István T.

Horváth - BMEVEKFM210

ENVIRONMENTALLY BENIGN AND CATALYTIC PROCESSES

KFM210
2020-21, Fall Semester, Friday 8:30 – 11:00

BIOCATALYSIS

István T. Horváth

Department of Chemical and Environmental Process Engineering

istvan.t.horvath@edu.bme.hu
istvan.t.horvath@att.net

SUSTAINABLE LIFE IN EARTH

Earth’s life-sustaining conditions are possible
because of its position in the solar system.

It is not too hot and not too cold – if the Earth

was any closer or further away from the Sun,
the planet could not support life.

Four Conditions for Life

1. Stable Temperature Range
2. The Importance of Water
3. The Importance of Gases
4. The Role of the Atmosphere

A stable temperature range (-50 to about +50 oC) allows life.

The average temperature has been between 10-20 oC for 3.5 billion years.

1
István T. Horváth - BMEVEKFM210

BIOCATALYSTS
Broadly defined as the use of enzymes or whole cells to increase
speed in which a reaction takes place but do not affects the
thermodynamics of the reaction.

High molecular weight globular protein or whole cells

Micrographs of R. oryzae cells.

Fukuda, H., Hama, S., Tamalampudi, S.,
Noda, H. Trends in Biotechn., 2008, 26, 668.

Operates at mild conditions, physiological pH and temperature.

ENZYMES
• Enzymes are protein molecules that catalyze
biochemical reactions
• The substance on which the enzyme acts is
known as the substrate

Models of some proteins – tertiary and quaternary structure.

2
István T. Horváth - BMEVEKFM210

ENZYMES

• 1825 Jon Jakob Berzelius discovered the catalytic effect of enzymes.

• 1926 James Sumner isolated the first enzyme in pure form.
• 1947 Northrup and Stanley together with Sumner were awarded the
Nobel prize for the isolation of the enzyme pepsin.

Berzelius Sumner Northrup Stanley

ENZYMES
• High molecular weight
proteins with masses
ranging from 10,000 to as
much as 2,000,000 grams
per mole
• Substrate specific
catalysts
• Highly efficient, increasing
reaction rates by a factor as
high as 108

3
István T. Horváth - BMEVEKFM210

NOMENCLATURE
• The earliest enzymes that were discovered have common names:
i.e. Pepsin, Renin, Trypsin, Pancreatin
• The enzyme name for most other enzymes ends in “ase”
• The enzyme name indicates the substrate acted upon and the
type of reaction that it catalyzes

Examples of enzyme names

• Glutamic Oxaloacetic Transaminase (GOT)

• L-aspartate: 2-oxoglutarate aminotransferase.

• Enzyme names tend to be long and complicated.

• They are often abbreviated with acronyms

ENZYME SPECIFICITY
• Enzyme specificity may be characterized as

1. Absolute: The enzyme catalyzes on one reaction

2. Group Specific: The enzyme acts only on molecules having a
particular functional group
3. Linkage Specific: The enzyme acts on a particular type of
chemical bond
4. Stereo-chemical Specific: The enzyme acts on a particular stereo
or optical isomer

• The action of an enzyme depends primarily on the tertiary and quaternary

structure of the protein that constitutes the enzyme.

• The part of the enzyme structure that acts on the substrate is called the
active site.

• The active site is a groove or pocket in the enzyme structure where the
substrate can bind.

4
István T. Horváth - BMEVEKFM210

COFACTORS

• Cofactors are other compounds or ions that enzymes require

before their catalytic activity can occur.
• The protein portion of the enzyme is referred to as the
apoenzyme.
• The enzyme plus the cofactor is known as a holoenzyme.

Cofactors may be one of three types

1. Coenzyme: A non protein organic substance that is loosely

attached to the enzyme
2. Prosthetic Group: A non protein organic substance that is firmly
attached to the enzyme
3. Metal ion activators: K+, Fe2+, Fe3+, Cu2+, Co2+, Zn2+, Mn2+, Mg2+,
Ca2+, or Mo2+

COFACTORS

• Enzymes have varying degrees of specificity.

