SlideShare una empresa de Scribd logo
1 de 27
Descargar para leer sin conexión
BY :
ROHINI YADAV
FERMENTATION TECHNOLOGY
Production of Amylase
Contents
• Introduction
• Types of amylase
• Production of amylase
• Recovery and purification
• Determination of enzyme activity
• Industrial application
Amylase
• Amylase is an enzyme that catalyses the hydrolysis of starch
into sugars.
• Amylase is present in the saliva of humans and some other
mammals, where it begins the chemical process of digestion
• These ezymes randomly cleave internal glycosidic linkages
in starch molecules to hydrolyze them and yield dextrins and
oligosaccharides
Types of Amylase:
α-Amylase
• α-Amylase is a hydrolase enzyme that catalyses the hydrolysis of
internal α-1, 4-glycosidic linkages in starch to yield products like
glucose and maltose.
• It is a calcium metalloenzyme i.e. it depends on the presence of a
metal co factor for its activity.
• There are 2 types of hydrolases: endo-hydrolase and exo-
hydrolase. Endo- hydrolases act on the interior of the substrate
molecule, whereas exo-hydrolases act on the terminal non
reducing ends .
• The substrate that α-amylase acts upon is starch. Starch is a
polysaccharide composed of two types of polymers – amylose and
amylopectin.
• Amylose constitutes 20-25% of the starch molecule. It is a linear
chain consisting of repetitive glucose units linked by α-1,4-
glycosidic linkage. Amylopectin constitutes 75-80% of starch and
is characterized by branched chains of glucose units.
• The optimum pH for activity is found to be 7.0.
β – Amylase
• β-Amylase is an exo-hydrolase enzyme that acts from the
nonreducing end of a polysaccharide chain by hydrolysis of α-1, 4-
glucan linkages to yield successive maltose units.
• It is unable to cleave branched linkages in branched polysaccharides
such as glycogen or amylopectin, the hydrolysis is incomplete and
dextrin units remain.
• During ripening of fruits, β-Amylase breaks down starch into maltose
resulting in the sweetness of ripened fruit.
• The optimal pH of the enzyme ranges from 4.0 to 5.5.
γ – Amylase
•
• γ-Amylase cleaves α(1-6)glycosidic linkages, in addition to
cleaving the last α(1-4)glycosidic linkages at the nonreducing end
of amylose and amylopectin, unlike the other forms of amylase,
yielding glucose.
• γ- amylase is most efficient in acidic environments and has an
optimum pH of 3.
•
Production of α-Amylase
Sources
• α-Amylase can be isolated from plants, animals or microorganisms.
• The enzyme has been isolated from barley and rice plants .
• It has been found that cassava mash waste water is a source of α-
Amylase which is active in wide range of pH and temperature .
• The most widely used source among the bacterial species is the
Bacillus spp. B. amyloliquefaciens and B. licheniformis
• Fungal sources of α-Amylase are mostly Aspergillus species and only
few species of Penicillium, P. brunneum , penicillium fellutanum
Production Methods :
There are mainly two methods which are used for production of α-Amylase
on a commercial scale.
These are :
1) Solid State fermentation.
2) Submerged fermentation
• Method used for microbes which require less moisture content for
their growth.
• The solid substrates commonly used in this method are, bran,
bagasse, and paper pulp.
• The main advantage is that nutrient-rich waste materials can be easily
recycled and used as substrates in this method.
1. Solid State Fermentation [SSF]
• employs free flowing liquid substrates, such as molasses and
broths.
• The products yielded in fermentation are secreted into the
fermentation broth. The substrates are utilized quite
rapidly,hence the substrates need to be constantly replenished
• SmF is primarily used for the extraction of secondary metabolites
that need to be used in liquid form .
2. Submerged Fermentation
[SMF]
Composition (g/l)
• Na2HPO4. 6g
• KH2PO4 3g
• NH4Cl 1g
• NaCl 0.5g
• CaCl2 0.15g
• MgSO4.7H2O 0.25g
• Casein hydrolysate 0.20g
• Yeast extract. 0.10g
• Starch 20g
Amylase Production Media
1. Carbon Source:
Common carbon sources used as substrates include maltose,
sucrose and glucose. A. tamarii actively synthesizes α-Amylase
when cultured on maltose, starch and glycogen under static
conditions. . The aim is to use substrates that are waste or by
products of other processes in order to make the process of enzyme
production environment friendly. One such substrate is oil cake.
Wheat bran has been used as substrate for α-Amylase production
by B.licheniformis and A.niger.
Substrate
2. Nitrogen Source
The nitrogen source used for production of α-Amylase may be organic or
inorganic. Few of the inorganic nitrogen sources include ammonium
sulphate, ammonium chloride and ammonium hydrogen phosphate. Most
