Ion Exchange Chromatography Lab Report

Box on right illustrates the peptide bond resulting from the condensation of both the amino acids. The box on the left illustrates the separation of the hydroxide group from glycine and the hydrogen atom from valine.

There are six variant forms of an enzyme; one is normal and five are mutant. The normal enzyme has glutamine at amino acid position 150, which is on the protein surface. Each mutant form has an amino acid substitution at position 150 as indicated. Which mutant form is most likely to have an increase

This technique separates Rubisco samples based on their size. The electrophoresis has a positive and a negative end. Positive charge proteins are loaded from the positive end and migrate towards the negative end. Negative charge proteins are loaded from the negative end and migrate towards the positive end (Sakthivel & Palani, 2016). The sample that contained the highest molecular weight of Rubisco will travel the shortest distance on the gel while the protein with the smallest molecular weight will travel the longest distance (Sakthivel & Palani, 2016). The size proportion of each Rubisco molecule correlates with the distance traveled. Rubisco will be in its purest form after running through SDS-page since each technique will increase the purity of the protein. If the salting out, the ion exchange and the SDS-page protein isolation techniques are performed on protein Rubisco, then it is purified and separated by solubility, charge, and size. The rationale of this experiment is to isolate the purest form of Rubisco so that it can perform carbon fixation at an optimal

When the pH is not at its optimum, the differing pH's will disrupt the bonding between the R groups of the amino acid causing its structure and the shape of the activation site to change

To prevent fluctuation in the pH, a solution known as a “buffer solution” was used in the experiment. Buffer solutions are mixtures of at least two chemicals which counteract the effect of acids and alkalis. Therefore, when a small quantity of alkali or acid solution is added the pH of the enzyme doesn’t change.

The pH of a solution is the measure of the concentration of charged Hydrogen ions in that given solution. A solution with a pH lower than seven is considered to be acidic. A solution with a higher pH is a base. It is very important for organisms to maintain a stable pH. Biological molecules such as proteins function only at a certain pH level and any changes in pH can result in them not functioning properly. To maintain these constant pH levels, buffer solutions are used. A buffer solution can resist change to small additions of acids or base’s. A good buffer will have components that act like a base, and components that act like an acid.

amino acid was weighed and placed in a 250-mL volumetric flask. About 100 mL of

purified. Through the different migration rates, compounds with a more negative charge will migrate down faster due to the positive electrode. The LDH isozymes also have different isoelectric points (pI) , giving a different net charge, therefore affects its migration rate. Two standards are placed into the agarose gel for comparison next to our own sample.

The titration curve of the unknown exhibited many characteristics, such as equivalence points, pKa of ionizable groups, isoelectric point, and buffer regions, that are particularly distinct to lysine. For unclear reasons, the pH during the titration did not reach the pH for pure 0.2 M NaOH nor 0.2 M HCl and normal equivalence points expected at two extreme ends of the titration curves for all amino acids were not observed. The titration of a phosphate buffer showed that the buffer capacity is directly proportional to the molarity of the buffer. However, our results showed that although the initial pH of the phosphate buffer was less than the pKa value, the measured buffer capacity was higher towards acid than base. The accuracy of the pH meter and calibration process was questioned under assumptions that the pH of the prepared phosphate buffer was actually above pKa.

Table 2: Consists of color extract taken from a red cabbage for a natural indicator. The pH reading that was measured by using the pH meter and the result of the pH reading to determine whether the solution was acidic or basic.

Purpose the purpose of this experiment was to perform test to detect the presence of carbohydrates, proteins, lipids, and nucleic acids. Explain the importance of a positive and a negative control in biochemical test. Use biochemical test to identify an unknown compound.

III. Materials and Methods Solution Descriptions: Solution A – Alkaline buffer Solution B – Substrate (0.003 M para nitrophenol-phosphate (pNPP)) Solution C – Enzyme ALP, low concentration Solution D – Enzyme ALP, high concentration Solution E – Solution B/dH2O. This solution will be used for the pH experiment.

An acid-base titration is the determination of the concentration of an acid or base by exactly neutralizing the acid/base with an acid or base of known concentration. This allows for quantitative analysis of the concentration of an unknown acid

Enzymes owe their activity to the precise three-dimensional shape of their molecules. According to the 'lock-and-key' hypothesis, the substrates upon which an enzyme fit into a special slot in the enzyme

     Liquid chromatography is an analytical technique that is useful for separating ions or molecules that are dissolved in a liquid phase. If the sample solution is in contact