4thhour

EnzymeQPOELab

Questions:
A. Whatisanenzymeandwhatdoesitdo?
B. Whatistheeffectoftemperatureonenzymerate

KnowledgeProbe:
Enzymesareatypeofproteinthatspeeduptheprocessofachemicalreactionby
loweringtheactivationenergy.Enzymesbindoneormoremoleculestogether,calledthe
substratesorreactants.Eachenzymehasaveryspecificcombinationofsubstratemolecules
thatitcanfit,thisshapeiscalledtheactivesite.Differentenzymescanperformdifferenttasks
somemayweakenchemicalbondswhileotherscreateorstrengthenchemicalbonds.There
aremanydifferenttypesofcatalaseswithawidevarietyofoptimumtemperature.The
optimumtemperatureisthetemperatureorrangeoftemperaturesthattheenzymeproduces
thegreatestamountofproduct.Ifthetemperaturethatanenzymeisworkingundergetstoo
muchhotterthantheoptimumtemperature,theenzymewillgettoolooseresultinginiteither,
notworkaseffectively,ornotworkingatall.Similarly,ifthetemperaturegettoocold,thenthe
enzymewillcondense,againleavingtheenzymetonotworkaseffectively,ornotatall.The
optimumtemperatureinenzymesmainlyrangesfrom1590degreesCelsius.Catalaseisan
enzymethatconvertshydrogenperoxide(H2O2)intooxygen(O2)andwater(H20).Catalase
isusedbyeveryknownanimalineveryorgan.Catalaseisparticularlyprominentintheliver
andcanevencontributetoethanolmetabolism.Becausecatalaseissouniversal,ithas
severaldifferentoptimumtemperaturesinsidevariousorganisms.Forexample,Penicillium
cyclopiumsoptimumtemperatureis15degreesCwhileThermusbrockianusbest
temperatureisat90degreesC.Somecatalaseenzymescanevenworkatfreezingorboiling
pointsbutitisveryrare.Thecatalasethathumansmostusehaveanoptimumtemperatureof
mostoften37degreesC.Forplants,thecatalaseusedhaveanoptimumtemperaturethat
varieswithinthespeciesofplantandtheuseofthecatalase.Theaverageoptimal
temperatureforallcatalaseenzymesisroughly33degreesC.

Prediction:
Wepredictthattheoptimaltemperaturefortherateofcatalasewillbethewarm
temperature(38.2degreesC).Thisisbecause,asthetemperatureincreases,therewillbe
moreenergyaddedtopotato,allowingtheactivesitetoconvertreactantsintoproductsata
fasterratewiththeampleamountofkineticenergy.Wepredictwarmnothotbecauseifthe
enzymegetstoohot,thenitwillbecomedysfunctional.

InvestigationPlan:
Theexperimentisdesignedtomeasureandobservetheimpactthattemperaturehas
uponenzymerate,specificallytheenzymecatalase.Tomeasurethis,weplacedspecific
amountsofhydrogenperoxide,water,foodcoloring,soap,andpotatoinagraduatedcylinder.
Thehydrogenperoxide(H2O2)isasubstratewhichwhenplacedtogetherwithcatalase(in
potato)createstheendproductofOxygen(O2)andwater(H2O).Bymeasuringtheinitial
volumeoftheliquidswiththepotato,andthevolumeafterfourminutes,weareableto
measurethedifferenceinvolume.Ifthecatalaseisworkingatafasterrate,thevolume
increasewouldbegreaterbecauseofmorewaterandoxygen(primarilybubbles).Thefood
coloringisintendedtomaketheliquidmorevisible,increasingmeasurementvalidity.The
soapisusedtodetertheamountofbubbles.Inthisexperiment,wearechangingthe
temperatureofthepotato,containingcatalase,thatisplacedinthegraduatedcylinder.In
ordertominimizeerror,andincreasethevalidityofourresults,therewereseveralvariables
wecontrolled.Theseincludedthetypeofsoap,typeoffoodcoloringused,sourceofwater,
amount/sizeofeachingredientused(potato,hydrogenperoxide,water,soap,andfood
coloring,beakersused(cleanedaftereachtrial),durationofexperiment,amountoftrialsper
temperature,andrelativedistancetemperaturesfromroomtemp.

Procedure:

1.
Gatherallmaterialsnecessaryandplacepotatoslicesintherefrigeratorandthe
freezer
2.
Usingthethreegraduatedcylindersfilleachwith1mLofwater,2mLofHydrogen
peroxide,2dropsoffoodcoloring(.15mL)and3dropsofdishsoapfrompipette(.15
mL)
3.
Usegaugetocutthepotatointo1cm/.7cmpieces(gaugecutswidthat.7cm).
4.
Placeoneroomtemperature(22.8degreesCelsius)potatosliceineachofthethree
graduatedcylinders,recordtheinitialvolumeofallthree,andstartthetimer.
5.
After4minutes,recordthevolumeofeach,includingthebubbles.
6.
Emptyandcleanthegraduatedcylindersusingthescrubberbrush
7.
Retrievethepotatoslicesfromtherefrigerator(6.8degreesCelsius),repeatsteps26
for3trialsatthetemperature.
8.
Retrievethepotatoslicesfromthefreezer(.8degreesCelsius),repeatsteps26for3
trialsatthetemperature
9.
Heatpotatoslicesto38.2degreesCelsius,repeatsteps26for3trialsatthe
temperature.
10.
Heatpotatoslicesto45.3degreesCelsius,repeatsteps26for3trialsatthe
temperature.


