Purified Pectinase Lab Report

In this lab or experiment, the aim was to determine the following factors of enzymes: (1) the effects of enzymes concentration the catalytic rate or the rate of the reaction, (2) the effects of pH on a particular enzyme, an enzyme known and referred throughout this experiment as ALP (alkaline phosphate enzyme) and lastly (3) the effects of various temperatures on the reaction or catalytic rate. Throughout the experiment 8 separate cuvettes and tubes are mixed with various solutions (labeled as tables 1,3 & 4 in the apparatus/materials sections of the lab) and tested for the effects of the factors mentioned above (concentration, pH and temperature). The tubes labeled 1-4 are tested for pH with pH paper and by spectrophotometer, cuvettes 1a-4a was tested for concentration and cuvettes labeled 1b-4b was tested for temperature in four different atmospheric conditions (4ºC, 23ºC, 32ºC and 60ºC) to see how the enzyme solution was affected by the various conditions. After carrying out the procedures the results showed that the experiment followed the theory for the most part, which is that all the factors work best at its optimum level. So, the optimum pH that the enzymes reacted at was a pH of 7 (neutral), the optimum temperature that the reactions occurs with the enzymes is a temperature of 4ºC or

In Conclusion pectinase would be the better enzyme to use, I found that the more pectinase used in the applesauce the more apple juice was produced. The hypothesis is supported by the data. The trial for pectinase produced 13ml of apple juice. The trial for cellulase produced 8ml. The trial for water produced 5ml .A reason why this lab could have not worked is because there may have been too much enzymes dropped or too much water dropped in the applesauce. A suggestion I would give to make this experiment more efficient would to be able to filter the applesauce and pay more attention to the time and be careful when the enzymes are dropped into the apple juice also by making the time longer and by doing more than 1 trial. Testing the enzymes

The use of multiple test tubes and Parafilm was used for each experiment. Catechol, potato juice, pH 7 phosphate buffer, and stock potato extract 1:1 will be used to conduct the following experiments: temperature effect on enzyme activity, the effect of pH on enzyme action, the effect of enzyme concentration, and the effect of substrate concentration on enzyme activity. For the temperature effect on enzyme activity, three test tube were filled with three ml of pH 7 phosphate buffer and each test tube was labels 1.5 degrees Celsius, 20 °C, and 60 °C. The first test tube was placed in an ice-water bath, the second test tube was left at room temperature, and the third test tube was placed in approximately 60°C of warm water. After filling the test tubes with three ml of the

Background and Introduction: Enzymes are proteins that process substrates, which is the chemical molecule that enzymes work on to make products. Enzyme purpose is to increase the rate of activity and speed up chemical reaction in a form of biological catalysts. The enzymes specialize in lowering the activation energy to start the process. Enzymes are very specific in their process, each substrate is designed to fit with a specific substrate and the enzyme and substrate link at the active site. The binding of a substrate to the active site of an enzyme is a very specific interaction. Active sites are clefts or grooves on the surface of an enzyme, usually composed of amino acids from different parts of the polypeptide chain that are brought together in the tertiary structure of the folded protein. Substrates initially bind to the active site by noncovalent interactions, including hydrogen bonds, ionic bonds, and hydrophobic interactions. Once a substrate is bound to the active site of an enzyme, multiple mechanisms can accelerate its conversion to the product of the reaction. But sometimes, these enzymes fail or succeed to increase the rate of action because of various factors that limit the action. These factors can be known as temperature, acidity levels (pH), enzyme and/or substrate concentration, etc. In this experiment, it will be tested how much of an effect

These results shown from this experiment led us to conclude that enzymes work best at certain pH rates. For this particular enzyme, pH 7 worked best. When compared to high levels of pH, the lower levels worked better. The wrong level of pH can denature enzymes; therefore finding the right level is essential. The independent variable was the amount of pH, and the dependent being the rate of oxygen. The results are reliable as they are reinforced by the fact that enzymes typically work best at neutral pH

If temperature of the water(enzyme environment) is increased to 35°C, then the enzyme activity will

Enzymes are the very successful machines of natural life. They are in charge of lowering so as to catalyze responses the enactment vitality for these responses. Enzymes are comprised of proteins interpreted from nucleic corrosive coding material, so they have particular duties inside of a life form. Beta-lactamase is a enzyme that separates beta-lactam anti-toxins like penicillin. In this lab, the viability of the beta-lactamase catalyst was tried at diverse temperatures. The beginning speculation of the trial was that enzymatic action would increment as temperature expanded, in spite of the fact that at 60 degrees Celsius the chemical would denature. This was somewhat reflected in the information. The chemical did denature at 60 degrees; in

Figure 3- The effect of pH on enzyme activity every twenty seconds for two minutes. Cuvette 2 had a pH of 8 (acidic), Cuvette 9 had a pH of 7 (neutral), and Cuvette 11 had a pH of 11 (basic). Interpretation The purpose of this experiment was to find out the effect of temperature, pH, and enzyme concentration on enzyme activity every twenty seconds for two minutes.

