- - - Please do NOT write on the Pre-Lab - - -

Enzyme Pre-Lab
Using the Enzyme worksheet and Enzyme lab handout answer the Pre-Lab questions the pre-lab must be omplete be!ore beginning the lab" #a kground$ %n this in&estigation' you will study se&eral !a tors that e!!e t the a ti&ity o! enzymes" Enzymes are made up o! amino a ids and ha&e optimal working onditions" The enzyme you will use is atalase' whi h is present in most ells and !ound in high on entrations in li&er and blood ells" (ou will use li&er as the sour e o! atalase" )atalase promotes the de omposition o! hydrogen pero*ide +,-O-. in the !ollowing rea tion$ -,-O-,-O / O-

,ydrogen pero*ide is !ormed as a by-produ t o! hemi al rea tions in ells" %t is to*i and would kill ells i! not immediately remo&ed or broken down" 0ead the entire lab be!ore answering the questions" U1E )O2PLETE 1ENTEN)E1333 4" %n the rea tion abo&e' what is the substrate5 6hat are the produ ts5 -" 6hat is the purpose o! an enzyme5 7" 8e!ine the !ollowing$ a. 9 ti&e site$ b. 1ubstrate$ . Enzyme-substrate omple*$ :" 8es ribe the ;Lo k and <ey= model" +Use a pi ture and des riptions. >" #ased on the ;Lo k and <ey= model' do enzymes get ;used-up=5 ?" 6hat are enzymes made o!5 @" 6hat are the sa!ety pre autions !or this lab5 A" 6hy do we need to use )LE9N test tubes !or ea h part5 B" #rie!ly des ribe the purpose o! ea h part o! the Enzyme lab" 4C" 6rite out the hemi al question !or this lab"

- - - Please do NOT write on this lab - - -

Enzyme Lab
Materials: 0aw li&er Enzyme solution on i e 8ropper Draduated ylinder Dlass rod 1traw Safety: 1a!ety goggles Procedure: 0E)O08 8ET9%LE8 L9# 0E1ULT1 +temperature hange' olor hange' i! bubbles were produ ed' measure the height o! the bubbles' time. )omplete ea h part entirely +in luding the questions. be!ore ontinuing PART 1: Mixing an enzyme and substrate 4" Obtain supplies$ - test tubes and test tube ra k" -" Put -C drops +4ml. o! hydrogen pero*ide in one test tube +you will use the other test tube in part -." +)LE9N 9LL 1P%LL1. 7" 9dd > drops o! sto k enzyme solution + rushed li&er. to the pero*ide" +#E 1U0E TO <EEP T,E ENF(2E 1OLUT%ON ON %)E. Note what happens inside the test tube" :" Geel the bottom o! the test tube and note your obser&ation" >" 8ump solution in the waste ontainer +NOT down the sink. and )LE9N the test tube +use test tube brushes." ?" 9nswer questions" PART : !s an enzyme or substrate c"anged #$used u%&' in reaction( 4" Obtain supplies$ Dlass stirring rod +make sure it does not roll o!! the table." -" Label the test tubes 9 and #" 7" Put a pie e o! li&er +the size o! a ri e grain. into the bottom o! test tube 9' using a glass stirring rod to push it to the bottom o! the test tube" :" Put the tube in a ra k and add :C drops +- ml. o! hydrogen pero*ide" Note what happens and re ord in the table in part -" >" 9llow the rea tion to ontinue until the bubbling stops' stirring gently" ?" Pour only the rea tion liquid into test tube #' do not trans!er o&er the le!t o&er li&er pie es use the glass rod to assist you" @" Put another small pie e o! li&er +the size o! a ri e grain. into test tube #" Note whether or not a rea tion o urs when using the ;used pero*ide= +%s it bubbling5." 0e ord your results in the table in part -" #oiling water 6arm water bath % e bath C"4 2 sodium hydro*ide C"4 2 hydro hlori a id ? test tubes 47 * 4CC mm 7E hydrogen pero*ide p, paper !or eps test tube ra k hot plate test tube holder