• One cofactor may serve many different enzymes.

5
István T. Horváth - BMEVEKFM210

ENZYME MECHANISMS

The basic enzyme reaction can be represented as follows:

E + S  ES  E + P

Enzyme Substrate Enzyme substrate Enzyme Product(s)

complex

The enzyme binds with the substrate to form the

Enzyme-Substrate Complex.
Then the substrate is released as the product(s).

ENZYME MECHANISMS

6
István T. Horváth - BMEVEKFM210

ENZYME MECHANISMS
• An enzyme-substrate complex forms when the enzyme’s active
site binds with the substrate like a key fitting a lock.
• The shape of the enzyme must match the shape of the substrate.
• Enzymes are therefore very specific; they will only function
correctly if the shape of the substrate matches the active site.

E +S  ES  E + P

 

ENZYMES AND REACTION RATES

• At low concentrations, an increase in substrate concentration

increases the rate because there are many active sites available to
be occupied

• At high substrate concentrations the reaction rate levels off because

most of the active sites are occupied

7
István T. Horváth - BMEVEKFM210

ENZYMES AND REACTION RATES

• The maximum velocity of a reaction is reached when the active

sites are almost continuously filled.
• Increased substrate concentration after this point will not increase
the rate.
• Vmax is the maximum reaction rate

• The Michaelis-Menton constant,

Km is the substrate concentration
when the rate is ½ Vmax

• Km for a particular enzyme with a

particular substrate is always the
same

EFEECT OF TEMPERATURE

• Higher temperature increases the number of effective collisions and

therefore increases the rate of a reaction.
• Above a certain temperature, the rate begins to decline because the
enzyme protein begins to denature.

8
István T. Horváth - BMEVEKFM210

EFFECT OF pH
• Each enzyme has an optimal pH at which it is most efficient
• A change in pH can alter the ionization of the R groups of the amino
acids.
• When the charges on the amino acids change, hydrogen bonding
within the protein molecule change and the molecule changes shape.
• The new shape may not be effective.

Pepsin is most efficient at pH 2.5-3, while Trypsin is efficient at a much higher pH.

INHIBITORS

• Enzyme inhibitors are substances which alter the

catalytic action of the enzyme and consequently
slow down or stop catalysis.

• There are three common types of enzyme inhibition

1. Competitive inhibitors
2. Non-competitive inhibitors
3. Substrate inhibition.

9
István T. Horváth - BMEVEKFM210

COMPETITIVE INHIBITORS

• Competitive inhibition occurs when

the substrate and a substance
resembling the substrate are both
added to the enzyme.

• The inhibitor blocks the active site

on the enzyme stopping its catalytic
action.

COMPETITIVE INHIBITORS

• Non-competitive inhibitors
deactivate the active site of the
enzyme.

• They alter the enzyme so that it

can no longer bind to the
substrate

10
István T. Horváth - BMEVEKFM210

EFFECT OF INHIBITORS ON
REACTION RATE

• For non-competitive
inhibitors Vmax is lower but
Km is the same.

• For competitive inhibitors,

Vmax is the same but Km is
increased.

SUBSTRATE INHIBITORS

• Substrate inhibition occurs when

excessive amounts of substrate are
present.

• Additional substrate sometimes

interferes with the ability of substrate
molecules to find active sites on
enzymes.

• In these cases the reaction velocity

decreases after the maximum velocity
has been reached.

11
István T. Horváth - BMEVEKFM210

INDUSTRIAL APPLICATION OF BIOCATALYSIS

Ethanol 15,450,000 t/year

Sodium glutamate 1,000,000 t/year
Citromsav 412,000 t/year
Lysine 118,450 t/year
Glucuronic acid 51,500 t/year
Penicillin 15,450 t/year

• Many other smaller volume products including chemicals (lactic

acid, maleic acid, amino acids, steroids, vitamins, anitibiotiocs,
food ingredients, detergents, paper, textiles and biofuels

• The enzymes market was 10 billion USD in 2019.

• Laundry enzymes are the largest industrial enzyme application –

washing at low temperature results in energy savings.

ETHANOL PRODUCTION

12