ENZYME IMMOBILISATION

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Rabiya kazi 12DT                               Core practical 3 (immobilized enzyme)        16/07/2007

ENZYME IMMOBILISATION

Aim -The purpose of this investigation is to break down the lactose found in milk, into its disaccharide sugars; glucose and galactose. (A disaccharide is made up of two individual monomers (monosaccharide) units joined together by a condensation

reaction).

In a normal digestive system all the food we eat is broken down this helps the body to absorb the food. Lactose is a natural sugar found in milk, it is a disaccharide carbohydrate (sugar) made up of two single sugar units (monomers) joined by a hydrolysis reaction (adding of water). The sugar lactose is not very sweet and is insoluble.  The enzyme lactase produced in our body breaks lactose into its disaccharide forms-glucose and galactose. Glucose is the only sugar that can be absorbed into the bloodstream. In people that are lactose intolerant they do not have the enzyme to this job or the enzyme does not function well and therefore it could cause indigestion problems.

In this investigation I will immobilise enzymes by entrapping them in a gel. I feel there are many advantages of using the immobilised enzymes over the free ones, some advantages are:

  • The enzymes can easily be separated from the mixture and so enzyme loss at the end of the experiment is minimised.

  • Because the enzyme can easily be removed it is easier to re-cycle it, therefore it can be re-used a number of times.

  • The reaction can be stopped by removing the enzyme.

  • The immobilised enzyme has more stability than free ones and so o reaction rate is better as the enzyme is more firm.

                                                                                           

           I will immobilise the enzyme by

                trapping them in an alginate gel.

Hypothesis-I predict that at the end of the experiment when I test the glucose strip, there should be a change in colour, and thus considerable amount of glucose should be present.

In the experiment the enzyme lactase will be the independent variable as this will determine the amount of conversion of lactose to glucose and galactose. Thus the glucose and galactose production will be dependant on this. I will measure the production at the end using a glucose test strip against a colour scale to see how much is present. Other variables such as the temperature I will keep constant so that they do not affect the investigation and results.

Apparatus- In order to carry out the experiment I will need to use various apparatus. Firstly I will need to make sure that I wear a laboratory coat to ensure safety and reduce the risk of any accidents. This is because the acids used could stain the clothes or hands.

     

I will need to get two 20 ml beaker. One will be needed to mix the alginate and lactase and the other will be needed at the end of the experiment when allowing the milk to drop. I will use a 20ml beaker because it is small and so it will allow me to make more accurate measurements. I will also need two 20cm measuring cylinder one to measure the calcium chloride and the other to collect the milk. I will use cylinders rather than as beaker as I feel it is easier to transfer the content.

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I will then need to acquire 2cm lactase enzyme to convert the lactose into glucose and galactose. Also 8cm of alginate solution will be needed, to mix with the lactase to make the beads. In addition I will need 20cm of calcium chloride, to drop the beads into.

A glass rod will be needed to stir the alginate and lactose solution to ensure that the liquids are properly mixed.

I will then need to get three 10cm syringe. One will be needed to obtain the alginate and another one will be needed to get the lactase. ...

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