Essay on Lactase Enzyme Cofactor

The study of sciences often involves the reaction of substances. It should be noted that some of the reactions take long for the end product to be achieved which eventually delays the activities influenced by the reaction. In this regard, the scientists and researchers have come up with ways of fastening the rate of these reactions by using catalysts. It is worth noting that enzymes are biological catalysts which are macromolecular in nature. According to Illanes (2008), a catalyst is a substance that acts on the reactants which are also referred to as substrates by lowering the activation energy involved in the process of specific reactions while remaining chemically unchanged at the end of the reaction. Besides, a catalyst can provide an alternative pathway that quickens the rate at which the products are formed. However, there are several factors that limit the optimality of an enzyme. Some of these factors include enzyme specificity, enzyme cofactor, pH, and temperature among others (Jasewicz and Wasserman, 1961).

Lactase is an enzyme that was primary used in this experiment. This enzyme is essential used in the breaking down of disaccharides into monosaccharides and is found in the small intestine, kidney and liver of mammals. This experiment is important in the real life situation for individuals suffering from the deficiency of lactase enzyme (Järvelä, Torniainen, & Kolho, 2009).

Purpose of the Experiment

There were two sections in this experiment where the first experiment incorporated the investigation of lactase enzyme specificity taking lactose and maltose as substrates. The second experiment involved the determination of cofactor of lactase enzyme.

Hypothesis

First Experiment

            The following statements represent the hull and alternative hypotheses that were to be tested in the experiment;

H0: There is no difference between the glucose produced when lactase enzyme is introduced into two solutions of lactose and maltose in equal amounts.

H1: There is a difference between the glucose produced when lactase enzyme is introduced into two solutions of lactose and maltose in equal amounts.

Second Experiment

Similarly, the hypotheses of the second experiment were;

H0: There is no difference in glucose production between the solution of EDTA and the solution containing distilled water.

H1: There is a difference in glucose production between the solution of EDTA and the solution containing distilled water.

Materials and Methods

Experiment 1: Lactase specificity

Two test tubes were labeled L and M where letters represent lactose and maltose respectively. After which, of milk was added to tube L using a pipette. On the other hand, the maltose solution was added to the tube M up to 1.0 mL which implies that the solution of the maltose added was. The two tubes were left for 10 minutes in the water bath at a temperature of 40oC. Lastly, the amount of glucose was determined by the help of a glucose chart using a glucose strip. Besides, the strip was allowed thirty seconds before any substantive comparison were attained. The results obtained were recorded and graphically presented using bar graphs.

Experiment 2:  Enzyme cofactor

It was essential to wear gloves and goggles because EDTA is a hazardous substance. Therefore, after taking safety measures at hand, EDTA solution having a volume of was added to the tube labeled EDTA. Three drops of milk was then added followed by the inversion of the tube to allow the mixing of the two solutions. Thereafter, 3 drops of lactase solution was added to the EDTA tube. The same procedures were repeated for the tube labeled control, but water was used instead of EDTA. The control and EDTA tube were then left for 10 minutes in a water bath of 40oC. The amount of glucose was determined by the help of a glucose chart using a glucose strip. It should be noted that the strip was allowed thirty seconds before any substantive comparison were attained. The experiment was repeated several times for this case after which the descriptive statistic was conducted and recorded in Table 1.

Results

Experiment 1

            From the first experiment, it was evident that the amount of glucose produced in the tube containing lactose was 10 times higher than the tube containing maltose. The former had the amount of glucose produced at 1000 mg/dL while the latter had 100 mg/dL. However, it is fundamental for the results to be represents either as pictorials or graphs for easy interpretation especially for people with less knowledge in statistics and experimental works.

Figure 1: Specificity of Lactase in maltose and lactose.

From figure 1, it can be seen that there was production of glucose in the tubes containing maltose and lactose. However, the amount produced greatly differs.

Experiment 2

            In this section, the distilled water was used as a control for the determination of the cofactor of lactase enzyme using EDTA. The results were obtained from after performing several experiments. Therefore, for this section, the sample size was taken as 49 experiments where the maximum values were 500 and 100 for distilled water and EDTA respectively. On the other hand, the minimum amount of glucose produced was 50 mg/dL for distilled water and 0 mg/dL for EDTA. From the sample, the means of the variables regarding the glucose produced in mg/dL were found to be 211.73 and 20.918 for distilled water and EDTA respectively. The p-value at 5% level of significance was obtained as. It is worth noting that the p-value was obtained from the t-value of 56.95505 using the excel spreadsheet like other measures stated above.

Descriptive Statistics

Control (diH2O)

EDTA

mean

211.73

20.9

standard deviation

104.47

40

Variance

450

100

Minimum value

50

2

Maximum value

500

100

Sample

49

49

Table 1: Descriptive statistics for experiment 2

Discussion/Conclusion

The difference observed in the Figure 1 is associated with the specificity of enzyme lactase. Indeed, enzymes and their substrates have specific active sites where the bind and fit exactly in the process of a reaction (Illanes, 2008). It should be noted that the stronger the chemical interaction formed between the amino acid from the enzyme and the substrate, the quicker the products will be formed.  In this case, a high amount of glucose was produce in lactose than in maltose because lactase is specific with lactose. On the other hand, the maltose did not react with the lactase enzyme leading to less production of glucose. Therefore, the null hypothesis is rejected since there is a difference between the glucose produced when lactase enzyme is introduced into lactase and maltose respectively. Besides, the enzymes break down their own substrates.

The descriptive statistics (see Table 1) shows that the enzymes are highly affected by the presence of cofactor. Besides, the null hypothesis for the second experiment is rejected since the p-value of  is less than 5% level of significance (Peck, Olsen, & Devore, 2015). Therefore, there is a statistical significance difference the amounts of glucose produced in in the solution containing distilled water and a solution of EDTA. In this regard, the enzymes will work well in an environment with cofactor elements unlike when the cofactors are absent (Illanes, 2008). This is because, from the sample involving EDTA, there was less production of glucose and in one experiment here was zero production of glucose. On the contrary, the tube containing distilled water which does not have an effect on the cofactor elements depicted a high level of glucose production throughout the sample.

References

Illanes, A. (2008). Enzyme biocatalysis: principles and applications. Springer Science & Business Media.

Järvelä, I., Torniainen, S., & Kolho, K. L. (2009). Molecular genetics of human lactase deficiencies. Annals of medicine, 41(8), 568-575.

Jasewicz, L., &Wasserman, A.E. (1961). Quantitative determination of lactase. Journal of Dairy Science 44.3, 393-400.

Peck, R., Olsen, C., & Devore, J. L. (2015). Introduction to statistics and data analysis. Cengage Learning.