commonly used organic sources of nitrogen include peptone, yeast
extract and soyabean meal.
The optimum process control parameters vary depending on the
microbial source, desired end product, method of fermentation
employed and many other such factors.
Process Parameters:
1.Temperature
• There are two temperatures that need to be in optimum
range during production.
• They are temperature for the growth of the microbial
source and optimum temperature at which maximum
production of enzyme takes place.
• The optimum temperatures for growth and α-Amylase
production were found to be 45°C to 46 °C and 50 °C,
respectively.
2. pH
• Optimum pH is a critical factor for the stability of enzyme produced.
Enzymes are pH sensitive and hence care must be taken to control
the pH of the production process.
• Pyrococcus furiosus produces α-Amylase which shows activity at
an optimum pH of 6.5–7.5
• Optimum levels of initial moisture content may vary depending on
the microbial source used.
• For fungal sources the moisture content required is less whereas
bacterial sources need more moisture content for high yield of the
enzyme.
• production of α-Amylase by Penicillium janthinellum the moisture
content was varied within a range of 20- 80% by varying the amount
of salt solution used in moistening the substrate particles.
Moisture
• This is a crucial factor in fermentation process.
• The enzyme activity increases with increase in incubation time till it
reaches the optimum duration. In most cases, the production of
enzyme begins to decline if the incubation time is further increased.
This could be due to the depletion of nutrients in the medium or
release of toxic substances.
• Bacillus subtilis, show high yield of alpha amylase after 48 hours of
fermentation
• Penicillium fellutanum isolated from mangrove rhizosphere soil. The
culture when incubated at 96 h, showed the maximum activity.
Duration of fermentation
• The desired enzyme produced may be excreted into the culture
medium (extracellular enzymes) or may be present within the cells
(intracellular enzymes).
• Depending on the requirement, the commercial enzyme may be crude
or highly purified. Further, it may be in the solid or liquid form.
• The steps involved in downstream processing i.e. recovery and
purification steps employed will depend on the nature of the enzyme
and the degree of purity desired.
Recovery and purification of
enzymes
• In general, recovery of an extracellular enzyme which is present in the
broth is relatively simpler compared to an intracellular enzyme. For the
release of intracellular enzymes, special techniques are needed for cell
disruption. Microbial cells can be broken down by physical
means(sonication, high pressure, glass beads)
• The recovery and purification steps will be the same for both
intracellular and extracellular enzymes, once the cells are disrupted
and intracellular enzymes are released. The most important
consideration is to minimise the loss of desired enzyme activity.
• Removal of cell debris:
• Filtration or centrifugation can be used to remove cell debris.
• Removal of nucleic acids : Nucleic acids interfere with the recovery
and purification of enzymes. They can be precipitated and removed by
adding poly-cations such as polyamines, streptomycin and
polyethyleneimine.
• Enzyme precipitation: Enymes can be precipitated by using salts
(ammonium sulfate) organic solvents (isopropanol, ethanol, and
acetone). Precipitation is advantageous since the precipitated enzyme
can be dissolved in a minimal volume to concentrate the enzyme.
• Liquid-liquid partition : Further concentration of desired enzymes can
be achieved by liquid-liquid extraction using polyethylene glycol or
polyamines.
• Separation by chromatography: there are several
chromatographic techniques for separation and purification of
enzymes. These include ion-exchange, size exclusion, affinity,
hydrophobic interaction and dye ligand chromatography. Among
these, ion- exchange chromatography is the most commonly used
for enzyme purification.
• Drying and packing: The concentrated form of the enzyme can be
obtained by drying. This can be done by film evaporators or freeze
dryers (lyophilizers). The dried enzyme can be packed and
marketed.
1. Dinitrosalicylic Acid Method (DNA)
2. Nelson-Somogyi (NS) method
3. Iodine Activity
Determination of enzyme activity
• Hydrolysis of starch into the extreme forming a less viscous starch
suspension.
• In bakers industry, converting starch in two smaller fermentable
sugars.
• In textile industry, used to removal of starch sizing agent for woven
fabric.
• Fuel alcohol production, converting starch into smaller fermentable
• sugars which are acted upon by East to produce alcohol.
Application of amylase enzyme
Thank you

