The Effect of PH on the Activity of Rennin in Milk Aim: To investigate and determine the effect of a change in PH on the activity

of enzymes Hypothesis: One the renin (enzyme) is added to the milk (substrate), the milk will curdle. ! will have the fastest as it is the o"timum reaction. Equipment: test tube rack Thermometer milk (full cream) #ettle water Test Tube $mls rennin (%unket) acids& hydrochloric acid, acidic acid and 'odium Hydro(ide PH of

Risk Assessment:
)isk * hydrochloric,acidic and sodium hydro(ide Prevention acid is a to(ic chemical with a variety of household and industrial uses. +t is a dangerous li,uid that should always be handled with e(treme caution and used only when necessary. -hen inhaled, hydrochloric acid causes irritation, coughing and choking. +t also causes nose, throat and res"iratory inflammation. To "revent this from ha""ening do not "ut the . solutions near you. lways kee" the lids closed and away from the edge of the table as it might s"ill and cause in%ury due to the glass. /reakage may occur allowing damamge to the skin and eye. To "revent breakage, carrying a test tube and thermometer should be always handled with care. 0ay cause burning and irritation to the skin. To "revent this the kettle with boiling should be handled with care, as it should be ke"t away from the body.

* Test tube,thermometer

* kettle (hot water)

Method: PH of 2 $) "re"are the e("eriment by getting the e,ui"ment ready. (hydrochloric acid, one test tube, test tube rack, milk 1$2m34 and thermometer) !) 0easure .2ml of milk with a measuring beaker and "our it into a test book .) "out water in a beaker and "ut the test tube with milk in water bath and "lace the thermometer in the milk to observe the milk tem"erature reaches .5 degrees. 6) 7sing the mortal and "estle grind a ,uarter of renin tablet and "ut it into the test tube. 8) dd 8 dro"s of hydrochloric acid in the test tube.

PH of 4 $) "re"are the e("eriment by getting the e,ui"ment ready. (acidic acid, one test tube, test tube rack, milk 1$2m34 and thermometer) !) 0easure .2ml of milk with a measuring beaker and "our it into a test book .) 0ake a water bath in a beaker and "ut the test tube with milk in the water bath and "lace the thermometer in the milk to observe when the milk tem"erature reaches .59:. 6) 7sing the mortal and "estle grind a ,uarter of renin tablet and "ut it into the test tube. 8) dd 8 dro"s of acidic acid in the test tube. PH of 7 $) "re"are the e("eriment by getting the e,ui"ment ready. (water, one test tube, test tube rack, milk 1$2m34 and thermometer) !) 0easure .2 m3 of milk with a measuring beaker and "our it in a test tube .) 0ake a water bath in a beaker and "ut the test tube with milk in the water bath and "lace the thermometer in the milk to observe when the milk tem"erature reaches .59:. 6) 7sing the mortal and "estle grind a ,uarter of a rennin tablet and "ut it in the test tube. 8) dd 8 dro"s of water in the test tube. PH of 10 $) "re"are the e("eriment by getting the e,ui"ment ready. (sodium hydro(ide, one test tube, test tube rack, milk 1$2m34 and thermometer ) !) 0easure .2 m3 of milk with a measuring beaker and "our it in a test tube .) 0ake a water bath in a beaker and "ut the test tube with milk in the water bath and "lace the thermometer in the milk to observe when the milk tem"erature reaches .59:. 6) 7sing the mortal and "estle grind a ,uarter of a rennin tablet and "ut it in the test tube. 8) dd 8 dro"s of sodium hydro(ide in the test tube. Results: PH PH of ! PH of 6 PH of 5 PH of $2 !5 seconds 8$ seconds $ minute and 82 seconds ;.82 minutes Time Taken

The results stated when the e("eriment was being "erformed it was observed that the addition of hydrochloric acid to the full cream milk causing the substrate to curdle even without the enzyme (rennin) in the solution . This slightly affected the results as it was difficult to determine the coagulation of the substrate. The o"timum Ph is !, "roven with the results as it only took !5 second for the milk to coagulate in com"arison to the other time records. s the Ph gets higher from its o"timum the enzyme starts to denature ("h hydrogen bons in the molecule break a"art which destroys the sha"e of the molecule). This is evident when the time taken for the 0ilk curdles becomes more delayed.

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<iscussion =uestions& a) >("lain why water was added as well as acid or alkali,when altering the "H cidity and alkalinity in the body are measured by the "H ("otential of hydrogen) scale. -ater has a "H of 5.2 and is considered neither acid or alkaline but neutral. b) -hich variables are controlled?

