Introduction
Starch-degrading enzymes like glucoamylase are gaining more importance among the industrial enzymes because of the importance of starch, sugars and other products in modern biotechnological era1. Amyloglucosidases also known as glucoamylase hydrolyzes α-1,4 and α-1,6 linkages and produce glucose as the sole end-product form starch related polymers. Glucoamylase have applications for dextrose production, confectionery, baking and in pharmaceuticals2-3. Due to the increasing demand for these enzymes in various industries, there is enormous interest in developing enzymes with better properties such as raw starch degrading amylase suitable for industrial applications and their cost effective production techniques4. Traditionally,
…show more content…
The sterile medium was inoculated with known volume of inoculum and incubated at different temperatures (25, 30 and 350C) for different periods (48,72 and 96h) of fermentation. After fermentation the mouldy substrate was mixed with distilled water (1:4 w/v), agitated at through cheese cloth followed by centrifugation at 20,000 rpm for 20 minutes. The clear supernatant was used as crude enzyme.
Assay of glucoamylase The assay of glucoamylase was carried out according to the method of Shazia Malik, Tehreema Iftikhar and Ikram Ul Haq13. One unit (U) of glucoamylase is defined as the amount that liberates 1 µmol of reducing sugar as glucose/ml/min under the assay condition.
One ml of the diluted enzyme extract was added to 1.0 ml of 5% soluble starch solution prepared in acetate buffer (pH 4.8). The enzyme substrate mixture was incubated at 60 0 C for one hour. Then 2 ml of Dinitrosalicylic acid reagent (DNS) was added to each test tube. The test tubes were placed in boiling water for 5 minutes and cooled to room temperature. The transmittance of mixture was observed at 540 nm on spectrophotometer. The transmittance was converted to mg of glucose from standard
Therefore, acid fat, lactose fermentation, mannitol fermentation were not needed to be performed, because they are selective to a specific thing. As a result, Unknown bacteria “W” was concluded to be gram stain positive, endospore positive, bacilli shape (rod shape), and arranged in chains (strepto-). Test Purpose Reagents Observation Results Gram Stain To determine gram reaction of bacteria.
Set the wavelength to 470 nm, this is to measure the tetraguaiacol. Set the spectrophotometer to zero by using a blank. The blank should contain 13.3 mL of distilled water, 0.2 mL of guaiacol, and 1.5 mL of enzyme extract in a clean test tube. After, transfer a portion of this mixture into a cuvette, cover the top of the cuvette with Parafilm and then place the cuvette into the spectrophotometer and set it to
The purpose of this experiment was to analyze the effects of the variables: temperature, pH, and enzyme concentration, on the enzymatic reaction rate of catalase and the level at which its products are released, measuring the rate of absorption using the indicator solution guaiacol and a spectrophotometer to develop a hypothesis of the ideal conditions for these reactions. My hypothesis is that the extremes in concentration, temperature and pH will negatively affect the Au rate. This experiment used 11 solutions contained in cuvettes. Each cuvette, once mixed, is placed in spectrophotometer and then a reading taken every 20 seconds. Cuvettes 1, 8, and 10 are used as blanks to zero out the spectrophotometer.
Fill each cuvettes with its respective solution. Turn on the spectrophotometer, so it can warm up then calibrate it to 0% absorbance. Put the corresponding extract blank and set the spectrophotometer to 100% transmittance, then calibrate it to 540 nm. Once catechol is added in the cuvettes, make sure the solution is mixed. Place carrot cuvette in the spectrophotometer and record the resulting transmittance.
Enzymes are a form of protein that lowers activation energy and speeds up reactions as a catalyst. They are made by the stringing together of an abundant amount of amino acids and folded into a specific shape for chemical reactions. Turnip Peroxidase is the enzyme used in this lab and is derived from the vegetable. Enzymes are not used up or permanently altered by their environment Peroxidases are found in a range of organisms and function to break down alcohol (H2O2) and creates byproducts of oxygen and water. In this experiment, the reducing agent guaiacol is added with the substrate, hydrogen peroxide, to create water and oxygen.
