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Asaaib Nijah Hussain
Al-Wadi International School, Jeddah
AS- level
ENZYME
IMMOBILIZATION
What exactly is Immobilization?!
• Immobilization, as you can quite see from the word, is when you
take away the tendency of a substance to move around
• Or simpler, it is when you trap an extract of enzyme- or an enzyme
solution- to use it for catalytic purposes.
• Or biologically, “Enzyme immobilization is the process of confining
enzyme molecules to a solid support over which a substrate is
passed and converted to products”
Hi to the story of immoblized enzymes!
• Immobilized enzymes were discovered in three steps—
• First in 1815 (empirical use in waste water treatment)
• Second step was discovered in 1960 (in production of L- amino acid)
• Third in 1985 – 1995 ( co-factor regeneration )
How to do it? Is it easy…?
There are quite a few steps which mght be easy and others tough however,
reading or memorizing the steps will not be as difficult as actually applying
them in a lab. There you go, these are the steps:
• Make an enzyme solution (lets say amylase from a saliva extract)
• Mix it well with a sodium alginate solution
• Fill it up in a syringe
• Add in drop by drop to a calcium chloride solution held in a beaker
• Leave the “balls” for about 10 minutes, filter, with distilled water and let
them dry in a sieve
• TADA! Your immobilized enzymes are ready to be used!
• Try them out in a starch solution (you can use a potato extract)
• Decant away the solution after 10 minutes and check for the starch using
iodine solution
• Do you see the blue black or the brown colour..? Brown right! Means the
enzymes are still functioning!
Lets have a look!
Why would one immobilize enzymes?
Now lets take a situation where we want to make our milk lactose free to help
you understand this easier
Mobilized lactase
Enzyme
solution
Lactose
containing
milk
Solution
containing active
lactase and
lactose-free milk
Now this enzyme
extract cannot be
reused and is
hence wastedWait for ten minutes
for reaction to take
place
Immobilized Lactase
Immobilizd
enzymes in
alginate beads
Lactose rich
milk
Leave for 10
minutes for
reaction to take
place
Reuseable
enzyme in
beads
Lactose
free milk
So as you can see, the advantages of
immobilization are:
• Enzye can be reused as it is no contiminated nor has it got to be
removed from the product which again is a long process, in that the
enzyme we want needs to be wasted…. Because the only way we
can remove it by denatureing the enzyme
• If fre enzymes are present in product to be used/consumed it may
cause allergies.
• Separtion of product is easier from the nzyme as mentioned earlier
• Enzyme is less likely to denature in the alginate mixture
• The enzyme also becomes more stable in a bound or solid form
• Lesser chances of enzyme denaturation
But what about the disadvantages??
• Immoblization requires more time effort and work
• Production of immobilized enzymes can be expensive
• Reaction rate decreases as there is an extra surface that needs to
be crossed by the substrate therefore a diffusional limitation is
faced and slowness is observed in the reaction
• The enzyme active may not be facing the outer side of the alginate
“balls” so the active site may be unreachable to the substrate
• Most immobilized enzyms may not work at their usual optimum
temperatures, actually, a bit higher temperature is required
therefore more energy is needed and can prove to be expensive
Oh! But where exactly is it used?
• Some of the commonly immobilized enzymes are
• Amino acylase – production of L-aminoacids from D, L-acyl amino acids
• Amylase – production of glucose from starch
• Invertase – splitting sucrase into fructose and glucose
• Aspartase – production of aspartic acid from fumaric acid
• Fumarase – synthesis of malic acid from fumaric acid
I understood…
Did u?
well….. I really hope u
did… ’coz I badly need my
marks!

