Enzymatic Activity of Salivary Amylase

Enzymatic Activity of Salivary Amylase Ong, Janela Rose I. Paguia, female horse Tricia C. Placente, Dax Daven A. Posadas, Grace Catherine A. 3Bio3-Group 8 Department of Biological Sciences, College of Science University of Santo Tomas, Espana, Manila 1008 solicit This sample aims to examine the enzymatic activity and specificity of salivary amylase depending on the changes in pH and temperature and determine the optimum temperature and pH of the amylase. EXPERIMENTAL In the enzymatic activity of salivary amylase, 1ml saliva, 9ml distilled water and 30ml of 0. % NaCl made up the enzyme rootage. One percentage starch in phosphate buffer pH 6. 7 was the buffered starch. The experiment was comprised of both parts. For the first part (effect of temperature), 2 ml of the enzyme solution was set in a large raise tubing and label as 4?. In a separate large political campaign tube, 2 ml of the buffered starch solution was added. Both test tubes were incubated for 10 legal proce eding in an ice bath with a temperature of 4?. The two test tubes were IMMEDIATELY mixed aft(prenominal) 10 minutes, and three-drops were QUICKLY taken from the sort.The three-drops of the concoction simultaneously added with two drops of one solution were dropped onto the first well of a spot plate and was labelled as the zero minute. Incubation should be continued and after a one-minute interval, three-drops of the mixture simultaneously added with two drops of single solution were dropped onto the instant well of a spot plate and was labelled as one minute. The test tube of the iodine solution should not be exposed. It should be covered with a cork and wrapped almost with aluminium foil. The uniform step of taking three-drops of the mixture simultaneously added ith two drops of iodine solution was repeated until a light yellow-colored solution was observed. cartridge holder should be noted. The first part of the procedure was not only cerebrate on 4? but also other temp eratures such as room temperature, 37? , 50? , 60? and 70?. All temperatures were incubated at the desired incubation temperature. The mutual of time (1/t, min-1) was plotted versus the temperature (T) and the optimum temperature of the amylase was determined. For the second part (effect of pH), 1 ml of acetate buffer with ph 4 and 1 ml of 2% unbuffered starch were mixed in a large test tube.In a separate large test tube, 2ml of the enzyme solution was added. Both test tubes were incubated for 10 minutes in a 37? water bath. The two test tubes were IMMEDIATELY mixed after 10 minutes and three-drops of the mixture were QUICKLY and simultaneously added with two drops of the iodine solution onto the first well of a spot plate. This was labelled as the zero minute. Incubation should be continued and after a one-minute interval, three-drops of the mixture simultaneously added with two drops of iodine solution were dropped onto the second well of a spot plate and was labelled as one minu te.The test tube of the iodine solution should not be exposed also and it should be covered with a cork and wrapped around with aluminium foil. The comparable step of taking three-drops of the mixture simultaneously added with two drops of iodine solution was repeated until a light yellow-colored solution was observed. Time should be noted. The same steps for the second part of the experiment should be followed for other pH such as 5, 6. 7, 8 and 10 using the appropriate buffer. The reciprocal of time (1/t, min-1) was plotted versus the buffer pH and the optimum pH of the amylase was determined.