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Polyphenol Oxidase Activity of Bananas 2
Polyphenol Oxidase Activity of Bananas 2
Polyphenol Oxidase Activity of Bananas 2
Introduction
An enzyme is a protein molecule that is a biological catalyst that speed up the rate of reaction
without itself being part of the products at the end of the reaction. Like most proteins, they
are synthesized by the ribosomes in the cell and they have some most remarkable
characteristics such as they are regulated from a state of low activity to high activity and vice
versa and also most enzymes act specifically with only one reactant called a substrate, to
produce products. The activity of enzymes is strongly affected by changes in pH and
temperature. Each enzyme works best at a certain pH and temperature, its activity decreases
at values above and below that point due to denaturation. For enzymes, denaturation can be
defined as the loss of enough structure rendering the enzyme inactive.( Banowetz , et al.,
1996).
Polyphenol oxidase is the enzyme that is found in almost all living organisms including
plants, animals and microorganisms. In plants, it is involved in defence mechanism. When a
plant gets a bruise or cut, certain phenolic compounds are oxidized in the presence of
oxygen to form a polymeric structure which prevents microbial contamination and its
catalytic action is connected to undesirable browning and off-flavour generation in stored and
processed foods.( Zhao ,et al.,2005)
Aim:
To examine how the temperature and pH level changes the activity of polyphenol oxidase and
also to determine the narrow range of temperature and pH values at which polyphenol
oxidase exhibits its optimum activity.
Results
Table 1: A tabular presentation of absorbance recorded at 450nm using different pH values.
pH
3.7
4.8
5.6
6.4
7.0
Absorbance (450nm)
0.808
2.231
1.682
1.300
1.254
Absorbance (450nm)
0.595
0.897
0.908
1.329
0.620
Discussion
Each enzyme has an optimum temperature at which it performs best.Most enzymes are active
between 10-40oC,the effect of temperature and pH on the enzymatic activity of polyphenol
oxidase was determined by measuring absorbance in varying temperatures and pH. The
temperature was ranging from 4C to 70C using catechol as a substrate (0.06M).From the
graph of absorbance against temperature drawn above it can be observed that at extreme
temperatures that is 4 C and 70C the absorbance value recorded is small indicating low
activity of polyphenol oxidase, this is because these extreme temperatures cause the native
folded structure of proteins to uncoil into random configuration. As a result, the protein loses
its biological enzymatic activity and this condition is referred to as denaturisation. The graph
produced a bell-shaped curve with the highest peak at 50C indicating the optimum
temperature for enzymatic activity. At 4C, enzymatic reaction of polyphenol oxidase occurs
slowly due to lack of energy and heat. As the temperature increases, its enzymatic also
increases up until the optimum temperature.( Gumport , et al.,1989).
Most enzymes are active only over a narrow pH range and have an optimal pH, at which
reaction is the fastest. An increase or decrease in pH also causes denaturation in enzymes,
thereby affecting their activity. From fig 2 above is a graph of pH versus absorbance at pH
3.7, the enzyme is in a too acidic environment to function. As pH decreases, certain amino
acids like aspartate and glutamate are protonated, causing them to lose their net negative
charge which consequently denatures the enzyme. From the graph it can also be noted that
the curve has the highest peak at pH 4.8 which suggest that the enzyme had the maximum
activity in the pH range 4.8-5.6, however in literature the optimum pH of polyphenol oxidase
is around 6.5. Deviation of the results obtained from the expected value can be attributed to
human error such as inaccuracies in measurement and timing during the experiment.( Alicia ,
et al.,2002)
In general, the activity of enzymes may be markedly changed by any alteration in pH, which
in turn, alters electrical charges on the enzyme. Changes in charge affect the ionic bonds that
contribute to the enzymes tertiary and quaternary structure, thereby changing the proteins
conformation and activity. Thus, pH-activity relationship of enzymes is dependent on the
amino acid side chains present in the enzyme.( Talwar and Srivastava , 2006).
Conclusion
Several factors affect the activity of enzymes. Among these are the temperature and pH. At
optimum levels of these factors, enzymes perform their function best. Optimum temperature
and pH differ from one enzyme to another.
References