Más contenido relacionado

La actualidad más candente

Citric Acid Production
Citric Acid ProductionCitric Acid Production
Citric Acid ProductionDinesh S
 
Media for industrial fermentation
Media for industrial fermentationMedia for industrial fermentation
Media for industrial fermentationNithyaNandapal
 
Glutamic acid fermentation
Glutamic acid fermentationGlutamic acid fermentation
Glutamic acid fermentationNOMI KhanS
 
Enzymes & their Production
Enzymes & their ProductionEnzymes & their Production
Enzymes & their ProductionMayur D. Chauhan
 
Industrial production of penicillin
Industrial production of penicillinIndustrial production of penicillin
Industrial production of penicillinNischitha R
 
Citric acid production
Citric acid productionCitric acid production
Citric acid productionPraveen Garg
 
strain improvement techniques
strain improvement techniquesstrain improvement techniques
strain improvement techniquesjeeva raj
 
Production of vitamin B12 using fermentation
Production of vitamin B12 using fermentationProduction of vitamin B12 using fermentation
Production of vitamin B12 using fermentationvijaysrampur
 
Industrial production of chemical acids glutamic acid
Industrial production of chemical acids glutamic acidIndustrial production of chemical acids glutamic acid
Industrial production of chemical acids glutamic acidEsam Yahya
 
Production of lipases and cellulase
Production of lipases and cellulaseProduction of lipases and cellulase
Production of lipases and cellulaseponkalaivani s
 
FERMENTERS( BIOREACTORS) AND THEIR TYPES
FERMENTERS( BIOREACTORS) AND THEIR TYPESFERMENTERS( BIOREACTORS) AND THEIR TYPES
FERMENTERS( BIOREACTORS) AND THEIR TYPESAYESHA KABEER
 
Production of tetracyclin and cephalosporin
Production of tetracyclin and cephalosporinProduction of tetracyclin and cephalosporin
Production of tetracyclin and cephalosporinSamsuDeen12
 
Aeration and agitation
Aeration and agitationAeration and agitation
Aeration and agitationeswar1810
 

La actualidad más candente (20)

Citric Acid Production
Citric Acid ProductionCitric Acid Production
Citric Acid Production
 
Media for industrial fermentation
Media for industrial fermentationMedia for industrial fermentation
Media for industrial fermentation
 
Glutamic acid fermentation
Glutamic acid fermentationGlutamic acid fermentation
Glutamic acid fermentation
 
Enzymes & their Production
Enzymes & their ProductionEnzymes & their Production
Enzymes & their Production
 
Industrial production of penicillin
Industrial production of penicillinIndustrial production of penicillin
Industrial production of penicillin
 
Amylase ppt
Amylase pptAmylase ppt
Amylase ppt
 
Citric acid production
Citric acid productionCitric acid production
Citric acid production
 
Fermentation technology
Fermentation technology Fermentation technology
Fermentation technology
 
strain improvement techniques
strain improvement techniquesstrain improvement techniques
strain improvement techniques
 
Production of vitamin B12 using fermentation
Production of vitamin B12 using fermentationProduction of vitamin B12 using fermentation
Production of vitamin B12 using fermentation
 
Design of a fermentor
Design of a fermentorDesign of a fermentor
Design of a fermentor
 
Industrial production of chemical acids glutamic acid
Industrial production of chemical acids glutamic acidIndustrial production of chemical acids glutamic acid
Industrial production of chemical acids glutamic acid
 
Production of lipases and cellulase
Production of lipases and cellulaseProduction of lipases and cellulase
Production of lipases and cellulase
 
Vitamin b12
Vitamin b12Vitamin b12
Vitamin b12
 
FERMENTERS( BIOREACTORS) AND THEIR TYPES
FERMENTERS( BIOREACTORS) AND THEIR TYPESFERMENTERS( BIOREACTORS) AND THEIR TYPES
FERMENTERS( BIOREACTORS) AND THEIR TYPES
 
Production of tetracyclin and cephalosporin
Production of tetracyclin and cephalosporinProduction of tetracyclin and cephalosporin
Production of tetracyclin and cephalosporin
 
Fermentation media
Fermentation mediaFermentation media
Fermentation media
 
Development of inoculum buildup
Development of inoculum buildup Development of inoculum buildup
Development of inoculum buildup
 
Aeration and agitation
Aeration and agitationAeration and agitation
Aeration and agitation
 
Production of amino acid by microorganisms.
Production of amino acid by microorganisms. Production of amino acid by microorganisms.
Production of amino acid by microorganisms.
 

