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Enzymes proteases.pptx
1. Semester – iv
Fermentation
& industrial
microbiology
Seminar topic
: PROTEASES
Presented by:
UMA.R
II PG MICROBIOLOGY
Submitted to:
Dr. S. VISWANATHAN
HEAD DEPARTMENT OF
MICROBIOLOGY
2. INTRODUCTION:
WHAT ARE ENZYME ?
Enzyme are the of protein that act as
biocatalyst in our body.
One of the most important roles of
protease is the hydrolysis activity (
breaking down ).
3. What are proteases ?
Proteases were the first enzymes to be commercialized ,
partly on account of the history of availability of the first
commercial enzymes and partly on account of need ,
since proteins are ubiquitous in nature and can be found
in a wide variety of consumer stains , for example , variety
of food stains as milk , egg and soya , blood , grass and
human body fluids.
Proteases constitute one of the most important groups of
industrial enzymes , capturing almost 60% of the total
enzyme market.
The vast variety of proteases , with their specificity of
action and application , have been exploited widely in the
detergent ,food , pharmaceutical , and leather tanning
industries.
This has been considered as eco-friendly because of the
appropriate producers of these enzymes for commercial
exploitation and non toxic and non pathogenic.
4. There are many different types of proteases
produced by an extraordinarily large number of
microorganisms, but in actual practice the enzymes
prepared commercially are of vary in limited number
of types and they are derived from very few
organisms.
DEFINITION;
A protease is an enzyme that catalyses
proteolysis , the breakdown of protein into smaller
polypeptides , oligopeptides or single amino acids.
They do this by cleaving the peptide bonds within
proteins by hydrolysis , a reaction where water
break bonds.
Proteases are from the hydrolyses family of
enzymes.
These enzymes are widely distributed in nearly all
plants, animal and microorganisms.
The partial digestion by proteases produces peptide
sequence that may have biological properties and
5. HISTORY :
1833- French chemist anselme payen
–disease.
1860-Louis Pasteur- fermentation
techniques with sugar.
1897-Edward Buchner- paper on
“study of yeast extract.”
1926- James B.Sumner –proved
urease and catalase (1937) were
protein.
1965- David Chilton Phillips-structure
of lysozyme by x-ray crystallography.
7. SOURCES:
Proteases have been identified among
the following sources because of their
important physiological roles
Microorganisms (Bacteria, fungi,
virus)
Plants
Animals
Humans
8. PROTEASES FROM
MICROBES:
Microorganisms are responsible for
approximately 40 percentage of the total
worldwide enzyme sales.
BENEFITS OF THE USE OF
MICROORGANISMS:
Ease of genetic manipulation and
extensive diversity.
Lower manufacturing costs.
Ease to extract and purify.
Rapid culture development and Short
generation time.
9. MICROORGANISMS:
More than 50 percentage of the
industrially important enzymes are
now produced from genetically
engineered microorganisms produced
by conventional mutagenesis ( UV or
Chemical exposure) or through
Recombinant DNA technology.
Most commercial neutral and alkaline
proteases are produced by members
o the genus Bacillus.
10. SUBSTRATE :
The type of proteolytic enzyme formed
may depend on the composition of the
medium.
Agricultural wastes like rice bran, paddy
straw and pigeon pea waste also used
for protease production.
Carbohydrate sources : Molasses,
Barley, Corn, Wheat and Starch
hydrolyses.
Protein: Meals of soybean, Cotton
seeds, peanut and whey, corn steep
liquor and yeast hydrolysis.
11. FERMENTATION:
For cultivation of the microorganisms,
especially bacteria, the submerged
fermentation is the preferred method.
However, fungi usually give higher
yields when cultured on solid media.
So, solid state fermentation continues
to play a role.
Many of the organisms excrete more
than one kind of protease.
12. SUBMERGED
FERMENTATION:
Submerged fermentation is the cultivation of
microorganisms in liquid nutrient broth, industrial
enzyme can be produce using this process.
This involves growing carefully selected
microorganisms (bacteria, and fungi) in closed
vessel containing a rich broth of nutrients (the
fermentation medium )and a high concentration
of oxygen. As the microorganism break the
nutrients ,they release the desired enzyme into
solution.
Due to the development of large –scale
fermentation technologies , the production of
microbial enzymes accounts for a significant
proportion of enzymes.
13. SOLID STATE FERMENTATION
:
Solid state fermentation (SSF) is another
method used for the production of enzymes.
Solid state fermentation involves the
cultivation of microorganisms on a solid
substrate, such as grains, rice and wheat
bran, sugar beet pulp and wheat and corn
flour.
Larger particles provides more effects
aeration and respiration, but there is a
reduction in the surface area.
SSF requires moisture to be present on the
substrate , for the microorganism to produce
enzymes.
17. PROTEASE RECOVERY:
First step in harvesting of enzymes from the
fermentation broth is removed of insoluble
products like microbial cells which generally
carried out by filtration or centrifugation.
The biomass is treated with lime to deactivate
the microorganisms and stabilize it during and
storage then can be used as a fertilizer.
The enzyme remaining in the broth are then
concentrated by various methods like
evaporation, membrane filtration or crystallization
to yield protease concentrate.
As an alternate recovery procedure , the amylase
can be precipitated from aqueous solution by the
addition of cold acetone, ethanol, isopropanol or
ammonium sulfate.
18. DRYING AND PACKAGING:
Certain application require solid
enzyme produces, so the powder
enzymes are made into granules to
make them more convenient to use.
Therefore , enzymes are dried by
using vacuum or spray draying.
The powder obtained is finally
packaged for sale.
20. RECENT ADVANCES AND
FUTURE PROSPECTS:
Qualitative improvement: low –
temperature screening methods, protein
engineering , r-DNA technology , novel cold –
active proteases.
Quantitative enhancement needs: stains
improvement, site-directed mutagenesis, and
standardizations the nutrients media for the
overproduction.
Extensive attempts to engineer cold- adapted
protease from subtilis in BPN have previously
been mode.
21. CONCLUSION:
The need for the protease enzymes is
constantly increase till now and there
will be a great need for industries
producing proteases.
Future of enzyme technology also
depends on the research and
developments sector in modifying a
microbial strain to tolerate various
physical conditions and produce a
good yield.
