Fuctose-Bisphosphate Aldolase Lab Report

One of the most significant reactions in Glycolysis is reaction one which involves the phosphorylation of glucose to form glucose-6-phosphate. Through the transfer of the hydrolysis of ATP, this supplies energy for the reaction and makes it essentially irreversible, having a negative free energy change, which allows for a spontaneous reaction in cells. Although the preparatory phase is energy consuming and uses up 2 ATP, the pay off phase synthesizes 4 molecules of ATP, with the transfer of 4e- via 2 hydride ions to 2 molecules of NAD+. Therefore, a net gain of 2 ATP is achieved through the glycolytic pathway alone. Following the glycolytic pathway, due to the absence of oxygen, as oxygen cannot be supplied fast enough to undergo aerobic respiration, the athlete will instead, undergo lactic acid fermentation. Lactic acid fermentation involves pyruvate that is formed from the glycolytic pathway to be reduced to lactate, with the aid of the enzyme, lactate dehydrogenase, while the coenzyme Nicotinamide Adenine Dinucleotide (NADH) is oxidised to NAD+. The product NAD+ then re-enters the glycolytic pathway in order to produce 2 ATP. This process of lactic acid fermentation produces 2 ATP for each cycle, and thus, rapidly supplies the body with a small amount of energy. However, with the buildup of lactic acid in the body, the athlete will eventually encounter the feeling of discomfort as this accumulation of lactate causes the body to

bonds and taking a glucose molecule off of the structure (Ball, Vialle, Alonso-Casajus, Duavillee, Munoz, Baroja-Fernandez, Moran-Zorzano, Eydallin, Pozueta-Romero, 2006). The conditions for the reaction of Phosphorylase and

4. Fructose is component of sucrose, normal table sugar, along with glucose. Whereas glucose is able to immediately enter into glycolysis, fructose is not. Fructose is broken down via fructokinase into fructose- 1-phosphate. Fructose – 1-phospate then gets converted into DHAP+ glyceraldehyde via aldolase B. DHAP+ glyceraldehyde is used in glycolysis to produce pyruvate that goes into the citric acid cycle to produce ATP

As seen in Figure 1, the EMP pathway begins with glucose, a six carbon monosaccharide. In the first reaction, a phosphate group is transferred from ATP via substrate-level phosphorylation via hexokinase to yield glucose-6-phosphate. In the second reaction, it undergoes an isomerization to yield fructose-6-phophate. In the third reaction, another phosphate group is transferred from ATP to yield fructose-1, 2-bisphosphate and is catalyzed by the important regulatory enzyme phosphofructokinase. At this point in the pathway, the molecule still contains six carbons. However, in the fourth reaction, the six carbon molecule is broken into two three carbon molecules: glyceraldehyde-3-phosphate and dihydroxyacetone-3-phosphate catalyzed by aldolase.

Background: Enzymes are proteins that increase the rate of reaction by lowering the activation energy. When an enzyme binds to a specific substrate, an enzyme substrate complex is formed, allowing for the reaction to take place over less time. Since the enzyme is recyclable, the reaction produces a product and the unaltered enzyme. Catalase is the enzyme used specifically for the decomposition of hydrogen peroxide. Furthermore, the amount of substrate used can be determined by measuring the amount of product formed in the reaction. If the amount of product formed over time is graphed, the graph will prove the rate of reaction to decrease over time as the availability for a substrate and enzyme to bind decreases. The initial velocity of an enzyme-catalyzed reaction is constant because the number of substrate molecules at the beginning of a reaction is typically vastly greater than the number of enzymes, causing the rate to be determined by the characteristics of the enzyme. When the substrate concentration decreases to a certain point, when enzymes and substrates do not easily collided in the specific manner to form the enzyme-substrate complex, the rate of reaction will decrease.

Introduction Enzymes are considered catalysts that encourage growth in chemical reactions of cells. Enzymes are not eternally transformed, nor do they modify the products and reactants between cells during a chemical process (Cooper, 2000). Lactase is an enzyme formed in the body that helps break down lactose, a carbohydrate sugar often found in milk, into simpler sugars. Without lactase, a person who has consumed lactose will present with bloating of the abdomen, stomach cramps, and have a feeling of the need to vomit (Itan et al., 2010). Lactose is broken down by lactase makes it easier for the body to digest the simple sugars.

