Lecture – 4 - 09.09.

2022
Biochemical Engineering - (CHE F421)
BITS Pilani
Pilani Campus
Enzymes

• Enzymes

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What are Enzymes

• 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 higher molecular

weight (15,000 < MW < several million daltons) that
act as catalysts

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Enzymes
• More than 2000 enzymes are known

• Enzymes comprised by largest and most specialized

group of protein molecules within the cell

• Proteins are polymers of a mixture of 20 plus different

amino acids subunits

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Enzymes

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Enzymes
1. Enzymes have primary structure given by sequence of
chain of attached amino acids

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Enzymes
1. Enzymes have primary structure given by the sequence of
chain of attached amino acids

2. Enzymes also have a secondary structure that forms

when amino acids chain twists itself into a three
dimensional configuration held together by hydrogen
bonding between amino acids and sulfur to sulfur
linkages that are common to some of the amino acids

3. Many proteins also have tertiary structure, in which

protein is folded back upon itself and again held
together by hydrogen bonding

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BITS Pilani, Pilani Campus
Enzymes

1. This tertiary structure is vulnerable, causing enzyme

denaturation or deactivation

2. Enzyme structure changes with physical environment:

especially temperature, pH and many chemicals that may
bind to protein and change its secondary and tertiary
structure

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Enzymes
Hence two important characteristics include:

• Specificity- Meaning that enzymes channels

the transformation of a chemical along a
desired pathway

• Rate of reaction that it catalyses – rate

refers to the speed at which desired
reaction occurs when enzyme is present

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Enzymes
1. For example – in waste waters, many reactions are
thermodynamically possible; but do not occur by
themselves at finite rates under normal conditions of
temperature and pressure

2. However, by having right enzymes in correct amounts,

microorganisms can have a pathway through which
chemical reaction occurs

Enzymes do not increase amount of energy released from

a given reaction but they minimize diversion of resources
(electrons, energy, and elements) into nonproductive
pathways
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Enzymes
• Some enzymes depend for their activity only upon their
structure as proteins

• Other requires, a non-protein component for their activity

When non-protein component is a metal ion, it is called

cofactor
• This group is either a cofactor, such as metal ions, Mg,
Zn, Mn, Fe, or
If non-protein component is organic, it is called coenzyme
• a coenzyme, such as a complex organic molecule, or
some vitamins
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BITS Pilani, Pilani Campus
Enzymes
• Co enzymes are bounded and a given coenzyme may
combine with different enzymes at different times

• Co enzymes often serve as intermediate carriers of small

molecules from one enzyme to another

• Co enzymes generally serve in transfer of electrons,

elements, or functional groups from one molecule to
another from one place in cell to another

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Enzymes
• Enzymes are named by adding the suffix -ase to the
end of the substrate,

• For example
1. Proteinase acts in hydrolysis of proteins to form amino
acids
2. Hydrolases – that split complex carbohydrates, lipids, and
protein polymers into their simpler building blocks through
water addition
3. Oxido-reductases – which are involved in oxidation and
reduction reactions, that provide cells with energy and are
essential in synthesis

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Enzymes

1. Major source of energy is provided through oxidation-

reduction reactions that involve transfer of electrons from
one molecule to another

2. Electron donor move electrons from one compound to

another

3. When such carriers are involved; initial donor is

represented as primary electron donor

4. And the final acceptor as terminal electron acceptor

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BITS Pilani, Pilani Campus
How enzymes work
1. Enzymes lower the activation energy of the reaction
catalyzed by binding the substrate and forming the
enzyme – substrate complex.

2. Enzymes do not affect the free - energy change or the

equilibrium constant

• For example, the activation energy for the decomposition of hydrogen

peroxide varies depending on the type of catalysis
• The activation energy of the uncatalyzed reaction at 20deg C is 18
kilocalories per mole (kcal/mol), whereas delta E value for chemically
active catalyst & enzymatically active catalyst; varies from 13 and 7
kcal/mole, respectively
• That is catalase; accelerates rate of reaction by certain factor
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How enzymes work

1. The molecular aspects of enzyme–substrate interaction

are not yet fully understood
2. This interaction varies from one enzyme–substrate
complex to another
3. Various studies using x-ray and Raman spectroscopy
have revealed the presence of the enzyme–substrate
(ES) complex

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• Interaction between the enzyme and its substrate is
usually by weak forces

• In most cases, van der Waals forces and hydrogen

bonding are responsible for the formation of ES
complexes

• Substrate binds to a specific site on the enzyme known

as active site

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1. Substrate is a relatively small molecule and fits into a
certain region on enzyme molecule, which is a much
larger molecule

2. Simplest model describing this interaction is the lock-

and-key model, in which the enzyme represents the
lock and the substrate represents the key

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Enzymes
• More than 2000 enzymes are known

• It has been shown that certain RNA molecules are also

catalytic, but the vast majority of cellular reactions are
mediated by protein catalysts

• RNA molecules that have catalytic properties are called

ribozymes

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Enzyme reactivity
• Enzyme reactivity towards substrate involves specificity &
kinetics

• Enzyme which is much larger positions; itself around

smaller substrate molecule, which just fits within 3 - D
protein structure
• Coenzymes, cofactors, co substrates are brought into contact in
such way that electrostatic or vibrational forces between them
reduce the activation energy and permit a given transformation
to take place at specific site of substrate molecule
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Enzyme kinetics
• Rate (or kinetics) of an enzyme catalyzed reactions is
governed by principle that govern other chemical reactions

• In many cases; rates of growth and substrate utilization by

microorganisms can be described by kinetics developed to
describe rate of transformation of a single substrate by a
single molecule

• Hence kinetic of enzymes catalysis are of interest for

describing individual reactions and often complex
biochemical processes

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Enzyme kinetics
• A mathematical model of kinetics of single substrate –
enzyme catalyzed reaction was developed by Henri in
1902 and by L. Michaelis and M.L. Menten in 1913.

• Kinetics of simple enzyme – catalyzed reactions are

referred as Michaelis – Menten kinetics or saturation
kinetics

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Enzyme kinetics
• A characteristic of enzymes reactions is substrate
saturation

Observed effect of substrate concentration on rate of

catalyzed reaction is given :

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Enzyme kinetics
• The qualitative features of enzyme kinetics are similar to
Langmuir – Hinshelwood kinetics

• These models are based on data from batch reactors with

constant liquid volume in which the initial substrate, [S0],
and enzyme, [E0], concentrations are known.

• More complicated enzyme - substrate interactions such as

multi substrate – multi enzyme reactions can take place in
biological systems

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• An enzyme solution has a fixed number of active sites to
which substrates can bind.

• At high substrate concentrations, all these sites may be

occupied by substrates or the enzyme is saturated.

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BLACK BOARD

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The Rapid Equilibrium
Assumption
• In this case, the maximum forward velocity of the
reaction is Vm.
• Vm changes if more enzyme is added, but the addition of
more substrate has no influence on Vm.
• K`m is often called the Michaelis–Menten constant, and
the prime reminds us that it was derived by assuming
rapid equilibrium in the first step.
• A low value of K`m suggests that the enzyme has a high
affinity for the substrate.
• Also, K`m corresponds to the substrate concentration,
giving the half-maximal reaction velocity.

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