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ENZYME ACTION - LACTASE
Enzymes are natures catalysts. They speed up reactions to make them useful for
metabolic pathways. This protocol describes the action of lactase on lactose, a disaccharide
found in milk. Lactase is a mammalian digestive enzyme. In humans, lactase is present in
infants but for many people, the gene that regulates its expression is switched off in early
childhood, leading to lactose intolerance.
Code: MC23.51
Name: Lactase
Description: Amber coloured liquid.
Storage: Store in the refrigerator at 4
o
C.
Shelf Life: When stored correctly, MC23.51 has a nominal shelf life of 12 months.
Safety: MC23.51 is rated as safe for food contact. Nevertheless, enzymes are
biologically active proteins and should be handled with care. Avoid direct
contact or inhalation.
Suggested Protocol
Condition the lactase and a quantity of full cream milk to the desired temperature in a
water bath. Take 50mL of the milk and add 1mL of lactase with stirring.
Lactase hydrolyses lactose into glucose and galactose, so you can monitor the progress of
the reaction by measuring the increase in the concentration of glucose. Use a glucose
detection strip such as Diastix (G10.41) for this.
Milk typically contains about 140mmol/L of lactose which will release an equivalent amount
of glucose. The colour of Diastix indicator strips becomes dark brown when exposed to a
glucose concentration in excess of 111mmol/L, so you can expect the Diastix to indicate a
glucose concentration greater than 111mmol/L.
Run a control alongside your test solution by substituting water for the lactase. It is also
useful to control for the presence of glucose in the lactase by adding 1mL of lactase to
50mL of distilled water and testing with Diastix.
The optimum reaction conditions for this form of lactase are:
pH 6.0
Temperature 40 - 45
o
C
Recommendations
Try conditioning the lactase and milk to a range of different temperatures. You should see
a significant slow-down in the rate of reaction as you cool the milk below 20
o
C. Lactase is
denatured and ineffective at temperatures above 65
o
C.
The content of lactose is often reduced in treated forms of milk such as UHT because the
treatment process causes some hydrolysis.
Check our website for information on how to make a reactor that removes lactose from
milk.
Please Note:
Batch-to-batch variations in substrate composition and enzyme activity can mean that the
suggested experiment might not work exactly as described in every situation.

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