Alpha Amylase
Biology 181
Ana Marti-Subirana
Identification of unknown a-Amylase through testing different temperatures and pH values to detect the absorbance of maltose.
Introduction:
Enzymes are biological catalysts, mainly proteins for this experiment, generated by an organism to speed up chemical reactions. They have active sites on which the substrate is attached, and then broken up or joined. For this experiment we are going to work with the enzyme a-amylase. Amylase is an enzyme that breaks starch down into sugar. Amylase is present in human saliva, where it begins the chemical process of digestion. Foods that contain starch like rice and potatoes. Amylase turns some of their starch into sugar in the mouth. The pancreas makes amylase (alpha amylase) to hydrolyze dietary starch into disaccharides and trisaccharides which are converted by other enzymes to glucose to supply the body with energy. Alpha amylase in this lab is used to hydrolyze starch to break it down into maltose. Purpose of this lab is to show that both pH temperature affect enzyme actions. The optimal temperature for the enzyme from Aspagillus is 37°C, and an acidic pH of 4-6 whereas, the enzyme from the hot spring bacteria is 100°C and acidic pH of 4-6. For the enzyme from the pig pancreas, optimal temperature is 37°C and a neutral pH of 7.
Material and Methods:
Part I: Effect of Temperature on a-amylase activity
To be recorded in a notebook are which a-amylase preparation to be used (A,B,C). One test tube labeled as “blank” other five test tubes as :“4°C,”23°C, ”37°C,”
“65°C,” and “100°C.” 1 ml of 1% starch and 1ml of pH 7 was added to each tube. The substrate of the reaction was starch. Of the indicated temperatures, one tube was placed in each. At room temperature the “blank” and 23°C tubes are to be left, furthermore, the 4°C tube to be placed on ice. Equilibrate the tubes at each temperature for 10 mins.100ml of the concentrated a-amylase stock solution was added to 9.9 ml of
Ana Marti-Subirana
Identification of unknown a-Amylase through testing different temperatures and pH values to detect the absorbance of maltose.
Introduction:
Enzymes are biological catalysts, mainly proteins for this experiment, generated by an organism to speed up chemical reactions. They have active sites on which the substrate is attached, and then broken up or joined. For this experiment we are going to work with the enzyme a-amylase. Amylase is an enzyme that breaks starch down into sugar. Amylase is present in human saliva, where it begins the chemical process of digestion. Foods that contain starch like rice and potatoes. Amylase turns some of their starch into sugar in the mouth. The pancreas makes amylase (alpha amylase) to hydrolyze dietary starch into disaccharides and trisaccharides which are converted by other enzymes to glucose to supply the body with energy. Alpha amylase in this lab is used to hydrolyze starch to break it down into maltose. Purpose of this lab is to show that both pH temperature affect enzyme actions. The optimal temperature for the enzyme from Aspagillus is 37°C, and an acidic pH of 4-6 whereas, the enzyme from the hot spring bacteria is 100°C and acidic pH of 4-6. For the enzyme from the pig pancreas, optimal temperature is 37°C and a neutral pH of 7.
Material and Methods:
Part I: Effect of Temperature on a-amylase activity
To be recorded in a notebook are which a-amylase preparation to be used (A,B,C). One test tube labeled as “blank” other five test tubes as :“4°C,”23°C, ”37°C,”
“65°C,” and “100°C.” 1 ml of 1% starch and 1ml of pH 7 was added to each tube. The substrate of the reaction was starch. Of the indicated temperatures, one tube was placed in each. At room temperature the “blank” and 23°C tubes are to be left, furthermore, the 4°C tube to be placed on ice. Equilibrate the tubes at each temperature for 10 mins.100ml of the concentrated a-amylase stock solution was added to 9.9 ml of