In the cytoplasma of the bacteria cell

BACTERIA’S CELL WALL SYNTHESIS

1. A chain of 5 amino acid bind to the NAM
2. Then, NAG bind to NAM forming a peptidoglycan precursors

In the periplasmic space

transpeptidase
3. This precursor cross the cell membrane to the periplasmic space
4. NAG of this precursors will bind to NAM of other precursors (cell wall acceptor) through
transglycosylation
5. Transpeptidase and Carboxylpeptidase bind to the peptide chain
6. Tanspeptidase (D-Ala-D-Ala) enzyme allow CROSS LINK between the peptide chain of
peptidoglycan precursors
7. Then D-ala carboxylpeptidase cleaves a peptide bond at carboxyl terminal end of peptide
and forming amide bond between the cross linking peptide chains

PBP has different types:

B-LACTAM ANTIBIOTICS DISRUPT
THE BACTERIA’S CELL WALL
SYNTHESIS
 Carboxylpeptidase n transpeptidase are transpeptidase / endopeptidase / transamidase
Function: reside at the cell membrane and perform construction, repairing, maintain cell wall
integrity, cell growth and cell division

a) Penicillin inhibition

B-lactam antibiotics kill bacteria by inhibiting this

enzymes, thus terminate the formation of
peptidoglycan (cell wall of the bacteria)

The b-lactam ring is (1) STERICALLY SIMILAR

to the D-Ala-D-Ala of the NAM pentapeptide
b) Transpeptidase cross-linking
 The PBPs mistakenly use the b-lactam as the
binding block during cell wall synthesis
*(block PBP for transpeptidase irreversible)

 This results in the ACYLATION of the PBPs,

prevents the enzyme to catalyze
transpeptidation and carboxypeptidation
reactions

 Disrupting the cell wall function to maintain

osmotic pressure – lysis - bactericidal
 SUBSTRATE OF THE PBPS a) b) c)

a) D-Ala-D-Ala of the NAM pentapeptide (at peptide chain)

b) Penicillin backbone

c) Cephalosporin backbone

The (2) amide bond in antibacterial is more the strain than amide bond in than its natural substrate  very unstable bond  PBP mistakenly attack here

BECAUSE B-LACTAM RING IS VERY REACTIVE CHEMICAL STRUCTURE INSTABILITY

The planar amide carbonyls are NOT VERY REACTIVE

Due to the reactivity (instability) of the b-lactam amide bond:
because of efficient delocalization of the nitrogen lone pair into
the carbonyl group
1. HYDROLYSIS
 The “V” shape (fused b-lactam ring  bicyclic) – look like a  Nucleophiles (Nu-) or b-lactamase (eg: Water & Alkaline
half-open book solutions)

 Inhibits resonance of the lactam nitrogen with its b-lactamase (b-lactamase acylase / penicillinase) are
carbonyl group (the lone pair IS NOT enzymes produced by penicillin resistant bacterias
DELOCALIZED into carbonyl)
 b-lactam become MORE REACTIVE and thus MORE Catalyze the hydrolysis of the b-lactam amide bond
SENSITIVE to nucleophilic attack compared to normal (ring opening) and inactivate b-lactam antibiotics to
planar amides
penicillioic acids before they can reach the PBPs
 This happens because of a highly strain bond energy of the
amide bond

1. Four-membered ring distorts the normal bond angles  Acidic solutions

for carbon (sp3-120o) [sepatutnye 90o utk 4member ring]
2. The presence of the sp2 carbonyl adds to this strain 2. A very complex reaction
3. Oxygen and nitrogen at b-lactam ring are
electronegative causing the carbon to become partially 3. Solutions of penicillins are buffered between pH 6.0-
+ve charge (enhance NU- attack) 6.8

Therefore, the b-lactam ring is easily hydrolysed (by

aqueous base and enzymes-e.g., PBPs