The Objective Measurement Of Alpha Amylase

There are many types of enzymes and each has a specific job. Enzymes are particular types of proteins that help to speed up some reactions, such as reactants going to products. One of them is the amylase enzyme. Amylases are found in saliva, and pancreatic secretions of the small intestine. The function of amylase is to break down big molecules of starch into small molecules like glucose; this process is called hydrolysis. Enzymes are very specific; for example, amylase is the only enzyme that will break down starch. It is similar to the theory of the lock

1) Amylase is utilized to break down starch molecules into more simple sugars for use by the body. It performs this function by hydrolyzing glycosidic linkages in the polysaccharide chain.

There is another digestive enzyme (other than salivary amylase) that is secreted by the salivary glands. Research to determine what this enzyme is called. What substrate does it act on? Where in the body does it become activated, and why?

In this lab experiment the action of the enzyme Amylase was observed on starch (the substrate). Amylase changed the starch into a simpler form, the sugar maltose, which is soluble in water. Maltose then breaks down the glucose chains of starch in the pancreas and intestines. Amylase is present in human saliva, and begins to act on the starch in the food while still in the mouth. Exposure to heat or extreme PH (acid or base) will denature proteins. Enzymes, including amylase, are proteins; if denatured enzymes can no longer act as a catalyst for the reaction. In the presence of potassium iodide, starch turns a dark purple color; however maltose does not react with I2KI. The rate of fading of starch allows a quantitative measurement of reaction rate.

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

Bacterial amylases operate at higher temperatures than do fungal amylases. Fungal amylases react rapidly at lower temperatures; fungal amylases are used as an agent for alcohol fermentation for grain (Underkofler et al, 1958). Fungal amylases is said to be denatured – change shape (Alberte et al, 2012), at high temperatures above 60° C and bacterial amylases on the other hand are stable and show little denaturing at temperatures up to 85°C 3 The question answered by the experiment is if the temperature is not within the range of the enzymes (fungal and bacterial amylase) optimal temperature (higher temperature) then will the enzymes denature and if the enzymes are placed in lower temperature from optimal the activity then will it slow down enough to stop all reaction, meaning each enzyme will not be operating efficiently. Knowing about a bacterial amylases and fungal amylases optimal temperatures are important for knowing which food products and industrial products it can be used on to conserve the product because then the producer knows about which products it can be incorporated into depending on the temperature it is manufactured at.

An enzyme is a protein that acts as a catalyst which reduces the activation energy needed for a chemical reaction. Without the presence of enzyme, cell reactions would take so long that they would detectable. During a reaction, in the presence of an enzyme, the substrate first creates a complex with the enzyme. While the substrate is a part of the complex, it’s converted into the product. Then, finally, the complex dissociates from the molecule which allows the release of the enzyme and formed product. An enzyme’s activity depends on a variety of conditions which includes the pH level and temperatures. Phosphorylase is an enzyme that catalyze the addition of a

To test enzyme activity, an experiment was conducted in the laboratory. In this experiment, the enzyme Amylase was chosen because it assists disassemble the polysaccharide starch. Starch is the main energy storage in plants. Human cells need the energy of stored in the starch to be introduced in the form of Maltose. Maltose is a simple sugar that can be used to generate the energy needed to power cellular work

Different factors will either motivate or denature the enzymes, for example, the pH, temperature, and salt concentrations are few of the aspects that feel denature the enzyme. Thus, adding pH buffer to amylase will obviously influence the enzyme function proportional to its addition to starch; this will be proved by the iodine test. The color of iodine will change into blue-black if denatured, and into orange-yellow if the enzyme is functioning at its optimum rate because the enzyme due to the digestion of starch. However, the optimum level of pH will vary on the type of amylase. The optimum pH level for alpha- amylase: 6.7 to 7; the optimum pH level for gamma amylase: 3^3.

Record observations: what color did the solution turn? Orange or blue/ black? This will tell whether the amylase is able to denature the starch solution Orange=Negative Blue/black= Positive

In this lab our group observed the role of pancreatic amylase in the digestion of starch and the optimum temperature and pH that affects this enzyme. Enzymes are located inside of cells that increase the rate of a chemical reaction (Cooper, 2000). Most enzymes function in a narrow range of pH between 5 through 9 (Won-Park, Zipp, 2000). The temperature for which enzymes can function is limited as well ranging from 0 degrees Celsius (melting point) to 100 degrees Celsius (boiling point)(Won-Park, Zipp, 2000). When the temperature varies in range it can affect the enzyme either by affecting the constant of the reaction rate or by thermal denturization of the particular enzyme (Won-Park, Zipp, 2000). In this lab in particular the enzyme, which was of concern, was pancreatic amylase. This type of amylase comes from and is secreted from the pancreas to digest starch to break it down into a more simple form called maltose. Maltose is a disaccharide composed of two monosaccharides of glucose. The presence of glucose in our experiment can be identified by Benedicts solution, which shows that the reducing of sugars has taken place. If positive the solution will turn into a murky reddish color, where if it is negative it will stay clear in our reaction. We can also test if no reduction of sugars takes place by an iodine test. If starch is present the test will show a dark black color (Ophardt, 2003).

Amylase is an enzyme that is in human’s saliva as well as the pancreas. Enzymes are biological catalysts that speed up a chemical reaction. They break down complex molecules into simple ones. In this case, amylase converts starches (complex molecule) into simple sugars. That is why foods like potatoes for example, may taste sweet to us, because they contain starch. The optimum pH for pancreatic amylase is the pH of 7. In the experiment I have used buffer solutions with the pHs of 2.8, 4 and 6.5. I have also used iodine and starch. Normally, iodine is orange-yellow, however when you add starch to it, the solution will turn blue-black.

The objective of this lab was to assess the effect that temperature and pH had on pancreatic amylase. Amylase is an enzyme involved in digestion and is secreted by both the pancreas and salivary glands (Pandol, 2010). Both salivary and pancreatic amylase display the same enzymatic activities through their role in the digestion of starch (Pandol, 2010). The goal of both salivary and pancreatic amylase is to hydrolyze starch thus breaking down the polysaccharide into a disaccharide such as maltose (Pandol, 2010). As a result, amylase breaks down these sugars making them more easily absorbed in the body during digestion (Pancreatic Cancer Action Network, 2015). Through this lab we learned about the role of pancreatic amylase and how this enzyme operates under certain conditions.

The dough must contain some alpha amylase to be able to digest the portion of starch that is amylopectin. If the alpha amylase is too much in the dough the starch will completely liquify. Alpha amylase attacks starch virtually wherever along its chains, producing smaller chains of different lengths. These chains can hold smaller units called dextrin’s that contain numerous units of glucose. Beta amylase can then digest these dextrin’s into maltose.

BREAKING DOWN STARCH USING SALIVARY AMYLASE  Caution:    You  will  be  using  a  Bunsen  burner  and  glassware  to  create  your  own  constant  water  bath.   Appropriate  caution  should  be  exercised  when  dealing  with  the  Bunsen  burner,  hot  water,  and  glassware.  Purpose:  Many plants store their energy in the form of starch, a polysaccharide made from repeating  units of the monosaccharide glucose.  Our bodies break down starch into the individual glucose units,  which are further metabolized into CO2 and water through the process of glycolysis—this is the process  we  commonly  call  digestion.    The  enzyme  amylase  is  present  in  our  saliva  and