This document discusses the production of various enzymes including amylase, protease, and lipase through microbial fermentation. It provides details on selecting suitable microorganisms such as bacteria and fungi to produce these enzymes. The document describes fermentation processes and conditions used for enzyme production as well as methods for recovering and purifying the enzymes. Key points covered include common microbes used to produce specific enzymes, formulation of fermentation media, factors affecting fermentation, and downstream processing techniques.
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
Protein engineering is the process of developing useful or valuable proteins. It is a young discipline, with much research taking place into the understanding of protein folding and recognition for protein design principles
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
Protein engineering is the process of developing useful or valuable proteins. It is a young discipline, with much research taking place into the understanding of protein folding and recognition for protein design principles
In the field of biotechnology there are many industrial applications that result in biotech products that we use everyday at home. Some of these are food science applications that utilize enzymes to produce or make improvements in the quality of different foods. In the dairy industry, some enzymes are required for the production of cheeses, yogurt and other dairy products, while others are used in a more specialized fashion to improve texture or flavour.
Isolation and Purification of Enzymes
Enzymes are unstable molecules with a definite physico chemical organization. Even a slight change in this organization reduces the activity of enzyme and sometimes the enzyme is totally inactivated.
Therefore, the enzymes have to be isolated under controlled conditions of pH, ionic strength and temperature. Since they are proteinaceous in nature, standard extraction and purification procedures for enzymes are the same as those used for proteins except that the activity of the enzyme is assayed at each of the following four steps of extraction and purification.
Purification of Enzymes - Enzyme purification involves three steps, electrophoresis. These three techniques described in the following text
1.Dialysis
2.Chromatography.
This PPT will provide the basic idea of Fermentation technology and it's use. The reference book is 'Pharmaceutical Biotechnology' by Giriraj Kulkarni.
VIRAL VACCINES
Since viruses are intracellular parasites they will grow only within other living cells.
Methods of viral vaccine production:
Cultivation of virus using free living animals
Fertile eggs
Tissue cultures
In the field of biotechnology there are many industrial applications that result in biotech products that we use everyday at home. Some of these are food science applications that utilize enzymes to produce or make improvements in the quality of different foods. In the dairy industry, some enzymes are required for the production of cheeses, yogurt and other dairy products, while others are used in a more specialized fashion to improve texture or flavour.
Isolation and Purification of Enzymes
Enzymes are unstable molecules with a definite physico chemical organization. Even a slight change in this organization reduces the activity of enzyme and sometimes the enzyme is totally inactivated.
Therefore, the enzymes have to be isolated under controlled conditions of pH, ionic strength and temperature. Since they are proteinaceous in nature, standard extraction and purification procedures for enzymes are the same as those used for proteins except that the activity of the enzyme is assayed at each of the following four steps of extraction and purification.
Purification of Enzymes - Enzyme purification involves three steps, electrophoresis. These three techniques described in the following text
1.Dialysis
2.Chromatography.
This PPT will provide the basic idea of Fermentation technology and it's use. The reference book is 'Pharmaceutical Biotechnology' by Giriraj Kulkarni.
VIRAL VACCINES
Since viruses are intracellular parasites they will grow only within other living cells.
Methods of viral vaccine production:
Cultivation of virus using free living animals
Fertile eggs
Tissue cultures
Production of secondary metabolites : enzymes which involves the upstream technological process
Introduction
History
Process involved
Contribution of different micro-organisms
Flowchart
Example: Methods Production of Amyalse in industrial view
AMYLASES AND PROTEASES ARE THE ENZYMES USED A LOT IN FOOD INDUSTRIES FOR THE PRODUCTION OF FOODS. THESE ARE SUPPOSED TO PRODUCE AT A LARGER QUANTITIES IN ORDER TO FULFILL THE DEMANDS FROM THESE INDUSTRIES, THE LARGE SCALE PRODUCTION OF THESE ENZYMES MUST BE CARRIED OUT. THIS METHOD OF LARGER PRODUCTION OF THESE ENZYMES ARE EXPLAINED IN THIS PRESENTATION.