<eep lab area )LE9N and O0D9N%FE8

A" Pour another -ml o! hydrogen pero*ide in test tube 9 to see i! the ;used li&er= is still rea ti&e" 0e ord your results in table in part -" B" 8ump solution in the waste ontainer +NOT down the sink. and )LE9N the test tube +use test tube brushes. 4C" Using the results and answer questions" PART ): !s an enzyme s%ecific for only one substrate( 4" Obtain supplies$ straw -" Put 7 ml o! distilled water in a test tube and are!ully blow into the water with a straw !or 4 minute" )arbon dio*ide +)O-. is produ ed in rea tions in li&ing ells" #lowing into the tube will add arbon dio*ide to the water whi h will !orm arboni a id" 7" 9dd a small pie e o! li&er to the test tube to see what e!!e ts the enzyme atalase +!rom the li&er. has on arbon dio*ide" 0e ord your results in the table in part 7" :" 8ump solution in the waster ontainer +NOT down the sink. and )LE9N the test tube +use test tube brushes. >" 9nswer questions" Part *: Effects of %+ on enzyme acti,ity 4" Obtain a total o! ? test tubes' label test tubes 9' #' )" 8o not label the other 7 -" 9dd - ml o! distilled water to test tube 9" Dently swirl the test tube" 7" 9dd - ml o! hydro hlori a id +,)l. to test tube #" Dently swirl the test tube :" 9dd - ml o! sodium hydro*ide +NaO,. to test tube )" Dently swirl the test tube" >" 9dd 4 ml o! sto k enzyme solution to test tubes 9' #' )" ?" )LE9N-UP 9LL 1P%LL1" #e sure not to splash any hemi als in your !a e" @" Use a small pie e o! p, paper and tweezers' tilt the test tube slightly to test the p, o! test tube 9' # and ) +use a new pie e !or ea h test tube." 0e ord results" A" To ea h o! the three unmarked tubes add 4 ml o! hydrogen pero*ide" B" 9T T,E 192E T%2E' pour the ontents o! the unmarked tubes into ea h o! the labeled test tubes" 4C" 0e ord your results" Note the relati&e rates o! ea h indi&idual rea tion" 44" 8ump solution in the waster ontainer +NOT down the sink. and )LE9N the test tube +use test tube brushes. 4-" 9nswer questions" Part -: Effects of tem%erature on enzyme acti,ity 4" Using your leaned test tubes' label three ;9=' ;#=' ;)=" 8o not mark the other three" -" To ea h o! the labeled tubes' add 4 ml o! sto k enzyme solution +li&er atalase." 7" 9T T,E 192E T%2E$ put tube 9 in boiling water bath +4CC H). U1E TE1T TU#E ,OL8E0 put tube # in warm water bath +about 7@ H). U1E TE1T TU#E ,OL8E0 put tube ) in an i e bath +C H). :" 6ait : minutes" 6hile you wait' add 4 ml o! hydrogen pero*ide solution to ea h o! the three unmarked test tubes" >" 9!ter : minutes' remo&e the tubes !rom their baths and put them in a test tube ra k" ?" 9T T,E 192E T%2E' add the ontents o! the unmarked tubes to ea h tube" 9' #' )" @" Obser&e the results and #E 1U0E TO NOTE T,E 09TE OG E9), 0E9)T%ON3 0e ord your results" A" )LE9N 9N8 0ETU0N ALL 1UPPL%E1 B" 9nswer questions"

Name$ IIIIIIIIIIIIIIIIII Period$ IIII

Enzyme Lab Results S"eet

0E)O08 8ET9%LE8 L9# 0E1ULT1 +temperature hange' olor hange' i! bubbles were produ ed' measure the height o! the bubbles' time. PART 1: Mixing an enzyme and substrate Obser&ation notes$

4" 6hat was the sour e o! the temperature hange5

-" 6hat is a possible e*planation !or the bubbling during the rea tion5 +6hat hemi al might be being released5.