Potatoinmicrowave,eitherheatedtoPotatoinrefrigerator,cooledto6.8Potatoisinfreezer,cooledto.8degreesC.
38.2degreesC,or45.3degreesC.degreesC.Potatowedgesarecooled.Potatowedgesarefroze.Potatoisinfor
Potatowedgesisheated.PotatoisinPotatoisinforvisualization.visulization
forvisualization.

Materials:
3graduatedcylinder
4beakers
4pipets
Stopwatch
Potatoes(Hot,warm,normal,cool,cold)
DishSoap
100mLWater
50mLhydrogenperoxide
Papertowels
Red/Orangefoodcoloring
Refrigerator/meansofcooling
Knife
Ruler
Thermometer
Scrubberbrush
PaperforDatacollection
Microwave/meansofheating
PotatoGauger

Observations
QualitativeData:
Thisexperimentwasdesignedtoproducequantitativedataratherthan
qualitativedata.Despitethis,therewasstillsomeimportantqualitativeobservations.One
importantvisualobservationwaswhichtemperatureswouldcontinuetoincreaseinvolume
evenafterthe4minutes.Inbothoftheextremecases,hotandcold,thevolumeincreaseda
littleinthe1stminutehowever,afterthisthevolumedidnotincreaseanymore.Onthe
oppositeside,thecoldtemperature,whichincreasedthemostoverall,continuedtoincrease
involumearound.25mLperminute.Thisshowsthatnotonlydidthecoldincreasetherate

themost,butthatitalsowasabletosustainthisrateforalongerperiodoftime.Whilethe
roomtemperatureandwarmcontinuedtoincreasealittle,itwasonlyataratearound.1mL
perminute.

QuantitativeData:

increase
involume
(mL)

Trial1

Trial2

Trial3

Average

Room
Temp
(22.8C)

1.15

1.2

1.1

1.15

Refrigerat
or(6.8C)

2.3

2.4

2.5

2.4

Freezer
(.8C)

0.4

0.5

0.3

.4

Warm
(38.2C)

0.8

1.2

1.3

1.1

Hot(45.3
C)

0.2

0.3

0.1

0.2

DataAnalysis

Thisgraphshowstheincreaseinvolume(mL)overthefourminutespanforeachof
thefivetemperature.Thegraphclearlyshowsthedifferencesinincreasesinthevarious
temperatures.Thisgraphwasusedbecauseitclearlyshowstherefrigeratedpotato(6.8
degreesCelsius)havingthegreatestincrease,whiletheextremehot(45.2degreesCelsius)
andtheextremecold(.8degreesCelsius)increasingtheleast.

Thisgraphshowshowthepercentageofofvolumeincreasevariedamongdifferent
temperatures.Thisgraphisusednotonlytoshowwhichtemperaturesvolumesincreased
comparedtotheothertemperatures,butalsoshowhowmucheachtemperatureincreased
comparedtowhatitstartedat.

volumeisinmL
Thisgraphisusedtoshowthefinalvolumeofeachtemperatureafterfourminutescompared
totheothers.Thisgraphisverysimilartothe1stgraphwhichshowsjusttheincreasein
volumebecausetheyallstartedat3.4mL.Wedecidedtostillkeepthisgraphbecauseit
representstheheightstheliquidswouldappearatifobservedafter4minutes.

Explanation
Question:
Whatistheeffectoftemperatureonenzymerate?
Claim:
Therateoftheenzymecatalaseinpotatoesisgreatestatacoldtemperature
around6.8degreesCelsius.Ifthetemperaturebeginsgettingeithermuchcolder,ormuch
warmerthanthat,therateofcatalasewilldecrease.Thetemperatureoftheenzymedirectly
impactstheenzymesrate
Evidence:
Throughoutthe15trialsconducted,the3trialsthatwereconductedwithacold
potato(6.8degreesCelsius),thevolumeincrease(mL)wasgreaterthananyoftheother12
trials.Theaveragevolumeincreaseforthecoldpotato(6.8degreesCelsius)wasmorethan
doubletheaveragetemperatureforanyoftheothertemperatures.Theaveragevolume
increaseforthecoldpotatowas2.4mL,whilethenexthighestvolumeincreasewas1.15mL
whichwasatroomtemperature.
Reasoning:
Thedatashowsaveryclearcorrelationbetweenthecoldtemperatureanda
highcatalaserate.Thisstatesthatinnature,catalaseworksatfasterratesincoldclimates
ratherthanwarmclimates.Theevidencesupportstheclaimbecauseifthetemperaturegot
toomuchwarmerfrom6.8degreesCelsius,thentheenzymewouldbecometoolooseto

