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Biology 251 (Human Physiology) - Enzymes: Temperature, pH and Specificity Lab

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Question;ONE SENTENCE FOLLOWING ABSTRACT THAT SUMMARIZES DATA RESULTS FOUND IN LAB THATSUPPORTS THIS PARAGRAPH BELOWBy learning how enzymes act as catalysts to lower the activation energy required for areaction to occur in both catabolic and anabolic processes we studied competitiveinhibition, negative feedback, and enzyme specificity using lactase on varioussubstrates as well as the effects of temperature and pH on the ability of lactase to breakdown lactose.HYPOTHESISIF/THEN ASSUMPTION IN A STATEMENT THAT CAN BESUPPORTED BY DATA IN LABPROCEDURESIn exercise one, we prepared a five percent sucrose solution by using the digital scaleto weigh 2.5 g of sucrose. We placed the sucrose into one of the plastic cups providedin the LabPaq. We labeled the cup 5% Sucrose with the marking pencil. Wemeasured 50 mL of distilled water with the graduated cylinder and added the water tothe 2.5 g of sucrose in the 5% Sucrose cup, we mixed it with a stirring rod until thesucrose dissolves. We cleaned the stirring rod thoroughly and dried it. We thenprepared a lactose solution by retrieving one lactase pill and another plastic cup. Wecrushed the lactase pill between two spoons over the plastic cup. We placed thecrushed pill into the plastic cup. We labeled the cup Lactase Solution. We measured200 mL of distilled water with the graduated cylinder and poured it into the plastic cupcontaining the crushed lactase pill, we mixed with a stirring rod until dissolved. Wecleaned the stirring rod thoroughly and dry it. We measured 50 mL of milk with thegraduated cylinder and poured it into a clean plastic cup. We cleaned the graduatedcylinder thoroughly and dried it. We obtained the well plate. We labeled five of thewells A through E using the marking pencil, and followed the directions in the stepsbelow for the contents of the wells. We put three drops of sucrose and three drops ofH2O in test tube A. We put three drops of milk plus three drops H2O in test tube B. Weput three drops of sucrose plus three drops of lactase in test tube C. We put threedrops of milk plus three drops of lactase in test tube D. We put three drops of glucoseplus three drops of distilled water in test tube E. We used a clean, long-stemmed pipetto add three drops of the sucrose solution to well A. We kept this pipet in the sucrosesolution cup to be used later in the exercise. We used a new, clean, long-stemmedpipet to add three drops of distilled water to well A. We kept this pipet in the rinsed outgraduated cylinder to be used later in the exercise. By using a clean, long-stemmedEnzymes: Temperature, pH and Specificity Lab3pipet, we added three drops of milk and three drops of distilled water to well B by usingthe distilled water pipet. We kept the milk pipet in its cup. We added three drops of thesucrose solution and three drops of the lactase solution to well C using the sucrose andlactase pipets. We kept the lactase solution pipet in its cup. We added three drops ofmilk and three drops of the lactase solution to well D using the milk and lactase pipets.By using the pipet containing the 20% glucose solution, we added three drops of theglucose solution and three drops of distilled water to well E using the distilled waterpipet. While we were waiting for the reaction to occur for five minutes, we obtained fiveglucose strips. We labeled them a, b, c, d, and e using the marking pencil. Refer toFigure 6. We dipped a test strip into the solution in well A for five seconds. We gentlyblotted the side of the test strip on a piece of tissue to remove excess solution. If thestrip was not blotted, the glucose concentration will not be accurateit will be tooconcentrated. We waited 3060 seconds for the color to develop, and then comparedthe test strip to the color. We noted that we must take a reading of the test strip at leastno later than the 60-second mark so it wouldnt be inaccurate. We recorded the glucoseconcentration for well A in Data Table 1. We noted that the next reading would producea zero. We tested the glucose concentration in the remaining wells labeled B-E. Werepeated these procedures using one new test strip per well. We recorded the glucoseconcentrations of each well in Data Table 1. We noted that the next reading wouldproduce a zero. We saved the sucrose solution, lactase solution, and milk for the nexttwo exercises. We kept the pipets in their corresponding liquids so that they were notcontaminated with the other solutions. We rinsed the contents of the well plate downthe drain and then thoroughly cleaned and dried the well plate for use in the nextexercise. In exercise two we prepared a boiling water bath. We filled a pot with waterabout 5 to 6 cm deep. We placed the pot onto the stovetop. We placed the metal testtube rack into the pot. We brought the water to a boil. We labeled three 13 x 100 mmtest tubes with the black marking pencil as indicated as, A. 5 mL lactase hot watertemperature, B. 