Enzymes are bio-catalysts and they catalyse the biochemical reactions both in vivo (inside the cell) as well as in vitro (in the test tube). They are highly specific to its substrate and have great catalytic power, i.e., they enhance the rate of reaction tremendously without being changed.
All enzymes are proteins with exception of some small group of catalytic RNA molecules called ribozymes. Like proteins, the molecular weight of enzymes ranges from about 2000 to more than one million Dalton.
Classification of proteinaceous enzymes based on composition:
Classification of proteinaceous enzymes based on function:
The International Union of Biochemistry (I.U.B) devised a system of classification
and identification of enzymes in terms of the reactions they catalyse. This relies on a numerical system (the EC number) to classify enzymes in groups according to the types of reaction catalysed and systematic naming that describes the chemical reaction involved. According to this commission, all enzymes are classified into 6 major classes and they are listed below: Enzyme nomenclature:
Due to the growing complexity of and inconsistency in the naming of enzymes, the International Union of Biochemistry set up the Enzyme Commission (EC) to address this issue. Within this system, all enzymes are described by a four-component Enzyme Commission (EC) number. The general form of EC number is:
EC X. Y. Z .N (X, Y, Z and N are the four numbers)
In the EC number the 4 numbers classify an enzyme according to class, subclass, sub-subclass and the last digit is the serial number of that enzyme within that sub-subclass. For example, the enzyme with the trivial name lactate dehydrogenase has the EC number 1.1.1.27, and is more correctly called l–lactate: NAD+ oxidoreductase. The first part of the EC number refers to the reaction that the enzyme catalyses or the class of the enzyme. The remaining digits have different meanings according to the nature of the reaction identified by the first digit. For example, within the oxidoreductase category, the second digit denotes the hydrogen donor and the third digit denotes the hydrogen acceptor. Thus lactate dehydrogenase with the EC number 1.1.1.27 is an oxidoreductase (indicated by the first digit) with the alcohol group of the lactate molecule as the hydrogen donor (second digit) and NAD+ as the hydrogen acceptor (third digit), and is the 27th enzyme to be categorized within this group (fourth digit).
How enzymes works
Enzymes are biocatalyst and like catalysts they increase the rate of the reaction by lowering the activation energy of a reaction, allowing it to achieve equilibrium more rapidly. Activation energy of any reaction determines the speed of the reaction and it is the difference between the energy of the transition state (T.S) and the reactants. Mechanism of enzyme action:
Active site: The active site of the enzyme is the region where substrate binds and forms ES or enzyme substrate complex.
Schematically the enzymatic reaction can be summarized as follows:
There are two models used to describe the way enzymes interact with substrates:
1. Fischer’s Lock and Key Model: In 1894, the introduction of Lock and Key Model for the substrate and enzyme interaction was proposed by Emil Fischer. According to this model, complementary structural features are present between enzyme and substrate, and the active site is pre-shaped to fit the substrate. The substrate can fit into its complementary site on the enzyme as a key fits into a lock. This results in the formation of an enzyme-substrate complex
2. Koshland’s Induced Fit Model: Daniel Koshland in 1958 proposed Induced
Fit Hypothesis. He suggested that the structure of a substrate may be complementary to that of the active site in the enzyme-substrate complex but not in the free enzyme. The interaction between the substrate and the enzyme induces conformational changes in the enzyme which aligns the amino acid residues or other groups for substrate binding, catalysis, or both. In this model the active of the enzyme is considered as flexible
Properties of Enzymes:
1. Enzymes are mostly proteins but some RNAs can also act as an enzyme and they are known as ribozymes. 2. Enzymes are colloidal in nature 3. Enzymes neither start the reaction nor change the equilibrium. They just increase the rate of the reaction and help to attain equilibrium early 4. Enzymes are highly specific for their substrate and the reaction they catalyzed 5. Enzymes are in no way transformed or used up in the chemical reaction 6. Enzymes are heat and pH sensitive. The optimum range of temperature is 25-35oC and pH is 6 to 8
Factors affecting enzyme action:
1. Temperature: The rate of an enzyme catalysed reactions increases with
the increase in temperature up to a maximum and then falls. When a graph is plotted between temperature versus enzyme activity, a bell-shaped curve is obtained. The temperature at which the maximum rate of reaction occurs is called the enzyme’s optimum temperature. The optimum temperature is different for different enzymes.
2. pH: Enzyme activity is also affected by pH. A plot of enzyme activity
against pH results in a bell shaped curve. Each enzyme has its unique optimum pH at which the rate of reaction is greatest. The optimum pH is the pH at which the activity of a particular enzyme is at a maximum. Many enzymes of higher organisms show optimum reaction rate around neutral pH (6-8).
3. Substrate concentration: The substrate concentration also influences
enzyme activity. As the substrate concentration increases the rate of reaction also increases. This is because the more substrate molecules will interact with enzyme molecules, the more products will be formed. However, after a certain concentration, further increase in substrate concentration will have no effect on the rate of reaction, since the substrate concentration will no longer be the limiting factor. At this stage, enzyme molecules become saturated and work at their maximum possible rate. Kinetics of enzyme catalyzed reaction:
A mathematical relation between substrate concentration and enzyme action
is known as the Michaelis–Menten equation. It is also helpful in determining the kinetics of the enzymes catalyzed reaction. This mathematical relation
is derived based on the following reaction scheme:
According to the above scheme; the enzyme first binds to the substrate and forms an enzyme-substrate complex (ES) and this step is reversible. Next the ES complex is converted to the enzyme and the product and this step is irreversible. Based on the above scheme the final form of the Michaelis–Menten equation is :
Where v=initial velocity Vmax=maximum velocity=K2[E]T ; [E]T= Total enzyme conc. The maximum velocity, Vmax, represents the turnover number of an enzyme. Turnover number is the number of substrate molecules converted into product by an enzyme molecule in a unit time when the enzyme is fully saturated with substrate. It is equal to the kinetic constant K2, which is also called Kcat . [S]= Substrate concentration; in this case [S] >>[E]; [E]= free enzyme concentration KM= Michaelis–Menten constant= K-1+K2/K1 A small KM indicates that the enzyme requires only a small amount of substrate to become saturated. Hence, the maximum velocity is reached at relatively low substrate concentrations. When [S]<< KM, v =(Vmax/KM )/[S], i.e., the reaction rate is directly proportional to substrate concentration. At high substrate concentration (when [S]>>KM), v =Vmax, i.e., reaction rate is maximum and independent of substrate concentration. When [S]=KM, then v=V max/2. Thus, KM is the substrate concentration at which half of the maximum reaction rate is obtained.
