYNTHETIC PENICILLINS: Although penicillin was indeed an amazingly

effective drug, improvement was still desired in two areas. First, after just a few
years of its widespread use, some of the S. aureus strains had become penicillin-
resistant. Second, penicillin G (like penicillin V) is essentially inactive against
Gram-negative bacteria.
Production of semisynthetic penicillins: The starting material, usually penicillin G,
is cleaved enzymatically by penicillin acylase to yield 6-aminopenicillanic acid. This
is then chemically acylated with acyl chlorides to produce various semisynthetic
compounds. When the R group contains an amino function, as in the synthesis of
ampicillin, this amino group must be blocked first and then de-blocked after the
acylation reaction.

GENETICS OF ANTIBIOTIC PRODUCTION: Most antibiotics are still produced by

fermentation or by the chemical modification of fermentation products. The
efficiency of production of antibiotics is determined largely by the genetic makeup
of the producing strains, and much effort has been spent on improving these
strains.
Traditional Method of Strain Improvement: The traditional genetic approach to
improving the yield of an antibiotic-producing organism depends entirely on
random mutagenesis and screening of high producers. Until recently, the
screening had to be done by growing each progeny clone in liquid media and
assaying for the antibiotic in the culture filtrate. Because such screening was
laborious and slow, only a small number of progeny could be tested in one
experiment.
Methods of Classical Genetics: Because the traditional approach to strain
improvement involves many steps of heavy, random mutagenesis, each step
introduced many unwanted mutations into the organism, and the overproducing
strain that resulted was invariably a weakened strain, one that grew poorly. In this
technique, bacterial or fungal cells are converted to protoplasts by dissolving the
cell wall with lytic enzymes. The membranes of two protoplasts are then fused
together by the addition of high concentrations of polyethyleneglycol. Then cell
walls are regenerated in the progeny in suitable protective media. Usually the
chromosomes from the two parental protoplasts undergo recombination, and the
redundant material is eventually discarded in the process of successive cell
division.
PROBLEM OF ANTIBIOTIC RESISTANCE: A unique feature of the antibiotic field is
the constant need to develop new agents to keep pace with the constant increase
in the frequency of resistant isolates. Because physicians must begin antibiotic
therapy before the causative microorganism is identified and its drug
susceptibility pattern is determined, if the frequency of resistance for a given drug
in any given pathogen species exceeds a certain level, they will essentially stop
using that drug: the drug thus will become “useless.”
BIOCHEMICAL MECHANISMS OF RESISTANCE:
1. Enzymatic inactivation of the drug: This is a common resistance
mechanism for antibiotics of natural origin, as we have seen with
aminoglycosides.
2. Mutational alteration of the target protein: Completely man-made
compounds, such as fluoroquinolones, are unlikely to become inactivated by
the enzymatic mechanisms.
3. Acquisition of genes for less susceptible target proteins from other
species: Scientists discovered, by sequencing the genes coding for the
targets of penicillin, dd-transpeptidase or penicillin-binding proteins, that
penicillin resistance among S. pneumoniae, which has become more
 frequent in recent years, is largely the result of the production of mosaic
proteins, parts of which come from other organisms.
4. Bypassing of the target: Vancomycin, a fermentation product from
streptomycetes, has an unusual mode of action.
5. Preventing drug access to targets: Drug access can be reduced by an active
efflux process, discovered first with tetracycline, or, at least in Gram-
negative bacteria, by decreasing the influx across the outer membrane
barrier.