The effect of Temperature on Invertase enzyme

Summary

This video explores how temperature affects the activity of the enzyme invertase in a sucrose solution. The experiment involves using various temperature settings, from refrigeration to boiling, to observe enzyme performance. Key materials include a spectrophotometer, test tubes, and DNS reagent. After incubating samples at different temperatures and measuring absorbance, results indicate that 50 degrees Celsius is the optimal temperature for invertase activity. The video provides a detailed step-by-step guide to conducting the experiment safely and effectively, making it a valuable resource for understanding enzyme kinetics.

Takeaways

• 🌡️ Temperature significantly impacts the activity of the enzyme invertase on sucrose solutions.
• 🍬 Invertase is derived from yeast and plays a role in candy production.
• 🔬 Essential equipment includes a spectrophotometer, test tubes, cuvettes, and safety gear like gloves and goggles.
• 🧊 Solutions are incubated at various temperatures: 4°C, room temperature (20°C), 50°C, and 100°C.
• ⏳ Each temperature group is incubated for 15 minutes before adding DNS (dinitro salicylic acid).
• 🔥 DNS reacts with glucose, and its reduction can be measured using a spectrophotometer.
• 🧪 Proper labeling of test tubes and cuvettes is crucial to avoid confusion during the experiment.
• 🔄 The spectrophotometer should be zeroed using a blank solution containing only sucrose and buffer.
• 📈 The optimal temperature for invertase activity is identified as 50°C based on absorbance readings.
• ⚠️ Safety precautions are necessary when handling corrosive substances and high-temperature equipment.

Q & A

What is the purpose of the experiment described in the video?

The experiment aims to investigate how temperature affects the activity of the enzyme invertase on a sucrose solution.

What source is invertase obtained from?

Invertase is obtained from yeast.

What safety equipment is recommended for this experiment?

Recommended safety equipment includes gloves, goggles, lab coats, and the use of a fume hood.

What equipment is needed to conduct the experiment?

Essential equipment includes a spectrophotometer, water baths, test tubes, glass cuvettes, and pipettes.

What is the first step in preparing the test tubes for the experiment?

The first step is to label the test tubes according to the temperature groups being tested.

What solutions are added to the test tubes during the experiment?

1 ml of sucrose solution, followed by a pH 5 buffer (1 ml for test tube 1 and 0.5 ml for test tube 2), and 0.5 ml of invertase suspension only to test tube 2.

At what temperatures are the test tubes incubated during the experiment?

Test tubes are incubated at 4°C, 20°C, 50°C, and 100°C.

Why is dinitro salicylic acid (DNS) added to the test tubes?

DNS is added to reduce glucose, and its reduced form is measurable and visible on a spectrophotometer.

What is the optimal temperature for invertase enzyme activity according to the results?

The optimal temperature for invertase enzyme activity is 50°C.

How is the spectrophotometer prepared for measuring absorbance?

The spectrophotometer is zeroed using a blank cuvette containing only sucrose and pH buffer before measuring the absorbance of the other samples.