Trypsin

algorithm with the retrieved sequence of Trypsin-1, accession no P07477.7 [1] (Appendix 1). Trypsin-1 is a 247 amino acid long protein [2] (Appendix 3) of the serine protease family, with a molecular mass of 26,558 Daltons [3] (Appendix 2). It is globular in shape and contains two regions of α-helices and multiple regions of β-sheets, as seen in Figure 1[4]. It contains disulphide bonds at residue positions 30/160, 48/64, 139/206, 171/185 and 196/220. The tyrosine-154 undergoes sulphation as a post translational modification. There are 22 negatively charged and 20 positively charged residues which contribute to a theoretical pI of 6.02 [2] (Appendix 3). Trypsin-1 is a zymogen. Residues…

INTRODUCTION Trypsin is a proteolytic enzyme, important for the digestion of proteins. Enzymes are biological catalysts for metabolic process in cells. A catalyst is a substance that changes the rate of a reaction. Enzymes act as catalysts for chemical reactions in the body. Enzymes are large proteins that are not consumed or changed in a chemical reaction. They are highly specific to the substrate - the substance an enzyme acts on - they act upon. When an enzyme is available with an empty…

Because of the enzymes reacting less frequently with substrate molecules at 22°C, larger molecules of Casein will still be present. As a result, the absorbance will be higher from light passing between the molecules. Continuing on, Temperature 22°C will have more absorbance than 32°C because we are moving closer to the enzymes optimal working temperature. For our upcoming experiment, the Trypsin will work best at 37°C as stated above. At 37°C, the level of absorbance will be the lowest because…

hypothesized that the optimum pH of trypsin is 8. Three different pH levels were tested, and each level consisted of three trials. The pH of 5 showed an average absorbance of 0.021±0.002, the pH of 8 showed an average absorbance of 0.167±0.030, and the pH of 11 showed an average absorbance of 0.088±0.001. Based on the standard deviation, there was a lack of variability between each trial for the pH levels of 5 and 11, yet the trials of the pH of 8 varied greatly compared to the other levels. …

for 30 min at room temperature. Then, trypsin/chymotrypsin- digested samples of KPNB1 obtained from the UV-crosslinking experiments with various concentrations of compound 1 were analyzed by nanoflow liquid chromatography and tandem mass spectrometry. The tandem mass spectra of the trypsin/chymotrypsin-digested KPNB1-compound 1 adducts showed 10 peptides coupled to compound 1 (Table S1, the Supplementary Information). Among these adducts, we found that the amounts of three of the products,…

This experiment is still under optimization, I keep losing cells after infection and this might be due to the addition of TPCK-trypsin (L-(tosylamido-2-phenyl) ethyl chloromethyl ketone). TPCK-trypsin helps the virus to infect the cell through cleaving the HA protein into HA1 and HA2. According to Lee et al the optimum TPCK-trypsin concentration that can be used to infect DF-1 cells with IAV is 0.1 mg/ml (Lee et al. 2008). I am planning to try different concentrations to see which one will work…

Trypsin is also known to act on itself resulting in both active and inactive forms of trypsin during storage (Sugumaran). In this experiment, affinity chromatography was used to extrapolate the active form of trypsin from the inactive form of trypsin. A column containing P-amino-bezaidine was used to form a bond with the active form of trypsin in the matrix (Sugumaran). Two assays were performed. The first assay was to determine if there was any active trypsin by employing…

Protease plays in important role in processing the protein for proteomic analysis. Only a few proteases that are useful for proteomic analysis. For instance, trypsin and chymotrypsin are often used to cleave proteins at selective amino acid residues. Trypsin cleaves on the C-terminal side of the lysine and arginine amino acids, whereas chymotrypsin cleaves after the aromatic amino acids. Trypsin exhibits higher activity and narrower specificity compared to chymotrypsin. When the lysine reside…

different types of enzymes on different substrates, and how pH influences the ability of enzymes to breakdown proteins. Starch and albumin were used as the substrate to test the enzymes amylase, pepsin, trypsin, and unknown enzyme T. Lugol’s solution was used to test for the presence of undigested starch, while Biuret solution and Ninhydrin solution were used to test for the presence of undigested protein and amino acids respectively. There were many key findings in this experiment, one of which…

of Ethiopia) evaluated in this study are well endowed with all of the essential nutrients required for human nutrition. Should also be cautioned as they have indicated high antinutrient content especially trypsin inhibitors and alkoloids. All the tested samples contained antinutrients in varying concentrations. Consumption of this plant with low concentrations of antinutrients may appear safe at face value, but it should be noted that consumption of these plant materials over a long period as…