Undigested Starch Lab

In this lab or experiment, the aim was to determine the following factors of enzymes: (1) the effects of enzymes concentration the catalytic rate or the rate of the reaction, (2) the effects of pH on a particular enzyme, an enzyme known and referred throughout this experiment as ALP (alkaline phosphate enzyme) and lastly (3) the effects of various temperatures on the reaction or catalytic rate. Throughout the experiment 8 separate cuvettes and tubes are mixed with various solutions (labeled as tables 1,3 & 4 in the apparatus/materials sections of the lab) and tested for the effects of the factors mentioned above (concentration, pH and temperature). The tubes labeled 1-4 are tested for pH with pH paper and by spectrophotometer, cuvettes 1a-4a was tested for concentration and cuvettes labeled 1b-4b was tested for temperature in four different atmospheric conditions (4ºC, 23ºC, 32ºC and 60ºC) to see how the enzyme solution was affected by the various conditions. After carrying out the procedures the results showed that the experiment followed the theory for the most part, which is that all the factors work best at its optimum level. So, the optimum pH that the enzymes reacted at was a pH of 7 (neutral), the optimum temperature that the reactions occurs with the enzymes is a temperature of 4ºC or

The use of multiple test tubes and Parafilm was used for each experiment. Catechol, potato juice, pH 7 phosphate buffer, and stock potato extract 1:1 will be used to conduct the following experiments: temperature effect on enzyme activity, the effect of pH on enzyme action, the effect of enzyme concentration, and the effect of substrate concentration on enzyme activity. For the temperature effect on enzyme activity, three test tube were filled with three ml of pH 7 phosphate buffer and each test tube was labels 1.5 degrees Celsius, 20 °C, and 60 °C. The first test tube was placed in an ice-water bath, the second test tube was left at room temperature, and the third test tube was placed in approximately 60°C of warm water. After filling the test tubes with three ml of the

Amylase experiment # 2 was done to see how the pH affected the efficacy of the enzyme. First we collected all of the materials that were necessary to make this experiment. We needed five clean test tubes, the following standard solutions, 1% Starch Solution pH 3,1% Starch Solution pH 5,1% Starch Solution pH 7,1% Starch Solution pH 9,1% Starch Solution pH 11

The purpose of this experiment was to find out the nutrient content of unidentified food samples and to use chemical reagents to see if the unidentified foods for sugar, starch, protein and fat (lipid) content. After comparing the color changes in each unknown substance then you will be able to see the amount of sugar, starch, protein or fat there is in the unknown substance.

6) See attached for the graph. Correlation appears to be negative with this data. It appears like the high temperature caused the enzyme to become denatured.

The results from the experiment determining the effects of pH on enzyme activity show that as the independent variable, pH, increases the dependent variable, percentage transmission, decreases. This is shown in the results as at the lowest pH, pH 4.0 the average percentage transmission is at its highest at 97%. At the highest pH, pH 8.0 the average transmission is 78.5%. This is also supported from the graph as it produces a negative gradient showing that as the percentage transmission will decrease with an increasing pH. This happens because the enzyme trypsin acts on the gelatine. Therefore as the pH increases towards the optimum pH more jelly will be broken down by the enzyme,

An experiment was performed to test how temperature variations affect enzymatic activity of the enzyme amylase. The results of the experiment will also determine the optimal temperature of the amylase enzyme. The results of the experiment provide evidence for determining the environments that the enzyme amylase would most likely be present. By determining the possible environments, one can predict what and how environmental factors will affect the enzyme amylase. Two forms of amylase (Bacterial - Bacillus licheniformis and Fungal - Aspergyllus oryzae) were combined with starch molecules at four different temperatures (0⁰, 25⁰, 65⁰, 85⁰ Celsius). The combination of starch and the amylase enzyme resulted in a visual chemical reaction that was recorded. The enzyme activity was recorded every two minutes, starting at 0 and ending at 10. The start time 0 served as the control group of the experiment. The results concluded that both bacterial and fungal amylase has an optimal temperature around 65⁰C. This was possible to determine by recording the color change of the spot plate wells. Amylase catalyzes efficiently at its optimal temperature which resulted in yellow spot plate wells. Enzymatic activity decreased when the temperature was less than 65⁰C, resulting in a green-brown well. The green wells indicated that starch wasn’t broken down completely and was still present. Temperatures greater than 65⁰C resulted dark-green wells which resembled the denaturing of

enzymes that will be used during this lab to test the ability of amylase to break down starch ,a

