Amylase And Its Functionality At Various Ph Level Abstract Enzymes can be denatured at various ph levels in which then affects the shape of the enzymes structure and reactivity. The altering of shape causes substrates to not bind in the active site (Scott Freeman, Micheal Harrington, Joan. C Sharp, 2009). Amylase is used as a catalytic enzyme to determine the time period to convert starch into glucose monomers and transport into the bloodstream at different ph levels. Methods involved are obtaining six test tubes (different ph levels) with mixture of dilute starch solution and amylase.
Starch indicator solution is used to determine if the starch have been converted to glucose using the catalytic substance amylase. Under the chemical conditions of ph 4, 8 and 10, the amylase has been denatured and the reaction has occurred slowly throughout the time frames. Ph of 6 and 7 worked as the dominant level of ph for the enzyme to catalyze the reactants to produce glucose monomers in a period of sixty minutes. The enzyme (amylase) is only favourable to certain ph levels, in which are 6 and 7.
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Ph levels other then the favourable ones denature the protein by changing its shape which results into a slow activity of the reactants binding to the active site on the enzyme being a critical danger to human cells. Introduction Enzymes play significant role in reactions as they minimize the activation energy for the reaction to proceed to create products. Amylase is a type of enzyme found in human saliva and pancreas secretions which catalyze the complex carbohydrates (starch, glycogen) to glucose (Laursen, A. E. , de Jong, M. , Victorio- Walz, L. , McCarthy, L. & Bostan, V. , Gilbride, K. , 2010). A change in either of the enzymes structures will denature overall causing the enzyme to function inactively. Under specific chemical and physical conditions, the bonds in enzymes can be broken apart. At certain ph levels, the structures of enzymes are altered and and at certain temperatures ,the bonds tend to break apart. Amylase been used as a catalyst for the reaction of starch to glucose, this product is then transported to the bloodstream. However enzymes contain side chains that are ready to lose or gain H+ ions (Laursen, A.
E. , de Jong, M. , Victorio- Walz, L. , McCarthy, L. , & Bostan, V. , Gilbride, K. , 2010). At low ph levels, the enzyme will gain H+ ions causing a shift in shape. At increased ph levels, the enzyme also changes shape due to the loss of H+ ions. Starch being a complex carbohydrate can be alterted to convert into glucose within a change of its catalyst (amylase). Specific ph will denature the enzyme (amylase) while others are in natural favour for the reaction to proceed. Material and Methods Obtained five test tubes labeled ph 4,6,7,8, and 10 at volume 2ml each.
Obtained starch solution and pipette 0. 2ml of 0. 5% starch solution into each of the five labeled test tubes of ph. Gathered starch indicator solution, ceramic mini-well plate and one tube of amylase solution. Placed the five labeled ph test tubes in 37 degrees Celsius water bath, to thermally equilibrate for five minutes. Labeled each well with ph 4, 6, 7, 8 and 10. Added one drop of starch indicator solution to five wells of the ceramic plate containing labels of ph. Took out the test tubes from water bath, and added 0. 02ml of 0. 01% of amylase to all five test tubes.
Used parafilm to cover the top of for all test tubes and inverted it to mix several times. Obtained (pipette) 2ml of solution of ph 4 test tube and transferred one drop of solution to the matched ceramic well (ph 4). Transferred remaining solution to the ph4 test tube. Observed colour from well of ph 4 and determined its colour intensity of 0 being no observable colour to 5 being most intense colour (dark blue). Recorded data on observation chart at time 0 minutes. Repeated previous steps for rest of the ph test tubes containing starch and amylase solution to observe colour intensity at zero minutes.
Placed test tubes back to the water bath to thermally equilibrate. Cleaned ceramic well plate by emptying contents and washing with water. Transferred one drop of starch indicator solution to the each of the labeled ph wells. Obtained test tubes from water bath ten minutes after put in water bath. Repeated similar steps to obtain colour intensity level for the time intervals 10 ,20, 30,45 and 60 min and recorded on observation sheet. Emptied all content in test tube to safe chemical disposal sink and washed ceramic plate.
Results (Results Cont. nd) Under the chemical conditions of ph 6 and ph 7, the amylase tend to worked best as shown in the line graph based on experimental observation. There was little indication of starch present as the colour intensity was rated to be on average around 1 during the 60 minute time frame. At ph 4 and 8 the colour intensity remained above 3 which signified a mass amount of starch present in the solution. Ph 10 showed similar results as ph 4 and 8 but had inconsistent patterns according to other ph levels.
The graph overall portrayed a natural selection for amylase to function between ph of 6 and 7 while other ph below or above tend to denatured causing no little activity of the amylase. Discussion The hypothesis stated to be that amylase will function best or denature at specific ph levels causing the reaction to be incomplete or complete. Based on experimental results, the appropriate level of ph was in the range of 6-7. The experiment is performed for the importance to test how fast the starch will convert into glucose monomers at different ph levels.
The cells in the human body require energy to function. These energy come from glucose which is a product of the catalytic reaction starch to glucose. Having thousands of reactions occurring in human body, amylase lowers the activation energy required for the conversion of starch to glucose to send it to the bloodstream. Without this such catalytic human cells wouldn’t be getting glucose as much as with a catalytic enzyme. Denaturing the structure of amylase will slow down its process which is seen critical as cells require significant amount of energy.
Denaturing the amylase will reduce the normal amount of sugar put into the bloodstream which then causes health complications. Ph 7 is a natural selection for the amylase to function as it does not alter the structure of shape and reactivity and therefore proceeds binding the substrates together. Other experiments similar to the experiment possessed, showed optimum level for amylase to function is at the ph of 7 ( Corneillus Bessler, Jutta Schmitt, Karl-Heinz Maurer, Rold-D Schmid. Jan. 1st. 2009). The amylase started to lose function at below or after the optimum given a certain period of time.
Certain experimental errors have occurred causing the observations to differ from identical experiments done. The pipette process could have been contaminated as one pipette was used to transfer contents. Not all ph test tubes were at exact identical temperatures as some may have cooled down at different temperatures. Future research can be studied upon how to improve the amylase to work at certain ph levels other then the normal. Works Cited: References: 1)Scott Freeman, Michael Harrington, Joan C.
Sharp (2009) Biological Science, Canadian Edition with MasteringBiology. Pearson Prentice Hall (ISBN 0-32-163220-6) 2)2) Laursen, A. E. , de Jong, M. , Victorio- Walz, L. , McCarthy, L. , & Bostan, V. , Gilbride, K. (2010). Ryerson university: biology i lab manual. Toronto: Alicos Copy Centre. 3)Corneillus Bessler, Jutta Schmitt, Karl-Heinz Maurer, Rold-D Schmid. Jan. 1st. 2009. Directed Evolution o f bacterial alpha-amylase: Toward enhanced ph-performance and higher specific activity [Electronic Version]. Protein Science, Volume 12, page 2141-2149.