Enzymes

 History  

In 1833, the active agent breaking down the sugar (Starch and
glycogen) was partially isolated and given the name diastase (now
known as amylase).

later, pepsin extracted from gastric juice (digest dietary protein) were
given the name ferments. these ferments are non-living materials
obtained from living cells (Justus von Liebig ), but Louis Pasteur and
others still maintained that ferments must contain living material.

Wilhelm Kuhne in 1878 proposed the name Enzyme , comes from the
Greek, enzume, meaning 'in yeast’.
 Enzymes  History  

Eduard and Hans Buchner in 1897, showed that sugar fermentation

could take place when a yeast cell extract was added even though no
living cells were present.

In 1926, James Sumner crystallized urease from jack-bean extracts.

 Naming  enzymes

Enzymes are traditionally named by adding the suffix '-ase’ to the

substrate except proteolytic enzymes e.g. trypsin.

The names of enzymes usually indicate the substrate involved.

Thus, lactase catalyses the hydrolysis of the disaccharide lactose
to its component monosaccharides, glucose and galactose:
 Naming  enzymes

Fumarase, for example, by analogy with lactase might be supposed to

catalyse a hydrolytic reaction, but, in fact, it hydrates fumarate to form
malate:

Transcarboxylase, indicate the nature of the reaction without

specifying the substrates

Substrates are methylmalonyl-CoA and pyruvate

 Naming  enzymes

Some names, such as catalase, indicate neither the substrate nor the
reaction (catalase mediates the decomposition of hydrogen peroxide).

Also, the names of many enzymes make clear the substrate and the
nature of the reaction being catalysed. For example, there is little
ambiguity about the reaction catalysed by malate dehydrogenase.
This enzyme mediates the removal of hydrogen from malate to
produce oxaloacetate:
 Naming  enzymes

Malate dehydrogenase

Malate dehydrogenase has been given different names

 Enzymes systematic classification

To name enzymes systematically a commission was appointed by the

International Union of Biochemistry (later re-named the International
Union of Biochemistry and Molecular Biology, IUBMB).

Enzymes can be classified using a numbering system defined by the

Enzyme Commission.
—
This system consists of a four digit number which classifies based on the
type of reaction the enzyme catalyzes
 Classes  of  Enzymes  

EC 1. Oxidoreductases – Transfer electrons (Redox reactions)

—
EC 2. Transferases – Transfer functional groups between molecules

EC 3. Hydrolases – Break bonds by adding H2O —

EC 4. Lyases – Elimination reactions to form double bonds

EC 5. Isomerases – Intramolecular rearangements —

EC 6. Ligases – Join molecules with new bonds

 Nucleoside Monophosphate(NMP) Kinases
 

Nucleoside Monophosphate Kinases

EC number Nucleoside Monophosphate Kinases    

1. NMP kinase is a transferase, or member form group 2

2. A member that transfers phosphate group belong to subgroup 7, so it is 2.7

3. Various groups can accept the phosphate group, if a phosphate is

acceptor it is 2.7.4

4. The final number designates the acceptor more precisely and

nucleoside monophosphate is acceptor and enzyme designation is
2.7.4.4
Factors affecting enzyme activity
 Factors  affec5ng  enzyme  ac5vity    
 

•  The  factors  which  affect  enzyme  ac5vity  are  :  

•  Temperature  
•  pH  of  the  solu5on  
•  Concentra5on  of  substrates    
•   Concentra5on  of  enzymes  
•  Ac5vators  or  inhibitor  molecules  
 Enzyme  ac5vity  and  temperature  
 increases  interac5on  B/w  E  and  S  
 
•  Increase  in  temperature/heat  increases  the  rate  of  an  enzyme  
catalyzed  reac5on.  Since    both  E  and  S  interact  with  each  other  
more  frequently.  
•  It  is  more  likely  that  the  molecules  of  S  will  slot  into  ac5ve  site  
of  E  leading  to  the  forma5on  of  products.  
•  But  increase  in  temperature  increased  enzyme  ac5vity  to  a  
certain  limit  (op5mum  temperature)  as  aJer  that  limit  enzyme  
ac5vity  decreased  with  increase  in  temperature  due  enzyme  
denatura5on  
Effect  of  temperature  on  enzyme  ac5vity  
 High  temperature  leads  to  Denatura5on  
temperature  >  40C  

 
•  Enzymes   are   proteins,   increase   of   temperature   above   the  
op5mum  level  (40  C)  causes  the  protein  to  lose  their  :  

1.  3  D  structure  and  folding                                                                                                                                                                          

2.  Breakage  of  bonds  B/w  the  func5onal  groups  of  AAs                          

3.  Change  of  shape  and  loss  of  enzyme  func5on                                                      

4.  The  op5mum  temperature  for  majority  body  enzymes  is  37C°  
Hos5le  temperature  –  Denatura5on  of  E  
    Increase  in  Substrate  conc.    increases  Enzyme  ac5vity  
up  to  an  op5mum  level  

 
•  Increase   of   substrate   increases   the   enzyme   ac5vity   up   to   an  
op5mum  level  as  more  molecules  of  S  have  the    chance  to:  
•     Bind  with  the  ac5ve  site  of  enzyme  
•  Once   E   is   saturated   with   the   substrate,   rate   of   reac5on  
doesn’t  increase  with  increase  of  enzyme  conc.  
•  Excess   of     S   molecules,   reduces   the   chances   of   finding   an  
ac5ve  site  and  no  further  increase  in  enzyme  ac5vity.  
  Increase  substrate  conc.  Increased  enzyme  ac5vity  
up  to  op5mum  limit  
 Increase  in  E  conc.Incereases  E  ac5vity  
up  to  an  op5mum  level  

 
•  This  effect  is  much  like  that  of  substrate  
•  Increase  in  the  enzyme  conc.  increases  the    rate  of  enzyme  
reac5on  up  to  an  op5mum  level  
•  Further    in  enzyme  conc.  has  no  effect  on  the  reac5on  ac5vity  
as  no  more  S  molecules  are  available    
•  However,  further  increase  in  the  substrate  molecules,  will  
certainly  increase  the  rate  of  reac5on  
Increase  in  E  conc.  increases  E  ac5vity  
up  to  an  op5mum  level  
  pH  and  ac5vity  of  enzyme  
up  to  an  op5mum  level  

 
•  The  pH  level  of  a  solu5on  can  also  affects  enzyme  ac5vity.  
 
•   Many  enzymes  can  only  work  within  a  narrow  range  of  pH.  
•  Enzymes  increases  rate  of  reac5on  only  when  its  ac5ve  site  
residues  have  specific  charges.        
•  Above  or  below  op5mum  pH  range,  can  lead  to  denatura5on  of  
enzyme.  
•  The  op5mal  pH  for  many  enzymes  is  7.0-­‐7.5,  but  this  is  not  
always  the  case  and  may  be  variable  
Enzyme  ac5vity  -­‐  pH  
 Hos5le  pH  and  E  ac5vity  –  the  mechanism  
S  non  covalent  bonds  with  E  

•  When  a  substrate  slots  into  the  active  site  of  an  enzyme  
•   It  forms  temporary  bonds  (non  covalent)  with  the  groups  at  
the  active  site  (Amino  acyl  groups)  
•  These  groups  are  the  func=onal  groups  of    AA  in  the  protein  
side  chains  and  have  important  effects  on  :  
•   the  shape  of  enzyme