As the muscles begin to exert force, in order to obtain quick energy, energy must be produced through an anaerobic pathway. The first step of carbohydrate metabolism is glycolysis where a chain of reactions take place to release the energy stored in glucose. Glycolysis is a ten reaction pathway, the first five reaction constituting of the preparatory phase and the final five constituting of the payoff phase. This process all occurs in the cytosol where the glucose molecule is split to form two molecules of pyruvate. The preparatory phase is the production of two molecules of glyceraldehyde-3-phosphate through phosphorylation of a glucose molecule. The consumption of 2 ATP molecule allows this phase to proceed. Reaction 1 is catalysed by hexokinase and is the phosphorylation of glucose to form glucose-6-phosphate. The phosphate group from ATP is transferred to glucose forming glucose-6-phosphate. This reaction is an exergonic reaction with a ΔG˚’=-16.7KJ/mol and the energy released by hydrolysis of ATP drives the reaction forward. Reaction 2 involves the isomerization of glucose-6-phosphate to fructose-6-phosphate. This reaction is catalyzed by phosphohexose isomerase and is an endergonic reaction with ΔG˚’=+1.7KJ/mol. In reaction 3, fructose-1-phosphate is further phosphorylated to produce fructose-1,6-bisphosphate. Second ATP is consumed in this endergonic reaction with ΔG˚’=

Although we might not always acknowledge this small proteins, enzymes have come to be revolutionary when it comes to our survival. This is because enzymes are an example of catalysts, which affect the rate at which a chemical reaction occurs by speeding up. Moreover, this is imperative for our survival because chemical reactions provide the energy required by our bodies to perform several life processes that allow us to carry out many of the activities we perform as part of our everyday life. Thus, an enzyme's ability to function properly has become a major factor in our body's ability to function properly and maintain homeostasis. Unfavorably, however, there are some factors that can negatively affect an enzyme's ability to

An enzyme is a protein that acts as a catalyst which reduces the activation energy needed for a chemical reaction. Without the presence of enzyme, cell reactions would take so long that they would detectable. During a reaction, in the presence of an enzyme, the substrate first creates a complex with the enzyme. While the substrate is a part of the complex, it’s converted into the product. Then, finally, the complex dissociates from the molecule which allows the release of the enzyme and formed product. An enzyme’s activity depends on a variety of conditions which includes the pH level and temperatures. Phosphorylase is an enzyme that catalyze the addition of a

In order for maximal activity of E. coli β-galactosidase requires both Mg2+ and Na+ (Huber). Another component of the experiment was the use of ONPG as a substrate, which is a colorimetric substrate that turns yellow when a reaction occurs. Some other substrates that are commonly used to react with this enzyme, are X-gal and PNPG (Juers). All of these substrates have different rate-determining consequences, which are all involved in the kinetics of the mechanism. On a molecular level the individual interaction between these substrates and the proteins in this enzyme differ.

This experiment was conducted to investigate the enzymatic activity of the enzyme aldolase by using different urea concentrations. Stronger solutions of denaturant in this case urea were used to increase the polypeptide subunit unfolding and the dissociation of the tetramer.

Aldolase B is classified as a Lyase, or an enzyme that catalyzes the breaking of various bonds means other than hydrolysis and oxidation. Lyase often forms a double bond or a new ring structure. Fructose Intolerance is caused by mutations in the aldolase B gene which is expressed in the liver (Malay, Allen, and Tolan). Its molecular compound is shown in Figure 2 and has two active site residues, Lysine and Glutamate, as well as several charged residues. The functional assignment is complex because of the proximity of the many charged amino acids found at the active site. Figure 3 shows where the substitution of Proline for Alanine occur at position 149 (Malay, Allen, and Tolan). The A149P substitution is known as AP-aldolase (Malay, Procious, and

Fructose-1, 6-bisphosphate is a key regulatory step in gluconeogenesis, as well as many other intracellular metabolic pathways . During, gluconeogenesis there is an important process in which there is a conversion of glucose to pyruvate which is known as glycolysis. This process will require three irreversible steps that have a very high negative free energy that is in the forward reaction. So, in order to have a conversion from pyruvate into glucose, the pathway will require the use of enzymes, which will allow the bypassing of these irreversible steps. One of the enzymes that is used in this process is called Fructose 1, 6-bisphosphatase (Kelley, 2006). This step is a very important step in gluconeogenesis, being that it needs to have fructose bisphosphatase to catalyze the conversion of fructose-1, 6-bisphosphate into fructose 6-phospahate, and inorganic phosphate, that without it can block the pathway. Its activity is high regulated by the levels of Adenosine Monophosphate, fructose 2, 6-bisphosphate and also citrate (Kelley, 2006). When deficiencies are present in this pathway and devoid of this conversion, glycerol into glucose, it will lead to fasting hypoglycemia, lactic acidosis and other physiological conditions. This enzyme is highly active within the liver and the intestines. Therefore, when the liver glycogen stores are no longer available, the physical properties of the body will fight for its homeostasis (Eren, 2013) by converting a three

Phosphomannose isomerase is a key enzyme for mannose metabolism converts mannose-6-phosphate into fructose-6-phosphate, which enters the glycolytic pathway after isomerization to fructose-6-phosphate, thus allows mannose to become a sole carbon source.

The purpose of this report is to discuss about Fructose 1,6-Bisphosphate Aldolase Class II in L. lactis. However, studies on several other bacteria like E. coli, Streptococcus sp. and other Lactococcus sp. will also be used as reference containing similar versions of this enzyme taking part in their cycles.