:ontrolled @ariables mount of milk, which was $2ml mount of enzyme (A rennin) mount of hydrochloric acid (8 dro"s) cidic acid (8 dro"s) 'odium hydro(ide (8 dro"s) concentration of renin solution Tem"erature (.5 degrees) time allowed for coagulation to occur c) +dentify the inde"endent and de"endent variables. +nde"endent variable * The different levels of PH of !, 6, 5,$2 <e"endent variable * :oagulation

d) :reate a list of each "H and how long it took to coagulate PH PH of ! !5 seconds

Time Taken

PH of 6 PH of 5 PH of $2

8$ seconds $ minute and 82 seconds ;.82 minutes

e) :reate a gra"h with "H as the (*a(is and relative enzyme activity ( ie. $Btime to curdle) on the y*a(is

f) <etermine the o"timum "H for the enzyme rennin. The o"timal "H for rennin is around $*!, as this closely resembles the acidic environment of the stomach where rennin is most commonly found.

The Effect of Temperature on the Activity of Rennin in Milk >nzymes are s"ecific in their function, meaning one kind of enzyme can only digest a "articular substance. The active site is where the reaction takes "lace. the substrate and the enzyme have sha"es that mean they can only fit each other, hence the s"ecificity. )ennin is an enzyme secreted by walls of the stomach. +t acts on a "rotein in milk causing it to "um" into a semi*solid curd.+t hel"s us to digest "rotein. >nzymes have o"timum tem"eratures, since rennin is an enzyme from the human stomach, its o"timum tem"erature would be around .5* .; :elcius or $22 C, and the o"timum "H would be $*.. +f the tem"erature is too low, the enzymes move around too slowly. Too high tem"erature, or incorrect "H would destroy the active site, denaturing them.

aim& To investigate the effect of tem"erature on the activity of the enzyme rennin. hy"othesis& The higher the tem"erature of the milk the faster the time it takes for the milk to curdle. n increase in tem"erature results in more kinetic energy of the enzyme and the substrate. Equipment: test tubes milk measuring cylinders thermometer large beaker several small beakers tablet distilled water Method: de!"ees 1) get your e,ui"ment ready (one test tube, test tube rack, milk 1$2m34, thermometer and rennin enzyme.) !) 7sing the mortal and "estle grind a ,uarter of a rennin tablet and "ut it in the test tube. .) Put the test tube in a test tube rack in the refrigerator. 6) Det the test tube out of the refrigerator without holding onto the bottom where the milk has coagulated for too long. 8) record the results #7 de!"ees $) get your e,iu"ment ready (one test tube, test tube rack, milk 1$2m34, thermometer and rennin enzyme.) !) 0easure .2 m3 of milk with a measuring beaker and "our it in a test tube .) 0ake a water bath and "ut the test tube with the milk in the water bath and "ut the thermometer in the test tube till it reaches .59: 6) 7sing the mortal and "estle grind a ,uarter of a rennin tablet and "ut it in the test tube 8) )ecord )esults 0$% $) Det e,ui"ment ready (one test tube, test tube rack, milk 1$2m34, thermometer and rennin enzyme). !) 0easure .2 m3 of milk with a measuring beaker and "our it in a test tube .) 0ake a water bath and "ut the test tube with the milk in the water bath and "ut the thermometer in the test tube till it reaches ;29: 6) 7sing the mortal and "estle grind a ,uarter of a rennin tablet and "ut it in the test tube 8) )ecord the results. Results

Tempe"atu"e $% #7$% 0$% did not coagulate $ min $2 sec

Time

<id not solidify (most likely denatured)

The o"timal tem"erature based on the results was .5 degrees :elsius as this is also the average body tem"erature as a result of this the milk coagulated. on the other hand the two e(treme conditions ;9: and ;29: did not coagulate at all as it denatures due to not having a o"timal tem"erature.
Pi&tu"e:

'is&ussion: $. 'uggest what ha""ened to the enzyme at ;2 degrees? The renin did not coagulate because the the enzyme denatures, as the enzyme denatures the bonds which hold the amino acids falls a"art. the enzyme denatures as it lost its sha"e and could not bind with the substrate (milk) !. -hat would ha""en to the one ke"t in the fridge if allowed to warm? -hat does this say about the enzyme? when the test tube is taken out of the fridge it will warm u" and the milk will reach the room tem"erature (.5 degrees), holding the test tube will warm the test tube ,uicker allowing coagulation occur. .. <etermine from your results the o"timum tem"erature for the enzyme rennin? +t was shown the o"timum tem"erature is the tem"erature that is the hottest "ossible without

denaturing t "roteins. .5 degrees :elsius is the hottest that a enzyme can function without being denatures. this was shown as the milk coagulated in only !5 seconds. 6,-hat is the "ur"ose of the milk only test tubes? The "ur"ose of the milk was for a controlled variable as this makes the e("eriment more valid and reliable. +t was also tested to com"are and contrast the differences between the different results. 8.Eame the inde"endent variable, the de"endent variable and the fi(ed or controlled variables. +nde"endent <ifferent tem"eratures ranges from !5 degrees :elsius to ;2 degrees :elsius <e"endent :oagulation :ontrolled mount of milk ($2 ml) mount of enzyme ($B6 rennin) Time allowed for coagulation to occur.

Eancy.Dorial