ENZYMES - INTRODUCTION - PART I

(Introduction - Historical aspects - nomenclature)

 

Q-1   What is the importance of enzymes in living systems?

 A-1   Enzymes are the reaction catalysts of the biological systems.

They have extraordinary catalytic power, often greater than that of synthetic catalysts. They have high degree of substrate specificity & accelerate specific chemical reactions. They function in an aqueous medium under very mild conditions of temperature& pH. Very few non-biological catalysts show all these properties.  

 

Q-2   If enzymes are not present in body what will happen?

 A-2   The biochemical reactions taking place in living cells at body temperature at sufficiently rapid in a regular order. Such reactions would have been extremely slow, had they not been catalyzed by enzymes, which are present in every living cell and can also act independently of the cell. (e.g. hydrolysis of starch by salivary amylase).

 

Q-3  Were enzyme actions were known in earlier time?

A-3  Yes, Biological catalysis was first recognized  & described in the early 1800 in studies of digestion of meat by secretions of stomach & conversion of starch into sugar by saliva & various plant extracts.  In 1850 Louise Pasteur concluded that fermentation of sugar into alcohol by yeast is catalyzed by” FERMENTS “& later named than enzymes, are inseparable from living yeast cells, a view that prevailed for many years.

 

Q-4  What was Buchner’s important discovery regarding enzyme? (Or fermentation?)

A-4  In 1897 Buchner showed yeast extract (cell free) could ferment sugar to alcohol. This encouraged biochemists to attempt the isolation of many different enzymes & to examine their catalytic properties

 

Q-5  What was the important discovery made by James Sumner?

 A-5 In 1926 James Sumner crystallized UREASE& found it is as a protein and postulated that all enzymes are proteins. This idea remained controversial for some time.

 

Q-6  What was contribution of J.B.S.HALDANE regarding enzyme?

A-6  During this period (1930) J.B.S.HALDANE wrote a treatise entitled enzymes, even though molecular nature of enzymes was not fully yet appreciated. It contained remarkable suggestions of weak bonding interactions and substrate catalyzed reaction.    

 

Q-7  What is an enzyme?

A-7  Enzymes are the reaction catalysts of the biological systems. They are protein in nature (With exception of small group of catalytic RNA molecules). Molecular weight ranges from 12000 to over million. They are specific in action e.g. Urease Their catalytic activity depends upon the integrity of their native protein conformation. (If enzyme is denatured or dissociated into subunits catalytically activity is usually lost)

The enzymes carry out transformation of molecules and also mediate transformation of energy e.g. PHOTOSYNTHESIS

 

Q-8  What are simple and complex enzymes?

A-8  Some enzymes require no chemical groups other than amino acids for activity, Enzymes composed of only protein are known as simple enzymes. Complex enzyme composed of protein plus a relatively small organic molecule, (which is required for enzyme activity)

 

 Q-9   What is apo-enzyme?

 A-9   Apo-enzyme is the protein part of an enzyme without any cofactors or prosthetic group that may be required for the enzyme to be functional. The apo-enzyme is catalytically inactive.

 

Q-10  What is coenzyme or prosthetic group?

A-10   A non-protein component of an enzyme, which is required for catalytic activity, is known as co-enzyme or prosthetic group.

 

Q-11  What is co-factor?

A-11  Is small organic or inorganic molecules that an apo-enzyme requires for its activity. E.g. Lysine oxidase copper is loosely bound

 

Q-12  What is the difference between prosthetic group and co-enzyme?

A-12. A coenzyme or metal ion that is COVALENTLY bound to the enzyme protein is called prosthetic group. For e.g. in the cytochromes, the heme prosthetic group is very tightly bound and requires strong acids to dissociate from its apo enzyme.

 

Q-13  Do some Enzymes contain vitamin derivatives?

A-12  YES, Many prosthetic groups and coenzymes are water-soluble derivatives of vitamins. It should be noted that the main clinical symptoms of dietary vitamin insufficiency generally arise from the malfunction of enzymes, which lack sufficient cofactors derived from vitamins to maintain homeostasis.

 

 Q-13 What is a holo enzyme?

 A-13 A complete catalytically active enzyme together with its coenzyme and/or metal ions called a holoenzyme.

                        Apo-enzyme  + co enzyme ==holoenzyme

 

Q-14  What is metalloenzyme and metal activated enzymes?

A-14  Enzymes require a metal in their composition (such as Fe+2,Mg2+. Mn2+,Zn2+ ) are known as metalloenzymes if they bind and retain their metal atom(s) under all conditions, that is with very high affinity. Those, which have a lower affinity for metal ion, but still require the metal ion for activity, are known as metal-activated enzymes.

 

Nomenclature

 

Q-15  How enzymes are named?

A-15  Many enzymes have been named by adding the suffix “-ase” to the name of their substrate or to a word or phrase describing their activity. e.g. UREASE catalyze hydrolysis of urea, MALTASE act on maltose, and DNA polymerase catalyze the synthesis of DNA.  Other enzymes such as PEPSIN and TRYPSIN have names they do not denote their substrates.

 

Q-16 What is IUB system?