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Enzyme immobilization- AS level

  • 1. Asaaib Nijah Hussain Al-Wadi International School, Jeddah AS- level ENZYME IMMOBILIZATION
  • 2. What exactly is Immobilization?! • Immobilization, as you can quite see from the word, is when you take away the tendency of a substance to move around • Or simpler, it is when you trap an extract of enzyme- or an enzyme solution- to use it for catalytic purposes. • Or biologically, “Enzyme immobilization is the process of confining enzyme molecules to a solid support over which a substrate is passed and converted to products”
  • 3. Hi to the story of immoblized enzymes! • Immobilized enzymes were discovered in three steps— • First in 1815 (empirical use in waste water treatment) • Second step was discovered in 1960 (in production of L- amino acid) • Third in 1985 – 1995 ( co-factor regeneration )
  • 4. How to do it? Is it easy…? There are quite a few steps which mght be easy and others tough however, reading or memorizing the steps will not be as difficult as actually applying them in a lab. There you go, these are the steps: • Make an enzyme solution (lets say amylase from a saliva extract) • Mix it well with a sodium alginate solution • Fill it up in a syringe • Add in drop by drop to a calcium chloride solution held in a beaker • Leave the “balls” for about 10 minutes, filter, with distilled water and let them dry in a sieve • TADA! Your immobilized enzymes are ready to be used! • Try them out in a starch solution (you can use a potato extract) • Decant away the solution after 10 minutes and check for the starch using iodine solution • Do you see the blue black or the brown colour..? Brown right! Means the enzymes are still functioning!
  • 5. Lets have a look!
  • 6. Why would one immobilize enzymes? Now lets take a situation where we want to make our milk lactose free to help you understand this easier Mobilized lactase Enzyme solution Lactose containing milk Solution containing active lactase and lactose-free milk Now this enzyme extract cannot be reused and is hence wastedWait for ten minutes for reaction to take place
  • 7. Immobilized Lactase Immobilizd enzymes in alginate beads Lactose rich milk Leave for 10 minutes for reaction to take place Reuseable enzyme in beads Lactose free milk
  • 8. So as you can see, the advantages of immobilization are: • Enzye can be reused as it is no contiminated nor has it got to be removed from the product which again is a long process, in that the enzyme we want needs to be wasted…. Because the only way we can remove it by denatureing the enzyme • If fre enzymes are present in product to be used/consumed it may cause allergies. • Separtion of product is easier from the nzyme as mentioned earlier • Enzyme is less likely to denature in the alginate mixture • The enzyme also becomes more stable in a bound or solid form • Lesser chances of enzyme denaturation
  • 9. But what about the disadvantages?? • Immoblization requires more time effort and work • Production of immobilized enzymes can be expensive • Reaction rate decreases as there is an extra surface that needs to be crossed by the substrate therefore a diffusional limitation is faced and slowness is observed in the reaction • The enzyme active may not be facing the outer side of the alginate “balls” so the active site may be unreachable to the substrate • Most immobilized enzyms may not work at their usual optimum temperatures, actually, a bit higher temperature is required therefore more energy is needed and can prove to be expensive
  • 10. Oh! But where exactly is it used? • Some of the commonly immobilized enzymes are • Amino acylase – production of L-aminoacids from D, L-acyl amino acids • Amylase – production of glucose from starch • Invertase – splitting sucrase into fructose and glucose • Aspartase – production of aspartic acid from fumaric acid • Fumarase – synthesis of malic acid from fumaric acid
  • 11. I understood… Did u? well….. I really hope u did… ’coz I badly need my marks!

Notas do Editor

  1. On title- “As my friend suggested that history can be divided in two syllables- hi and story- so lets say hi to the story of immobilized enzymes”
  2. Draw n show the syringe beaker drawing like those of serial dilutions (with the arrows n all) , describe shape of balles with small tail Mention that use syringe to get exact sizes of the alginates balls. Mention how to chk for starch on tiles
  3. Skip the waiting parts Comment on apparatus- how subs can be used
  4. Try to explain adv here itself cuz next slide is all points n they shd not be reading….
  5. Skip if mentioned adv at pre slide
  6. Ont read… just say in any order BUT DON’T READ
  7. There r many more but major uses are: _ _ _ _ Invertase, as ms salma had mentioned b4 in bio mols