Similar a Production of amylase

Microbial Alpha-amylase production (Basics).pdf
Microbial Alpha-amylase production (Basics).pdfMicrobial Alpha-amylase production (Basics).pdf
Microbial Alpha-amylase production (Basics).pdfShahjahan Kabir
 
enzymes-151002080055-lva1-app6892.pptx
enzymes-151002080055-lva1-app6892.pptxenzymes-151002080055-lva1-app6892.pptx
enzymes-151002080055-lva1-app6892.pptxFatmaGomaa11
 
enzymes-151002080055-lva1-app6892 (1).pdf
enzymes-151002080055-lva1-app6892 (1).pdfenzymes-151002080055-lva1-app6892 (1).pdf
enzymes-151002080055-lva1-app6892 (1).pdfPratikShinde189184
 
Amylase production
Amylase productionAmylase production
Amylase productionPraveen Garg
 
Enzyme Production.pptx
Enzyme Production.pptxEnzyme Production.pptx
Enzyme Production.pptxDrSABINAK
 
Presentation of Enzyme- lipase.pptx
Presentation of Enzyme- lipase.pptxPresentation of Enzyme- lipase.pptx
Presentation of Enzyme- lipase.pptxmuskanmahajan24
 
5. Production of enzyme in biotechnolgy.pptx
5. Production of enzyme in biotechnolgy.pptx5. Production of enzyme in biotechnolgy.pptx
5. Production of enzyme in biotechnolgy.pptxHarshadaa bafna
 
Ergot alkaloids.pptx
Ergot alkaloids.pptxErgot alkaloids.pptx
Ergot alkaloids.pptxVed Gharat
 
Enzymes and its applications
Enzymes and its applicationsEnzymes and its applications
Enzymes and its applicationseswar1810
 
ApplicationsofEnzymesMADhotre.pptx
ApplicationsofEnzymesMADhotre.pptxApplicationsofEnzymesMADhotre.pptx
ApplicationsofEnzymesMADhotre.pptxSwetha464750
 
1830_developmentofmedia.pdf
1830_developmentofmedia.pdf1830_developmentofmedia.pdf
1830_developmentofmedia.pdfJasonWillardM
 
Citric acid production
Citric acid productionCitric acid production
Citric acid productionRABIYAZAFAR5
 
Industrial Microbiology of Molds
 Industrial Microbiology of Molds Industrial Microbiology of Molds
Industrial Microbiology of MoldsDuithy George
 

Similar a Production of amylase (20)

Microbial Alpha-amylase production (Basics).pdf
Microbial Alpha-amylase production (Basics).pdfMicrobial Alpha-amylase production (Basics).pdf
Microbial Alpha-amylase production (Basics).pdf
 
enzymes-151002080055-lva1-app6892.pptx
enzymes-151002080055-lva1-app6892.pptxenzymes-151002080055-lva1-app6892.pptx
enzymes-151002080055-lva1-app6892.pptx
 
enzymes-151002080055-lva1-app6892 (1).pdf
enzymes-151002080055-lva1-app6892 (1).pdfenzymes-151002080055-lva1-app6892 (1).pdf
enzymes-151002080055-lva1-app6892 (1).pdf
 
Amylase production
Amylase productionAmylase production
Amylase production
 
AMYLASE (M).pptx
AMYLASE (M).pptxAMYLASE (M).pptx
AMYLASE (M).pptx
 
Enzyme amylases
Enzyme amylasesEnzyme amylases
Enzyme amylases
 
Enzyme Production.pptx
Enzyme Production.pptxEnzyme Production.pptx
Enzyme Production.pptx
 
Production of Glutamic acid.pptx
Production of Glutamic acid.pptxProduction of Glutamic acid.pptx
Production of Glutamic acid.pptx
 
Presentation of Enzyme- lipase.pptx
Presentation of Enzyme- lipase.pptxPresentation of Enzyme- lipase.pptx
Presentation of Enzyme- lipase.pptx
 
5. Production of enzyme in biotechnolgy.pptx
5. Production of enzyme in biotechnolgy.pptx5. Production of enzyme in biotechnolgy.pptx
5. Production of enzyme in biotechnolgy.pptx
 
Ergot alkaloids.pptx
Ergot alkaloids.pptxErgot alkaloids.pptx
Ergot alkaloids.pptx
 
