Factors Affecting Enzyme Activity: Lactose Intolerance
Lactose is the primary carbohydrate in milk and is commonly known as milk sugar. Lactose comprises of almost 8% of human milk and almost 4% of cow’s milk. Lactose intolerance is a condition in human beings where they lack enzyme lactase since they have trouble in absorbing and digesting lactose. The goal of this experiment is to-:
- Learn how substrate concentration, pH and enzyme concentration can affect the enzyme activity in human body.
Data Variable (Procedures)
In this experiments, o-nitrophenyl-β-D-galactopyranoside (ONPG) would be used as a substitute to lactose since it is similar in shape and size and can bind at the active site of lactase enzyme. The procedures for the experiment are as follows-:
- Record: First, record the lactase brand, the expiration and the number of units of the tablet.
- Weigh: Second, weigh each lactase tablet and note down whether it is 3,000 FCC units/tablet or 9,000 FCC units per tablet.
- Grind: Third, grind the tablet in a mortar and pestle it until it is finely grounded.
- Suspend/ Dissolve: Grind the lactase tablet in approximately 5 mL of 0.01 M PO4 buffer. The solution would come out to be cloudy due to the undissolved binder present in it.
- Dilute: Add 0.2 mL enzyme suspension in about 5.8 mL 0.01 M PO4.
- Attach the SpectoVis to the computer via the USB cable and turn on the computer
- Once the Logger Pro 3.7 icon starts running, calibrate the spectrophotometer.
- The next step is to click on the configure spectrophotometer tab and select the Absolute v/s Concentration is the collection mode.
- Enter the sample number and then proceed to click ok.
- Repeat the steps 4 and 5 till the sample are read by the spectrophotometer.
- In order to analyse the data export it into an excel file.
If the liquid shoes colour then it must be become of the presence of some component that absorbs light. The concentration of the substance that absorbs light is directly proportional to the absorbance. Thus, it can be said that the more the reaction rate of the solution the more the yellow-coloured o-nitrophenolate ion would be produced. Thus, we can say that absorbance is directly proportional to the enzyme activity.
In this experiment, the absorbance would be plotted on the y-axis while each factor that affects the enzyme activity would be plotted on the x-axis.
The Resulting Graphs are given below-: