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biological reactions.
Most remarkable biomolecules due to their
extraordinary specificity and catalytic power
Far greater than those of man-made catalysts
Application
Enzymes
Uses
Food processing
Amylase, protease
Baby foods
Trypsin
Brewing industry
Fruit juices
Dairy industry
Meat tenderizers
Cellulases, pectinases
Rennin, lipase, lactase
papain
Starch industry
Amylase, glucoisomerases
Paper industry
Biofuel industry
Detergent industry
Photographic industry
ficin
1.Isolation of microbes.
2.Screening of microbes.
3.Fermentation.
4.Increase the yield of the enzymes.
1.Spontaneous mutation:
Occur spontaneously at the rate of 10-10 and 10-15 per generation and per
gene.
Occur at low frequency and hence not used much in industrial strain
improvement.
2. Induced mutation:
The rate of mutation can be increased by various factors and agents called
mutagens.
ionizing radiations (e.g. X-rays, gamma rays)
non-ionizing radiations (e.g. ultraviolet radiations)
various chemicals (e.g. mustard gas, benzene, ethidium bromide,
Nitrosoguanidine-NTG)
Steps involved:
Preparation of desired DNA
Insertion of desired DNA into vector DNA
Introduction of recombinant DNAs into host cells
Identification of recombinants
Expression of cloned genes
Enzyme Technology
Enzyme technology is concerned with the application of enzymes
as tools of industry, agriculture and medicine
Enzymes are biological catalysts that fulfil their role
by binding specific substrates at their active sites
This specificity is one property of enzymes that
makes them useful for industrial applications
The value of using enzymes over inorganic catalysts in the
technological field is their efficiency, selectivity and specificity
Enzymes are able to operate at room temperature, atmospheric
pressure and within normal pH ranges (around 7)
all of which create energy savings for industry
Enzymes possess specifically shaped active sites for reacting with
one specific substrate thereby generating pure products
free from unwanted by-products
Enzymes are biodegradable and, unlike many inorganic
catalysts, cause less damage to the environment
The micro-organisms
(such as yeast) are really used as
a source of enzymes during the
manufacture of these products
of biotechnology
Many industrial processes now make use of pure sources of enzymes, i.e.
the enzymes have been ISOLATED from the micro-organisms before use
Non-recombinant Sources
GRAS
Bacillus
protein is secreted into fermentation medium
easier purification
Aspergillus
Yeast
Recombinant Sources
Most industrial enzymes are produced recombinantly
Why?
A. Higher expression
B. Higher purity (% protein:other junk)
C. cheap
D. can engineer protein
E. can express enzymes which are found
in pathogenic organisms
Protein engineering
Make oxidation resistant
make enzymes tolerant of processes used in industry
less substrate specificity
more thermostable
more stable in presence of detergent
MODIFICATION possible
application of genetic
engineering to improve
the microbial strain
INDUSTRIAL SCALE
FERMENTATION
PRODUCTION OF PECTINASE
Evaporate to concentrate
the enzyme
Aspergillus niger is grown in
a fermenter with a source of
nitrogen, with sucrose as the
carbon source and the substrate
pectin to stimulate pectinase
production by the fungus
Pure, powdered
pectinase
Enzymes in Biotechnology
Enzymes are used in industrial processes and as analytical
reagents in medicine
Thermostability and an ability to
withstand extremes of pH are
essential properties for enzymes used
in many industrial processes
Immobilisation of enzymes is an important technique used
in industry as it enables economical operation of a process
and protection of enzymes during their use
Because of their sensitivity and specificity, enzymes are
used as analytical reagents in systems such as the detection
of glucose in human blood and urine
Immobilised Enzymes
The costs associated with the use of enzymes for industrial
purposes can also be reduced by immobilising the enzymes
Starch Paste
Dextrins
(short chains
of glucose
molecules)
Glucose
2H2O + D
No
glucose
Biosensors
Biosensors are electronic monitoring devices that make use of
an
enzymes specificity and the technique of enzyme
immobilisation
Amplifier
Read-out
Transducer
Immobilised
enzymes bind
with specific
molecules
even when they
are present
in very low
concentrations
The enzyme
reaction brings
about a change
that is converted
into an electrical
signal by a
transducer
The electrical
signal is amplified
and gives a
read-out on a
small display
screen
Biosensors
A biosensor has been developed for detecting
glucose in the blood of diabetics
Amplifier
Transducer
Glucose
molecules
in the blood
Glucose
oxidase
Glucose oxidase
oxidises any glucose
present in the blood to
release electrons these
are detected by the
transducer and converted
into an electrical current
2. Vinegar production:
To make vinegar, wine is slowly pored over oak
chips in a tall tower, open to the air.
Bacteria (Acetobacter) on the wood oxidise the
ethanol in the wine and turn it into ethanoic acid
or vinegar, giving out a great deal of heat as well.
If the vinegar is made from fermented raisins and
stored in oak vats (similar to the solera system
used for making sherry) then the sweet, highlyprized Balsamic vinegar is made mainly around
Modena in Italy.
3. Yoghurt production
Milk goes sour within a few hours in the hot
conditions common in the Middle East.
If stored in a leather bag and mixed with a
suitable starter culture, however, it rapidly
turns into yoghurt, which will keep for
several days.