Enzymes and its applications
Enzymes and its applicationsEnzymes and its applications
Enzymes and its applications
 
Primary Metabolites PHARMACOGNOSY-I.pptx
Primary Metabolites  PHARMACOGNOSY-I.pptxPrimary Metabolites  PHARMACOGNOSY-I.pptx
Primary Metabolites PHARMACOGNOSY-I.pptx
 
Primary Metabolites FROM THE PLANT AND ANIMALS.pptx
Primary Metabolites FROM THE PLANT AND ANIMALS.pptxPrimary Metabolites FROM THE PLANT AND ANIMALS.pptx
Primary Metabolites FROM THE PLANT AND ANIMALS.pptx
 
ApplicationsofEnzymesMADhotre.pptx
ApplicationsofEnzymesMADhotre.pptxApplicationsofEnzymesMADhotre.pptx
ApplicationsofEnzymesMADhotre.pptx
 
1830_developmentofmedia.pdf
1830_developmentofmedia.pdf1830_developmentofmedia.pdf
1830_developmentofmedia.pdf
 
Development of media
Development of mediaDevelopment of media
Development of media
 
Fermentation.pptx
Fermentation.pptxFermentation.pptx
Fermentation.pptx
 
Citric acid production
Citric acid productionCitric acid production
Citric acid production
 
Industrial Microbiology of Molds
 Industrial Microbiology of Molds Industrial Microbiology of Molds
Industrial Microbiology of Molds
 

Último

Alternative system of medicine herbal drug technology syllabus
Alternative system of medicine herbal drug technology syllabusAlternative system of medicine herbal drug technology syllabus
Alternative system of medicine herbal drug technology syllabusPradnya Wadekar
 
Applied Biochemistry feedback_M Ahwad 2023.docx
Applied Biochemistry feedback_M Ahwad 2023.docxApplied Biochemistry feedback_M Ahwad 2023.docx
Applied Biochemistry feedback_M Ahwad 2023.docxmarwaahmad357
 
SCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptx
SCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptxSCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptx
SCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptxROVELYNEDELUNA3
 
Physics Serway Jewett 6th edition for Scientists and Engineers
Physics Serway Jewett 6th edition for Scientists and EngineersPhysics Serway Jewett 6th edition for Scientists and Engineers
Physics Serway Jewett 6th edition for Scientists and EngineersAndreaLucarelli
 
An intro to explainable AI for polar climate science
An intro to  explainable AI for  polar climate scienceAn intro to  explainable AI for  polar climate science
An intro to explainable AI for polar climate scienceZachary Labe
 
CW marking grid Analytical BS - M Ahmad.docx
CW  marking grid Analytical BS - M Ahmad.docxCW  marking grid Analytical BS - M Ahmad.docx
CW marking grid Analytical BS - M Ahmad.docxmarwaahmad357
 
Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...
Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...
Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...Sérgio Sacani
 
Human brain.. It's parts and function.
Human brain.. It's parts and function. Human brain.. It's parts and function.
Human brain.. It's parts and function. MUKTA MANJARI SAHOO
 
Intensive Housing systems for Poultry.pptx
Intensive Housing systems for Poultry.pptxIntensive Housing systems for Poultry.pptx
Intensive Housing systems for Poultry.pptxHarshiniAlapati
 
soft skills question paper set for bba ca
soft skills question paper set for bba casoft skills question paper set for bba ca
soft skills question paper set for bba caohsadfeeling
 
TORSION IN GASTROPODS- Anatomical event (Zoology)
TORSION IN GASTROPODS- Anatomical event (Zoology)TORSION IN GASTROPODS- Anatomical event (Zoology)
TORSION IN GASTROPODS- Anatomical event (Zoology)chatterjeesoumili50
 
Role of herbs in hair care Amla and heena.pptx
Role of herbs in hair care  Amla and  heena.pptxRole of herbs in hair care  Amla and  heena.pptx
Role of herbs in hair care Amla and heena.pptxVaishnaviAware
 
3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...
3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...
3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...PirithiRaju
 
geometric quantization on coadjoint orbits
geometric quantization on coadjoint orbitsgeometric quantization on coadjoint orbits
geometric quantization on coadjoint orbitsHassan Jolany
 
Pests of ragi_Identification, Binomics_Dr.UPR
Pests of ragi_Identification, Binomics_Dr.UPRPests of ragi_Identification, Binomics_Dr.UPR
Pests of ragi_Identification, Binomics_Dr.UPRPirithiRaju
 
