THE INFLUENCE OF ENZYME PRECENTAGE TO SPEED OF REACTION

By : MUTI ATIKA FURI (103204003)

International Biology of Education Mathematics and Science Faculty SURABAYA STATE UNIVERSITY 2012 2013

CHAPTER I INTRODUCTION 1.1. BACKGROUND In body every organisms always happens molecular solution, from the bigger size of molecule become smaller molecule (catabolism) and the creation of big molecule from the small molecule (anabolism) or generally it called metabolism. Metabolism process is controlled by a specific protein is called enzyme. Enzyme is a protein which have catalyzed activity. Metabolism process can run faster and slower. So that its needed a catalyzer to speed up the metabolism reaction. But the rate of enzymatic reaction in cell is determined by temperature, pH, substrate concentration, product concentration , time, and be affected by enzyme also. One of the enzyme in plants is amylase. Amylase enzyme can hydrolyze an amilum or starch to glucose in several steps, are amilodectrine forming of amilum, and become eritrodectrin to acrodectrin to the glucose. Amylase is produced in seeds or leaves. Amylase activity in the reaction is influence by anorganic salt, pH, temperature, and light. Based on the background above, we can arrange a report in tittled The influence of enzyme percentage to speed of reaction. 1.2. PROBLEM FORMULATION The problem formulation for this practicum is : How the influence of enzyme precentage to speed of reaction? 1.3. OBJECTIVE The objective from the experiment is : To know the influence of enzyme percentage to speed of reaction.

CHAPTER II LITERATURE REVIEW In the organisms body there are a millions chemical reaction which have simultaneous that called metabolism. Metabolism process have a goal to produce the energy for any activity of organism, beside that metabolism have a goal to forming the structural components of organism. Metabolism process divides into two process, are catabolism and anabolism. Catabolism is a molecular solution from the bigger molecule to the smaller molecule, but anabolism is a molecular forming from the bigger molecule to the smaller molecule. Largely, biochemical reaction which happens inside of body catalyzed by a specific protein called enzyme. Enzyme is a bigger protein molecule, folded in the crease so that the group of amino acid will be formed of active side. This enzyme has a function as catalisator in reaction. Enzyme was spread in all of cell of living things and have many amount, but not all of cell have a number and kinds of enzyme that same. The general characteristic of enzyme, are :
1. Have a specific characters. It means that one enzyme just can match to one

substrate. 2. Colloid, large surface area, hydrophyl. 3. Can be reacted with anothers compound, base or acid.
4. Really sensitive to the factors that can caused the damage or enzyme

denaturation, are pH, temperature, etc. 5. As biocatalysator. In small amount, it can be increasing the reaction rate without changes the balancing of reaction. 6. Enzyme is not participate to the reaction. 7. Have a big molecule. Some of enzyme consist of polypeptide, and not consist chemical group beside amino acid residue. But, another enzyme need chemical components added for its activity. These components is called cofactor. Cofactor is an anorganic molecule as

like ions Fe2+, Mn2+, Zn2+ or some complex organics molecule as coenzyme. Some enzyme needed coenzyme or one more metal ion for its activity. In several enzyme, coenzyme or metal ion only bounded weakly in short time. But another enzyme in this compounds, its bounded strongly or permanently that called prosthetic group. Enzyme which have a perfect structure and catalyzes actively together with the coenzyme or metal groups that called holoenzyme. Coenzyme and metal ion have a characteristic that stable when the heating process, but the part of enzyme protein that called apoenzyme denatured by the heating. The works of enzyme is specificly, its mean that the conversion of certain substance be needed the enzyme also. The speed of reaction which have catalyzed by enzyme was influenced by some factor, as : a. Temperature. Enzymatic reaction speed will be increasing while the increase of temperature process till the optimum point, is 37C. After the temperature was high, the increasing of temperature will be decrease the reaction speed it caused the damage of enzyme protein in high temperature. b. pH Enzymatic reaction speed will be higher with the pH increasing until the optimum point is 7. In optimum point, the increasing of pH will be decrease the reaction speed because the lower or higher pH, so the enzyme protein will be crack. c. Substrate concentration The increasing of enzyme concentration that constant, the increasing of substrate concentration will be increase a reaction speed until the optimum point. d. Enzyme concentration The increasing of enzyme concentration causes reaction speed till optimum point. After the high point, the increasing the enzyme concentration can

causeds the reaction speed become constant because the substrate was limited. e. Product concentration As more as many product which have been formed, the reaction speed will be decrease, the substrate concentration and the hoarding of product in several enzyme, product become a resistor for enzyme activity.

CHAPTER III RESEARCH METHOD 3.1. VARIABLE RESEARCH

Manipulation variable Controle variable amount of KI-I2.

: enzyme levels (0%, 25%, 50%, 100%) : the kinds of enzyme, the kinds of substrate, the

concentration of substrate, substrate volume, time range for dropping, and the

Response variable

: the reaction speed and colour changes.

