Laccase Of Catechol

The Effect of Different Concentrations of the Enzyme Catechol Oxidase on the Rate of Benzoquinone Production When Mixed with Pure Catechol

An Enzyme is a protein that in essence speeds up biological reactions. So that would mean that a Catalase is an enzyme reaction that decomposes hydrogen peroxide to water and oxygen. It is primarily found in the liver and it is important in protecting the cell from damaging oxidative reactions.

In order to see the effects of pH and temperature on the enzymatic reaction of catechol oxidase when separated from potato tissue. We used a spectrophotometer to measure how much blue light energy is absorbed by benzoquinone. Benzoquinone is a product of catechol when it has been oxidized by different temperatures and pHs. We hypothesized that the benzoquinone absorbance rate would be faster when the pH added to the cuvettes were greater than the pH of the potato tissue. The pH of the potato tissue was pH 6. Our results show that pH 7 had the faster absorbance rate, slightly slower at pH 4, and slowest at pH

In this experiment, the 5mM catechol (substrate) reacted with catechol oxidase in the presence of 5 different pH buffers mentioned above. This experiment was used to measure the buildup of the colored product, benzoquinone, to observe the change in the absorbance of the mixture in a spectrophotometer at a wavelength of 486 nm. Ithypothesized that since each enzyme has an optimal pH and that the enzymes are proteins, the enzyme activity will increase with pH level and will be at its highest at pH 7, which is water. As seen in figure 1, absorbance is low pH 2 because catechol oxidase activity was minimal at low pH concentration due to the catechol oxidase denaturing in the acidic solution. The catechol oxidases also denatured at high pH concentration such as pH 11, which is a basic solution, lowering catechol oxidase activity and absorption. Catechol oxidase activity was highest at pH 8 making it an optimal pH for catechol oxidase to catalyze the reaction and create more product which in turn increased the absorption of blue

Catechol, in the presence of oxygen is oxidized by catechol oxidase to form benzoquinone (Harel et al., 1964). Bananas and potatoes contain catechol oxidase that acts on catechol which is initially colorless and converts it to brown (Harel et al., 1964). In this experiment, the effect of pH on the activity of catechol oxidase was conducted using buffers ranging from pH2 to pH10. Two trials were conducted due to the first trial results being altered by an external factor. The results were acquired by taking readings every 2 minutes for 20 minutes from a spectrophotometer and then recorded on to the table. The data collected in the table were then made into graphs to illustrate the influence of pH on the catechol oxidase catalyzed reaction. After analysis, the data revealed that pH did have a significant influence on the enzyme as recorded by absorbance per minute. However, the data was collected was not accurate due to external factors, thus the results are debatable and should be experimented again for validation.

Enzymes are types of proteins that work as a substance to help speed up a chemical reaction (Madar & Windelspecht, 104). There are three factors that help enzyme activity increase in speed. The three factors that speed up the activity of enzymes are concentration, an increase in temperature, and a preferred pH environment. Whether or not the reaction continues to move forward is not up to the enzyme, instead the reaction is dependent on a reaction’s free energy. These enzymatic reactions have reactants referred to as substrates. Enzymes do much more than create substrates; enzymes actually work with the substrate in a reaction (Madar &Windelspecht, 106). For reactions in a cell it is

Lactose is a sugar that can be put into smaller molecules, glucose and galactose. Lactose is when you are not able to digest milk and dairy meaning that the enzyme lactase that breaks down lactose is not functioning properly. ONPG was used as a substitute for lactase because even though it is colorless it helps show enzyme activity by turning yellow. This experiment measured the absorbance ONPG when exposed to lactase within an environment of different salinity’s. The enzyme, lactase, was obtained by crushing a lactaid pill and then was added into four cuvettes. ONPG and salt solution of different concentrations were added and their levels of absorption was measured by a spectrophotometer. The results showed that higher salt concentrations have a lower level of absorption. There were 4 cuvettes and within those cuvettes that solutions within them were being tested and the results showed the more salt solution added with the lactase the lower the absorbance. The less salt solution there was a higher rate of absorbance. The data supported the hypothesis that with increasing NaCl concentration there would be a decrease in enzyme activity.

In order to generate a bicyclic lactone in this experiment, a Diels-Alder adduct was produced. The bicyclic lactone to be generated was cis-1,3,3a,4,5,7a-Hexahydro-5-methyl-3-oxo-4-isobenzofuran-carboxylic Acid and was produced using a Diels-Alder reaction. The product was also analyzed quantitatively using percent yield. To prepare the Diels-Alder adduct 0.40 g of 2,4-hexadien-1-ol was added to a flask, then 5.00 mL of toluene and 0.40 g of maleic anhydride were added to the flask in that order. The mixture was warmed and stirred to induce a reaction. The reaction progress was monitored using a TLC plate with 30:70 hexane used as the mobile phase and silica gel as the stationary phase. The TLC plate revealed a new spot for the crude product, indicating the reaction had begun.

The enzyme that will be used in this experiment is lactase. Lactase is an enzyme that breaks down a sugar found in milk called lactose into glucose and galactose. Lactase is found in the small intestine, and is naturally produced by the body. If someone’s body does not produce the lactase properly, or the lactase itself doesn’t act properly, that person would be considered lactose intolerant. This means your body would not be able to digest milk properly, and you could suffer symptoms such as cramping, diarraea, painful gas, and nausea after having some sort of dairy. If one would was lactose intolerant, but wanted to have some sort of dairy product, they could take a lactase supplement. The lactase in the supplement will break down the lactose

With an important level of lactate and a prosperous protons gradient, we can see a

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 effects on Inhibitors on Catechol Oxidase. Valerie Rodriguez, 2014, Functional Biology, Texas State University, San Marcos, TX 78666. This experiment showed the relationship between a competitive inhibitor benzoic acid with a substrate catechol oxidase. Benzoic acid and catechol oxidase compete to attach to an active site in an enzyme. In this experiment cuvettes and a spectrophotometer were used to measure different amounts of benzoic acid and catechol oxidase to determine how the competitive inhibitor benzoic acid affected the catechol oxidase and what reaction it gave off towards it. The reaction time was recorded for a total of 1200 seconds to determine how the toxin benzoic acid reacted to the catechol oxidase and if it sped up or

The data in experiment one showed the darkest color in the tube containing the substrate catechol, which was scaled at a 10, and showed no change in any of the other tubes containing the other substrate, which were scaled at 0, showing that the enzyme only reacted with the catechol substrate. This supported the hypothesis that the enzyme would be specific to only one type of substrate, in this case the substrate was catechol, this is known as enzyme specificity. The active site of the catechol oxidase enzyme has as structure that is able to remove the hydrogen of the catechol to create benzoquinone. This is because the oxygen atoms in the catechol oxidase are electronegative enough to pull the hydrogen atoms away from the oxygen atoms present in the catechol.

pH along with temperature and concentration is a limiting factor of enzyme activity. Enzymes, which are proteins, have a distinct pH range in which they work most effectively. Beyond or below this specific pH, enzyme activity begins to diminish and eventually the enzyme is rendered useless. The optimum pH of an enzyme is the pH at which it is most effective and causes the fastest rate of reaction(source). This decrease in enzyme activity beyond the optimum pH is attributed to the change in shape of the active site(explained earlier) of the enzyme. This change in shape can be brought upon when the surrounding pH moves away from the optimum pH, and it can prevent the enzyme from binding with the substrate.

Do not put the lactase pill in your mouth. Unless your doctor prescribes you, you must not eat it. This could lead to an allergic reaction.