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ACTION AND SPECIFICITY OF ENZYMES Action and enzyme specificity Summary- In the first instance you want to analyze the action of enzymes in different media, with different substrates and after having gone through some processes. Thus checking its specificity, its reactivity and finally comparing the action of an enzyme and an inorganic catalyst. Key words— Catalyst, Reactions, Enzyme, Protein, Catecolasa.

I.

A. Action of the Catecolaa:To carry out this procedure we used several tubers. In this first case it was the potato where we macerated 7 grams of it; With 40 ml of water to obtain the catecolase, we used several test tubes each with different concentrations, all in order to see the action of the catecolase as shown in figure 1.

Substrate,

INTRODUCTION

Enzymes are biological catalysts which are in charge of accelerating the reaction speed of some metabolic processes, for each reaction there is an enzyme which will do its catalytic action in the reaction. Enzymes consist of an active site which is responsible for generating the binding with the substrate and reacting this metabolic process, but the enzyme is a protein compound which can also be denatured and cancel the usefulness of this enzyme. On the other hand, there are inhibitors which are hindering the reaction; These bind to the enzyme preventing any catalytic process.

Figure 1. Potato maceration

OBJECTIVES 1. Observe the action of enzymes and show the effect that heat and heavy metals have on them. 2. Make the comparison between an enzyme and an inorganic catalyst and demonstrate the specificity of an enzyme. II.

MATERIALS AND METHODS

In the previous practice we gave ourselves the task of using three methods to see the action and specificity of the enzymes; It started with:

Reception Date: 02/22/2016

After maceration, the potato extract obtained is filtered. 6 test tubes are listed to make the following dilutions.

Tube # 1: 5ml of the substrate (catechol solution) and 5ml water are added. Tube # 2: 5ml of water and 5ml of potato extract are added Tube # 3: 5ml of potato extract and 5ml of catechol solution are added. In tube # 2 we add water and catecolase, in which none reacted because the tubes did not find the enzyme with its respective substrate, on the contrary, in tube # 3 we added catecolase and the catechol solution in which it reacted as expected, giving a brown color compared to the previous 2 tubes

Figure3. Test tubes 4,5 and 6

In tubes 4,5 and 6 the enzyme is denatured and inhibitors such as ascorbic acid are added. Finishing the reaction, we conclude that only the potato contains the catecholase enzyme because it only reacted when binding with the catechol substrate. B. Substrate Specificity:It was possible to appreciate the specificity of the enzymes where the enzyme (Catecolasa) and 5 ml of phenol were added to a test tube, which has almost the same chemical structure as catechol, unlike phenol that only has one (OH) and catechol 2, making clear the specificity of the protein. We confirm this by comparing test tube # 3 of the previous method with the tube of this reaction.

Figure2. Test tubes 1, 2 and 3 Tube # 4: 5 ml of boiled potato extract and 5 ml of catechol Tube # 5: 5ml of potato extract, 10 drops of Silver Nitrate (AgN3) and 5ml of catechol solution are a T of a

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Figure 4. Test tubes 3 and 7

C. Comparison of an enzyme and an inorganic catalyst:Finally we use this method to compare the action of the enzymes of an inorganic catalyst on a different substrate (hydrogen peroxide), the bubbles being the evidence to determine if the relationship occurred or not. 6 test tubes were used. In the first 3 tubes there is little presence of bubbles as shown in figure 5.

Figure6. Inorganic catalyst

III.DISCUSION

Figure5. Comparison of an enzyme In tubes 4, 5 and 6 the test is done with an inorganic catalyst such as manganese dioxide, which upon contact with water and different temperatures generates a reaction, forming a greater amount of bubbles than when the enzyme was used as seen in figure 6.

In the catecolase action at the beginning of the process we had to macerate and separate the potato extract to join it to a catechol solution generating various reactions, at a given moment ascorbic acid is used: (Ascorbic acid is a colorless, odorless crystal, solid, soluble in water, with an acid flavor. It is an organic acid, with antioxidant properties, from sugar) to see what effects changes in lto reaction showing us solor a different coloration; if we compare el ascorbic acid with e l lemon juice and we got to lA conclusion that ascorbic acid is vitamin C. while citric acid is the one that comes from all citrus. When the catechol solution and water mix, there is no reaction present that can alter your environment. Catechol is made up of (C6H6O2)and phenol for (C6H6O);its differences is that catechol is benzene with two hydroxyl groups, while phenol is benzene with one hydroxyl. When we talk about the specificity of an enzyme it can be absolute or relative, since it can only act on one type of substrate, the enzyme is said to show absolute specificity for the substrate. That is the case of succinic dehydrogenase, which is specific for succinate, or L-glutamic dehydrogenase, specific for glutamate. If the enzyme can act on substrates with

very similar structures, the enzyme is said to show relative specificity for the substrate. L-amino acid oxidase, for example, can catalyze the oxidation of different L-series amino acids. This characteristic of some enzymes can be exploited in some clinical settings. For example, in patients intoxicated with methanol, ethanol is used in treatment. The enzyme can bind to either of the two alcohols (relative specificity), but has 10 to 20 times more affinity for ethanol, which favors the oxidation of ethanol over the oxidation of methanol. Avoiding the oxidation of methanol to promote its elimination without being transformed is very important, since the metabolic oxidation of methanol produces metabolites that are very dangerous for the body, such as formaldehyde and formic acid. Poisons such as mercury bichloride or corrosive sublimate and mercury oxycyanide have been widely used as locally acting antiseptics; mercurious chloride or calomelanes, used as a laxative, can become sublimated by the action of alkalis and chlorides in the body; organic derivatives are used as diuretics and have reduced toxicity. Mercury used in industry is a cause of mercury poisoning at work; it penetrates by respiratory (vapors and mercurial salts) and digestive; many of these salts are strongly caustic and act as such at contact sites. Evidence that the potato has an enzyme called catalase 1. Catalase is found in cells of animal and plant tissues. Tissue function is necessary because during cellular metabolism, Hydrogen peroxide is a residue of the cellular metabolism of many living organisms and has, among others, a protective function against pathogenic microorganisms, mainly anaerobes, but given its toxicity, it must rapidly transform into less dangerous compounds. This function is carried out by this enzyme that catalyzes its decomposition into water and oxygen. Despite the fact that hydrogen peroxide is relatively stable at room temperature, many substances act as catalysts for its decomposition. These can be biological catalysts like enzymes, which are protein molecules. Or, be inorganic catalysts, of which there are a wide variety of forms such as transition metals, alkalis and metal oxides. Daylight also favors the decomposition of hydrogen peroxide,

IV.CONCLUSIONS

In the practice that took place on February 22, 2016, we made different processes for the action of catecolase and the comparison of an enzyme and an organic catalyst Enzymes, unlike inorganic catalysts, catalyze specific reactions. However there are different degrees of specificity. Since catalase is chemically a protein, we can denature it by subjecting it to high temperatures. By losing the tertiary structure, it will also lose its function and consequently its catalytic function. By carrying out this experiment we were able to clearly observe the catalase enzyme in potatoes and how different factors such as temperature or pH influence, and also the reactions that occur with the different reagents used. Enzymes are biocatalysts that serve to accelerate processes, they are substances of a protein nature that catalyze chemical reactions whenever thermodynamically possible. The importance of an enzyme is that it accelerates the reactions in the vegetables, the enzymes constitute the main catalyst of the undesirable reactions of the browning of the enzymatic type which makes the fruits or vegetables ripen much faster.

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