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FACTORS AFFECTING ENZYME ACTIVITY

-Temperature -pH -Substrate concentration -Enzyme concentration

Temperature

Enzymes have an optimum temperature at which they work fastest. For mammalian enzymes this is about 40C, but there are enzymes that work best at very different temperatures, e.g. enzymes from the arctic snow flea work at -10C, and enzymes from thermophilic bacteria work at 90C.

The rate is not zero at 0C, so enzymes still work in the fridge

(and food still goes off ), but they work slowly. Enzymes can even work in ice, though the rate is extremely slow due to the very slow diffusion of enzyme and substrate molecules through the ice lattice. Above the optimum temperature the rate decreases as more and more of the enzyme molecules denature. The thermal energy breaks the hydrogen bonds holding the secondary and tertiary structure of the enzyme together, so the enzyme (and especially the active site) loses its shape to become a random coil. The substrate can no longer bind, and the reaction is no longer catalysed. At very high temperatures this is irreversible. Remember that only the weak hydrogen bonds are broken at these mild temperatures; to break strong covalent bonds you need to boil in concentrated acid for many hours.

pH
1.Most enzyme are affective in a narrow pH range only 2.The optimum pH is the particular pH at which the rate of reaction is fastest 3.Deviation from the optimum pH decrease the rate of reaction because bonds maintaining the tertiary shape of the enzyme are broken

pH
4.The active site loses its shape and the enzymesubstrate complex can no longer be formed. The enzyme is denatured 5.Unlike the effect of temperature on enzymes, the effect of pH is normally reversible. Restoring the pH to the optimum level usually restore the rate of reaction

Figure 4.2 The effect of pH on some digestive system

6.Pepsin functions most effectively in an acidic medium at a pH of about 2. Pepsin is found in the stomach where the conditions are acidic 7.Trypsin fuctions most effectively in an alkaline medium at about pH 8.5. It is found in the duodenum where conditions are alkaline

Eksperimen
Pernyataan masalah : Apakah kesan pH ke atas aktiviti

enzim? Hipotesis: Pepsin bertindak balas paling berkesan dalam keadaan berasid. Pembolehubah dimanipulasikan : pH larutan Pembolehubah dimalarkan : Suhu larutan Pembolehubah bergerak balas : Penguraian substrat oleh enzim.

Radas dan bahan : Tabung uji; picagari; corong turas;

termometer; jam randik; bikar; penunu bunsen; kertas pH; tungku kaki tiga; kasa dawai; ampaian albumen; larutan pepsin 1 %; asid hidroklorik, HCl 0.1 M; larutan natrium hidroksida, NaOH 0.1 M.

Prosedur : 1. bahan albumen telur(bahagian putih telur) di sediakan

2. 3. 4. 5. 6. 7. 8. 9.

dengan menambah 1 gram albumen telur yamg kering ke dalam 100cm3 air. Campuran itu kemudiannya dipanaskan Tiga tabung uji dilabelkan P,Q,R. 5 ml albumen dan 1 ml larutan pepsin dimasukkan dalam setiap tabung uji. 5 titis air suling dimasukkan dalam tabung uji P. 5 titis larutan hidroklo ric asid dimasukkan dalam tabung uji Q 5 titis NaOH(natrium hidroksida) dimasukkan dalam tabung uji R Kesemua tabung uji direndamkan di dalam kukus air pada suhu 37oC selama 20 minit. Setiap tabung uji diuji dengan kertas pH Isi kandungan tabung uji itu direkodkan.