Thermonuclear explosions on neutron stars reveal the speed of their jets
Thermonuclear explosions on neutron stars reveal the speed of their jetsThermonuclear explosions on neutron stars reveal the speed of their jets
Thermonuclear explosions on neutron stars reveal the speed of their jetsSérgio Sacani
 
Pests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdf
Pests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdfPests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdf
Pests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdfPirithiRaju
 
Pests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdf
Pests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdfPests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdf
Pests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdfPirithiRaju
 
Basic Concepts in Pharmacology in molecular .pptx
Basic Concepts in Pharmacology in molecular  .pptxBasic Concepts in Pharmacology in molecular  .pptx
Basic Concepts in Pharmacology in molecular .pptxVijayaKumarR28
 
001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...
001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...
001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...marwaahmad357
 

Último (20)

Alternative system of medicine herbal drug technology syllabus
Alternative system of medicine herbal drug technology syllabusAlternative system of medicine herbal drug technology syllabus
Alternative system of medicine herbal drug technology syllabus
 
Applied Biochemistry feedback_M Ahwad 2023.docx
Applied Biochemistry feedback_M Ahwad 2023.docxApplied Biochemistry feedback_M Ahwad 2023.docx
Applied Biochemistry feedback_M Ahwad 2023.docx
 
SCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptx
SCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptxSCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptx
SCIENCE 6 QUARTER 3 REVIEWER(FRICTION, GRAVITY, ENERGY AND SPEED).pptx
 
Physics Serway Jewett 6th edition for Scientists and Engineers
Physics Serway Jewett 6th edition for Scientists and EngineersPhysics Serway Jewett 6th edition for Scientists and Engineers
Physics Serway Jewett 6th edition for Scientists and Engineers
 
An intro to explainable AI for polar climate science
An intro to  explainable AI for  polar climate scienceAn intro to  explainable AI for  polar climate science
An intro to explainable AI for polar climate science
 
CW marking grid Analytical BS - M Ahmad.docx
CW  marking grid Analytical BS - M Ahmad.docxCW  marking grid Analytical BS - M Ahmad.docx
CW marking grid Analytical BS - M Ahmad.docx
 
Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...
Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...
Legacy Analysis of Dark Matter Annihilation from the Milky Way Dwarf Spheroid...
 
Human brain.. It's parts and function.
Human brain.. It's parts and function. Human brain.. It's parts and function.
Human brain.. It's parts and function.
 
Intensive Housing systems for Poultry.pptx
Intensive Housing systems for Poultry.pptxIntensive Housing systems for Poultry.pptx
Intensive Housing systems for Poultry.pptx
 
soft skills question paper set for bba ca
soft skills question paper set for bba casoft skills question paper set for bba ca
soft skills question paper set for bba ca
 
TORSION IN GASTROPODS- Anatomical event (Zoology)
TORSION IN GASTROPODS- Anatomical event (Zoology)TORSION IN GASTROPODS- Anatomical event (Zoology)
TORSION IN GASTROPODS- Anatomical event (Zoology)
 
Role of herbs in hair care Amla and heena.pptx
Role of herbs in hair care  Amla and  heena.pptxRole of herbs in hair care  Amla and  heena.pptx
Role of herbs in hair care Amla and heena.pptx
 
3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...
3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...
3.2 Pests of Sorghum_Identification, Symptoms and nature of damage, Binomics,...
 
geometric quantization on coadjoint orbits
geometric quantization on coadjoint orbitsgeometric quantization on coadjoint orbits
geometric quantization on coadjoint orbits
 
Pests of ragi_Identification, Binomics_Dr.UPR
Pests of ragi_Identification, Binomics_Dr.UPRPests of ragi_Identification, Binomics_Dr.UPR
Pests of ragi_Identification, Binomics_Dr.UPR
 
Thermonuclear explosions on neutron stars reveal the speed of their jets
Thermonuclear explosions on neutron stars reveal the speed of their jetsThermonuclear explosions on neutron stars reveal the speed of their jets
Thermonuclear explosions on neutron stars reveal the speed of their jets
 
Pests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdf
Pests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdfPests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdf
Pests of cumbu_Identification, Binomics, Integrated ManagementDr.UPR.pdf
 
Pests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdf
Pests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdfPests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdf
Pests of wheat_Identification, Bionomics, Damage symptoms, IPM_Dr.UPR.pdf
 
Basic Concepts in Pharmacology in molecular .pptx
Basic Concepts in Pharmacology in molecular  .pptxBasic Concepts in Pharmacology in molecular  .pptx
Basic Concepts in Pharmacology in molecular .pptx
 
001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...
001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...
001 Case Study - Submission Point_c1051231_attempt_2023-11-23-14-08-42_ABS CW...
 

