Authentic Scientific Research in regard to Lactose Intolerance

Enzymes play a quintessential position in the digestion of the food molecules like carbohydrates, proteins, and lipids in the human body. Understanding the factors that have an effect on the enzymatic activities is essential for comprehending some of the troubles that are related to the problems encountered when some meals is consumed. Lactose intolerance is an example of the problem that outcomes when there is no proper and sufficient digestion of carbohydrates. An actual scientific research is important in producing the knowledge that is useful for grasp how lactose intolerance occurs in the body when human beings consume dairy products. The purpose of this laboratory test is to explore how the environmental factors of pH, temperature, and attention affect the rate of carbohydrate digestion with the aim of understanding how lactose intolerance occurs in individuals. To achieve the goal of the study, the lab session is divided into four sections with different objectives set to experiment the effects of pH, temperature, and concentration (enzyme and substrate). This paper is, therefore, a laboratory report showing the methods and outcomes of the experiment. The materials used for each of the tests include the conventional laboratory apparatus while the reagents used in the experiments include Benedict’s reagent, pH buffer solutions (1, 4.4, 9.2, and 13) enzyme solution, and sucrose (substrate). The procedures for each the experiments vary significantly with the variations indicated in the conditions of pH, concentrations, and temperatures being made to test the effects of the specificity of the factors.

The outcomes are primarily demonstrated in the rates of reactions observed in each of the conditions. The effects of substrate concentrations are seen in the fact that the highest rate of reaction (1) is attained at the enzyme concentration of 0.5mL while the further increase of the concentration leads to a constant rate of reaction (0.6). The volumes of the substrate also has significant effect on the speed of reactions with the lowest volumes (0 and 1mL of sucrose) showing a reaction rate of zero respectively. On the other hand, the highest volume of sucrose shows the highest rate of reaction. It can thus be concluded that the substrate and enzymes concentrations have a significant effect on the enzymatic actions. Findings on the impact of temperature indicate that the optimum point for the enzymatic reaction is at the temperatures 30OC and 60OC that are both 0.6. The rate at the ice bath is 0.4 while there is no reaction at the boiling point and hence the relationship between temperature and enzymatic activities. Similarly, a connection is also noted between pH and the rate of the reactions. pH towards neutral indicates highest rates of reaction. The speed of the reaction at pH 4.4 is 0.8 while at pH 9.2 the rate is 0.8 while there is no reaction at the pHs 1 and 13. In retrospect, it can be seen that the enzymatic reactions are affected by the aspects of temperature, concentration, and pH. The concept of the lactose intolerance can thus be explained regarding the factors that influence the enzymatic digestion processes.

Keywords: Enzymes, temperature, pH, concentration, lactose intolerance

Introduction

Background

Enzymes play critical roles in the human body as they enhance the essential chemical reactions in the body. Enzymes are utilized in the digestion of the carbohydrates, proteins, and lipids into simple sugars, amino acids and fatty acids respectively through the various metabolic pathways. The reactions occurring inside each and every cell of the living organisms are made faster due to the presence of the enzymes that make the reactions to proceed at faster rates (Palmer & Bonner, 2007). The enzymes are protein in nature and are affected by environmental aspects like temperature, pH and concentration. Most enzymes work best in temperatures of between 20oC and 40oC. It can thus be observed that there is an increase of the enzymatic activities with the rise of temperature until the optimum level is attained after which the enzymes are denatured. On the other hand, the lower temperatures inactivate the enzymes and hence the slow reactions. The concentration of either the enzyme or substrate affects the rate of reactions. On the same note, it can be observed that most enzymes perform optimally at neutral pH.

Some of the food consumed are in the form of carbohydrates and may be contained substances like bread, sugars, potatoes, wheat, and others. Forms in which carbohydrates exists include monosaccharides (glucose, fructose, and galactose), disaccharides (sucrose, lactose, and maltose, and polysaccharides (starch and cellulose). Maltose is the product of the digestion of starch while lactose is the major sugar in the milk. Various types of enzymes are responsible for the digestion of the different forms of carbohydrates in the digestive systems. For instance, enzyme maltase digests maltose to glucose through the process of hydrolysis. Other examples of the enzymes include the sucrase and lactase that catalyzes the hydrolysis of sucrose and lactose into simple sugars respectively. Industrial production of enzymes through biotechnological processes allows the provision of the essential products that can be utilized for catalyzing of biochemical reactions in the instances that enzymatic activities are inhibited or slowed.

Purpose of the Experiment

Since the enzymatic activities are subject to the mentioned environmental factors, it can be noted that the vital reactions in the body can be affected leading to abnormal reaction rates in the body systems. Carbohydrate digestion is one of the enzymatic reactions that can is influenced by the aspects of pH, concentration, and temperature (Palmer & Bonner, 2007). This purpose of this experiment is to study how the environmental factors of pH, temperature, and concentration affect the rate of carbohydrate digestion with the aim of understanding how lactose intolerance occurs in individuals.