UNIT-5 Protein Engineering: Brief introduction to protein engineering,Use of ...Shyam Bass
UNIT-5 6th Sem B.PHARMA PHARMACEUTICAL BIOTECHNOLOGY)
Protein Engineering: Brief introduction to protein engineering, Use of microbes in industry, Production of enzymes-general considerations, Amylase, Catalase, peroxidase, Lipase Basic principles of genetic engineering
BY- SHYAM BASS
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
2. ⦁ Many microbes synthesize and excrete large
quantities of enzymes into the surrounding
medium. Using this feature of these tiny
organisms, many enzymes have
produced commercially. These
been
include
Amylase, Cellulase, Protease, Lipase,
Pectinase, Streptokinase, and many others.
⦁ Enzymes are extensively used in food
processing and preservation, washing
powders, leather industry, paper industry and
in scientific research.
4. ⦁ In early days, animal and plant sources largely contributed to
production of enzymes.
⦁ Even now, they act as the major source for certain enzymes
⦁ Animal organs and tissues are very good sources for enzymes
such as lipases, esterases and proteases.
⦁ Some plants are excellent sources for certain enzymes-papain
(papaya), bromelain (pineapple)
5. ⦁ The most important limitations are the difficulty in
isolating, purifying the enzymes and the cost factor.
⦁ For this very reason, microbial production of enzymes is
preferred.
⦁ Microbes are the most significant and convenient sources
of commercial enzymes
⦁ They can be made to produce abundant quantities of
enzymes under suitable growth conditions.
⦁ Microbes can be cultivated by using inexpensive media
and production can take place in a short period.
6. ⦁ Its easy to select microbes for the production of specific type of
desired enzymes.
⦁ Recovery, isolation and purification processes are easy with
microbial enzymes than that with animal or plant sources.
⦁ In fact, most enzymes of industrial applications have been
successfully produced by microbes. Various fungi, yeast and
bacteria are employed for this purpose.
8. ⦁ SUBMERGED FERMENTATION:-
1. It employs free flowing liquid substrates, such as molasses and
broth
2. The products yielded in the fermentation are secreted into the
fermentation broth.
3. This method is suitable for those microbes such as bacteria that
require high moisture content for their growth.
4. The sterilization of the medium and purification process of the
end products can be done easily.
5. Also the control of process parameters like temperature, pH,
aeration, oxygen transfer and moisture can be done
conveniently.
9. ⦁ SOLID STATE FERMENTATION:-
1. Method used for microbes which require less moisture content
for their growth
2. The solid substrates commonly used in this method are bran,
bagasse and paper pulp
3. The main advantage is that, nutrient-rich waste materials can be
easily recycled and used as substrates in this method.
4. It requires simple instruments over SMF, higher concentration of
products with less effluent generation
10. AMYLASE
⦁ Amylases are important hydrolase enzymes. It usually
degrades complex polysaccharide molecules such as starch
into glucose.
Amylase
⦁ Starch Glucose
🞂
🞂
Present in saliva of humans (for digestion)
This enzyme randomly cleave internal glycosidic linkages in
starch molecules
⦁ Hydrolysis of starch with amylase results in formation of
dextrin and then disaccharide maltose (2 glucose
molecules) and finally glucose.
⦁ Types of amylases:-α-amylase, β-amylase, γ-amylase
⦁ The substrate that α-amylase acts upon is starch.
⦁ Starch is composed of 2 polymers- amylose (20-25% of the
starch) and amylopectin (75-80% of starch).
⦁ Amylose is broken down to give maltotriose and maltose
molecule
⦁ Amylopectin is broken down to give dextrin and glucose
molecule
11. ⦁ Primary source of β-amylase are the seeds of
higher plants and sweet potatoes.
⦁ BACTERIAL SOURCE OF α-Amylase:- Bacillus
subtilis, B. stearothermophilus, B. licheniformis, B.
amyloliquefaciens
⦁ Carbon source:- maltose, sucrose, glucose
⦁ Nitrogen source;- Inorganic nitrogen sources such
as ammonium sulphate, ammonium chloride,
ammonium hydrogen phosphate, Organic sources
such as soyabean meal, peptone, yeast extract
12. ⦁ Bacillus licheniformis has been used
industrially for the production of α-amylases.
⦁ The fermentation of bacterial amylases is
accomplished in submerged culture at neutral
pH and at a temperature of 30-40ºC.
⦁ Cereal meal and starch rich medium are used
along with an organic source of nitrogen.
