Enzyme

INTRODUCTION

 Special kinds of protein molecules: long chain of amino

acids bound by peptide bonds
 produced by living cells
 absolutely essential as catalysts in biochemical reactions.
i.e., controlling the metabolic processes, converting
nutrients into energy, breaking down food materials, etc
 to catalyze the making and breaking of chemical bonds.
 Enzymes are specific, versatile, and very effective
biological catalysts, resulting in much higher reaction
rates as compared to chemically catalyzed reactions
under ambient conditions.
 Enzymes are usually proteins of high molecular weight
(15,000 < MW < several million daltons).
 Some protein enzymes require a nonprotein group for
their activity. (Mg, Zn, Mn, Fe, or a coenzyme)
HOW ENZYMES WORK

 Enzymes lower the activation energy of the reaction catalyzed by binding the
substrate and forming an enzyme–substrate complex.
 Enzymes do not affect the free-energy change or the equilibrium constant
 increase the rate of reaction without themselves
undergoing permanent chemical changes.
 Decrease activation energy
 enzymes are released again after a reaction ceases
and can continue in another reaction
 This processes cannot go forever  limited stabilities
and, slowly, they become inactive
Characteristics of enzymes
 High activity. Increase rate by reducing Ea
 Selectivity.Active sites only to a specific
substrates; may be unable to recognize the
substrate minor changes
 Controllability by the amount of substrate & by
other factors (temperature, pH, etc.) active
site
 Environmentally friendly  lower by-product
produced. Natural substrates  natural products
Enzim Action

“Lock and Key”

products
substrate

enzyme
Different Catalyst Inorganic And Enzyme

catalyst inorganic ENZYME

H, OH, P Protein

EA EA

Specific

Low Heat Resistance

Selection of enzyme
 Cost benefit (adding value or reducing production cost)
 Availability, consistency, and quality support (reputation of
suppliers)
 Activity (specific substrate alteration by pH, ions,
temperature, and inhibitors)
 Ability to modify a reaction’s quality (quality measurement
and understanding of an enzyme can control its activity more
precisely)
Cofactor
 A cofactor is a non-protein chemical compound
 cofactors can be considered "helper molecules" that aid in biochemical
changes.
Cofactor Function

 conversion of three-dimensional structures of

enzyme.
 conversion of three-dimensional structures of
substrate.
A stabilizer of protein enzyme.
 Carrier molecule for CO2, CH3, NH2 and H
Factors for the determination of enzyme
activity
 Theequation reaction catalyzed by the
enzyme.
 Cofactor’s need.
 Substrate and Cofactors concentration.
 The optimum condition of enzyme.
Effect of substrate
Effect of pH
Effect of temperature
Applications in industry

 Carbohydrases: production of corn syrups from

starch (glucoamylase); conversion of cereal
starches into fermentable sugars in malting,
brewing, distillery, baking industry (amylase).
 Proteases: meat tenderizers (bromelin, papain,
ficin).
 Lipases: Flavor production in chocolate and
cheese.
-amylase

 -amylase enzyme is also called endoamylase or

endoglucanase. And the systematic name is  -D-
1,4 glucan glucanohydrolase.
 Hydrolysis of starch by -amylase produces
glucose, maltose, and a small amount of dextrin
derived from the amylopectin.
 produced by animals, plants, microorganisms
 Microorganisms produce  -amylase: Bacteria genus Bacillus like
Bacillus cereus, B. amyloliquefaciens, B. licheniformis, B.
subtilis, B. polymyxa, B. stearothermophilu and fungi
Aspergillus, Rhizopus, Mucor, Candida, Neurospora.

 The highest -amylase enzyme is produced by Bacillus

amyloliquefaciens, B. licheniformis, dan Aspergillus oryzae
Application  -amylase in industry
 Starch industry, -amylase from Bacillus, on starch liquefaction
process to produce maltose, glucose, fructose.
 Alcohol industry,  -amylase from Bacillus and from Aspergillus,
on liquefaction and saccharification process of starch.
 Brewing, amylase from Bacillus, for processing barley, additive
liquefaction.
 Feed industry,  amylase from Bacillus, improves the use of
enzymatically processed barley in poultry
 The sugar industry, -amylase from Bacillus, refines the
filterability of sugar cane juice through the breakdown of
starches in juice.
Glucoamylase
 Glucoamylase systematic name is -1,4-glukan
glukohidrolase.
 This enzyme can hydrolyze maltose into
glucose.
 Microorganisms produce glucoamylase:
Aspergillus niger, A. oryzae, A.amori, Rhizipus
niveus, R. delemar, R. javanicus, and R.
formosaensis.
Protease
 Protease is the second most important industrial enzyme
after amylase.
 Proteases are enzymes that catalyze the process of
protein hydrolysis into amino acids.
 The protease enzymes used in detergents are mainly
produced by strains of Bacillus (Bacillopeptidase).
 The producers are: Bacillus strains such as B.
licheniformis, B. amyloliquefaciens, B. firmus, B.
megaterium, and B. pumilus; Streptomyces strains such
as S. fradiae, S. griseus, and S. rectus; and fungal strains
such as Aspergillus niger, A. sojae, A. oryzae, and A.
flavus.
Application protase in industry

 Detergent industry
 The enzyme based detergent formula was
first released in the 1960s (protease serin
alkalin dari Bacillus licheniformis).
 leather tanning industry
The advantages
 An enzyme catalyst highly specific; reducing the number
of side reactions and by-products. A great variety of
enzymes exist, which can catalyze a very wide range of
reactions.
 The rate of an enzyme-catalyzed reaction is usually much
faster
 the rates of reaction are easily controlled by adjustment
of incubation conditions
 Only require a small amount
 mild conditions of temperature and pH
The disadvantages
 Sensitive or unstable molecules require more care
 Expensive
 insome products, enzymes must be inactivated or
removed after processing which adds to the cost of
the product
 Likeother proteins, enzymes may cause allergic
responses
 usuallycoated or immobilised on carrier materials to
reduce the risk of inhalation, reduce cost.
Enzyme Kinetics
 Kinetics of simple enzyme-catalyzed reactions are often referred to as
Michaelis–Menten kinetics or saturation kinetics.
the rate of product formation is

The rate of variation of the ES complex is

Since the enzyme is not consumed, the conservation equation on the enzyme yields
Substituting eq.
Michaelis Menten Equation
 Km

• Km is the [S] at ½ Vmax

• Km is a constant for a given enzyme
• Small Km means tight binding; High
Km means weak binding