7" The sto k enzyme solution was made by rushing a pie e o! li&er in a small amount o! water" This released many mole ules o! enzyme + atalase. into the water" The rea tion mi*ture in your test tube ontained hydrogen pero*ide +the substrate !or the rea tion. and this enzyme atalase" #elow are three possible e*planations$ a. The enzyme' atalase' and the substrate' hydrogen pero*ide' rea t together to !orm a new produ t and both hanged during the rea tion" b. The substrate' hydrogen pero*ide' is hanged during the rea tion' but the enzyme' atalase' is un hanged" . The enzyme' atalase' is hanged during the rea tion' but the substrate' hydrogen pero*ide' is un hanged" 6hi h o! the abo&e e*planations do you think is orre t5 E*plain why"

PART : !s an enzyme or substrate c"anged #$used u%&' in reaction( 1tep J Tube : 9 Enzyme 1our e Gresh pie e o! li&er 1ubstrate 1our e - ml o! pero*ide 0esults

? @

# 9

Gresh pie e o! li&er ;Used= li&er +9.

;Used= pero*ide +9.

+rea tion liquid !rom 9.

- ml !resh pero*ide

4" 8oes the pero*ide substrate +used pero*ide. hange a!ter the rea tion5 ,ow do you know' use e&iden e !rom the lab5

-" 8oes the enzyme +used li&er. hange a!ter the rea tion5 ,ow do you know' use e&iden e !rom the lab5

7" %! le!t open in the sunlight' a bottle o! hydrogen pero*ide would slowly break down into water and o*ygen" Under these onditions' it takes a long time !or the rea tion to o ur" On the other hand' when atalase is present' this rea tion takes pla e almost immediately" 6hat does this tell you about the !un tion o! an enzyme5

PART ): !s an enzyme s%ecific for only one substrate( 1tep J Tube 7 Enzyme 1our e 1mall pie e o! li&er 1ubstrate 1our e )arboni a id 0esults

Using your data olle ted in parts 4-7 answer the questions below$ 4" 6rite out the !ormula !or the breakdown o! hydrogen pero*ide in the ell"

-" 6hat role does atalase play in keeping the ell in a healthy en&ironment5

7" %s this enzyme !ound only in the li&er5 +,int$ i! you put hydrogen pero*ide on a ut !inger' it bubbles.

:" )ompare your results !or the a tion o! the li&er atalase on the two substrates +hydrogen pero*ide in part and arboni a id. you ha&e tested so !ar" 8id the enzyme atalase ha&e an e!!e t on the arboni a id +!ormed when you blew bubbles into the water.5 #ased on your results' is an enzyme substrate spe i!i 5 E*plain"

Part *: Effects of %+ on enzyme acti,ity Tube 9 # ) )ontents 8istilled water / 4 ml enzyme ,)l / 4 ml enzyme NaO, / 4 ml enzyme p, 0esults

4" 6rite out an e*planation about the possible e!!e ts o! p, on enzyme a ti&ity"

-" 6hat e!!e t does a high p, +base. ha&e on the enzyme a ti&ity5 Use e&iden e to support your answer"

7" 6hat e!!e t does a low p, +a id. ha&e on the enzyme a ti&ity5 Use e&iden e to support your answer"

:" 6hat e!!e t does a neutral p, ha&e on the a ti&ity o! the enzyme5 Use e&iden e to support your answer"

>" 6hi h p, is optimal !or this rea tion to o ur5 ,ow does this relate to what we know about the bodyKs normal p, le&el5 Use e&iden e to support your answer"

?" Draph p, &ersus enzyme a ti&ity rate +slow' medium' !ast." Label the a id' base and neutral p, ranges and use an arrow to show the optimum rate and p, le&el !or this rea tion"

!ast 0ate o! 0ea tion medium slow

? p,

4C

4-

4:

Part -: Effects of tem%erature on enzyme acti,ity Tube 9 # ) )ontents 4 ml enzyme / 4 ml hydrogen pero*ide 4 ml enzyme / 4 ml hydrogen pero*ide 4 ml enzyme / 4 ml hydrogen pero*ide Temp 4CC H) 7@ H) C H) 0esults

4" 9t whi h temperature was the optimal !or enzyme a ti&ity5 Use e&iden e to support your answer"

-" Draph the enzyme a ti&ity rate +slow' medium' !ast. &ersus temperature" Use an arrow to point out the area o! the graph showing the optimal rate and temperature at whi h the rea tion o urred"

!ast 0ate o! 0ea tion medium slow

-> >C @> Temperature H)

4CC

7" 6hat is the e!!e t o! the temperature on enzyme a ti&ity and how does this relate to the normal body temperature o! mammals5 Use e&iden e to support your answer"