5 mL lactase cold water temperature, and C. 5 mL lactase roomtemperature. We stirred the lactase solution (from Exercise 1) with the lactase solutionpipet. We used the pipet to squeeze 5 mL of the lactase solution into the graduatedcylinder. We poured the 5 mL of lactase solution into test tube A. By using the test tubeclamp, we carefully placed test tube A into the test tube rack in the boiling water bath.We recorded the time that the test tube was placed into the water bath into Data Table2. We placed test tubes B and C in separate clean plastic cups to hold them upright.We measured and poured 5 mL of lactase solution into test tube B, and placed the cupwith the test tube in the freezer. We measured and poured 5 mL of lactase solution intotest tube C, and placed the cup with the test tube on the counter at room temperature.We left the test tubes A and B at the different temperatures for 15 minutes. While wewaited for fifteen minutes we measured the room temperature solution by placing thethermometer into test tube C for 1 minute. We recorded the temperature in our labreport. We removed, washed, and dried the thermometer. After the fifteen minuteEnzymes: Temperature, pH and Specificity Lab4period we carefully placed the thermometer in test tube A (the boiling water bath) for 1minute. We recorded the temperature in our lab report. We removed, washed, anddried the thermometer. We placed the thermometer in test tube B (located in thefreezer) for 1 minute. We recorded the temperature in our lab report. We removed,washed, and dried the thermometer. We labeled the three wells in the well plate: A, Band C as illustrated in Figure 8. We prepared the well plate by adding three drops ofmilk to three different wells leaving one empty well between the milk wells. We labeledthree, clean long-stem pipets: A, B and C. We retrieved 12 glucose test strips. Welabeled four strips A, four strips B, and four strips C. We used the test tube holderclamp to carefully remove test tube A from the rack in the boiling water bath. We placedtest tube A in a plastic cup. We removed the test tube B from the freezer. We notedthat if the solution was too solid to draw into a pipet, we would have to wait 3060seconds for the solution to thaw. By using pipet A, pull a small amount of lactasesolution out of test tube A. We added three drops of the solution to well A. By usingpipet B, we pulled a small amount of lactase solution out of test tube B. We added threedrops of the solution to well B. By using pipet C, we pulled a small amount of lactasesolution out of test tube C. We added three drops of the solution to well C. We waited 5minutes for reactions within the wells to complete. We dipped a test strip into thesolution in well A for 5 seconds. We blotted the test strip with a piece of tissue toremove excess solution. We waited 3060 seconds for the color to develop and thencompared the test strip to the color chart shown in Figure 7. We recorded the glucoseconcentration into Data Table 2. We noted that we must take a reading of the test stripat least no later than the 60-second mark so the results wouldnt be inaccurate. Wetested the glucose concentration in the remaining wells B and C, following the sameinstructions given in the previous steps. We used one new test strip per well. Werecorded the glucose concentration into Data Table 2. After 10 minutes, we dipped anew glucose test strip into each of the three wells. We recorded the glucoseconcentration into Data Table 2. After 15 minutes, we dipped a new glucose test stripinto each of the three wells. We recorded the glucose concentration into Data Table 2.After 20 minutes, we dipped a new glucose test strip into each of the three wells. Werecorded the glucose concentration into Data Table 2. We thoroughly cleaned and driedthe well plate so it is ready for the next exercise. In exercise three we placed the wellplate onto our work surface. We labeled four wells A, B, C and D. We left an emptywell between each labeled well. We put 2 drops of buffer pH 3.5 plus 2 drops of lactaseplus 2 drops of milk in test tube A, we put 2 drops of buffer pH 5 plus 2 drops of lactaseplus 2 drops milk in test tube B, we put 2 drops buffer pH 6.8 plus 2 drops of lactaseplus 2 drops of milk in test tube C. We put 2 drops of buffer pH 11.5 plus 2 dropslactase plus 2 drops of milk in test tube D. We put on our gloves and wore them duringthis exercise. We used scissors to cut off the tips of the pipets that contain the pHsolutions. We set the pipets in a clean plastic cup, bulb-end down. We put two drops ofEnzymes: Temperature, pH and Specificity Lab5pH 3.5 buffer into well A. We put two drops of pH 5 buffer into well B. We put two dropsof pH 6.8 buffer into well C. We put two drops of pH 11.5 buffer into well D. We addedtwo drops of the lactase solution to wells A through D. We added two drops of milk toeach of the wells A through D. We waited 10 minutes for reactions to occur. During the10-minute waiting period, we labeled four glucose test strips A, B, C and D. We thencarefully dipped a test strip into the solution in