The standard curve was plotted for FeSO4 with concentration range of 0.2-1mM, and there was a strong correlation between FeSO4 concentration and antioxidant capacity (R²= 0.992) and the equation was Y=0.1958X-0.2452. The standard curve displayed a linear trend between 0.2 to 1mM FeSO4. There was strong correlation between pectin concentration and antioxidant activity for all the pectin samples tested (R2=0.9778, 0.9885, 0.9742 and 0.9954), for the native, 100W, 200W and 400W degraded pectin respectively. Sonicated pectin had increased antioxidant activity with 400W treated pectin having 43% relative FRAP activity at 4mg/mL, the same concentration of native pectin had a lower relative FRAP activity 16.4% compared to FeSO4. There was generally increased antioxidant activity with increasing sonication power applied. The results are consistent with the previous observation by Pokora et al., [66], who reported that enzymatic hydrolysis of egg yolk protein and white protein improved their radical scavenging (DPPH) capacity, ferric reducing power, and chelating of iron activity. Native pectin is a complex molecule with complex side group structure, and during sonolysis the large molecule is depolymerized yielding low degree of polymerization pectin , thus exposing prior hidden functional groups and creating functional groups at the scission sites, e.g. carbonyl groups. The reducing agents mostly act as hydrogen/electron atom donors thus

The enzyme activity is affect by its surroundings especially when pH, temperature, and the concentration of activator and inhibitor are involved. When the pH rise or drops out of the enzyme stretch in tolerance, the structure of the protein can be alter or in other worlds denatured. Most of the enzyme in the bodies of humans

Introduction: Enzymes are protein catalysts facilitating the conversion of substrates into products (Alexander and Peters, 2011). They go through a whole chemical reaction which starts off with the substrate and then ends up with a product. The only way this reaction can be adjusted or not even work is if they end up going through some sort of affect which only temperature and pH levels can do determining the environment. When enzymes are in an environment that is too acidic or alkaline, their chemical properties, sizes and shapes can become altered (Magher, 2015) Chemical modification of proteins is widely used as a too; to maintain a native conformation, improving stability (Rodriguez-Cabrera, Regalado, and Garcia-Almendarez, 2011) In this experiment, four trials were conducted and recorded every 15 seconds for 5 minutes in order to calculate the optimum levels and IRV.

It can be inferred that as sodium chloride, a strong electrolyte, dissolves completely in aqueous solutions, pervading the solution with free sodium and chlorine ions, interferes with the binding of catechol oxidase to the substrate, buffered catechol oxidase. As enzymes are proteins, charged R-groups interact with the oppositely charged ions of salt, altering the enzyme’s shape and rendering it unable to bind. According to (__), the chlorine anion is responsible for the change in enzyme activity. The ions act as noncompetitive inhibitors, binding to sites other than the active site. Because enzyme shape is so specific to its function, even slight interactions are likely to considerably lower its activity. Solutions more concentrated than 20% significantly drop enzyme activity and bring it almost to a halt (Ming Hui Fan et al.,2005). On the other end, our data reflects that too low of a salt concentration may also affect the viability of the enzyme. A 2.5% salt solution exhibited a decrease in activity (Fig. 1) which leads one to believe that the natural salt content of potatoes likely lies around 5%, the optimal concentration

Intro/Background: An Enzyme is an organic element (proteins and RNA molecules) that performs like a catalysts and assist in intricate reactions that can happen anytime in life. Enzymes accelerate the rate of all the chemical reactions that happen in cells. Enzymes are very important by the fact that they help in the human body by helping out with digestion and with your metabolism. Enzymes are choosy catalysts and this means that they only speed up certain reactions. The elements that can affect and not make the enzyme function properly is when there’s a temperature change, and the bonds that connect the protein’s uncommon shape are disturbed and some of these bonds can be broken apart and then changes the shape of the active site. The active site of an enzyme will change a different shape and it also gathers molecules together or it can break them apart. The role of the catalysis is when the substrate is altered, it is then broken down or mixed with some other molecule to make something new. The duty of the catalyst is a element that escalates the rate of a chemical reaction by lessening the amount of energy needed to start that reaction. The Product is when the enzyme let 's go and it turns back to normal. It then prepares to start another reaction but the enzyme will not change and so the substrate is no longer the same. The enzyme pectinase has a responsibility to rift down the central part of the plant cell walls. The pectinase enzymes are proteins so accelerate the

9). (-1) represents the low concentration level and (+1) represents the high concentration level for each component. The average of results of pectinase activity and protein content are presented in (table 13).The main statistical analysis of this experiment is shown in (table 14).The main effect of each variable upon enzyme activity and protein content was estimated and represented graphically in (figure 12, 13) respectively Show that the highest pectinase activity was recorded for trial no. 6 while the lowest activity was recorded for trials No. 7 and 1.

Enzymes are biological catalysts which speeds up chemical reactions without being used up. There are three main factors which affect enzyme activity, and determines if their rate of reaction which are; sufficient energy (temperature), the pH, and the concentration. When there is a reaction, there must be energy around, the higher the energy, the faster the reaction because the particles will move faster therefore the chance of the particles to collide into each other will increase. However, if the temperature is too high or too low, the enzyme would denature and it will no longer work. This is similar to the pH; each enzyme will have a different optimal pH (a pH which an enzyme works best at) but if the pH is too high or too low, it will change shape and denature. Lastly as the concentration increases, so does the rate of reaction since more concentration means that there are more particles to react with. “To have a chemical reaction, there must be a least amount of energy which is called activation energy and enzymes help lower the activation energy therefore the speed of the reaction will increase”. People made wine for a long time without the help of enzymes in the winemaking process, but now pectinase is used around the winemaking society which enhances the process of wine making, however, there are limitations to use this enzyme. In this essay, the conditions that pectinase will need to be in will be analysed, and it’s affects economics and society.