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 + S 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 Nicotinic acid Riboflavin Thiamine Pantothenic acid Biotin

Coenzyme NAD FAD Thiamine PP Coenzyme A Biotin

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