Abstract
Three distinct variables were theorized and examined in the parameters on the outcome of the productivity of the lactase enzyme. The assessment of the temperature, substrate composition and the pH were reviewed in this experiment. The pH values were recorded over time. The effect of the pH on the lactase production in addition to the qualities of the substrate identity and the temperature were documented. In consideration of the findings of the experiment, it was discovered that a pH between 5 - 7 is the most suitable pH for the production of lactase. In this lab report, the works of Farkhauser (2009), Melton (2001), O’ Connell and Walsh (2010), Pope et al. (1998) and Sharma et al. (2007) were explored
Essential Conditions for Enzyme Lactase
Introduction
The enzyme that is designated as lactase is applied in the food manufacturing industry in order to decompose lactose into the fundamental sugars of galactose and glucose. The deficits in the production of lactase in the body deter the decomposition of lactose and have the outcome of lactose intolerance. Lactose intolerance has the potential of deriving diarrhea, indigestion and abdominal cramps when dairy products are ingested. In the application of lactase in the food, manufacturing processes, the milk acquires a more viscous characteristic in order to develop the manufacture of products that include ice cream (Farkhauser, 2009; Melton, 2001, O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
A number of enzymes are recognized as proteins that decrease the activation energy. These processes are required in order to initiate the chemical reactions that are integral to the perpetuation of life. The enzymes experience modifications in their form in relation to the environment that is changing. The changes in form influence the changes in the productivity of the enzyme. There are a number of actors that influence the development of the enzymes. These factors are temperature pH and the substrate identity (Farkhauser, 2009; Melton, 2001, O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
In this experiment w the temperature, substrate identity and the pH were examined. The examinations that were consumed in order to review the influences of the substrate identity, temperature and the pH on the comprehensive productivity of the enzyme that is s designated as lactase. The more elevated the amount of the glucose that is created, the more elevated the production of the lactase enzyme. The strips that are applied for the testing of the pH will be applied in order to document4e the concentration of glucose (Farkhauser, 2009; Melton, 2001, O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
Methods
A lactase solution was prepared by pulverizing a lactase tablet and by actively blending it with fifty milliliters of water that was distilled. The solids were filtered and the solution that remained that was labeled as lactase was examined. One percent solutions of maltose, sucrose and sugar lactose were applied by dissolving one gram of the substrate in 100 ml of water which was distilled. These solutions were labeled. In each of the trials, one milliliter of the enzyme and ten milliliters of the water were applied. Considering the exception of the pH trials, there were no additional buffering agents that would be needed. The concentration of the glucose was examined with the pH testing strips subsequent to the enzyme n mixing for an interval of fifteen minutes. The temporal periods that were applied in mixing were documented in order to facilitate the timing of the assessment. The solutions that were used were discarded in the drainage in addition to a substantial quantity of water. The used testing strips were discarded in the trash.
Substrate Activity and pH
There were three distinct buffering solutions that were provided. One milliliter of each of the buffering solutions was inserted into distinct beakers. The lactose was added and was blended. The timing imitated when all of the beakers possessed lactose. The glucose concentration was reviewed and recorded subsequent to fifteen minutes. The pH that manifested the most elevated amount of enzyme activity was discovered to be a pH of between 5- 6 (Farkhauser, 2009; Melton, 2001, O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
Substrate Activity and Temperature
The lactose was placed in separate testing tubes from the lactase solutions. The testing tubes were placed into an ice water bath in order to refrigerate. The testing tubes were facilitated in reaching a nearly consistent thermal quality. The temperature was recorded. The enzyme and the sugar were placed together. The testing tubes were regressed to the ice water bath for refrigeration. The glucose concentration was documented subsequent to a period of fifteen minutes. The procedure was repeated in a container of water with a thermal quality of 45°C. The concentration of the glucose was documented subsequent to a fifteen minute temporal interval. The actual temperature of the solutions was recorded. The Lactase solutions that included t that excluded the sugar were inserted into a container of water that reached its ebullition. The testing tubes were allowed to sit in the container of water that had an ebullient quality for fifteen minutes. The tube was extracted and permitted to refrigerate. The enzyme was aggregated to the lactose solution. The glucose concentration was recorded after fifteen minutes. The effect of the varying temperature was recorded in a table (Farkhauser, 2009; Melton, 2001. O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
Specificity with Regards to the Substrate
The sucrose and the maltose solutions were examined at a thermal characteristic of room temperature in the same manner that the lactose was reviewed. The lactase was added and the glucose levels were assessed after fifteen minutes. The influence of the lactase on the sucrose and the maltose was recorded.
Table1.
