enzyme’s activity might get influenced owing to several factors such as; pH, temperature etc. How pH affects the action of enzyme it maybe understood through addition of acid and base [JC2] (in the form of food)in reaction mixture orby encouragingdirect reaction between enzyme and foodstuff(Eswar et al., 2008).
Experiment: Effect of low pHon Enzyme’s activity
Date
Introduction
Enzyme’s activity might get influenced owing to several factors such as; pH, temperature etc. How pH affects the action of enzyme it maybe understood through addition of acid and base [JC2] (in the form of food)in reaction mixture orby encouragingdirect reaction between enzyme and foodstuff(Eswar et al., 2008).
Objective
Recognition of impactof low pHon theactivity of enzyme.
Material Required
Yeast,H2O2, Fresh meat (fish) means; acidic food, Fresh vegetable (potato) means[JC3] ; alkaline food, Scaled Beaker and Dropperand so on.
Hypothesis
Low pHcan affect enzyme’sactivity.[JC4]
Hypothesis Testing
Catalase enzyme reacts on its substrate in the following manner:
H2O2=H2O+1/2O2↑
If pH could affect the activity of an enzyme then any transformation introduced in it might influence the level of oxygen tootherefore; the procedure related with creation of bubble mightgetaffectedas well because, it depends on oxygen’s level.
Method
The experimentwas performed in the following way: Reaction of yeast was promoted on Hydrogen peroxide in the presence of food.Here we need to include both types of food means; acidicand alkaline with the intention that, impactof acidic as well as basic both types of pH on enzyme’s action might be scrutinized. Owing tocatalase (present in yeast), oxygen gas was releasedfrom H2O2in the form of bubbles. Now, bymeasurement of level of bubbles,enzyme’saction on food itemscould be observed effortlessly. Here are the details of protocol followed by me:
- 1 cm. cube of meat was added [JC5] in to a 100 ml graduated cylinder.
- Addition of 2cm3 of yeast(a source of catalase enzyme) was also done into the same cylinder.
- Then I introduced 2 ml. of Hydrogen peroxide into a boiling tube.
- Cylinder and boiling tube both were kept in the water bath at 250C[JC6] .
- Afterwards, I poured Hydrogen peroxide from boiling tube into the cylinder.
- Further, I noted the volume (initial volume) of fluid present in the cylinder in an immediate mode and recorded the same.
- After 2 minutes again I noted the volume (final volume) and recorded it.
- Initial volume was subtracted from final volume so that, an idea about the volume of foam could be gained.[JC7]
- Further, I repeated the same procedure by replacing the acidic food from alkaline one
(1 cm. cube of potato).[JC8]
- On the basis of data obtained from above experiment,I prepared a table and graph that have been presented below.
Result
Collection of Data
Collection of data can be done with the help of graduated cylinder.
Table 1: Effect of acidic and basicconditions on Enzyme’sactivity
| Tube | Food | Initial volume | Final volume | Bubbling (foam level) Final volume -Initial volume |
| 1 | Potato (basic) | 25.00 ml. | 28.40 ml. | Bubbling Present, volume= 3.40ml. |
| 2 | Fish- meat (acidic) | 25.00 ml. | 26.20 ml. | BubblingPresent, volume= 1.20ml. |
Graph
Activity of
Enzyme
0__1__2__3__4__5__6__7__8__9__10__11_
pH
Discussion
Data Analysis
Data analysis might be performed through havingan idea about altered pH condition and level of oxygen generated.
Observation
With the aid of datagained via experiment, we might identifydiminished oxygen generationduring acidic conditionbecause ofreduced activity of enzyme, but in basic situation,enzymatic activity is superiorso; augmentationin foam’screationmay be observed here.In addition, according to thegraph, maximum activity of enzyme couldbe viewed at pH=8.
Thus; my hypothesis that acidic fluid can affect enzyme’s activity (adversely here) could be establishedand can be acknowledgedtoo. It reveals the requirement of establishment of proper pH condition all the way through the enzymatic reaction in order that,utmostprofitcould be attained from it(Machida et al., 2000).
Conclusion
Low pH can affect enzyme’s activity, and this impact might be due tomodification in the ionization of amino acid residues and alteration of the non-covalent bonds present in enzyme.
References
Eswar N, Eramian D, Webb B, Shen MY, Sali A. (2008). Protein structure modeling with Modeller. Methods Mol Biol. 426: 145–159.
Machida S, Ogawa S, Xiaohua S, Takaha T, Fujii K, Hayashi K. (2000). Cycloamylose as an efficient artificial chaperone for protein refolding. FEBS Lett. 486: 131–135.
Cristal,
I am sorry you have done this experiment, but it has to be redesigned and redone. Both the fish and potato contain catalase, so you have not actually tested the effect of pH on enzyme function, but rather you tested how the amount of catalase present affects enzyme function.
You need to give your independent, dependent and controlled variables. You need experimental controls (positive and negative).
Look at my comments, ask me questions, and revise accordingly. Please do not repeat the experiment until I have approved your outline.
50/100pts
[JC2]Remember the project asks you to test the effect of acids not pH in general.
[JC3]Meat, potato and fish all contain catalase in them, which will confuse your results. Please just you yeast as your catalase source. Can you think of acids you have in your house? Perhaps in the form of vinegar and citrus juices? These will not contain catalase in them.
[JC4]Please be specific – will acids increase or decrease enzyme function?
[JC5]Meat has catalase in it. So this will confuse your results.
[JC6]WE have already tested the effect of temperature on enzyme function, so you should be doing this entire experiment at room temperature.
[JC7]It is better to use balloons to capture the oxygen produced or to measure the height of the foam with a ruler.
[JC8]No bases in this experiment! Potatoes are not a good option.
[JC9]Did you draw this graph? If not it needs to be cited.
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