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The Effect of Concentration, pH, and Temperature of Salivary Amylase on the Rate of Starch Disappearance

The Effect of Concentration, pH, and Temperature of Salivary Amylase on the Rate of Starch Disappearance. By: Caroyln Gee, Helen Zhang, Amanda Kim, Katie Senter . Results. The Effect of % Amylase Concentration on the Time of Starch Disappearance .

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The Effect of Concentration, pH, and Temperature of Salivary Amylase on the Rate of Starch Disappearance

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  1. The Effect of Concentration, pH, and Temperature of Salivary Amylase on the Rate of Starch Disappearance By: Caroyln Gee, Helen Zhang, Amanda Kim, Katie Senter

  2. Results

  3. The Effect of % Amylase Concentration on the Time of Starch Disappearance  ~Graph 1 depicts the data collected from the first experiment, which tested the Effect of % of Amylase Concentration on the Time of Starch Disappearance. ~As depicted, the average time in seconds of starch disappearance lessens as the % concentration of amylase increases. 

  4. The Effect of pH on the Time of Starch Disappearance ~Graph 2 depicts the data collected from the second experiment, which tested the effect of pH on Starch Disappearance. As depicted in the graph, when the pH was low (more acidic), the time required for the disappearance of starch increased.  ~Likewise, when the pH was high (more basic), the time required for the disappearance of starch increased as well. +Note: After 9  minutes, the experiment was stopped because it was such a long time and the amylase may have lost its shape and stopped functioning properly

  5. Discussion

  6. Effect of % Amylase Concentration on Time of Starch Disappearance Hypotheses - Rejected or Supported? 1.) Increased amylase concentration will decrease time of starch disappearance. SUPPORTED ~Evident by the trend line in graph 1, time before starch disappearance decreases as amylase concentration increases.  When the concentration is .25% amylase, the average time for starch disappearance was 16.6 seconds, whereas when the amylase concentration was only about .03%, 223.6 seconds were required for complete starch disappearance

  7. It makes sense that as the enzyme concentration increases, the time of starch disappearance will decrease because there are more chances for substrates to react with a higher concentration Since the rate of reaction is increased, the time it takes for the starch to disappear decreases Other research also supports the data collected--this graph shows the same trend--as enzyme concentration increases, the time to starch disappearance decreases Do the results make sense? http://www.und.nodak.edu/dept/jcarmich/101l ab/tests/test1.html

  8. Relevance in Reality • Concentration of amylase in saliva is 94 x 10^3 U/L (4) • It makes sense that the concentration would be very low because digestion of starch only starts in the mouth with salivary amylase, and finishes later in the digestive system

  9. The Effect of pH on the Time of Starch Disappearance Hypotheses - Rejected or Supported? 2.) As pH nears the optimum temperature for amylase function, time of starch disappearance will decrease. SUPPORTED ~In graph 2, with a pH of 6.8, the amylase took only approximately 30 seconds to break down all the starch.  However, the trend line for the graph is shaped like a parabola with 30 seconds as the minimum value - with a pH of 10, the starch took an average of 8 minutes to disappear, and with a pH of 2, at least 9 minutes was required for the starch to disappear +Note: After 9  minutes, the experiment was stopped because it was such a long time and the amylase may have lost its shape and stopped functioning properly

  10. Do the Results Make Sense? • It makes sense that the time of starch disappearance be the least at pH 6.8 because as shown by other researchers, the optimum activity of amylase is at a pH level around 6 • Since the activity level is at an optimum, it will take less time for the starch to react with the amylase and disappear • Likewise, at pH 8 there is less amylase activity, but the time of starch disappearance increased • If the pH is too high or low, the enzyme would become denatured, causing the decrease in activity and increase in time of starch disappearance  (7) <http://genesdev.cshlp.org/>.

  11. Relevance in Reality • The pH of the human esophagus ranges from about 5-6, and according to the graph on the previous slide and from collected data, salivary amylase functions best between pH levels of 5 and 7, with maximum effect at a pH level of about 6 (6) • Thus, the lab data collected is logical because amylase cleaved starch into simple sugars at a faster rate the closer the pH of the amylase-starch mixture was to 6.8

  12. Sources of Error and how future experiments can be improved

  13. Sources of Error • In Experiment #1, in one of original pieces of data, it took 90 seconds for starch to disappear when mixed with a .125% amylase solution. When the starch was mixed with a .0625% amylase solution, however, it took 50 seconds for starch disappearance. This outlier may be due to the amylase-starch mixture not being mixed properly with the iodine for the test that resulted in the 90 second starch disappearance.  • Furthermore, in Experiment #1, one lab group reported that the .0625% amylase solution took 180 seconds to break down all of the starch, whereas the other groups recorded 50-70 seconds for complete starch elimination •  These strange outliers can possibly be eradicated in future experiments if one participant supervised the iodine and amylase-starch mixture to make sure it was properly mixed in between each 10 second interval.

  14. Sources of Error continued... • There was a time lapse between when the amylase and starch was mixed and when the mixture was added to the iodine (error with experiment that cannot be completely prevented). • However, the time lapse can be improved in future experiments by setting a certain amount of time before the amylase and starch mixture is added to the iodine or by starting the timer the moment amylase is mixed with starch.

