Oct 02, 2017 term paper 2

Investigate the effect of different temperatures on the activity of pepsin (enzyme) with HCL and egg white?

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Biology: HCL Egg

Question:

Investigate the effect of different temperatures on the activity of pepsin (enzyme) with HCL and egg white?

Answer:

Evaluation

The egg white suspension is cloudy because of the presence of solid albumin particles suspended in water. After addition of hydrochloric acid it is seen that the solution is clear because the particles have been dissolved. So, from in-vitro analysis it can be said that the pepsin enzyme dissolves protein inside the stomach, in the presence of hydrochloric acid, at optimum temperature. While performing the experiment the data accuracy and the source of error should be identified and need to be improved. In this experiment to check standard enzymatic activity, four different temperatures have been selected. In this analysis four different temperatures are considered below 10◦C, 23◦C, 37◦C and 60◦C. Enzymes are denatured above 45-50◦C which means the activity of that particular enzyme is damaged by boiling. Then for individual temperature an average has been taken out of three trials. The result showed that enzymatic activity more than 10◦c and 60◦c contain the highest range of variation. To check the accuracy of data trial number need to be increased, which confirms data accuracy. If we consider the result analysis in terms of point estimation the chances of error is high hence we consider the data analysis in terms of interval estimation. To reduce chance of error the interval time between two consecutive trials should be minimized. This will ultimately reduce the deviation in results in each of this trial.

Based on this study, thus it can be said that protein is not dissolved by boiled enzyme. Egg white or albumin is proteineceous in nature and the experiment suggests that pepsin can digest this protein. But, based on this experiment a generalized conclusion should not be made, where it should not be stated that all enzymes will dissolve protein at same temperature and condition. If color change takes longer time that means reaction rate is slow, so temperature is either less or more than the optimum temperature. On the other hand if color change takes less time that means reaction rate is fast and temperature is optimum. Error represents the possibility of deviation of the standard result. In this experiment, considering the variables if we say that the range shows for 37◦c is standard then as per the formula, presence of minimum 25% error is possible and possibility is greater for more than 37◦c.  In this graph the X-axis presented as temperature and Y-axis presented as the time taken for the reaction. From this graph it is obvious to mention that temperature at different degrees affect enzymatic activity and the rate of reaction in solution. The most important part is the change of solution color which indicates the reaction rate and time. The graph illustrates that at 10◦C the time taken for completion of reaction is more (7minutes), means the rate of reaction is less; comparatively at 23◦C the time taken for completion of reaction is less (1.2minutes); at 37◦C the time taken for completion of reaction is 0.4minutes and at 60◦C the time taken for completion of reaction is 6.7minutes, which is again increased. Therefore, it can be stated that at optimum temperature the enzyme reaction rate is faster and hence the time required is minimum. In this condition we can say that time is equally proportional to the rate of reaction. In both the extreme cases, less and above optimum temperature the reaction rate will increase.  On the other hand, solution color change is considered as a good presentation of enzyme activity. The solutions change its color when all protein particles inside the solution get saturated completely by the enzyme.

To minimize the random error trial frequency need to be maximized and also there should be a proper time balance between each trial.

Application

Enzymes break down complex molecule (for example: carbohydrates, proteins and fats) into simple form. This process takes place during food digestion inside the intestinal tract.  Enzymes are responsible for energy release and storage, respiration and many more physical mechanisms (Ikemoto, Kurahashi and Shi, 2012). Each and every enzyme has their own optimum pH, which is the most favorable condition for them to work (Lee and Song, 2010). Enzymes speed up the reaction rate (catalysis) or slower the reaction rate (catalysis) at a particular pH (Purich, 2002). Defective enzyme or deficiency in enzyme activity can result in different hereditary diseases or metabolic disorders. Therefore, it is important to understand enzyme activities, their substrates, rate of reaction along with optimum environment (Frey and Hegeman, 2007). As different enzymes work well at different temperatures and pH, they are used commercially, for example: antibiotic synthesis, preparation of washing powders. Some domestic products utilize enzymes to increase the chemical reaction, for example: enzymes are used in washing powders to breakdown fat or protein strains on clothes or tenderization of meat (Knutsen and Liberatore, 2010). Therefore, understanding the enzyme activity, enzyme-substrate complex, rate of reaction are of utmost importance. Particularly, this experiment helps us to understand the optimum temperature for enzyme pepsin is 37◦c with hydrochloric acid, where the substrate is albumin (Mueller, 2006). This understanding might further help in enzyme modification and research for further applications in different industries.

References

Frey, P. and Hegeman, A. (2007). Enzymatic reaction mechanisms. Oxford: Oxford University Press.

Ikemoto, T., Kurahashi, I. and Shi, P. (2012). Confidence interval of intrinsic optimum temperature estimated using thermodynamic SSI model. Insect Science, 20(3), pp.420-428.

Knutsen, J. and Liberatore, M. (2010). Rheology Modification and Enzyme Kinetics of High Solids Cellulosic Slurries. Energy Fuels, 24(5), pp.3267-3274.

Lee, S. and Song, W. (2010). Surface modification of polyester fabrics by enzyme treatment. Fibers and Polymers, 11(1), pp.54-59.

Mueller, R. (2006). Biological degradation of synthetic polyesters—Enzymes as potential catalysts for polyester recycling. Process Biochemistry, 41(10), pp.2124-2128.

Purich, D. (2002). Enzyme kinetics and mechanism. San Diego: Academic Press.


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