Adaptation And Regulation Of Fatty Acid Composition: The Effect Of Enzyme Concentration, PH And Temperature

Materials and methods

Discuss about the Adaptation And Regulation Of Fatty Acid Composition.

Enzymes are found in different parts and the body tissues are some places which contain some of these enzymes. In the body, these enzymes play a critical role in energy metabolism. To function effectively, the enzymes are expected to function at some key conditions (Enger, Ross & Bailey, 2012). By definition, enzymes are micromolecular biological catalysts which accelerate key processes (Kuby, 2001). The enzymes are able to act on different substrates and convert them into different molecules of known products. More than 5,000 biochemical reactions are known to be catalyzed by the enzymes. Most of the enzymes are proteins while other are RNA molecules (Stivers & Rutgers University, 2011). In terms of structure, enzymes are mostly three-dimensional structures. Temperature and PH levels are some of the key conditions which affect the functionality of the enzymes. Enzyme activity is the rate of conversion, which the enzymes are able to implement during their action. Pyrophosphatese enzyme is found in tissues and like all other enzymes, it is affected by PH and temperature change. Inorganic pyrophosphatase, which is also referred to as pyrophosphate phosphorlyase E.C. 3.6.1.1 has been previously purified to achieve homogeneity (Li, 2010 and Symposium, 2009). Investigation of the conditions which are able to affect the functionality of the enzyme is important. When dissolved in water, the enzyme is able to produce inorganic phosphate. The hydrolysis process is a perfect process which is used to purify the enzyme and therefore can be used to determine the level at which different conditions affect its functionality.

Enzymes activity, which is the enzyme reaction velocity of the pyrophosphatase enzyme, is affected by key factors such as enzyme concentration, pH level on medium of action and temperature of the medium (Kim et al., 2011). Changing these factors is the best way to determine the level at which the enzymes are affected by different conditions which they operate. Pyrophosphatase is relative stable enzyme and therefore its reactivity can be well measured. In addition, the enzyme is soluble and this makes it easy to measure its reactivity (Piazza, Dieckmann & University of Waterloo, 2016). In addition, the enzyme is not permanently attached to any of the sub-cellular structures. Therefore the enzyme can be well purified by the key methods which will be applied in this experiment. In order to measure the reactivity of the enzyme, will be determined through the measurement of the amount of inorganic phosphate (Pi) which will be produced in the hydrolysis of the pyrophosphatase (PPi) as in the figure below;

Results

The functioning of enzymes on different conditions has not been intensively carried out and therefore it is important to analyze these conditions and the way the affect the functioning of the enzymes (Dixon, 2014).

The experience will be carried to determine the reactivity of the enzyme at different reactants. Some of the materials which will be used include;

• Pyrophosphate
• MgCl₂
• Triethanolamine buffer (TEA) of pH 7.2
• Enzyme pyrophosphatase
• PO₄
• H₂O
• Acid-molybdate

First, the tubes were labeled. Then different reactants were added with reaction of the inorganic phosphate reacting with molydbate acid. Standard curve for the experiment will be provided first. The mixture will be well mixed and different reactants added in each of the experiments. A gap of 10 minutes will be allowed to ensure that the reactants are able to develop the preferred results.

[PO4]mM	Abs1	Abs2	Mean Abs	Range
0	0	0	0	0
0.2	0.047	0.047	0.047	0
0.4	0.099	0.1	0.0995	0.001
0.6	0.152	0.151	0.1515	0.001
0.8	0.203	0.202	0.2025	0.001
1	0.283	0.28	0.2815	0.003

Increasing the PO₄ concentration is able to influence and increases the absorbance reaction. Decreasing concetration level decreases the molarity of the acid in each case and therefore reducing the absorbance rate. Water molecules decrease means that the concentration of the acid is able to decrease the velocity of action for the enzyme. The reagents are able to change due to the decrease on the enzyme concentration level. In relations, the water and acid concentrations can be described to be inversely related. As the water concentration reduces, the acid concentration is able to increase. Phosphate concentration is able to increase gradually with reducing water level.

abosrbance	tempreture
tempreture	abs1	abs2	average abs	eange
4	0.027	0.094	0.0605	0.067
30	0.093	0.105	0.099	0.012
50	0.296	0.303	0.299	0.007
70	0.358	0.384	0.371	0.026
100	0.171	0.201	0.186	0.03
100	0.05	0.051	0.0505	0.001

The amount of temperature has great effect on the enzyme action. The increased enzyme is able to lead to increase with amount of temperature present for action. The velocity reaction of the enzyme can therefore be concluded that it increases with increase with temperature level. This part of the experiment is set up at low temperature and temperature is gradually increased and change of the reaction velocity of the enzyme is monitored. Nevertheless, the enzyme velocity action has a maximum level at which it has to affect the temperature of the enzyme. Therefore, temperature concentration is able to influence the reaction velocity of the enzymes. The enzyme concentration is able to determine the ability of the condition to affect the enzyme action.

Ph	Absorbance at 620		avrage Abs	Range
4	0.026	0.03	0.028	0.002
5	0.045	0.044	0.045	0.001
7	0.073	0.073	0.073	0
8.5	0.062	0.064	0.063	0.001
10.5	0.025	0.029	0.027	0.002

PH level is able to affect the functionality of the enzymes. High level of acids and high levels of bases is never the conducive environment for the action of the enzymes. That is why the curve reaches a maximum point and then lowers down.

The enzyme is affected by different conditions. These conditions determine the level of action which is experienced each time the specific enzymes get into action (Owen, Punt, Stranford, Jones, & Kuby, 2013). First, the concentration of the enzyme, temperature and pH are some of the common key conditions which the enzymes are able to experience during their action. These conditions are prone of changes and this means that the action curve for the enzyme changes when each of these factors changes (Sherwood, 2015). The experiment carried out was able to demonstrate clearly that these conditions are able to affect the action curve and velocity for the PO₄ enzyme.

