EFFECT OF TEMPERATURE ON ENZYME

EFFECT OF TEMPERATURE ON ENZYME

                                                                 Introduction

Enzymes are biological molecules that help in speeding up the rate of all the chemical reactions that usually take place within cells (Arsalan and Younus, 2018). They are essential for life since enzymes serve a wide variety of functions in the body. Therefore, aiding in digestion and metabolism. The aim of the experiment was to find out how to change in temperature (℃) has an effect on the production of maltose (the activity of amylase in reaction). The independent variable that was used in the experiment was the temperature (℃), and the dependent variable was maltose concentration (mg/ml). The investigation is essential since it will able to give insight into how the increase and decrease in temperature aid in digestion.

If temperature increases, then maltose concentration will increase until a particular optimal temperature, at which point maltose production will decrease. The prediction of the experiment was if amylase activity is observed at rising temperatures, then the concentration of maltose will increase.

Materials and Methods

For the experiment, groups were to be 5 test tubes with 100 µL of amylase and start at respective temperatures of 30 ℃, 37 ℃, 45 ℃, 65 ℃, and 85 ℃. The control groups that were to be blank while using the spectrophotometer were to be set up like a tube with starch, but no amylase at room temperature (℃).

The reagents, materials, and equipment that were used during the experiment include 5% starch solution, water, amylase, 6 test tubes, micropipettes, DNS reagent solution, hot plate, spectrophotometer, b cuvettes, environmental chamber, glass pipette, plastic pump, and test tube rack.

Six test tubes were labeled as B, 2, 3, 4, 5, and 6. 450 µL of 5y starch and 450 µL of water were added in all test tubes. The addition of 100 µL of water to tube B. Test tube one was placed on a rack for 10 minutes (at room temperature). 100 µL of amylase was added to tubes 2 to 6, which was staggered by 30 seconds. Tubes 2, 3, 4, 5 and 6 were placed in env chamber at respective temperatures of 3 ℃, 37 ℃, 45 ℃, 65 ℃ and 85 ℃ for a period of 10 minutes. Then the addition of 1000 µL of DNS to all tubes by using a micropipette. The tubes were placed in a boiling water bath for 5 minutes. Using a 10 ml glass pipette and an automatic gun, I added 8 ml of deionized water. Thereafter I calibrated spectrophotometer using tube B. I measured absorbance of all tubes at 540nm and recorded in table 1. Finally, the estimation of maltose concentration basing on a previously determined standard curve.

Results

            The findings in the experiment for the concentrated maltose and its absorbance at 540 nm, concentrated maltose (ml) C₂ of 0.1, 0.2, 0.3, 0.4, 0.5 showed the absorbance at 540 nm as 0.462, 0.870, 1.256, 1.478 and 1.676 respectively. The results from temperature against concentrated maltose showed that at three ℃, 37 ℃, 45 ℃, 65 ℃, and 85 ℃ for a period of 10 minutes the concentration of maltose was as follows 0.208, 1.112, 1.530, 0.932 and 0 respectively. The findings of the experiment are represented in the tables below

Table 1                                                                              Table 2

Graph presentation of the effect of temperature on Maltose concentration

Graph representation of the effect of maltose concentration to absorbance at 540 nm

It can be concluded from the experiment that if temperature increases, then maltose concentration will increase until a certain optimal temperature, at which point maltose production will decrease as illustrated in the experiment the maltose concentration increases as temperatures increase until at 45 ℃ when we have a fall in the concentration of maltose. The experiment also illustrated that an increase of maltose concentration leads to an increase in its absorbance at 540 nm.

References

Arsalan, A., & Younus, H. (2018). Enzymes and nanoparticles: Modulation of enzymatic activity via nanoparticles. International journal of biological macromolecules, 118, 1833-1847.