Succinate Dehydrogenase Lab Report

If feeding efficiency and reproduction have a direct correlation, and a population started with equal proportions of individuals with each of three feeding types, metal spoon, metal knife, and plastic fork, the frequency of the population with metal spoons as their feeding structure will increase in the next generation. While the frequency of metal knifes and plastic forks will decrease. Furthermore, since the organisms with the metal spoon feeding structure have a higher fitness level, this population will evolve by natural selection to a point where the metal spoon phenotype will be in abundant. While the organisms with metal knifes and plastic forks phenotypes will decrease in frequency due to the lack of reproduction. Eventually, if this population persist overtime, most of the organisms, if not all, will have the metal spoon phenotype, while very few, if not any, will have the metal knife or the plastic fork phenotype.

One gram of liver was sliced into pieces, then 10ml of homogenization buffer was added to it. The mixture of the liver and homogenization buffer was then placed into a homogenizer to make the liquid slurry. Once the slurry was made it was placed into a 50ml falcon tube to then be placed in the centrifuge. The slurry was centrifuged for 2 minutes at 2000rpm. Once the spin in the centrifuge was complete, the slurry had separated, the most dense particles to the bottom of the tube forming sediment and the lighter (liver succinate dehydrogenase enzyme) also known as the supernatant. The supernatant was extracted from the falcon tube and placed into a test tube. The tube was then kept at a low temperature, (in an ice bath) until it was required for use.

In our experiment we will use differential centrifugation to isolate the mitochondria of cauliflower and then assay SDH activity using a fixed time assay. We will then measure protein content in our fractions and calculate specific activity and total activity of our fractions.

3. Explain your prediction for the effect Na+ Cl- might have on glucose transport. In other words, explain why you picked the choice that you did. How well did the results compare with your prediction?

The procedure of this lab is to determine if liver and potato cells contain natural buffers that resist large change in pH as 1. NaCl or 1. NaOH are added to the solution.

In Exercise 1, diaminofluorene is used to determine the hemoglobin concentration in the daphnids. A higher hemoglobin concentration is indicated by a darker blue color. A spectrophotometer was used to determine the absorbance at 610nm. When measuring the absorbance levels a blank is necessary to have a zero reference, the blank is the “starting point” for the measurements of the sample (re-word). The blank consists of 10µL of diaminofluorene(DAF), 50µL of hydrogen peroxide, and 0.5mL of PBS. The PBS acts as a buffer in this experiment. The cuvette with the sample of daphnids consisted of 10µL of DAF, 50µL of hydrogen peroxide, and 0.5mL of the sample of Daphnia. In Exercise 2, the Pasteur pipette was used to obtain the sample of Daphnia. The depression slide used in this experiment isolated the daphnid, cotton was used to keep the daphnid still while the heartbeat was counted. The ocular micrometer on the microscope allows the tail spine length to be measured accurately, as well as using an ocular magnification table.

Understanding the activity of enzymes in different muscle types can aid greatly in obtaining more information about other processes such as metabolism of the tissues (Anderson et al., 2012). There are many different methods in order to achieve this information based in two different major categories but the most convenient method is one called continuous assay. This process includes the use of a spectrophotometer to continuously monitor the assay. This method allows for an easy way to calculate the initial rate of reaction as one can establish time points easily. In the lab performed the continuous method was used in order to determine the measurement of the activity enzyme succinate dehydrogenase (SDH). SDH is most commonly found in mitochondria. It is present in many different muscle tissues including the heart, red, and white tissue. In the following lab the enzyme was tested and measured in these three muscle types in the Oncorhynchus mykiss in order to determine which type contained the highest and the lowest activity. This enzyme is involved in multiple processes such as involved in both the Krebs cycle and the electron transport chain (ETC) (Smith, 2014). Its role in

|(2 marks) | |c. |State two symptoms that may have led the veterinarian to suspect that George was suffering from a thyroid hormone |

Enzymes are catalysts that function to speed up reactions; for example, the enzyme sucrose speeds up the hydrolysis of sucrose, which breaks down into glucose and fructose. They speed up reactions but are not consumed by the reaction that is taking place. The most important of the enzyme is the shape as it determines which type of reaction the enzyme speeds up. Enzymes work by passing/lowering and energy barrier and in doing so; they need to bind to substrates via the active. Once they do, the reaction speeds up so much more quickly than it would without the enzyme. Coenzymes and cofactors aid the enzyme when it comes to binding with the substrate. They change the shape of the active site so the substrate can bind properly and perform its function.

2. We measured 1 mL of turnip peroxidase (the enzyme) and 3 mL of neutral buffer (pH corresponding to the test tube number i.e. pH 5 in test tube 5) with a syringe and disposed it into tubes 3, 5, 6, 7, 8, and 10

I learned that anaerobic is an organism or tissue that is living in the absence of air or oxygen while aerobic is involves the organism or tissue receiving and requiring air. Furthermore I learned about the anaerobic cellular respiration that uses an electron acceptor rather than oxygen to complete metabolism using electron transport-based chemiosmosis. Also in this reading I learned about fermentation which is an anaerobic process in which energy can be released from glucose even though oxygen is not available.

3. Water is a precious natural resource in Florida and it is said that supplies are dwindling. Desalinization (removing salt from) ocean water has been recommended as a solution to the problem. Why shouldn’t we drink salt water? Explain.

Protein purification is a process that can be employed to separate a single protein from a larger starting material which may be anything from an organ to a cell. Isolating a purified protein from a larger fraction enables further analysis such as determination of amino acid sequence, potential biological function, and even evolutionary relationship. (Cuatrecasas 1970) In this experiment, the enzyme lactate dehydrogenase will be purified, this enzyme is found extensively in human cells and catalyzes the conversion of lactate to pyruvate, an essential part in energy production. LDH is a key part of anaerobic energy production especially within glycolysis in which LDH catalyzes the conversion of the reverse reaction, pyruvate to lactate, generating NAD+ from NADH, reproducing the oxidized form of the coenzyme which can be used for oxidative respiration. (Markert 1963) Due to the fact that number of purification steps correlates with the purity of the protein multiple purification techniques will be used to isolate a pure form of LDH. LDH will be isolated from a larger “cytosol” fraction collected from a homogenized rat liver in a previous fractionation exercise. Of the procedures that will be used to isolate and purify proteins from a larger fractionate are a set of techniques collectively known as chromatography. These techniques all have the same premise, in that they consist of a stationary phase, also known as the

The first step was to obtain the White Rat and to tie it in the supine position, anterior surface facing up in side the dissection pan. To tie the animal, we used butcher’s twine and secured the front and hinds legs using a “lasso”

We could go even depeer in the explanation of metabolism reaction but this is just an