Objectives

To accomplish the purpose of the experiment, four goals are set to be met in this trial.

1. To investigate the impact of the enzyme concentration on the rate of enzymatic activity
2. To study the effect of the substrate concentration on the rate of enzymatic activity
3. To explore the impact of temperature on the rate of enzymatic activity
4. To investigate the impact of pH on the rate of enzymatic activity

Hypotheses and Variables

Hypotheses are formulated for each of the listed objectives and are based on the four major parts of the laboratory activity.

Part I:

The increase in the enzyme concentration leads to the rise in the concentration of the glucose, which indicates the associated increase in the rate of reaction.

Independent Variable: Enzyme concentration

Dependent Variable: rate of reaction

Part II:

The increase in the substrate concentration leads to the rise in the volumes of the glucose, which indicates the associated increase in the rate of reaction

Independent Variable: Substrate concentration

Dependent Variable: rate of reaction

Part III

The rise in temperature leads to increase the rate of reaction from 0oC to 60oC while the rate is slowed with the boiling.

Independent Variable: Temperature

Dependent Variable: rate of reaction

Part IV

The rate of reaction (concentration of glucose) is high towards the neutral pHs while the rates are lower at extreme pHs.

Independent Variable: pH

Dependent Variable: rate of reaction

Materials and Methods

Materials

The reagents used in the experiments include Benedict’s reagent, pH buffer solutions (1, 4.4, 9.2, and 13) enzyme solution, and sucrose (substrate). Other parts of the materials include the laboratory apparatus that are useful in assisting in conducting the experiments like test tubes, hot plates, measuring cylinders, stop the clock and others.

Procedures

The procedures used differed from one section to the other with the variations observed based on the intended goal of the part. In part, I, 5 test tubes labeled 1 through 5 were each filled with 2mL of 1% sucrose and then 2mL pH buffer was added into each of the test tubes. Designated amounts of enzyme solution and water were then added into each of the test tubes as indicated in table 1. The test tubes were then put in a half full beaker with water and then placed on a hot plate. The set up was allowed to sit at room temperature to allow for the hydrolysis of the sucrose for five minutes after which 3mL of Benedict's reagent was added into each of the test tubes. All the test tubes were then placed in water bath for 2-3 minutes with the respective color changes indicated in table 3

Table 1: Designated amounts of water and enzymes added in each test tube

Part II

All the procedures in part I was followed in part II apart from the designated amounts of the substrate (sucrose) and water added. Another significant difference between the procedures in parts I and II is noted in the sense that the amount of the enzyme added is constant in part II.  Table 2 shows the number of substances added into each of the test tubes. The test results from part II is as indicated in table 4

Table 2: Designated amounts of water and substrate added in each test tube

Part III

Four test tubes were labeled 1A through 4A. Additional four test tubes were labeled as 1B through 4B. 2mL sucrose and pH 4.4 buffers were added into test tubes 1A, 2A, 3A, and 4A and then placed into designated temperature baths as 0oC (ice water), 30oC, 60oC, and 100oC (boiling) respectively for 15 minutes. In test tubes 1B through 4B, 3mL of the enzyme was added and placed into the temperature baths as in the case of 1A, 2A, 3A, and 4A for 15 minutes. The contents of 1B were then added into 1A then the contents of 1AB were placed in the designated water bath. Similar procedures were done to 2A and 2B, 3A and 3B, and 4A and 4B. 3mL of Benedict’s reagent was then added to the combinations of 1AB, 2AB, 3AB, and 4AB and boiled for three minutes. The results were recorded in table 5.

Part IV

Four test tubes were labeled 1 through 4. To test tube #1, 4mL pH 1 buffer and 1mL enzyme solutions were added. To test tube #2 pH 4.4 buffer and 1mL enzyme solutions were added. To test tube #3 pH buffer and 1mL enzyme solutions were added. To test tube #4 9.2 pH 13 buffer and 1mL enzyme solutions were added. The contents of all the test tubes were then mixed and then let to sit for 5 minutes at room temperature. 2mL of sucrose was then added into each of the test tubes and thereafter allowed to sit for 5 minutes after which 3mL of the Benedict reagent was added into the mixtures and then boiled for 3 minutes. The results indicating color changes were then recorded in table 6.

Calculations

Calculations were conducted to obtain the rate of reaction in each of the experimental sections. The following equation was used to calculate the reaction rate.

Reaction rate =Assigned Unit Number5

The assigned unit number is the value obtained corresponding to the color derived from Benedict’s test while five is the reaction time.

Results

Part I

Table 3: Results of the effect of enzyme concentration on the rate of enzymatic reaction

Results in table 3 show that there is no reaction when the volume of the enzyme is 0. The highest rate of reaction is attained at the enzyme concentration of 0.5mL. A constant rate of reaction is achieved at 0.6 with the burnt orange color observed for concentrations 1mL, 1.5mL, and 3mL respectively. The trend obtained is represented in the graph in figure 1.

Figure 1: Effect of enzyme concentration

Part II