⦁ After 10-20 hours, formation of α-amylase
starts and continues for another 100 hours
⦁ pH must be below 6 during fermentation to
prevent the denaturation of α-amylase.
13. ⦁ SELECTION OF MICROORGANISM:-Aspergillus
niger, A. oryzae, A. kawachii
⦁ They produce amylase and secrete acid hence
easily degrades complex raw material.
⦁ SELECTION OF RAW MATERIALS:-fungi cannot grow
on natural media, so we provide synthetic or
artificial media.
CHEMICALS QUANTITY USED
Corn starch 24g/L
KCl 0.2g/L
Na2HPO4 4.7g/L
CaCl2/CaCO3 1g/L
MgCl2.6H2O 0.2g/L
STEPS INVOLVED IN PRODUCTION OF FUNGAL AMYALSE
14. ⦁ FERMENTATION PROCESS
Fermenter is filled with the raw material or
fermentation medium
Inoculate mycelium of A. niger
Maintain temperature at 30ºC-45ºC
After 72 hours recovery process is carried out.
15. ⦁ RECOVERY PROCESS
Filtrate contains amylase, synthetic media and mycelia of
fungi
By filtration, remove mycelia
Amylase is separated from synthetic media by adding
ammonium hydroxide which forms precipitate with amylase
By filtration collect the precipitate (amylase+ammonium
hydroxide)
Precipitate is subjected to crystallization at 4ºC
Amylase in form of crystals are separated and collected
16. ⦁ Fungal amylase differ from bacterial amylases by low
pH 4-5.
⦁ The production of fungal amylase is carried out in
submerged culture with selected strains
solid-substrate culture and sometimes in
of
Aspergillus.
⦁ Fermentation medium used remains the same as in
the case of bacterial α-Amylase but the
concentration of glucose inhibits the formation of
amylase, therefore the concentration of glucose is
kept low.
⦁ Fungal amylases- used in manufacture of baked
products and production of maltose rich syrups
17. LIPASE
⦁ It is an enzyme that catalyzes the breakdown of
most triglycerides into fatty acids and glycerol
(lipolysis).
⦁ Lipase enzyme is usually found naturally in
pancreatic juice and stomach.
⦁ They also control the volume of fat in body that is
synthesized and burned by reduction of adipose
tissue.
⦁ Lipase can be purified or extracted from plant,
animal, yeast, bacteria and fungal sources.
18. ⦁ Uses:-used in detergents (laundry) due to the wide use of
washing machine,
-fat and oil processing (inexpensive and less needed lipid
can be converted into greater value fat), PUFAs (Poly Unsaturated
Fatty Acids) used remarkably as pharmaceutical, nutraceutical
etc.
-obtained by using microbial lipases from plant, and animal
lipids.
19. ⦁ Selection of microorganism
⦁ Formulation of medium
⦁ Production process
⦁ Recovery and purification of enzymes
20. ⦁ SELECTION OF MICROORGANISM
1. Bacterial source:-Pseudomonas aeruginosa,
Staphylococcus caseolyticus, Bacillus coagulans,
Bacillus subtilis, Bacillus stearothermophilus
2. Fungal source:-Aspergillus niger, Candida
rugosa, Candida utilis, Penicillium citrinum,
Penicillium restrictum, Candida cylindracea
Candida cylindracea was grown on yeast agar
medium and maintained at 4ºC. A loop full of cells
from freshly grown culture (agar slant) of Candida
cylindracea was transferred to flask. Flask was then
incubated at 30ºC on a rotary shaker at 200 rpm
for 36 hrs.
21. ⦁ FORMULATION OF MEDIUM:-medium
containing flask was sterilized in an
autoclave at 121ºC for 20 min
CHEMICALS REQUIRED QUANTITY (g/L)
KH2PO4 6
MgSO4.7H2O 1
Urea 4
FeCl3.6H20 10 mg
Inositol 0.4 mg
Thiamine hydochloride 0.2 mg
Biotin 0.8 mg
Glucose 10 mg
Distilled water 1L
22. ⦁ PRODUCTION PROCESS:-
1. The medium used in preparing the inoculum is the
same as the production medium. The only difference
is the use of glucose as the carbon source.
2. Palm oil can be used as the main carbon source in
production medium but not used in the inoculum
preparation medium
3. 50g of sterile palm oil was transferred aseptically to
the sterilized medium in the fermentor
4. 10% of the inoculum was added to the total volume.
