Lysozyme Activity

Tables 2,3,4,5 and 6 show that as duration increased the absorbance also increased for each pH. The solution in the conical flask became darker (yellow) in time this is because the substrate, p-nitrophenyl phosphate was catalysed by acid phosphatase, releasing Nitrophenolate anion. It was the Nitrophenolate anion giving off the yellow colour; the presence of this feature increases the absorbance rate. The addition of sodium hydroxide distorted the shape of the enzyme making it no longer effective in its function.

The purpose of this experiment was to extract and purify LDH enzyme from chicken muscle tissue using a variety of techniques including homogenization, ammonium sulfate precipitation, dialysis, and affinity chromatography. Activity and Protein assay were used to track the overall amount of LDH present in the samples.

In order to examine the variation in sensitivity of lysozyme, gelatin, and BSA, the absorbance reading was obtained using the Bradford and BCA assay

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The same solution of 0.5 ml BSA was then added from test tube 1 to the test tube 2 after being properly mixed, and from test tube 2 the solution was being added to test tube 3, and so forth all the way up to test tube 5, with the same exact procedure. From the last tube, we then disposed the 0.5 ml solution. After above procedures, we now labeled another test tube “blank”; 0.5 ml blank distilled water was purred into the tube with the serial dilution of 1:10. We also had a tube C labeled “unknown” with the same 0.5 ml of solution. And after adding 5ml of Coomassie Blue to each tube (1-5) and to the blank, the result of absorbance was read at 595 nm.

3) Waiting 5 minutes allowed the enzymatic reaction to proceed for sufficient time and break down the starch.

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.

What other conditions that may affect the action of enzymes? Substrates could affect the action of the

The reason for the plan of this experiment is not only to find out which brand has better absorbency, but

The rate of enzyme activity of Lactase. The change in absorbance is key in determining the optimal salt concentration. The enzyme activity was measured by the use of the Spectrophotometer. Over the course of the experiment, the lowest concentrations showed the largest change in absorbance.

according to the temperature but it will be faster in bacterial than in fungal, 100% hydrolysis

After the substrate solution was added, five drops of the enzyme were quickly placed in tubes 3, 4 and 5. There were no drops of enzyme added in tubes 1 and 2 and in tube 6 ten drops were added. Once the enzyme solution has been added the tubes were then left to incubate for ten minutes and after five drops of DNSA solution were added to tubes 1 to 6. The tubes were then placed in a hot block at 80-90oC for five minutes. They were then taken out after the five minute period and using a 5 ml pipette, 5 ml of distilled water were added to the 6 tubes and mixed by inversion. Once everything was complete the 6 tubes were then taken to the Milton Roy Company Spectronic 21 and the absorbance of each tube was tested.

The absorbance is measured using a Plate reader and a Standard curve is generated. Also, the different types of pipetting techniques are assessed in this Assay.

The aim of this investigation was to determine the effect of ethanol on the membrane permeability using Beta vulgaris. Beta vulgaris contains red pigments called betalain sequestered in vacuoles. The cell membrane is generally impermeable to betalain as this pigment is relatively large and cannot pass through the membrane by diffusion. (123HelpMe.com, 2015) However, by increasing the permeability of the cell membrane, betalain can leach out of the cell and colour the liquid red. The colour intensity of the solution due to leakage of betalain is proportional to the membrane permeability. To quantify the colour intensity, the light absorbance of the solutions containing a Beta vulgaris cube were measured by a spectrophotometer. These measurements were used to analyse the membrane permeability. (Flinders University, 2015)

The products produced by the first reaction were used as a substrate for the second. In this case the enzyme used NADH, which resulted in the decreased absorbance due to the NADH oxidation to NAD+. In addition, the spectrophotometer was used as a measuring device to follow the change in absorbance of the NADH molecules at 340nm.