  15. How can this experiment be improved? ~ The iodine should be mixed immediately with the starch-amylase mixture so that the time it takes for the starch to disappear is most accurate.  ~ There should be a given shade of the iodine when the time should be stopped. This would improve consistency and accuracy of time of starch disappearance because it would clarify what to look for.

  16. Graph of Amylase Function Regarding Temperature (9)http://www.google.com/imgres?imgurl=http://www.pharmainfo.net/files/images/stories/CTBT/EffectOfIncubationTemperatureonamylasea ctivity.jpg&imgrefurl=http://www.pharmainfo.net/articles/production-%25CE%25B1-amylase-agricultural-byproducts-humicola-lanuginosa-solid-state-fermentation&usg=__fXPlL43cX4jVnCfIy9kF1DHbWwA=&h=281&w=450&sz=15&hl=en&start =0&zoom=1&tbnid=60w53oI8fZYsOM:&tbnh=125&tbnw=200&prev=/images%3Fq%3Damylase%2Btemperature%2Boptimum%26um%3D1%26hl%3Den%26safe%3Dactive%26sa%3DN %26biw%3D986%26bih%3D816%26tbs%3Disch:1&um=1&itbs=1&iact=rc&dur=375&ei=PjC3TN3oNYT6lwfRqIS7DA&oei=PjC3TN3oN YT6lwfRqIS7DA&esq=1&page=1&ndsp=16&ved=1t:429,r:3,s:0&tx=108&ty=86&safe=active

  17. Starch Disappearance at 80, 37, 22, and 4 degrees Celsius • According to other experiments, the time of starch disappearance is least at a temperature of 37 degrees Celsius • This makes sense since the "normal" temperature of human body when taken in mouth is 37 degrees Celsius  (8) http://www.associatedcontent.com/article/8 679/the_effect_of_tem perature_on_amylase_pg2.html?cat=5

  18. Digestive Application

  19. Source: found in pancreas ~Active in small intestine, especially in small animals where pH is around 8 (2) which is close to the pH level of the small intestine, and works best at normal body temperature (95-99.5 degrees Fahrenheit) (3) ~The substrate for this enzyme is protein (2) ~Major products include protein fragments and peptides (2) Trypsin and chymotrypsin are structurally very similar, although they recognize different substrates (1) : ~Trypsin acts on lysine and arginine residues (1) ~Chymotrypsin acts on large hydrophobic residues such as tryptophan, tyrosine and phenylalanine (1) Trypsin and Chymotrypsin

  20. Bibliography 1) "European Bioinformatics Institute | Homepage | EBI." European Bioinformatics Institute | Homepage | EBI. N.p., n.d. Web. 18 Oct. 2010. <http://www.ebi.ac.uk/>. 2) Starr, Cecie, and Ralph Taggart. Biology: The Unity and Diversity of Life. 9 Har/Cdr ed. New York: Brooks/Cole Pub Co, 2001. Print. 106, 734-735 3) "Normal Body Temperature: Rethinking the Normal Human Body Temperature." Harvard Health Publications. Web. 18 Oct. 2010. <http://www.health.harvard.edu/press_releases/normal_body_temperature>. 4)"Laboratory #5-Enzymes."  University of North Dakota.  Web. 19 Oct. 2010. <http://www.und.nodak.edu/dept/jcarmich/101lab/lab6/lab6.html>. 5) Auvdel, M. J., "Amylase Levels in Semen and Saliva Stains," Journal of Forensic Sciences, JFSCA, Vol. 31. No. 2. April 1986, pp. 426-431. <http://projects.nfstc.org/workshops/resources/literature/Amylase%20Levels%20in%20Semen%20and%20Saliva%20Stains.pdf>. 6) "RIVM - Human, Mouth." RIVM - Home. 17 Dec. 2007. Web. 19 Oct. 2010. http://www.rivm.nl/interspeciesinfo/intra/human/mouth/  7) "Genes & Development ." Genes & Development . N.p., n.d. Web. 19 Oct. 2010. <http://genesdev.cshlp.org/>. 8) “The Effect of Temperature on Amylase” Wed 19 Oct. 2010 http://www.associatedcontent.com/article/8679/the_effect_of_tem • perature_on_amylase_pg2.html?cat=5

  21. Bibliography Continued (9) “Effect of Incubation on Temperature on Amylase” Fri Oct 15 2012 http://www.google.com/imgres?imgurl=http://www.pharmainfo.net/files/images/stories/CTBT/EffectOfIncubationTemperatureonamylasea ctivity.jpg&imgrefurl=http://www.pharmainfo.net/articles/production-%25CE%25B1-amylase-agricultural-byproducts-humicola-lanuginosa-solid-state-fermentation&usg=__fXPlL43cX4jVnCfIy9kF1DHbWwA=&h=281&w=450&sz=15&hl=en&start =0&zoom=1&tbnid=60w53oI8fZYsOM:&tbnh=125&tbnw=200&prev=/images%3Fq%3Damylase%2Btemperature%2Boptimum%26um%3D1%26hl%3Den%26safe%3Dactive%26sa%3DN %26biw%3D986%26bih%3D816%26tbs%3Disch:1&um=1&itbs=1&iact=rc&dur=375&ei=PjC3TN3oNYT6lwfRqIS7DA&oei=PjC3TN3oN • YT6lwfRqIS7DA&esq=1&page=1&ndsp=16&ved=1t:429,r:3,s:0&tx=108&ty=86&safe=active

  22. Thank You! Questions?

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