function.Also,ifthetemperaturebecametoomuchcolderthan6.8degreesCelsius,thenthe
enzymewouldcondensesofarthatitwouldntfunctionproperly.Anexplanationforwhythe
optimaltemperatureisat6.8degreesCisthatwhenpotatoesareplanted,andintheearly
stagesofgrowth,thegroundmayhavetoendurefrostandacoldground.Sincecatalaseis
prominentinpotatoes,theenzymewouldhavetofunctionproperly,andeffectivelyatthis
temperature.Analternateexplanationfortheresultsoftheexperimentisthatoneofour5
temperatureswerenotcloseenoughtocatalasesactualoptimaltemperature.Bynothitting
thispoint,6.8degreesCmaybethebesttemperatureofthe5ratherthantheactualbest
temperature.

Evaluation
A.

Whatarethesourcesoferror?

Somepossiblesourcesoferrorwouldbethatthepotatoessatoutovernightwithholespoked
throughthem.Thismayhavecausedthepotatoestodryout,skewingthedata.Inaddition,
thebeakersweusedweremarkedoffevery.2mL.Theseimprecisemeasurementsmadeit
impossibletotakeanydatamorespecificthan.1mL.Also,therewasashortperiodoftime
afterwemeasuredthetemperature,andwhenweplacedthepotatointhegraduatedcylinder.
Thisshortperiodoftimemayhaveallowedthepotatotoeitherthaworcooloff.Lastly,we
couldhavedonemoretrialswithmoretemperaturestodetermineamorepreciseoptimal
temperature.

B. Whatwouldyoudodifferentlynexttime?
Ifweweretodothisexperimentagain,wewouldhavetakenmoredatawithagreatervariety
oftemperatures.Byonlyhaving5temperatures,wehaveageneralideaofcatalases
preferedtemperaturehowever,weareunsureofamoreprecisetemperaturethatitmay
prefermore.

C. Howconfidentareyouinyourresults?
Weareconfidentinourresults.Weran15trialstotal,butwouldhavelikedtoranmore,with
moretemperaturesandindividualtrialsiftherewasmoretimeallotted.Havingmore
temperatureswouldbridgethegapbetweenroomtemperatureandthewarm/cold.Even
thoughonlythreetrialswereranpertemperature,thedatacollectedwasveryconclusive.In
all15trialstherewerentanysevereoutliers,withthemajorityofthedatacoincidingwiththe
restofthedata.Inaddition,weworkedhardtocontroloutsidevariablesleadingustobe
confident.

D. Whatsurprisedyou?
Wewereverysurprisedthatthepreferredtemperaturewascoldratherthanwarmoreither
roomtemp.Wewereexpectingtoseecatalaseperformbestinawarmtemperaturesince
catalaseisusedbyhumans,whichareatawarmtemperature.Wealsoexpectedittobe
warmerbecauseofthegreateramountofenergysurroundingtheenzyme.Despiteourshock,
ourdatawasveryconclusiveinshowingthecoldertemperaturebeingthebest.

E. Whatwouldyourpredictionbeifyouconductedtheinvestigation
again?
Wepredictthattheoptimaltemperatureforcatalaserateisacoldtemperature(around6.8
degreesC).Thisisbecausecatalaseisprominentinpotatoes,whichareplantedwhilethe
groundisstillcold,andmayhavetoendureseverefrosts.Becauseofthepotentialforthis
coldtemperature,catalasewillpreferitoverothertemperatures.

F. Whatquestionwouldyouliketopursuenext?

Buildingoffofthisexperiment,wewouldlikeresearchhowthetemperaturethatcatalase
workedfastestat,comparedtootherenzymes.Bycomparingthisdatatootherenzymes,we
willgainageneralideaofwhattemperaturetheaverageenzymeconvertsreactantsinto
productsis.

Application:
Onewaythatthiscanbeappliedtoreallifeisifyouwantedtogrowsomepotatoesina
greenhouse,thenthisexperimentwouldgiveyouatemperaturerangetoproducethebest
potatoeswiththemosteffectiveenzymes.Secondlyitcangiveyouarangeoftemperatures
thatarebestforgrowingsoitcangiveyoutheclimateforanareatogrowthebestenzyme
richpotatoes.Lastlyifyouhaveanareathathasbeencontaminatedwithhydrogenperoxide
orsomethingverysimilarthenyoucanplantpotatoesatthespecifictemperaturetocreatethe
bestreactionanddecontaminatethearea.

http://www.answers.com/Q/What_is_the_optimum_temperature_for_catalase
http://www2.vernier.com/sample_labs/BWV06ACOMPenzyme_action_O2.pdf
http://www.biologyonline.org/biologyforum/about3820.html