LYSOZYMES

BY: FAIZA RASHID

SADAF MEHREEN
 Lysozyme
• Lysozyme is also known
as muramidase or N-
acetylmuramide glycanhydrolase.
• Lysozyme is an antimicrobial
enzyme produced by animals that
forms part of the innate immune
system.
• It is an enzyme that target bacterial
cell wall.
 Discovery
• Lysozyme was discovered by Alexander Fleming in 1921 when he demonstrated
that his own nasal mucus had the ability to inhibit the growth of a certain strain
of bacteria in culture.
• He realized that this was largely due to the action of a protein within the mucus
that caused the bacterial cells to lyse or break apart.
• Hence, he named the protein lysozyme.
• In a 1922 publication, he reported its activity in hens' egg white, tears, saliva,
sputum, and nasal secretions.
• In a subsequent study, Fleming in collaboration with V. D. Allison detected
lysozyme in human blood serum, saliva, milk, and a wide variety of other fluids.
 Where is lysozyme found?
• It is a protein occuring in animals, plants,
bacteria and viruses.
• Human lysozyme is expressed in the
mucous membranes of the nasal cavity
and tear ducts.
• It is also found in saliva, tears, milk,
cervical mucus, leukocytes, and kidney
tissue.
• The majority of the lysozyme used in
research is purified from hen egg whites.
 Structure of lysozyme
• The primary structure of lysozyme is a
single polypeptide containing 129
amino acids.
• In physiological conditions, lysozyme is
folded into a compact, globular
structure with a long cleft in the protein
surface.
• This cleft is the active site involved in
binding to the bacterial carbohydrate
chain and subsequently cleaving it.
 Function of lysozymes
• Lysozyme is naturally found in body secretions like tears, saliva, breast milk and
mucus.
• It is also stored in tiny sacs within cells that form part of the innate immune
system.
• It is present in macrophages—the large cells that literally eat up bacteria and in
neutrophils, a type of white blood cell.
• Lysozyme is an important part of the innate immune system because it breaks
up (digests) components of the cells walls of bacteria.
• In other words, lysozyme acts as an anti-bacterial enzyme. It uses at least two
different mechanisms to kill bacteria—it digests the bacterial cell wall and it
punches holes into that cell wall, causing the contents to leak out of the
bacteria.
 Function of lysozymes
• In vitro, lysozyme is generally considered effective against some Gram-
positive bacteria, but ineffective against Gram-negative bacteria.
• However, lysozyme, perhaps indirectly, can affect Gram-negative
bacteria in vivo.
• Due to these antimicrobial properties, lysozyme has been used
effectively in the food industry.
• For example, it has been used in the cheese industry to prevent late
blowing.
• Lysozyme has also been used as a preservative for other fresh foods,
including controlling meat spoilage.
 Action of lysozymes on bacterial
membranes and walls
• The main component of bacterial cell wall is peptidoglycan, which is composed of alternating
units of N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM), linked by β-(1,4)
bonds.
• Gram-positive bacteria have a single lipid membrane surrounded by a cell wall composed of
a thick layer of peptidoglycan, while in Gram-negative bacteria the cell wall consists of a thin
layer of peptidoglycan placed between the cytoplasmic and outer lipid membranes.
• Lysozyme can kill bacteria through 2 mechanisms.
1. Lysozyme causes bacterial cell lysis via targeted hydrolysis of bacterial cell walls, which are
critical for the resistance of bacteria to osmotic stress.
2. Lysozyme can also kill bacteria independently of PG hydrolysis through a mechanism
involving its cationic nature. Cationic killing of bacteria may involve the formation of pores by
lysozyme on the bacterial cell membrane.
 The active site
of lysozyme
accommodates
up to 6
consecutive
sugars through
6 subsite
contacts,
annotated A-F.
Lysozyme
hydrolyzes the
β-1,4 glycosidic
bond between
the NAM at
subsite D and
the NAG at
subsite E.
1. Targeted hydrolysis of bacterial
cell walls
• Lysozyme is an enzyme that cleaves peptidoglycan in bacterial cell walls by catalyzing
the hydrolysis of β-(1,4) linkages between the NAM and NAG saccharides.
• In this enzymatic reaction, the residue Glu-35 acts as an acid, donating a proton to
the glycosidic bond, generating an oxonium that is then stabilized by a covalent bond
with Asp-52. The addition of a water molecule completes the hydrolysis and
regenerates the protonated form of Glu-35.
• This damages the peptidoglycan integrity. The progressive rupture of bacterial cell
wall by lysozyme gives rise to cells called protoplasts or spheroplasts, on which there
is a continuous inflow of water from the external medium leading to an increase in
pressure that finally results in cellular lysis.
• Lysozyme disruptive effect on the bacterial cell wall has helped to develop and
improve new antibiotics.
 2. Mechanism involving its
cationic nature
• Lysozyme can also kill bacteria independently of
Peptidoglycan hydrolysis through a mechanism involving its
cationic nature.
• Because the bacterial envelope is negatively charged,
lysozyme may have an enhanced charge-mediated
attraction for the bacterial surface that is proposed to lead
to a catalytic-independent mechanism of bacterial killing.
• Cationic killing of bacteria may involve the formation of
pores by lysozyme on the bacterial cell membrane.
• (A) A newly synthesized (B) Lysozyme hydrolyzes the β-1,4
Peptidoglycan monomer glycosidic bond between the NAM of 1
consists of a disaccharide, monomer and the NAG of the adjacent
NAG linked to NAM with monomer. Lysozyme hydrolysis of PG
an attached peptide stem,
leads to cell wall instability and bacterial
and the NAM is anchored
to the membrane via a cell death.
lipid carrier (grey).
Monomers are added to a (C) Lysozyme can also kill bacteria
growing chain through the independently of Peptidoglycan hydrolysis
action of through a mechanism involving its cationic
glycosyltransferases nature. Cationic killing of bacteria may involve
(green). the formation of pores by lysozyme (red
cylinders) on the bacterial cell membrane.