The hydrolytic and degradative reactions were done next. A starch plate was inoculated to determine if the unknown organism was able to hydrolyze starch. Then an identical procedure using urease broth was done to examine if the organism was able hydrolyze urea to ammonia. The tryptone degradation test was used to determine if the organism was able to break down tryptone into indole. The phenylalanine deaminase test was performed last, which determined the ability of the organism to produce

In part II of the lab six small glass tubes were obtained in a test tube rack. Ten drops of distilled water were then added to test tube 1, five drops to tubes 2-4, and no drops in tubes 5 and 6. Five drops of 0.1M HCl were added to test tube 5 and five drops of 0.1M NaOH to test tube 6. Five drops of enzyme were then added to all tubes except tube 1. Tube 3 was then placed in the ice bucket and tube 4 was placed in the hot bucket at 80-900C for five minutes, the remaining tubes were left in the test tube rack. After the five minutes five drops of 1% starch was added to every tube and left to sit for ten minutes. After ten minutes five drops of DNSA were then added to all the tubes. All the tubes were then taken and placed in the

During these experimental procedures, the implication of multiple different temperatures on fungal and bacterial amylase was studied. In order to conduct this experiment, there were four different temperatures used. The four temperatures used were the following: 0 degrees Celsius, 25 degrees Celsius, 55 degrees Celsius, and 80 degrees Celsius - Each temperature for one fungal and one bacterial amylase. Drops of iodine were then placed in order to measure the effectiveness of the enzyme. This method is produced as the starch test. The enzyme was tested over the course of ten minutes to determine if starch hydrolysis stemmed. An effective enzyme would indicate a color variation between blue/black to a more yellowish color towards the end of the time intervals, whereas a not so effective enzyme would produce little to no change in color variation. According to the experiment, both the fungal amylase and bacterial amylase exhibited a optimal temperature. This was discovered by observing during which temperature and time period produced a yellow-like color the quickest. Amylase shared a similar optimal temperature of 55 degrees Celsius. Most of the amylases underwent changes at different points, but some enzymes displayed no effectiveness at all. Both amylases displayed this inactivity at 0 degrees Celsius. At 80 Celsius both the enzymes became denatured due to the high temperatures. In culmination, both fungal and bacterial amylase presented a array of change during it’s

At the end of three minutes the absorbance level was only 0.234. The hot temperature seems to have slowed the enzyme’s ability. The other group’s ice bath reading started at a low 1.326 it then grew rapidly over thirty seconds to the 2.4 range that the room temperature was also at. The cold temperature seemed to slow the enzymatic activity. The difference between the cold and room temperatures can be observed in graph 1

2cm of a solution was tested and added 2 cm of 10% of potassium hydroxide solution and the test tube was shaked.

Hydrolysis is a chemical reaction that uses water to split complex molecules. The water molecule H2O is split in the mechanism of hydrolysis, hydrogen cations and hydroxide anions. When the enzyme catalyzes its reaction inside the cell, it is referred to as intracellular hydrolases. When the enzymes secreted from the organism to catalyze reactions outside the cell, it is referred to as exoenzymes or extracellular hydrolases. Four hydrolysis reactions are discussed in the report, bile esculin hydrolysis, starch hydrolysis, urea hydrolysis and casein hydrolysis. Bile esculin hydrolysis determines whether an

The experiment was carried out in two parts, reaction rate verses temperature and reaction rate verses pH. For reaction rate verses temperature five test tubes were labelled with temperatures, E 0°C, E RT, E 37°C, E 60°C and E 100°C for the enzyme alpha-amylase. Another five test tubes were also labelled with the temperatures S 0ºC, S RT, S 37ºC, S 60ºC and S 100ºC for the substrate 1% Starch and NaCl.