A-16  Sometimes the enzyme may have two or more names, or two different enzymes have the same name. Because of such ambiguities, and ever increasing number   of newly discovered enzymes a system for naming and classifying enzymes has been adopted by inter national agreement, International Union of Biochemistry (IUB-system)

 

Q-17  How enzymes are classified according to IUB system?

A-17  According to IUB system enzymes are grouped in six major classes.

Each with sub classes based on the type of reaction catalyzed. Systemic classification of enzymes based on numbering system is used. Each enzyme is assigned a code number or EC (enzyme commission number) four-digit classification number and a systematic name, which identifies the reaction catalyzed.

 

Q-18  According to IUB system which enzymes are included in group 1?

A-18  OXIDO-REDUCTASE      (EC-1)

Enzymes of this group add or remove hydrogen atoms during the catalysis. They include dehyrogenases & oxidases, and are mostly concerned with biological oxidation. 

Dehydrogenases removes H+ from substrate in the presence of H+ acceptor group. e.g. Lactate Dehydrogenase. Enzymatic action can be represented by:                            

                       AH2 +B  à A + BH2

OXIDASES – transfer two electrons from the donor to oxygen resulting usually in hydrogen peroxide formation (H2O2) e.g. Glucose oxidase. CYTOCHROME oxidase produces H2O rather than H2O2. Oxygenases catalyze the incorporation of oxygen into a substrate.

 

Q-19  According to IUB system which enzymes are included in group 2?

A-19  TRANSFERASES    (EC-2)

These enzymes transfer functional groups between donors and acceptors.

The AMINO,  ACYL, PHOSPHATE, ONE CARBON and GLYCOSYL are the major groups that are transferred. E.g.

A— X+ B    à     A+ B—X

A. METHYL group---àe.g. Transmethylase

B. ALDEHYDE or KETONIC group e.g. Transaldolase or transketolase.

C. ACYL GROUP      e.g.Aceyltransferase

D. SUGAR GROUP e.g.       Glucosyltransferase

E. AMINO-KETO GROUP- Aminotransferase or transaminases

F. KINASES are specialized trnsferases that regulate metabolism by transferring phosphate from ATP to other moleculs e.g. Hexokinase:

                           ATP +Glucose -----àG-6-P+ ADP

 

Q-20  According to IUB system which enzymes are included in group 3? 

A-20  HYDROLASES    (EC-3)

A special class of transferases in which the donor group is transferred to water. The generalized reaction involves the hydrolytic cleavage of C-O, C-N, O-P and C-S bonds. In other words enzymes, which add, water to the substrate and hydrolyze or decompose it to give products.

              A—B + H2O à AH+ BOH

1. LIPASES-------e.g. Glycerol ester hydrolase

2. PHOSPHATASES------e.g.Glucose-6-Phosphatase

3. CHOLINE ESTERASE hydrolysases acetylcholine

4. PEPTIDASES-----hydrolyses peptides

5. NUCLEASES     e.g.nucleotidase, nucleosidase

6. CARBOHYDRASES e.g. Amylase act on amylose Lactase, Maltase

7. Enzymes acting on C—N linkage   Urease converts urea into ammonia, Asparginase. Glutaminase, Arginase

 

Q-21  According to IUB system which enzymes are included in group 4?

A-21  LYASES   (EC4)

 Lyases add or remove water, ammonia, or carbon dioxide from the substrates.

                           A-B + X-Y   ---- >  A-X  +  B-Y

1. DECARBOXYLASE removes CO2 from a orb keto acids or aminoacids.

2. Carbonic anhydrase

3. Cysteine desulfurase

 

Q-22  According to IUB system which enzymes are included in group5?

A-22  ISOMERASES    (EC5)

A hetrogeneous group of enzymes catalyze transfer of groups within molecules to yield isomeric forms e.g. isomerazition of!  1) Optical isomers 2) geometrical isomers

                                                  A-----à A’

1. Epimerases or Racemases catalyze inversion at asymmetric carbon atoms

2. MUTASES involve intramolecular transfer of a group such as a phosporyl

3. CIS-TRANS ISOMERASE e.g. all trans retinene isomerase

 

Q-23  According to IUB system which enzymes are included in group 6?

A-23  LIGASES   (EC6)

TO liagate means to bind, Formation of C-C, C-S, C-O& C-N bonds by condensation reactions. These enzymes carry out synthetic reactions where two molecules joined at the utilization of a “high energy phosphate bond of ATP. ”

                                REACTION:     A+B---à A-B

1. Pyruvate carboxylase   Pyruvate+CO2+ATP-----àOxaloacetate+ ADP+Pi

2. GLUTAMINE SYNTHETASE

3. Acetyl Co A CARBOXYLASE

 

Q-24  The following enzyme is classified as EC –2.7.1.1.  What does this mean?

A-24  The name of the enzyme catalyzing following reaction is ATP:glucose phospo transferase. Its enzyme classification number (E.C.number) is 2.7.1.1  

            ATP+ D-Glucose---àD-Glucose-6-phosphate+ ADP  

The first digit (2) denotes the class number (transferase)

The second digit (7) denotes sub-class - phospo transferase;

The third digit  (1)-Hydroxyl group as an acceptor;

The fourth digit (1) D-glucose as the phosphate group acceptor.

When the systematic name of an enzyme is long or cumbersome, a trivial name may be used in this case Hexokinase.