Production of amylase

  • 3. Contents • Introduction • Types of amylase • Production of amylase • Recovery and purification • Determination of enzyme activity • Industrial application
  • 4. Amylase • Amylase is an enzyme that catalyses the hydrolysis of starch into sugars. • Amylase is present in the saliva of humans and some other mammals, where it begins the chemical process of digestion • These ezymes randomly cleave internal glycosidic linkages in starch molecules to hydrolyze them and yield dextrins and oligosaccharides
  • 5. Types of Amylase: α-Amylase • α-Amylase is a hydrolase enzyme that catalyses the hydrolysis of internal α-1, 4-glycosidic linkages in starch to yield products like glucose and maltose. • It is a calcium metalloenzyme i.e. it depends on the presence of a metal co factor for its activity. • There are 2 types of hydrolases: endo-hydrolase and exo- hydrolase. Endo- hydrolases act on the interior of the substrate molecule, whereas exo-hydrolases act on the terminal non reducing ends .
  • 6. • The substrate that α-amylase acts upon is starch. Starch is a polysaccharide composed of two types of polymers – amylose and amylopectin. • Amylose constitutes 20-25% of the starch molecule. It is a linear chain consisting of repetitive glucose units linked by α-1,4- glycosidic linkage. Amylopectin constitutes 75-80% of starch and is characterized by branched chains of glucose units. • The optimum pH for activity is found to be 7.0.
  • 7. β – Amylase • β-Amylase is an exo-hydrolase enzyme that acts from the nonreducing end of a polysaccharide chain by hydrolysis of α-1, 4- glucan linkages to yield successive maltose units. • It is unable to cleave branched linkages in branched polysaccharides such as glycogen or amylopectin, the hydrolysis is incomplete and dextrin units remain. • During ripening of fruits, β-Amylase breaks down starch into maltose resulting in the sweetness of ripened fruit. • The optimal pH of the enzyme ranges from 4.0 to 5.5.
  • 8. γ – Amylase • • γ-Amylase cleaves α(1-6)glycosidic linkages, in addition to cleaving the last α(1-4)glycosidic linkages at the nonreducing end of amylose and amylopectin, unlike the other forms of amylase, yielding glucose. • γ- amylase is most efficient in acidic environments and has an optimum pH of 3. •
  • 10. Sources • α-Amylase can be isolated from plants, animals or microorganisms. • The enzyme has been isolated from barley and rice plants . • It has been found that cassava mash waste water is a source of α- Amylase which is active in wide range of pH and temperature . • The most widely used source among the bacterial species is the Bacillus spp. B. amyloliquefaciens and B. licheniformis • Fungal sources of α-Amylase are mostly Aspergillus species and only few species of Penicillium, P. brunneum , penicillium fellutanum
  • 11. Production Methods : There are mainly two methods which are used for production of α-Amylase on a commercial scale. These are : 1) Solid State fermentation. 2) Submerged fermentation
  • 12. • Method used for microbes which require less moisture content for their growth. • The solid substrates commonly used in this method are, bran, bagasse, and paper pulp. • The main advantage is that nutrient-rich waste materials can be easily recycled and used as substrates in this method. 1. Solid State Fermentation [SSF]
  • 13. • employs free flowing liquid substrates, such as molasses and broths. • The products yielded in fermentation are secreted into the fermentation broth. The substrates are utilized quite rapidly,hence the substrates need to be constantly replenished • SmF is primarily used for the extraction of secondary metabolites that need to be used in liquid form . 2. Submerged Fermentation [SMF]
  • 14. Composition (g/l) • Na2HPO4. 6g • KH2PO4 3g • NH4Cl 1g • NaCl 0.5g • CaCl2 0.15g • MgSO4.7H2O 0.25g • Casein hydrolysate 0.20g • Yeast extract. 0.10g • Starch 20g Amylase Production Media
  • 15. 1. Carbon Source: Common carbon sources used as substrates include maltose, sucrose and glucose. A. tamarii actively synthesizes α-Amylase when cultured on maltose, starch and glycogen under static conditions. . The aim is to use substrates that are waste or by products of other processes in order to make the process of enzyme production environment friendly. One such substrate is oil cake. Wheat bran has been used as substrate for α-Amylase production by B.licheniformis and A.niger. Substrate
  • 16. 2. Nitrogen Source The nitrogen source used for production of α-Amylase may be organic or inorganic. Few of the inorganic nitrogen sources include ammonium sulphate, ammonium chloride and ammonium hydrogen phosphate. Most commonly used organic sources of nitrogen include peptone, yeast extract and soyabean meal.