3.2. RESEARCH MATERIALS AND INSTRUMENTS

Tools : Porcelain mortar and pestle (1pieces), reaction tube (8 pieces), measuring cup 10 mL (1 pieces), centrifuge (1 pieces), drip cup (1 pieces), pippete (3 pieces), rubbing alcohol (1 pieces).

Materials : mung bean, amilum solution 4%, aquades, KI-I2 solution, phosphate sitrate buffer with pH 5,6 10 mL.

3.3. METHOD 1. Prepare the mung bean seeds that a day-old, waste of skin.
2. Weigh 20gr of mung bean, then adding 20 mL of phosphate sitrate buffer 20

mL until the mung bean were smooth (its destroyed in porcelain mortar and pestle). 3. Enter the mung bean solution into the reaction tube, take in centrifuge during 5 minutes. Then take a supernatant liquid of mung bean, enter to the another test tube. This liquid is considered as amylase enzyme 100%. 4. Make an enzyme with 0 %,25 %, 50 %. 5. Prepare the test tube, filled it with 5 mL of enzyme solution 100 % then added 2 mL of amilum solution 1 %. Mix it unti it mixed.

6. Drops 1 drop the mixture of amilum solution with the enzyme in drip cup then

we test it with 1 drop of KI-I2 solution. Record the the time needed. 7. In every 2 minutes, do the testing until the discoloration appears. 8. Record the result, n do it also in 50 %, 25 %, and 0 % enzyme levels also.

CHAPTER IV RESULT AND DISCUSSION 4.1. RESULT 1. Table Table of the influence of enzyme percentage to speed of reaction Time range every minutes 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ ++++ ++++ ++++ ++++ +++ +++ +++ +++ +++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ +++++ ++++ ++++ ++++ ++++ ++++ Enzyme concentration in 100 % 2 50 % 25 % 0%

19 20 21 22 23 24 25 26 27 28 29 30

+++ ++ ++ ++ ++ ++ +

+++ +++ +++ +++ +++ +++ ++ ++ ++ +

++++ ++++ +++ +++ +++ +++ +++ +++ ++ ++ ++ +

2. Histogram
34 32 30 28 26 24 22 20 18 16 14 12 10 8 6 4 2 0

2 minutes to (average)

25

50

100

Enzyme concentration (%)

The histogram of influenceof enzyme percentage to speed of reaction

4.2. DATA ANALYSIS Based on the result observation, we can see that the enzyme concentration influence to the enzyme percentage to speed of reaction. The faster reaction was happened in 100 % enzyme concentration. Its just need 25 times of dropping. The colour was changed quickly than others. The slower reaction was happened in 0 % enzyme concentration. The color its doesnt changes yet until 30 times of dropping. In the 25 % enzyme concentration, the colour was changed in 30 times of dropping. But in the 50 % enzyme concentration, the colour was changed in 28 times of dropping, its faster than 25 %.

4.3. DISCUSSION The colour that got from the KI-I2 testing in amilum solution which added with enzyme 100 % dark blue, violet, blue, green, and yellow. The first step colour (dark blue) is a step when the amylase enzyme was starting to work. For the next colour, its mean that amylase enzyme was active to solved amilum become glucose. Phosphate sitrate buffer have a function to preserve the pH for enzyme amylase, so that amylase doesnt destroyed. The other function is as buffer solution, it means so that the enzyme will be actively to working and not destroyed in optimum condition and keeping the condition in still base. The factors which influence the enzyme working, are : temperature, pH, substrate concentration, enzyme concentration, time, product concentration.

CHAPTER V CONCLUSION The enzyme concentration have influence to the reaction rate of changes amilum to glucose. Higher enzyme concentration so the reaction rate will be faster. In reaction which have a less enzyme, the reaction will be slow and need a long time.

BIBLIOGRAPHY Sasmita Mihardja, Dradjat. 1996j. Fisiologi Tumbuhan. Bandung ITB. Soerodikosoemo, Wibisono dkk. 1993. Anatomi dan Fisiologi Tumbuhan. Jakarta : Departemen Pendidikan dan Kebudayaan. Sri Rahayu, Yuni dkk. 2008. Petunjuk Praktikum Fisiologi Tumbuhan. Surabaya Soewardiati. 1991. Biologi Umum. Surabaya : Unipress IKIP Surabaya.

ATTACHMENT

Figure weight of

1.

Measurement Figure 2. After 2 minutes to Figure 3. Centrifuge mung bean by 27th, there is changing colour (Author Documentation,2012) of solution. (Author Documentation,2012)

using scales (Author Documentation,2012)

Figure

4.

process

make Figure 5. 25%, 50% and Figure 6. After 2 minutes to 100% enzyme concentration 3rd, still there isnt changing after adding iodine (Author Documentation,2012) colour (Author Documentation,2012)

extract sprout. (Author Documentation,2012)

Figure 9. the changing colour Figure 7. Extract sprout (Author Documentation,2012) Figure 8. Extract sprout after of solution centrifuge process. (Author Documentation,2012) (Author Documentation,2012)