TITLE: PRODUCTION OF

ALKALINE PROTEASE USING

COW DUNG
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PRESENTED BY:
• AFIFA SHAKEEL
• MAIRA SAJID
• WALI UR REHMAN BUKHARI
• NOMAN IQBAL
PRESENTED TO:
DR. IFTIKHAR ZEB
SLIDES BY:
ALISHA ZAFAR

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 Introduction of alkaline protease
Uses of the bioproduct
Feed stock
TABLE OF Formation of bioproduct
CONTENTS: Adjustment of the sources
Scope in Pakistan
Optimization of process parameters
Purification of protease

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DETERGENT INDUSTRY:
Improve washing performance
for use in domestic laundering to
solutions for cleaning contact
lenses.

Protease digests on stains during

fabric washing due to:
Food
Blood
Other body secretions

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FOOD INDUSTRY:
• The modification of properties of
food proteins
• Proteases play an important role in
the production of soy sauce and
other soy products.
• Primarily used in cheese
manufacturing
• Tenderization of meat.

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LEATHER INDUSTRY:
• Alkaline proteases with elastolytic and
keratinolytic activity can be used in
leather-processing industries.
• Soaking
• Dehairing
• Batting process

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WASTE
MANAGEMENT:
• Helps in clearing pipes
clogged with hair-
containing deposits
• Degrading waste keratinous
material from poultry
refuse.

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SILVER RECOVERY:

• Bioprocessing of used x-ray or

photographic films .
 contain 1.5-2.0% silver by weight
in their gelatin layer.

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FEEDSTOCK; COW DUNG:
• Cow dung was obtained as a feed
stock.
• Cow pats or cow manure, is the
waste product of bovine animal
species.
• Undigested residue of plant matter,
passed through the animal's gut.
• The resultant fecal matter is rich in
minerals.
• Also used to produce biogas to
generate electricity and heat.

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PRETREATMENT OF FEEDSTOCK:

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ISOLATION OF
PROTEOLYTIC
MICROORGANISM:

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IDENTIFICATION OF POTENTIAL
ISOLATES:

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OPTIMIZATION OF PROCESS PARAMETERS

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 PURIFICATION OF PROTEASE
• Lyophilization the enzyme with 92.7 % recovery
of activity was achieved.

• Dialysis: for dialysis 86.8 %

• Ultrafiltration: for ultrafiltration 93.6% of yield,

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SDS PAGE & ZYMOGRAPHY:

Silver staining was

SDS was carried out using
performed to visualize
11% polyacrylamide gel.
protein bands.

Zymographic analysis was

performed by the enzyme In the SDSPAGE, the
pattern of proteins, purified enzyme migrated
obtained by zymogram with as a single band with an
1% casein substrate and apparent molecular weight
detected using Coomassie of 38.5 kDa.
brilliant blue R-250.

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 The protease that was obtained,
its activity was characterized
based on following 3 criteria:

CHARACTERIZATION
OF PROTEASE Effect of pH
ACTIVITY:

Effect of temperature

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 THE MAXIMUM ENZYME THE MAXIMUM ALKALINE PROTEASE
PRODUCTION WAS FOUND IN PRODUCTION WAS OBSERVED AT PH
MEDIUM CONTAINING STARCH, 10 WHICH BEST COMPLEMENTED
PEPTONE, BEEF EXTRACT, CACO3. IDEAL BACILLUS SUBTILIS ACTIVITY.

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ADJUSTING
CARBON
SOURCES FOR Maltose,
sucrose as
These other
sources were
It was
performed to
But only
served to
ALKALINE well as some adjusted with
other sources cow dung as
optimize the
process to its
provide
evidence that
PROTEASE of carbon
were used to
a substrate as
well
peak
efficiency.
cow dung is
sufficient as
PRODUCTION: test the best
possible
a substrate.

protease
production
amounts.

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• Gelatin is found to be an optimal
nitrogen support for the fermentation
process.
• The best possible temperature found
for the process was to maintain it at
50 oc

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ALKALINE PROTEASE AS BIO DETERGENT IN VARIOUS PAKISTANI INDUSTRIES:

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 Second largest exporter industry of Pakistan after
textile

LEATHER
Produces about 6 million hides and 36
INDUSTRY million skins annually

OF
Lacks in product quality as compared to
PAKISTAN: other foreign countries

Mainly due to local lacking

Dehairing, tanning,
appropriate chemicals
etc.
needed for process. Such as:

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 It will help in Locally available
reducing costs and eco-friendly
Improves leather alternative to
texture. dehairing
Advantage of using Alkaline chemicals.
protease in Pakistan leather
industry:

Help in efficient
Also fend off and
dehairing
reduce some off
process of hides
the
without
environmental
excessively
concerns.
damaging them.
 There are various detergent manufacturing
companies in Pakistan which use different enzymes
in their products.

ALKALINE
PROTEASE The bio-detergent manufactured by this process allows
for a great source of protease which can be easily
USE IN utilized in laundry detergent manufacturing.

DETERGENT
INDUSTRY OF These enzymes obtained from the bioprocess have
high potential for protein related stain removal in high

PAKISTAN: alkaline conditions.

As the enzymes would be derived from a cheap locally

available source they would help in reducing cost of
detergent manufacture process.
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 Cow dung is a
cheap and easily
available
feedstock in
Pakistan that can
be used to
produce alkaline
proteases.

The bio detergent

Excessive cow
would also be
dung that would
helpful in being
be dumped in the
an additive to
rivers be used to
laundry
generate a
detergents as well
revenue stream
CONCLUSION for protein stain
removals.
instead.

These alkaline
proteases offer a
Versatile kind of
cheap and eco-
enzymes that can
friendly
be utilized in the
alternative to
many industries
dehairing
of Pakistan
chemicals used in
leather industry.
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REFERENCES:

Research Report on Pakistani Leather Industry by Ravi Magazine. Jillani Hashmi, Ghulam & Dastageer, Ghulam & Saqib Sajid,
https://www.ravimagazine.com/research-report-on-pakistani-leather- Muhammad & Ali, Zubair & Malik, Muhammad & Liaqat, Ihtsham.
industry/ (2017). Leather Industry and Environment: Pakistan Scenario.
International Journal of Applied Biology and Forensics. 1. 20-25.

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