Enzyme Immobilization

Asaaib Nijah HussainAl-Wadi International School, JeddahAS- levelEnzyme 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 aroundOr 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

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Hi to the story of immoblized enzymes!Immobilized enzymes were discovered in three stepsFirst 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 )

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 enzymes3

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 solutionFill it up in a syringeAdd in drop by drop to a calcium chloride solution held in a beakerLeave the balls for about 10 minutes, filter, with distilled water and let them dry in a sieveTADA! 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 solutionDo you see the blue black or the brown colour..? Brown right! Means the enzymes are still functioning!

Draw n show the syringe beaker drawing like those of serial dilutions (with the arrows n all) , describe shape of balles with small tailMention that use syringe to get exact sizes of the alginates balls.Mention how to chk for starch on tiles4

Lets have a look!

Skip the waiting partsComment on apparatus- how subs can be used5

Why would one immobilize enzymes? Now lets take a situation where we want to make our milk lactose free to help you understand this easierMobilized lactase

Enzyme solution

Lactose containing milk

Solution containing active lactase and lactose-free milk

Now this enzyme extract cannot be reused and is hence wasted

Wait for ten minutes for reaction to take place

Immobilized Lactase

Immobilizd enzymes in alginate beadsLactose rich milk

Leave for 10 minutes for reaction to take place

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Reuseable enzyme in beadsLactose free milk

Try to explain adv here itself cuz next slide is all points n they shd not be reading.7

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 enzymeIf fre enzymes are present in product to be used/consumed it may cause allergies.Separtion of product is easier from the nzyme as mentioned earlierEnzyme is less likely to denature in the alginate mixtureThe enzyme also becomes more stable in a bound or solid formLesser chances of enzyme denaturation

Skip if mentioned adv at pre slide8

But what about the disadvantages??Immoblization requires more time effort and workProduction of immobilized enzymes can be expensiveReaction 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 reactionThe enzyme active may not be facing the outer side of the alginate balls so the active site may be unreachable to the substrateMost 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

Ont read just say in any order BUT DONT READ9

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 acidsAmylase production of glucose from starchInvertase splitting sucrase into fructose and glucoseAspartase production of aspartic acid from fumaric acidFumarase synthesis of malic acid from fumaric acid

There r many more but major uses are: _ _ _ _Invertase, as ms salma had mentioned b4 in bio mols10

I understoodDid u?well.. I really hope u did coz I badly need my marks!