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Liver Enzyme Lab Purpose: How will enzyme activity be affected by manipulating independent variables? Background Information: In each individual cell of a human there are many chemical reactions taking place, performing the necessary functions for being a large, complex, multicellular organism. This is relatively easy to understand. How do these reactions occur? This is not so easy to understand. Chemical reactions involve the breaking and reforming of chemical bonds between molecules (substrates or reactants), which are transformed into different molecules (products). Some chemical reactions can occur spontaneously. Meaning, the chemical reaction can occur without any added energy or intervention. While certain chemical reactions occur spontaneously, other chemical reactions cannot and instead, involve specific metabolic pathways for the reaction to happen. Enzymes are biological protein catalysts. They help to increase the rate of chemical reactions. Enzymes are most often proteins and their three-dimensional (3-D) shape is important to their catalytic activity. Because of their 3-D shape, enzymes are highly specific for the reactions they catalyze. Enzyme activity and effectiveness is influenced by many factors. First, enzyme activity may be negatively affected by changes in temperature. Normally, enzymes work at a particular temperature optimum and if the temperature is either too high or too low, the enzyme activity is reduced. Since enzymes are proteins, enzyme activity is also affected by changes in pH (measure of acidity). If the pH changes are too drastic, enzymes can become denatured and their activity is reduced or eliminated. Lastly, enzyme activity can be affected by concentration. Concentration refers to how much of the enzyme is present. In this lab, you will be conducting a series of experiments to look at how enzyme activity is affected by changes in temperature and pH. The enzyme used in this experiment is called, catalase. Catalase is found in many cells but is in highest levels in the liver because the liver functions to break down toxins present in the blood. Further, catalase is responsible for breaking down hydrogen peroxide (H 2 O 2 ) and produces oxygen gas and water. As the reaction occurs, bubbles form, indicating that hydrogen peroxide is being broken down in water and oxygen gas. The more active the enzyme, the more oxygen (and water) it produces. The specific chemical reaction that we will be studying is the decomposition of 6% hydrogen peroxide into water and oxygen gas. The formula for the reaction is as follows: 2H 2 O 2 Catalase 2H 2 O+O 2 Hydrogen Peroxide Water Oxygen PRELAB Questions: You do not have to write out the questions, BUT you do have to answer in complete sentences. 1. What are the four macromolecules and which macromolecule are enzymes? 2. What is the function of enzymes and why are they important for our cells? 3. Write down the equation for today’s lab and do the following: highlight the reactant(s) above yellow. Highlight the product(s) in the equation above green. 4. What indication will we have that the enzyme is working (Hint: Look at the products, what would have these together look like?) 5. What three factors will affect the enzyme activity?

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LiverEnzymeLabPurpose:Howwillenzymeactivitybeaffectedbymanipulatingindependentvariables?BackgroundInformation:Ineachindividualcellofahumantherearemanychemicalreactionstakingplace,performingthenecessaryfunctionsforbeingalarge,complex,multicellularorganism.Thisisrelativelyeasytounderstand.Howdothesereactionsoccur?Thisisnotsoeasytounderstand.Chemicalreactionsinvolvethebreakingandreformingofchemicalbondsbetweenmolecules(substratesorreactants),whicharetransformedintodifferentmolecules(products).Somechemicalreactionscanoccurspontaneously.Meaning,thechemicalreactioncanoccurwithoutanyaddedenergyorintervention.Whilecertainchemicalreactionsoccurspontaneously,otherchemicalreactionscannotandinstead,involvespecificmetabolicpathwaysforthereactiontohappen.Enzymesarebiologicalproteincatalysts.Theyhelptoincreasetherateofchemicalreactions.Enzymesaremostoftenproteinsandtheirthree-dimensional(3-D)shapeisimportanttotheircatalyticactivity.Becauseoftheir3-Dshape,enzymesarehighlyspecificforthereactionstheycatalyze.Enzymeactivityandeffectivenessisinfluencedbymanyfactors.First,enzymeactivitymaybenegativelyaffectedbychangesintemperature.Normally,enzymesworkataparticulartemperatureoptimumandifthetemperatureiseithertoohighortoolow,theenzymeactivityisreduced.Sinceenzymesareproteins,enzymeactivityisalsoaffectedbychangesinpH(measureofacidity).IfthepHchangesaretoodrastic,enzymescanbecomedenaturedandtheiractivityisreducedoreliminated.Lastly,enzymeactivitycanbeaffectedbyconcentration.Concentrationreferstohowmuchoftheenzymeispresent.Inthislab,youwillbeconductingaseriesofexperimentstolookathowenzymeactivityisaffectedbychangesintemperatureandpH.Theenzymeusedinthisexperimentiscalled,catalase.Catalaseisfoundinmanycellsbutisinhighestlevelsintheliverbecausetheliverfunctionstobreakdowntoxinspresentintheblood.Further,catalaseisresponsibleforbreakingdownhydrogenperoxide(H2O2)andproducesoxygengasandwater.Asthereactionoccurs,bubblesform,indicatingthathydrogenperoxideisbeingbrokendowninwaterandoxygengas.Themoreactivetheenzyme,themoreoxygen(andwater)itproduces.Thespecificchemicalreactionthatwewillbestudyingisthedecompositionof6%hydrogenperoxideintowaterandoxygengas.Theformulaforthereactionisasfollows:

2H2O2Catalase2H2O+O2 HydrogenPeroxideWaterOxygenPRELABQuestions:Youdonothavetowriteoutthequestions,BUTyoudohavetoanswerincompletesentences.

1. Whatarethefourmacromoleculesandwhichmacromoleculeareenzymes?2. Whatisthefunctionofenzymesandwhyaretheyimportantforourcells?3. Writedowntheequationfortoday’slabanddothefollowing:highlightthereactant(s)aboveyellow.

Highlighttheproduct(s)intheequationabovegreen.4. Whatindicationwillwehavethattheenzymeisworking(Hint:Lookattheproducts,whatwouldhave

thesetogetherlooklike?)5. Whatthreefactorswillaffecttheenzymeactivity?