Discussion

First, from the standard curve, the concentration of the enzyme is important in determining the amount of molecules which are present for the action (Dingee, 2010). Increasing the amount of particles of the enzyme means that an increase in velocity of action. The curve presented is able to show that increasing the concentration of the PO₄ enzyme, the absorbance rate is able to increase as well. The result in the concentration increase shows that there is a direct correlation on the enzyme activity and the concentration of the enzymes. The increase in concentration means that the enzyme has mole molecules par liter to carry out the activity.

The curve result shows that the absorbance rate increases with increase of the concentration rate of the enzyme. From the simple explanation, the molarity of the enzyme is able to indicate the amount of particles which are presence in a specific mass of the enzyme (Watson, 2014). Increasing the molarity and maintaining the volume of the enzymes means the concentration increases. This shows that there are many particles which are therefore to take part in the acceleration of the activity of the enzyme. The absorbance rate, which is an indicator of the action of the enzyme therefore, is able to increase with the amount of particles which are presence for action.  Therefore, it can be concluded that in order to achieve high velocity reaction for the enzymes, high concentration is required. The amount of particle which available for reaction is usually increased when concentration is high and this increases the rate of reaction for the enzyme. In addition, the range which is minimal for the experiment is able to indicate that there are key factor which as well affect the reaction of the enzyme. In order to achieve effective PO₄ enzyme action, high concentration is therefore required. This provides the more particles which are needed for it to act and therefore increase the velocity reaction of the enzyme.

The experiment was also carried to determine the effect of pH change as an environment and condition where the enzyme acts. The pH level is able to increase from low acidic conditions to high basic levels (In Bathaie & In Tamanoi, 2015). The middle part of the pH is usually at 7, which is neither acidic nor basic. Different enzymes have different environments which they act well. Some will act well at the acidic conditions, other acting well at neutral conditions while others will act well act basic conditions (Tamanoi, Stafforini & Inoue, 2015).). Like the other enzymes, PO₄ will as well act perfectly as some key conditions. The absorbance rate, which indicates the velocity of reaction for the enzyme is clearly affected by ph condition according to the result of the experiment.

From the results and curve, the action of the PO₄ enzyme increases with the increase of pH level. This increase reaches the peak at the middle of the pH level and then it starts to decline. Therefore at the acidic condition, the velocity of enzyme reaction is able to increase. The peak of the curve means that the maximum action of the enzyme is reached. Maintaining this level of the pH means that the maximum action of the enzyme will be achieved. Projecting downwards, the maximum action point at which the PO₄ acts is at pH 7.0. This is the neutral point of the pH indicator. Therefore in order to achieve the maximum action for the PO₄ enzyme, a neutral condition is required. The near neutral condition will as well favor the action of the enzyme and the extreme ends will provide the least reaction of the enzyme. The extreme ends of the pH are able to reduce the activity of the enzymes and thus unfavorable for the enzyme action. In addition, it can be seen from the point where the extreme ends are able to offer conditions which damage the action molecules of the enzymes which are responsible for the action of the enzyme.

Lastly, this experiment was able to analyze the absorbance rate of the enzyme considering the different temperatures. Like the other conditions, the temperature level plays a critical role in the velocity action curve for the different enzymes (Svendsen, 2016). Different enzymes are able to act well at specific temperatures and this is the same with the PO₄ enzyme. The action of the enzyme depends the level at which the different enzyme components are able to allow their maximum action (In Page, 2004). For instance, low temperatures may freeze the enzyme components therefore making the enzyme components inactive. This will in turn slow the velocity action for the enzyme. Moreover, extreme high temperatures conditions may as well offer unfavorable conditions for the enzyme components to act. Each enzyme components are able to have specific temperature at which they will have their maximum activity and action.

From the results in this experiment, the absorbance rate for the PO₄ is increasing with the increase with temperature. The low temperature which is at 4^oc has the lowest action of the enzyme. This is an indicator that the temperature is able to freeze the molecules of the enzyme and therefore making them inactive to act resulting to the low velocity. The temperature is gradually increased up to 70^oC and them reduces as the 100^oC is reached. In turn, the velocity action of the enzyme is able to increase with the increase with the temperature and decrease the extreme increase of the temperature. Therefore the maximum point of action for the PO₄ enzyme is at around 70^oC.  This temperature is able to provide the maximum action of the enzyme and therefore offering the highest velocity of action. Moreover, the high temperatures can be seen to damage the molecules and therefore making them inactive. This means that the enzyme molecules will be unable to perform their duties and act according due to the high temperatures. This will lower the action of the enzyme.

Conclusion

In conclusion, it is clear from the experiment that the different conditions are able to affect the action of the enzymes. From the experiment, I was able to learn that the enzymes are able to operate effectively at certain conditions. Additionally, different enzymes are able to act different differently when these conditions are changes. PO₄ was able to show that these conditions are essential for the maximum action of the enzyme. First, the concentration increase of the enzyme is critical for the enzyme action. Increasing the concentration is able to increase the action and velocity of action. Secondly, the experiment was able to show that too much temperature is able to lower the action of the enzyme. Likewise, low temperature also lowers the action of the enzyme. Under this the perfect temperature must be analyzed for each enzyme to achieve the maximum action. For instance the experiment was able to show that PO₄ acts well at temperature around 70^oc. lastly, the pH level is able to affect the enzyme activity curve. PO₄ is affected by acidic and basic conditions of the pH. The favorable condition of this enzyme is achieved at neutral level and it is the point where the highest activity is achieved.

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