CHEM 212 LAB REPORT 3 2023| PORTAGE LEARNING|WITH PROCEDURES, EXPLANATION AND FINAL RESULTS.HIGHLY VERIFIRD Experiment #: Lab 3 Title: Enzymatic Activity of Lactase. Purpose: The purpose and goals of this experiment are 1. Isolate an enzyme. The student should be able to explain how to isolate an enzyme and be able to isolate an enzyme in the lab. The student should also be able to explain and perform 2. Assay enzyme activity spectrophotometrically, 3. Investigate the temperature effect on enzyme activity. 4. Assess thermal denaturing conditions on enzyme activity. The student should be able to explain, recall, and perform all these goals after learning them in the current lab. Procedure: • The isolation of lactase o Procedure: ▪ Step 1: Put on appropriate lab equipment based on what solutions and chemicals will be used in the lab that they are performing. Ex: Gloves, Lab coat, Closed toe shoes, Protective glasses, etc. Also have the appropriate materials needed for completion of the lab. ▪ Step 2: Take one pill from a lactase packet. And crush pill into powder form in a mortar and pestle. ▪ Step 3: Place powder into plastic test tube and add 10 mL of phosphate buffered saline. ▪ Step 4: Place plastic test tube on vortex machine, place on machine until solution is fully mixed. Which should be around 30 seconds to 1 minute. ▪ Step 5: Using a pipette, take 1 mL of solution and place into microcentrifuge tube and place into centrifuge. ▪ Step 6: Set to 10,000 revolutions per minute, set timer for a minute. Make sure there is a counterbalance in centrifuge for proper spinning. ▪ Step 7: Remove liquid by setting pipette 500 mL, then place liquid in tube labeled extract. • Initial qualitative lactase activity assay o Procedure: ▪ Step 1: Put on appropriate lab equipment based on what solutions and chemicals will be used in the lab that they are performing. Ex: Gloves, Lab coat, Closed toe shoes, Protective glasses, etc. Also have the appropriate materials needed for completion of the lab. ▪ Step 2: Pipette 390 mL of phosphate buffer saline to a microcentrifuge tube. ▪ Step 3: Pipette 100 mL of ONPG to solution tube.
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