ENZYMES AND SUBSTRATES

Rondang R. Soegianto
2009

Enzyme = Functional protein

Has all the properties of a protein
(macromolecule, heat, pH)
Functions as biocatalyst

Biocatalyst:
- substrate specific
- reaction specific
- can be regulated in the body
Inorganic catalyst
- resistant to heat
- not specific to certain substr/reaction
- Pt, Zn, H+

Cellular enzymes exist in compartments

Carry out activities in organelles, membranes
and cytosol
Enzymes in blood for coagulation process

Enzymes catalyze conversion of substrates

into products
E

ES

E + P

E interacts with S to for ES complex

Mechanism of interaction ;
1. Emil Fischer
Lock and Key Model
2. Koshland
Induced fit model
Enzyme undergoes conformational change

Active site, substrate site, catalytic site

Specific area on enzyme surface that binds
to substrate to form E-S complex, before
enzyme reaction takes place

Why are enzymes essential to body

Homeostasis?
- At body temp. biochem. processes
have to proceed fast enough to
meet needs of the body
- Enzymes (unlike inorganic catalysts)
can be regulated in the body

Inborn Error of Metabolism is caused

enzyme abnormality

What is enzyme activity?

Enzyme activity measured by velocity of
enzyme reaction (v)
v = amount of product per time unit
(sec. minute)
Also
v = remaining substrate per time unit

FACTORS THAT AFFECT REACTION RATE

A.Substrate Concentration
v = velocity
Vm = maximum velocity
When enzyme conc kept constant,
v will >> with increasing [S]
Vm reached when enzyme molecules
are saturated with S

B. Temperature
Temperature >>

v increased

Heat will cause increase of kinetic energy

Incr collision of E and S ES P
Vm is reached at optimum temp.
Raising temp beyond opt.temp. will damage
enzyme protein enz activity decreased

C. Acidity, pH
Enzymes have specific optimum pH
Pepsin ~ pH = 1
Trypsin ~ pH = 7-8
Enzymes denature at high or low pH
relative to optimum pH

ENZYME REGULATION
A. Competitive Inhibition
- Similarity between S and I
- S and I compete for active site
- Enzyme reaction inhibited when I binds
to active site no P formation

E + I EI E + I
- E not destroyed.
After release, can react with substrate
- Excess substrate eliminates effect of I
- Enzyme reaction can reach Vmax
Example: Inh of succinate DH by malonate

B. Non-competitive Inhibition
-

Inhibitor very different from S

I can bind on E outside of active site
I causes E denaturation
Ex. : Hg, Pb

ZYMOGEN, PROENZYME
- Active site located in the interior of E molecule
- Can be exposed to the surface upon activation
- Trypsinogen Trypsin
Zymogen prevents self digestion of tissues on its
way from site of formation to site of E reaction.

COENZYMES, COFACTORS
Non-protein molecule needed for enzyme activity
A. Metal ions: Zn2+, Fe2+, Mg2+
B. Organic molecules = Coenzymes
Derivatives of vitamin B
- Holoenzyme + Enzyme + Coenzyme (Cofactor)
- Apoenzyme = Protein portion of holoenzyme

Vitamin

Coenzyme

Nicotinic acid
Riboflavin
Thiamine
Pantothenic acid
Biotin

NAD
FAD
Thiamine PP
Coenzyme A
Biotin

Activity
Oxid-Reduct
Oxid-Reduct
Decarboxylation
Activation of FA
CO2 binding