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Effect of Lead Ions On Bacterial and Fungal Amylase

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EFFECT OF LEAD IONS ON BACTERIAL AND FUNGAL AMYLASE Amylases are enzymes, which hydrolyse starch into Maltose. There are two types of amylase: Alpha amylase that degrades starch, which is a polysaccharide, into fragments 10 glucose residues long. Beta Amylase breaks these down into maltose, which is a disaccharide of two glucose molecules joined together. Both enzymes work by hydrolysis of the glycosidic link in carbohydrates. Enzyme amylase are produced from different sources but in my experiment I will be concerned with Bacterial and fungal amylase. Heavy metal ions cause inhibition of enzyme. Inhibition of an enzyme means either to slow down the rate of reaction, to increase the rate of reaction or both. Inhibitors can slow the rate of reaction in various ways. One way is that it competes with the normal substrate for binding of active sites. . Once it is attached to an enzyme molecule it prevents the normal substrate from doing so. This is known as a competitive inhibition. A quite different inhibition would be is when an inhibitor such as lead ions can attach itself permanently to the active site of the enzyme thereby excluding any possibility of normal substrate taking up its rightful space. So in this case the inhibitors are not competing with each other so this is known as non-competitive inhibition. Inhibition is not confined to be not only confined to substances, which combine with the active sites of enzyme. Some inhibitors combine with the other parts of an enzyme molecule altering the shape in such a way that substances no longer fit with the active sites. ...read more.


Apparatus required Incubated flask - Doing this experiment in normal test tubes will create temperature problems. In order to measure the rate of enzyme effectively use incubated flask in which temperature of enzyme will be controlled in all experiments. Buffers- In this experiment I will need a Citrate buffer. This is a mixture of citric acid and disodium phosphate in appropriate proportions to give the required PH. These two chemicals are able to absorb either added acid or added base by reacting and effectively neutralizing the environment. Buffers around PH 6.5 will be needed. Dropper bottle of starch solution- we will place five droppers full of starch every experiment we do and all of these size of drops have to be equal. Dropper bottle of lead chloride- this experiment is concerned with increased concentrations of lead ions so firstly I will begin by adding one drop of solution to both bacterial and fungal amylase Pipette- this will be used to draw out the exact volume and starch of into incubated flasks. They are very accurate for measuring right volume of liquid to have fair rate of reaction. Dropper Bottle of glucose solution- this would enable to see whether Glucose is being produced in the experiment. Stop clock - I need to know what time I have added Enzymes into each test tubes. There is only a limited amount of time available for this experiment. As soon as enzyme is put into the test tubes I will start the stop clock and then after a certain amount of time I will measure the concentration of glucose formed. ...read more.


Record your results. Does starch contain any detectable sugar? Depression D is the standard for glucose. 3. I will then Place about 0.5 ml of saliva in G, and then add 2 droppers full of starch solution. Let set for 10 - 15 minutes. Now add 1 dropper full of GLUCOSE TEST SOLUTION to G. I would then Record my results. What has the Amylase done to the starch solution? I will then record the results. 4. Place 0.5 ml of saliva into depression F, add 4 drops of GLUCOSE TEST SOLUTION and note the colour change. Would the solution contain any glucose? Risk assessments and hazards Handling of both enzymes must be extreme care if there are to work properly in an experiment. There will be some toxic materials used in this experiment. Inhalation can cause breathing problems and cause damage to the body. If enzymes are in powdered form then it can cause eye problem. Wear goggles to give protection to your eyes. About harmful liquid solutions getting into your eyes, which can be very irritant so rinse your eyes immediately. Some solutions could also spill in your skin. If this happens rinse your skin immediately. You can reduce the risk by wearing disposable gloves. It is important to maintain cleanliness because minor impurities may ruin a good bio-chemical experiment. It is also very important to label different containers so we do not get the solutions mixed up. This involves less risk as it does not involve anything being heated up so less chance of skin burn. Ethical implications It would not be ethical to use a source of enzymes from animal cell sources. ...read more.

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**Plan only. This plan is lacking in a number of key aspects that would allow it to gain a good grade at A level. The written method cannot be followed and the methodology used to measure the rate of reaction is very unclear.
To improve
Research and Rationale
There is background research but the information discussed is a limited and is very confused in parts. Additional carefully selected references would help inform the project.
Planning The project could be improved by listing all the key variables in a table and then discussing their control methods and likely effect on the experiment if they are not controlled well. The choice of apparatus needs to be justified and the design of the experiment explained. The written method is lacking in key details such as volumes and concentrations. The methodology itself is questionable but if the steps were written in a clear, concise and logical way then it would be much easier for the reader to follow.
A risk assessment should be included.

Marked by teacher Stevie Fleming 26/07/2013

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