Testing for the Presence of Macromolecules in McDonald’s Happy Meals Clayton Wagoner MST Biology White 4 duPont Manual High School Introduction Carbohydrates, lipids, proteins, and nucleic acids are organic molecules found in every living organism. These macromolecules are large carbon based structures. The macromolecules are assembled by joining several smaller units, called monomers, together through a chemical reaction called dehydration synthesis. The resulting polymer can be disassembled through the complementary process called hydrolysis.
Sucrase activity increases with increasing sucrose concentration Materials and Methods Effect of pH on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2. Independent Variable pH 3. Controlled Variables temperature, amount of substrate (sucrose) present, sucrase + sucrose incubation time Effect of Temperature on Enzyme Activity 1.
LABORATORY REPORT Activity: Enzyme Activity Name: Natalie Banc Instructor: Elizabeth Kraske Date: 09.22.2016 Predictions 1. Sucrase will have the greatest activity at pH 6 2. Sucrase will have the greatest activity at 50 °C (122 °F) 3. Sucrase activity increases with increasing sucrose concentration Materials and Methods Effect of pH on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2.
Proteins have an important role in the body and must be ingested on a regular basis. They provide the body with the capacity to repair skeletal muscle tissue and to synthesize hormones and enzymes among other activities. In this experiment, we wanted to determine the organic molecules found in each brand of protein supplements and the amount of starch present. A protein supplement should contain amino acids and proteins; however, some supplements contain other molecules that are used as fillers, thus reducing the concentration of the protein.
Enzymes speed up chemical reactions enabling more products to be formed within a shorter span of time. Enzymes are fragile and easily disrupted by heat or other mild treatment. Studying the effect of temperature and substrate concentration on enzyme concentration allows better understanding of optimum conditions which enzymes can function. An example of an enzyme catalyzed reaction is enzymatic hydrolysis of an artificial substrate, o-Nitrophenylgalactoside (ONPG) used in place of lactose. Upon hydrolysis by B-galactosidase, a yellow colored compound o-Nitrophenol (ONP) is formed.
Introduction In class, a series of experiments were performed that pertained to the enzyme known as catalase, which converts hydrogen peroxide into oxygen. Due to peroxide being toxic to the tissues of both plants and animals, both possess the enzyme catalase, which breaks into two non-toxic compounds: water and oxygen gas. Enzymes are proteins that react to certain substrates to create a product, and continue doing so afterwards. Methods and Materials To test reactions between catalase and hydrogen peroxide, groups of three to four people were formed.
In this experiment , we can prove that the temperature, pH and salt are the factors that will affect the structure and function of the enzyme as it is a kind of protein . Therefore, there may be an influence on the activity of enzyme which substrates cannot be binded on the active site if the amylase in too high or low ph and temperature and excess salt environment . On the other hand optimum ph and temperature and suitable salt concentration may favour the amylase activity . Reference : 1.2016, May 08). Effects of pH on Amylase Activity.
5 water bath were set up each to10 °C. (5 were used do the experiment faster) 5 cm3 of starch solution were added into the 5 test tubes that were labeled test tubes. Then 5 cm3 of amylase enzyme was added into the other 5 test tubes that were labeled. Put one of the starch solution test tube (preferably the one labeled 1) and one of the test tube containing amylase into the water bath (10 °C).
ABSTRACT: The purpose of the experiments for week 5 and week 6 support each other in the further understanding of enzyme reactions. During week 5, the effects of a substrate and enzyme concentration on enzyme reaction rate was observed. Week 6, the effects of temperature and inhibitor on a reaction rate were monitored. For testing the effects of concentrations, we needed to use the table that was used in week 3, Cells.
In order to utilize casein, bacteria cells secrete proteolytic exoenzymes (amylases, proteases, pectinases, lipases, xylanases and cellulases) outside of the cell that hydrolyze the protein to amino acids. The amino acids can then be used by cells after crossing the cell membrane via transport proteins [169]. Starch hydrolysis test is used to differentiate bacteria based on their ability to hydrolyze starch with the enzyme α-amylase or oligo-l, 6-glucosidase. These enzymes hydrolyze starch by breaking the glycosidic linkages between the sugar subunits. It aids in the differentiation of species from the genera Corynebacterium, Clostridium, Bacillus, Bacteroides, Fusobacterium and members of Enterococcus [170].