5. Samples of 50ml was withdrawn aseptically at regular
time interval for analysis.
6. Temperature=28-33ºC, pH=6-7, stirring
speed=500rpm
23. ⦁ RECOVERY AND PURIFICATION OF ENZYMES:-
1. Removal of debris by filtration
2. After fermentation the extract was
precipitated with ammonium sulphate
3. Ultrafiltration was used to concentrate the
sample
24. PROTEASE
⦁ These are a group of enzymes belonging to a class of hydrolases
whose catalytic function is to hydrolyze peptide bonds of proteins
⦁ They are also called proteolytic enzymes
⦁ USES:-for cleaning contact lenses to remove large variety of stains
of food, blood and body secretions
-in the manufacture of cheese
-meat tenderization (reduce toughness of meat)
-to correct lytic enzyme deficiency syndrome
-treatment of industrial waste:- keratinase used as depilatory
to remove hair from drains
25. ⦁ SELECTION OF MICROORGANISM
⦁ FORMULATION OF MEDIUM
⦁ PRODUCTION PROCESS
⦁ RECOVERY AND PURIFICATION OF ENZYMES
26. ⦁ SELECTION OF MICROORGANISM
1. Bacterial source:- Bacillus licheniformis, B.
amyloliquefaciens, B. stearothermophilus
2. High carbohydrate level in the medium
stimulates protease production
27. ⦁ FORMULATION OF MEDIUM
1. Media used contains ground barley as the
carbon source
2. Starch level is limited
3. Protein hydrolysates or sodium glutamate is
used as the nitrogen source
28. ⦁ PRODUCTION PROCESS
1. The medium has a pH of 6.5-7.5 and incubation
temperature of 37ºC
2. Fermentation proceeds for 3-5 days
Preserved inoculum Inoculum development
Inoculation tank Fermenter Cell disruption
Filtration (to remove debris) Remove nucleic
acids Salt treatment Cool storage
Final purification (chromatography etc.)
Fitration
Freeze
drying
*During fermentation, insoluble protein of the
medium is partially hydrolysed by boiling in dilute
acid/enzymatic treatment.
29. ⦁ RECOVERY AND PURIFICATION OF ENZYMES
1. Culture is filtered
2. Aqueous portion is concentrated by
evaporation at reduced pressure and at
temperature not less than 40ºC
3. Recovery can also be carried out by
precipitation.
30. ⦁ SELECTION OF MICROORGANISM
1. Various fungi produce protease such as Aspergillus oryzae, A.
sojae, A. niger, A. wentii, Mucor pusillus, Mucor miehei,
Mucor delemar, Amylomyces rouxii, Endothia parasitica
31. ⦁ FORMULATION OF MEDIUM
1. The fungus is grown on wheat bran under
fermentation conditions similar to those for
amylase production
2. Trace elements used in Inoculation medium:-
CHEMICALS REQUIRED QUANTITY
Fe(NH4)2(SO4)2 1 mg
ZnSO4 1 mg
MnSO4 0.5 mg
CuSO4 0.08 mg
CoSO4 0.1 mg
H3BO3 0.1 mg
Distilled water 1 L
32. ⦁ Medium for inoculum preparation (g/L):-
⦁ Medium for production (g/L):-
Chemicals required Quantity
Casein hydrolysate 5
Soya protease digest 2
Yeast extract 2
Soluble starch 10
D-mannitol 5
Trace element 1 ml
FeSO4.7H2O 15 mg
Chemicals required Quantity
Soyabean meal 30
Glucose 10
NaNO3 3
Skim milk 10
KH2PO4 0.5
MgSO4.7H2O 0.25
33. ⦁ PRODUCTION PROCESS
1. Endothia parasitica is employed for the
production of this enzyme in lab scale
2. A 5% inoculum from 96 hrs flask is utilized for the production
in a stirred, aerated vessel for 48 hrs at 28ºC.
3. Optimum pH is 4-5
4. Proteases from Aspergillus oryzae and A. sojae are produced
by solid substrate fermentation.
34. ⦁ RECOVERY AND PURIFICATION OF ENZYMES
1. After growth the harvest can be dried at 50ºC or less
2. Protease can also be extracted by water followed by
addition of alcohol for precipitation and dried at 55ºC