Non-mechanical

Method
BY: KUO GAROL SARONG
 3
Enzymatic methods 2
 Use of digestive enzyme decomposes the microbial
cell wall.
 Used enzyme depends on microbe.

It
Lysozyme is hydrolyzes
commonly β- 1-4-
used enzyme glucosidic
to digest cell bonds in
wall of gram the
positive peptidogly
bacteria. can
 Digestive Substance Product
juices and digested formed

digestive enzyme
enzymes

Saliva Starch Maltose

Amylase

Digestive enzymes speed up the Gastric juice Proteins

Protease (pep
Partly
digested
breakdown (hydrolysis) of food sin) and
hydrochloric
proteins

molecules into their ‘building acid

block’ components. Pancreatic Proteins Peptides and

juice Fats amino acids
Proteases emulsified by Fatty acids
(trypsin) bile and glycerol
Lipases Starch Maltose
Amylase

Intestinal Peptides Amino acids

enzymes Sucrose Glucose and
Peptidases (sugar) fructose
Sucrase Lactose (milk Glucose and
Lactase sugar) galactose
Maltase Maltose Glucose

Bile from the Fats globules Fat droplets

liver
Bile salts
Cell wall
A microbial cell wall is a well-
defined polymeric matrix
located just outside the plasma
membrane of a cell.
Cell walls can be composed
of polysaccharides, proteins, li
pids, or a combination of these.
Cell walls provide increased
mechanical strength and
resistance to lysis by osmotic
shock (water moving into the
cell).
Gram-positive Bacteria
The cell wall consists
of several layers of
peptidoglycan.
Running
perpendicular to the
peptidoglycan sheets
is a group of
molecules
called teichoic acids.
Gram-negative Bacteria
The cell wall is
composed of a single
layer of peptidoglycan
surrounded by a
membranous
structure called
the outer membrane.
It contains a unique
component, lipopolys
accharide (LPS or end
otoxin), which is toxic
to animals.
Enzymatic methods
(in addition)
• Enzymes such as beta(1-6) and beta(1-3) glycanases,
proteases and mannase can be used to disrupt the
cell wall.
• This method is particularly useful for isolating the cell
without the wall (protoplast).
• Enzymes commonly used for degradation of cell wall
of yeast and fungi include different cellulases,
pectinases, xylanases and chitinases.
Enzyme Functions
pro·te·ase group of enzymes that break the long chainlike
molecules of proteins into shorter fragments
(peptides) and eventually into their
components, amino acids
Cellulase is any of several enzymes produced chiefly by
. fungi, bacteria, and protozoans that catalyze
cellulolysis, the decomposition of cellulose and
of some related polysaccharides

Pectinase is an enzyme that breaks down pectin, a

polysaccharide found in plant cell walls

xylanases class of enzymes which degrade the

linear polysaccharide beta-1,4-
xylan into xylose, thus breaking
down hemicellulose, one of the major
components of plant cell walls .

Chitinases are enzymes that degrade chitin. It contribute

to the generation of carbon and nitrogen in
the ecosystem
Disadvantages
 The enzyme’s high price
and limited availability
limits their utilization in
large scale processes
The added enzyme may
complicate downstream
processing (e.g.
purification).
Disadvantages
 Drawbacks could be minimized by immobilization
of enzymes
Immobilization of enzymes
Enzyme immobilization may be defined asa process of
confining the enzyme molecules to a solid support over
which a substrate is passed and converted to products
An immobilized enzyme is one whose movement in space
has been restricted either completely or to a small limited
region