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Investigate the use of immobilized lactase to produce lactose-free milk

Lactose Tolerance Test: When your body breaks down lactose, it releases
sugar into your blood. This tests how much sugar is in your blood. After you fast,
a small sample of blood is taken. Then, you drink a liquid that is high in lactose.
Two hours later, you give another blood sample. Lactose causes blood sugar
levels to rise, your blood sugar levels in this sample should be higher. If you're
lactose intolerant, you'll have just a low rise in blood sugar and symptoms.


A simple answer from yahoo answers

The advantage is that the immobilized lactase (which is bound to very small
beads, typically) can be used over and over, instead of having to add fresh
lactase to each batch of milk. The milk is passed over the column and comes out
lactose-free; the column is then cleaned and ready for more milk.

It's also useful in case anyone drinking the milk is allergic to lactase. Lactic acid
test can be used to determine whether the milk is lactose-free or not.

The Lactic acid test

Just titrate a fixed quantity of milk with dilute NaOH of known concentration.
Maybe 25cm
3
of 0.1 mole NaOH solution. The lactose in milk is constantly being
converted into Lactic acid. Keep adding milk from the burette into the conical
flask containing NaOH solution. Keep adding milk until the solution becomes
colorless (white because of the milk).

If the milk doesnt have lactic acid, then it cant have lactose.


The official procedure from Nuffield Biology

Using a bioreactor to reduce the lactose content of milk
The following procedure may be used to investigate lactose reduction of
skimmed milk using
immobilised lactase enzyme.

Health and safety
Wear eye protection. Before carrying out any practical work a risk assessment
must be done.
Some people are allergic to lactase solution; avoid getting on the skin or rubbing
eyes after handling.

Equipment and materials
0.5 cm3 lactase enzyme solution (available from Philip Harris or NCBE)
[Irritant]
10 cm3 of 2% sodium alginate solution
100 cm3 of 1.5% calcium chloride solution
skimmed milk (not UHT, 25 cm
3
for each test)
Diastix (one for each test)
small beakers (25 or 50 cm
3
)
stirring rod
150 cm3 beakers
nylon gauze or net curtain (1-3 cm
2
) If glass wool is used, forceps are
needed for handling it
tea strainer
spatula
10 cm
3
graduated syringe
25 cm
3
pipette
2 cm
3
pipette or syringe
test tubes
boiling tubes
ice (if requested)
insulating material (if requested)
water baths at several temperatures
thermometers
stop watch






Investigating enzymes used in laundry detergents

1 A suitable dilution of detergent is around 5%, that is 5 cm
3
of detergent per 100
cm
3
. This is calculated from the typical detergent pack suggestion of 100 cm
3
of
detergent in a medium soil wash and machines using around 20 litres of water
for the wash cycle.

2 Provide portions of enzymes to add to known volumes of the detergent, or
make up solutions of enzymes (protease only, lipase only, amylase only) in non-
bio detergent. Students mix these to make enzyme cocktails, following their
investigation plans.

NCBE supplies five detergent enzymes individually or in a pack of five. They
supply lipase (Lipex) in 100 cm
3
bottles. The data sheets in the packs
recommend suitable amounts of enzyme to use as a starting point for any
investigation.

They also provide information about the enzyme including likely optimum
temperature for enzyme action. NCBE recommend testing any protocol with
different concentrations of enzyme before use with students, because enzyme
activity in different situations and after storage is hard to predict. Use the data
sheet provided with the enzyme to guide your decisions.

3 Web research into the source and nature of the fibres in different fabrics will
reveal that some are plant products (varieties of cellulose fibres), some are
synthetic polymers, and others are of animal origin (protein fibres). With their
knowledge of enzymes, students should be able to work out which laundry
enzymes are likely to have a negative effect on silk and wool.
4 Students may devise their own stains, but could include cooking oils, fresh
grass, milk, egg yolk, chocolate sauce, rich gravy, fruit juice. Encourage students
to be sparing in how they stain the fabric, especially if investigating fatty stains at
low temperatures.





Procedure

SAFETY: Rinse any splashes of enzyme solutions from the skin quickly and
thoroughly. Wear eye protection when handling enzyme solutions. Clean up spills
with a damp cloth before they dry. Avoid raising dust into the air with
detergents and powdered enzymes. Avoid rubbing eyes after handling solutions,
washed fabrics etc.

Preparation

(a) Make up detergent and enzyme solutions

(b) Prepare fabrics to test

Investigation

(c) Fabrics will need to be stained, mixed with detergent and/or enzyme solutions
at a certain temperature for a certain time, agitated in some way, rinsed and
left to dry before assessing the stain-removing effects of the detergent/ enzyme
mixes.