The Lactaid in its genuine form and the tablet form were pulverized and blended with 50 mL of distilled of water. The lactase that was undissolved was filtered. The same solution that possessed the undissolved lactase was applied to the 1% solutions of maltose, sucrose and sugar lactose. These solutions were prepared by dissolution of one gram of the substrate in 100 ml of water that had been distilled. Subsequently, one milliliter of the enzyme was added to ten milliliters of the sugar. In the first trial 10 milliliters of lactase and one milliliter of enzyme was used. The second trial one milliliter of the enzyme and ten milliliters of the sucrose were applied. In the third trial one milliliter of enzyme and milliliters of the lactase was applied.
Discussion
In the empirical examination, the variables that were reviewed were substrate specificity, pH and temperature. There was a change in the concentration of the lactase in the distinct temperature pH and substrate specificities. The initial hypothesis was that as the concentration was enhanced, the absorption level of the glucose would be enhanced. The second hypothesis was that the optimal pH would facilitate absorption at a pH of five. Lactase is an enzyme that is restricted by a membrane that is encountered in the epithelial cells that are located in the jejunum of the smaller intestines in humans. The lactase enables the digestion of disaccharide lactose. The enzyme adheres to the lactose into two distinct monosaccharides which are galactose and glucose by means of a process of being hydrolyzed (Farkhauser, 2009; Melton, 2001, O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
The deficiency of decomposing lactose is encountered primarily in adults. The causal attribute of the deficiency is the lack of lactase that is located in the smaller intestines. The phenomenon afflicts about 75% of the population. The symptoms are the sensation of discomfort in the ingestion of lactose. The best manner of assisting those who are lactose intolerant is to provide products that are developed commercially. These products are lactase powder and drops that facilitate the introduction of the lactase into the body. The introduction of lactase into the body enables the decomposition of lactose in the smaller intestines that facilities the decomposition of the lactose. These commercially developed products that compensate for the lactase deficiency (Farkhauser, 2009; Melton, 2001, O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
These products enable those who suffer from lactose intolerance to effectively utilize the monosaccharides that are formed. This category of product could prove to be Instrumental to the members of the poor population with regards to relieving a portion of the symptoms of the starvation and challenges of malnutrition that are presently experienced. The extra strength tablets that possess lactase that are composed of (ONPG) ortho nitro phenyl galactose can be applied as indicators for the activities of the enzymes. The buffers may be applied in order to vary the pH quality and the thermal conditions may also be varied in order to ascertain the potential activity of the enzyme (Farkhauser, 2009; Melton, 2001, O’Connell and Walsh, 2010; Pope et al., 1998; Sharma et al. 2007).
Conclusion
In the event that the pH and the temperature conditions are not within a specific range, it becomes very difficult for an enzyme to thrive. All proteins react distinctly has a distinct reaction when it is presented in a new environment. The concentration of the lactase and the optimal ranges of pH are reviewed in this experiment, the primary characteristic that was not reviewed in detail was the function of time. The lactase enzyme was the catalysts of the reaction were adapted to endure for approximately eight minutes. A potential change that could be made was to lengthen the amount of time that the experiment endured.
In the application of the lactase tablets, they are most effective in the food articles that possess a pH of 7 and a have a thermal characteristic that is within room temperature and the average body temperature. This is significant with regards to ensuring that the lactase enzyme is an effective agent in the decomposition of lactose and is able to function and derive its optimal benefits. In the absence of harm of the consumer, they would not have the identical results and attain relief from the undesired secondary effects of lactose ingestion when they are lactose intolerant. These findings may seem to be reasonable in consideration of the environment where lactase is normally found. The lactase cells normally thrive in the small intestines. Perhaps the distinctions in temperature could be detailed by the distinctions of temperature at which milk is normally storied in refrigerated conditions and the manner that its can find an equilibrium temperature subsequent to its entry into the human digestive system. There were some errors that were performed while the first day of the experiment was being conducted. These mistakes could be attributed to the unfamiliarity with many of the instruments that were being used. The subsequent collection of experiments was much smoother which caused a reduction in the potential margins of error. A potential field of future research could be to examine the distinct categories of lactose products at distinct temperatures which introducing lactase drops or powder in order to observe if these products have an enhanced enzyme production at the distinct temperature conditions.
References
Farkhauser, D. (2009). Lactase pH optimum. University of Cincinnati Clermont College.
Melton, T. J. (2001). A modification of a lactase experiment by use of commercial test strips. J Chem Wisconsin, 78(9): 1243.
O’ Connell, S. and Walsh, G. (2010). A novel acid stable acid active galactosidase potentially suited to the alleviation of lactose intolerance. Applied Microbiology and Biotechnology, 86(2): 517- 524.
Pope, S. R., Tolleson, T. D., Williams, R. J., Underhill, R. D. and Deal, S. T. (1998). Working with enzymes- Where is lactose digested? J Chem Wisconsin, 75(6): 761.
Sharma, S., Kumar, A., Chaudhury, R. Pudir, C. and Sehgal, N. (2007). Lactose biosensor based on lactase and galactose oxidase immobilized in polyvinyl format. Artificial Cells, Blood Substitutes and Biotechnology, 35(4): 421- 430.
