Investigating the affects of immobilised lactase on lactose.

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Investigating the affects of immobilised lactase on lactose

The aim of this investigation is to analyse the affect immobilising lactase will have on lactose.

The equipment needed to carry out the experiment includes:

Large syringe, rubber tubing and screw clamp

Plastic gloves apron and safety glasses (important when dealing with glass wool and calcium chloride, gloves are needed as both are irritant)

Clamp stand, clamp and bosses

Glass beakers - 1 x 500cm3, 3 x 250cm3

Filter funnel

Glass wool

Balance

Sodium Alginate

Distilled water

Calcium Chloride solution

Hot water bath

Test tube rack, boiling tubes

Lactase enzyme, Benedict’s reagent

Milk

Sieve

Method:

Firstly weigh 4g of sodium alginate, dissolve this into 100cm3 of distilled water in a beaker, and stir thoroughly.  Mix 2cm3 of lactase into 8cm3 of cooled sodium alginate solution in a test tube.  Using a stirring rod release droplets of the alginate-enzyme mixture into 1.5% calcium chloride solution, one drop at a time.  Using a sieve separate the beads from the calcium chloride.  Rinse the beads with distilled water.  Push a small amount of glass wool into the barrel of the syringe.  Using a spatula pack the syringe with the beads.  Pour the milk into the syringe until all the beads are engulfed by it.  Collect 1cm3 of milk from the tube at the end of the syringe every five minutes.  Add 1cm3 of Benedict’s reagent to the milk and heat in the hot water bath for about 5 minutes.  Repeat the experiment every 5 minutes until concordant results are achieved (this should take about 6 or 7 tests)

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Diagram

The following diagram shows the equipment as it is set up to allow the lactase beads to break down the lactose in the milk:

Results:

Conclusion:

The purpose of immobilising enzymes before they are used to catalyse a reaction is so that they can be reused i.e. they are not lost in the solution.  We immobilised lactase enzyme by dissolving it in sodium alginate gel. This is one of the more widely used practises in industry as it is very efficient and effective.  We ...

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