well A for 5 seconds. We blotted the teststrip with a piece of tissue to remove excess solution. We waited 3060 seconds for thecolor to develop, and then compared the test strip to the color chart shown in Figure 7.We noted that we must take a reading of the test strip at least no later than the 60second mark so they results wouldnt be inaccurate. We recorded the glucoseconcentration into Data Table 3. We tested the glucose concentrations in the remainingwells B-D, following the same instructions given in the previous steps, we used a newtest strip per well. We recorded the glucose concentration in Data Table 3.OBSERVATIONSPROVIDE DATA FOR DATA TABLESData Table 1: Glucose ConcentrationWellsabcdeConcentration of GlucoseEnzymes: Temperature, pH and Specificity Lab6Data Table 2: Presence of glucose in wells indicating lactase activity atvarious temperaturesWellTime (min)510152051015205101520abcConcentration ofGlucoseData Table 3: Glucose in wells a-d indicates enzyme activity at various pHlevelsWellabcdpH3.55.06.811.0Concentration of GlucoseRESULTSANSWER QUESTIONSQuestions:A. What determines a persons ability to digest lactose?B. Which of the wells showed a positive result for glucose? Explain the results.Enzymes: Temperature, pH and Specificity Lab7C. Explain why testing for glucose is used to determine the activity of the enzymelactase.D. Explain the experimental conditions for the five different wells.Questions:A. Graph the effect of temperature on the activity of the enzyme lactase.B. What happens when an enzyme is boiled? Is this effect reversible?C. Based on your experiment results, what is the optimal temperature for lactasefunction?D. Explain what happens as far as the effectiveness of the enzyme at the freezingtemperature.Can this effect be overcome when the temperature rises?Questions:A. Graph the data placing glucose concentrations on the y-axis and the pH values onthe x-axis.B. What was the effect of pH on the enzyme lactase? Is this true for all enzymes?Questions:Faith discussions promote an atmosphere of fellowship andcommunity (Rom 15:7). Please read the discussion topicbelow and a post your comments. Comment and responsesare a minimum of a paragraph consisting of 3-5 lines.Answer from mostly a human physiology standpoint but anysupport from scriptures or faith would be additionallybeneficial.Read Ps 31:10: My life is consumed by anguish and my years by groaning, my strengthfails because of my affliction and my bones grow weak. This verse describes howdespair and other negative feelings take a toll on our physical well-being. There aremany hypothalamic disorders that result in physical maladies such as wasting away(cachexia), obesity, sleep disturbances, dehydration, as well as a wide range ofemotional disorders which over time causes the body great harm. Similarly, in Eccl 8:1:Who is like the wise? Who knows the explanation of things? A persons wisdomEnzymes: Temperature, pH and Specificity Lab8brightens their face and changes its hard appearance. Solomon describes how anxiety,anger, sorrow and frustration can harden ones face, but enjoyment of life and attainingwisdom will reverse these conditions.1. Discuss how stress can causes excessive and prolonged cortisol release and thedisease state that may result from this.2. What character in the bible suffered from severe sleep disturbances? Might thisbe related to a hormonal imbalance? Previously, we discussed the viscera of theabdominopelvic cavity. Splanchnon is the Greek word used to describe variousanatomical, physiological and emotional states. The viscera in theabdominopelvic cavity have its own blood and nerve supply, aptly named thesplanchnic nerves and the splanchnic circulation. The splanchnic nerves are partof the autonomic nervous system (ANS) and so the organs they innervate are notdirectly under our voluntary control. However conscious emotional states(anxiety, fear, happiness, etc.) can influence nervous output to the organs. Inaddition, the adrenal medulla has the same embryological organ as thesympathetic division of the ANS, however it releases epinephrine andnorepinephrine as hormones (instead of neurotransmitters) that circulatethroughout the blood stream contacting many organs. Interestingly, in atheological context, splanchnon in the bible is related to emotions and is alwaysthe responder. The Kardia refers to aspects of the mind and its control overemotions. Thus, the Kardia is considered the originator.3. Discuss the connection between mind and body, and how being in tune withGods will make us feel comfortable and confident as opposed to how we feelwhen out of fellowship with the Lord.CONCLUSIONSUMMARIZETHESEOBJECTIVESWITHBIGPICTUREIDEA/12SENTENCESPEROBJECTIVE.To learn how enzymes workTo understand enzyme specificityTo relate enzyme activity to temperature, pH, and concentrationTo understand the role of enzymes in digestion of lactoseEnzymes: Temperature, pH and Specificity Lab9REFERENCESMarieb & Hoehn (2010). Human Anatomy and Physiology.San Francisco, CA: PearsonBenjamin Cummings

 

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