  • 17. The optimum process control parameters vary depending on the microbial source, desired end product, method of fermentation employed and many other such factors. Process Parameters: 1.Temperature • There are two temperatures that need to be in optimum range during production. • They are temperature for the growth of the microbial source and optimum temperature at which maximum production of enzyme takes place.
  • 18. • The optimum temperatures for growth and α-Amylase production were found to be 45°C to 46 °C and 50 °C, respectively. 2. pH • Optimum pH is a critical factor for the stability of enzyme produced. Enzymes are pH sensitive and hence care must be taken to control the pH of the production process. • Pyrococcus furiosus produces α-Amylase which shows activity at an optimum pH of 6.5–7.5
  • 19. • Optimum levels of initial moisture content may vary depending on the microbial source used. • For fungal sources the moisture content required is less whereas bacterial sources need more moisture content for high yield of the enzyme. • production of α-Amylase by Penicillium janthinellum the moisture content was varied within a range of 20- 80% by varying the amount of salt solution used in moistening the substrate particles. Moisture
  • 20. • This is a crucial factor in fermentation process. • The enzyme activity increases with increase in incubation time till it reaches the optimum duration. In most cases, the production of enzyme begins to decline if the incubation time is further increased. This could be due to the depletion of nutrients in the medium or release of toxic substances. • Bacillus subtilis, show high yield of alpha amylase after 48 hours of fermentation • Penicillium fellutanum isolated from mangrove rhizosphere soil. The culture when incubated at 96 h, showed the maximum activity. Duration of fermentation
  • 21. • The desired enzyme produced may be excreted into the culture medium (extracellular enzymes) or may be present within the cells (intracellular enzymes). • Depending on the requirement, the commercial enzyme may be crude or highly purified. Further, it may be in the solid or liquid form. • The steps involved in downstream processing i.e. recovery and purification steps employed will depend on the nature of the enzyme and the degree of purity desired. Recovery and purification of enzymes
  • 22. • In general, recovery of an extracellular enzyme which is present in the broth is relatively simpler compared to an intracellular enzyme. For the release of intracellular enzymes, special techniques are needed for cell disruption. Microbial cells can be broken down by physical means(sonication, high pressure, glass beads) • The recovery and purification steps will be the same for both intracellular and extracellular enzymes, once the cells are disrupted and intracellular enzymes are released. The most important consideration is to minimise the loss of desired enzyme activity. • Removal of cell debris: • Filtration or centrifugation can be used to remove cell debris.
  • 23. • Removal of nucleic acids : Nucleic acids interfere with the recovery and purification of enzymes. They can be precipitated and removed by adding poly-cations such as polyamines, streptomycin and polyethyleneimine. • Enzyme precipitation: Enymes can be precipitated by using salts (ammonium sulfate) organic solvents (isopropanol, ethanol, and acetone). Precipitation is advantageous since the precipitated enzyme can be dissolved in a minimal volume to concentrate the enzyme. • Liquid-liquid partition : Further concentration of desired enzymes can be achieved by liquid-liquid extraction using polyethylene glycol or polyamines.
  • 24. • Separation by chromatography: there are several chromatographic techniques for separation and purification of enzymes. These include ion-exchange, size exclusion, affinity, hydrophobic interaction and dye ligand chromatography. Among these, ion- exchange chromatography is the most commonly used for enzyme purification. • Drying and packing: The concentrated form of the enzyme can be obtained by drying. This can be done by film evaporators or freeze dryers (lyophilizers). The dried enzyme can be packed and marketed.
  • 25. 1. Dinitrosalicylic Acid Method (DNA) 2. Nelson-Somogyi (NS) method 3. Iodine Activity Determination of enzyme activity
  • 26. • Hydrolysis of starch into the extreme forming a less viscous starch suspension. • In bakers industry, converting starch in two smaller fermentable sugars. • In textile industry, used to removal of starch sizing agent for woven fabric. • Fuel alcohol production, converting starch into smaller fermentable • sugars which are acted upon by East to produce alcohol. Application of amylase enzyme