Keputusan: Hanya tabung uji P kelihatan jernih. Ampaian

albumen boleh dilihat dengan jelas dalam tabung uji Q dan R

Tabung pH Isi uji


P
Q R

Bentuk bahan
Permulaan eksperimen Akhir eksperimen Bahan putih terhasil Larutan yang bersih/jernih Bahan putih terhasil

7 2 9

Albumen + pepsin + Bahan putih terhasil Air suling


Albumen + pepsin + Asid hidroklorik Albumen + pepsin + Natrium hidroksida Bahan putih terhasl Bahan putih terhasil

Perbincangan:
Tabung uji P,Q dan R direndam di dalam kukus air

pada suhu 37oC kerana suhu ini merupakan suhu optimum bagi enzim untuk bertindak balas dengan substrat. Pada akhir eksperimen, isi kandungan dalam tabung uji P menjadi jernih kerana albumen telah diuraikan oleh protein. Pepsin menguraikan albumen dalam keadaan berasid. Kesimpulan : -Aktiviti enzim dipengaruhi oleh pH -hipotesis diterima

Substrate Concentration
FACTOR EFFECTING ENZYME ACTIVITY

APA ITU SUBSTRAT??


Permukaan atau bahan atau dari mana kehidupan organisma, tumbuh, atau mendapat khasiat Bahan pada yang satu tindakan enzim

LOW SUBSTRATE CONCENTRATION

HIGH SUBSTRATE CONCENTRATION

Few substrate molecule Increase in subs. Conc. Will not alter the rate of reaction

More substrate molecule than enzyme molecule Increase in subs. Conc. Mean substrate molecule

ITS ALL ABOUT SUBSTRATE CONCENTRATION

It has been shown experimentally that if the amount of the enzyme is kept constant and the substrate concentration is then gradually increased, the reaction velocity will increase until it reaches a maximum. After this point, increases in substrate concentration will not increase the velocity (delta A/delta T).

DOES SUBSTRATE CONCENTRATION AFFECT ENZYME ACTIVITY?

When substrate concentration increases, enzyme activity increases. This is due to the increased amount of substrate in one place so it is more likely that a substrate molecule and enzyme collide. More successful collisions will occur higher rate of reaction.

EXPERIMENT
Study the effect of substrate concentration on the activity of

Salivary Amylase

PROBLEM WHAT ARE THE EFFECT OF SUBSTRATE CONCENTRATION ON SALIVARY AMYLASE VARIABLE MANIPULATED CONCENTRATION OF STARCH SUSPENSION RESPONDING TIME TAKEN FOR HYDROLYSIS OF STARCH TO BE COMPLETED CONSTANT ENZYME CONCENTRATION
HYPOTHESIS RATE OF ENZYMATIC REACTION INCREASE WITH THE INCREASE IN SUBSTRATE CONCENTRATION UNTIL IT REACH A MAXIMUM RATE

RESULT

Test tube

Conc. Of starch suspension %

Time taken sec min

Rate of reaction

A
B C D

0.1
0.2 0.3 0.4

240
240 240 240

4.0
4.0 4.0 4.0

0.025
0.050 0.075 0.100

E
F

0.5
0.6

300
360

5.0
6.0

0.100
0.100

RESULTS
GRAPH OF REACTION RATE AGAINST STARCH SUSPENSION CONCENTRATION

0.12
0.1

0.08
0.06

0.04
0.02

0
0.1 0.2 0.3 0.4 0.5 0.6

DISCUSSION

of reaction refer to the substrate concentration that has been catalysed per unit Rate of reaction increase in concentration of starch suspension until a maximum point. At this point, further increase the in starch concentration does not increase the rate of reaction anymore Enzyme reaches it saturation point at 0.4% of substrate concentration At this substrate concentration, the rate of reaction does not increase even though the substrate concentration is increased because the concentration of the enzyme has became a limiting factor. The time taken to hydrolyse the starch completely at low substrate concentration is constant, since the enzyme does not yet saturated. However, when the saturation of starch suspension increase, the time taken for the hydrolysis of starch to be completed also increase as the enzyme molecules become saturated.

Rate

CONCLUSION
The rate of enzymatic reaction increase with the increase in substrate concentration until it reaches a maximum rate. The hypothesis is accepted.

Order

Rate Equation

Comments rate is independent of substrate concentration rate is proportional to the first power of substrate concentration rate is proportional to the square of the substrate concentration rate is proportional to the first power of each of two reactants

zero first second

rate = k rate = k[S] rate = k[S][S]=k[S]2

second

rate = k[S1][S2]

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