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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!
Editor's Notes
- 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”
- 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
- Skip the waiting parts Comment on apparatus- how subs can be used
- Try to explain adv here itself cuz next slide is all points n they shd not be reading….
- Skip if mentioned adv at pre slide
- Ont read… just say in any order BUT DON’T READ
- There r many more but major uses are: _ _ _ _ Invertase, as ms salma had mentioned b4 in bio mols