Introduction:

I-nzymcs arc proteills that arc uscd as c.ltalvsts in biochcnical rcactions. A ciital)/st is sortrelhing that controls thc rltc ofa
feaction without being changcd thenrsclvcs. Olicn cnzyrnes arc tiscd to speed lrp the ratc of a reactioD. Flowever, thcle arc
somc lactors that can lirrit horv rvcll an enzyrue rvill rvork. Trvo ofthc Drain lactors thall can allcct enzynlc rates ate
ternperature and the pH olthe solution the cnzyrne is in. Iflhe tempemturc ofthe solution is too lorv, cnzyrnes may not bc
able to lower the activation energy enough to allow thc rcactiou to continuc. 'fhe enzyrle is unaffcctod by thc lower
temperalure. There is just not enottgh energy in tbe environrnent for the cherllical [eaction to continuc. I1'the tentpcrature
or pH ofthe solution is too bigh, the cnzyme rvill becomc denatured, or changc its chcmical shape, so the enzyne is unable
to altach to the substlate aud bfeak it apart.

'I'his investigation
enzyrne function.

will look

at the rate an enzymc is able to catalyzc a substrate and the effects ofbeing dcnatured has on

Procedure:
Paft A: Rate ofProduct Formation
1. Make 5 piles of40 toothpicks in each pile.
2. In groups of3, one person will be the enzyn toothpickase, one person rvill clean/remove broken toothpicks, and tho
third pelson will calcuiate the numbel oftoothpicks broken for each pile.
3. When the instructor tell the class when to begin, the nzyrne (toothpickase) will begin to break the substrate
(toothpicks) one at a tine starting at the first pile. When the instructor shouts "switch", the enzlane wil
IMMEDIATELY move to the next pile and contirue to break toothpicks.
4. The instructor will call out "switch" after l0 seconds,30,60, 120, and l80seconds.
5. Once toothpickase has moved to the next pile, student 2 should remove the broken pieces from the desk, and student 3
should begin calculating the uumber oftoothpicks broken.
6. At the end of 180 seconds, the data should be recorded in the table provided and the rates ofenzyme fihction will be
calculated.
7. Gmph data collected in Tables I
Rate of enzyrre activity

Table

nurnber of toothpicks brol<en

Tirrre (at each pile)

Total Time
(sec)

0
10

30

(additional 20 sec)
60

(additional 30 sec)
120
(additional 60 sec)
180

(additional 60 sec)

Toothpicks
metabolized
(broken) on
each plate

Rate of
enzylne

activity

Total
Toothpicks
Metabol rzed
(Broken)
over tirne

l.[1 l]: ii,,lc ol l),]t:r rcJ j ./]trtr'.

i.
2.

7.

i\1rkc 5 pilcs of40 bolhpicl(s in cach pile.

in gfoups ol'j, ona pelson rvill be the enzyrne toothpicl(ase, one pelson lvillclean/rcrnove blokcn toothpicl(s, and the
thifd ircfson will cl1laulate thc nltlnbcr oftoothpicl<s broken fbr each pilc.
Studcnt l, tootllpici(ase, rvill bccome detratuled by taping lheir thur)rbs clo\\,r to lhcir first lingcr on cach hand.
Whcn thc instnrctor tell the class rvhen to begin, the enzryme (toodlpickase) rvill begin to bleal( the substlatc
(toothpicks) onc at a tilne starti|g at the flrst pile. When the instnlctor shouts "swi1clt", tlrc cnzyme rvill
IlvlM EI)lATIll,Y rlova to thc next pile and continue to break toothpicks.
'lhc instlLrctor will call out "switch" alter l0 seconds,30, 60, 120, and 180 scconds.
Once toothpickase has rnoved to the next pile, studcrrt 2 should rernove thc brol(en pieccs florn the desk, and student 3
should begin calculating the number oftoothpicks brokn.
At thc end of 180 seconds, the data should be recorded in the table providcd and the rates ofenzyule fulction lvill be

8.

calculated.
On tlte satl]e graph fi-om Part A, add data fio]n Parl

L
4.
5.
6.

B.

Create a legend to ideDtily the data fi'orn Pafts A and B.

Part C: Rate of Enzymes with temperature change

9. Make 5 piles of40 toothpicks in each piJe.
10. In groups of3, one petson will be the enzyme toothpickase, one person will clean/remove broken toothpicks, and the
third person will calculate the number oftoothpicks broken for each pile.
11. Student l, toothpickase, will becorne refrigerated by placing their hands in ice water for 30 seconds..
12. When the instfuctor tell the class when to begin, the enzyme (toothpickase) will begin to break the substrate
(toothpicks) one at a tlre starting at the first pile. When the instructor shouts "switch", the eDzynle will
IMMEDIATELY move to the next pile and continue to break toothpicks.
13, The instructor will call out "switch" after l0 seconds, 30, 60, 120, and 180 seconds.
14. Once toothpickase has moved to the next pile, studnt 2 should remove the broken pieces from the desk, and student 3
should begin calculating the number oftoothpicks broken.
15. At the end of 180 seconds, the data should be recorded in the table provided and the rates ofenzyme function will be
calculated.
16. On the same graph fi-om Pan A, add data from Part B. Create a legend to identi$/ the data from Pafts A and B.

Table 2

Total Time
(sec)

l0
30

(additional 20 sec)
60

(additionai 30 sec)
120

(additional 60 sec)
180

(additional 60 sec)

Toothpicks metabolized
(broken) on each plate

Rate of enzyme activity

Total Toothpicks
Metabolized (Broken)
over time

Denatured

Cold

Denatured

Cold

l)enatured

Card

Paft B

Part C

Part B

Part C

Parl B

Part C

Analysis and Conclusions:

On a shcet ofgraph paper, graph youl rcsults ofthc total lumbcf oftoolhpicks br-okcn olcr
the nunrber oftoothpicks broker aud "X" axis are the tirne interyals.

tillc.

Your

"Y"

axis shoulcl bc

l)uring r'vhich activity was the greatest antoLlut oftoothpicl(s able to bc btoken?

2.

Which tirne had the highest rate ofcnzyme activity?

3.

Why rvoLrld the rates of enzyrre activity cltzltge cluring the trial?

4.

What effect did havingthe enzytne becoure denatured have on the el'f-ectiveness of the enzynre?

5.

What eft-ect did having the enzyrne being placed in cold water have on its effectiveness?

6.

Which parl of the enzynre toothpickase would be the active site?

7.

What was the substrate that the enzylne reacted with?

What effect would it have on the enzylne rate if the broken toothpick pieces where not rentoved during the trials?
Explain your answer.

9.

What would happen to the enzyme rate if the time continued for 360 seconds? Would it speed up, stay the same or
slow down? Explain your answer?

10. Does

a lower temperature change/denature the enzyme?

1. Using the infonnation we learned fi'orn this experiment, why do you think we place foods in refrigerators?

12. What does a higher temperature or pH do to the enzyme? Explain the

change?