Introduction to

Biochemical
Reaction Systems
Aim
To provide an introductory overview of
how biological systems can be used by
Chemical Engineers

Learning outcomes of the

lecture.
- Recognise the roles of a biochemical engineer .
- Demonstrate an understanding of the main
characteristics of a biological process.
- Enzyme Kinetics

Learning outcomes of the

lecture.
-

Demonstrate an understanding of the kinetics of enzymecatalyzed reactions.

Explain the lock and key and induced fit model of

enzymatic action.

Demonstrate an understanding of the mechanism of enzyme

action.

Understand the Michaelis-Menten equation and ability to apply

the Michaelis-Menten equation to the analysis of enzyme
kinetics.

An overview of
Biochemical Engineering
Bioprocess/Biochemical Engineering is related with the
design, development, implementation and operation of
engineering processes required to facilitate biomanufacture and bio-processing at all scales. Requires
multidisciplinary understanding drawn from:

Chemical Engineering.
Microbiology.
Biochemistry.
Genetics.
Medics.

Bioprocess operations make use of microbial, animal and

plant cells and components of cells such as enzymes to
manufacture new products and destroy harmful wastes.

Biological Process Characteristics

The concentrations of the principal participants in
bioreactions may often be very low indeed. A few may
be in kg m-3, usually it is g m-3, even only mg m-3.
Because of this, and the characteristic time scale of
biological growth being in minutes and hours,
bioreactors are often very large indeed; with capacities
often in hundreds of tonnes, a few even of thousands.
Scale-up of such facilities is not easy - compromise is
needed between competing requirements in mass and
heat transfer, homogeneity, controllability and
practicality in construction and operation.

Applications
Chemical Industry
Production of bulk chemicals and solvents such as ethanol, citric acid,
acetone and butanol.
Synthesis of fine specialty chemicals such as enzymes, amino acids, alkaloids
and antibiotics.

Food Industry
Production of bakers' yeast, cheese, yogurt and fermented foods such as
vinegar and soy sauce.
Brewing and wine making.
Production of flavors and coloring agents.

Medicine

Development of novel therapeutic molecules for medical treatments.

Diagnostics.
Drug delivery systems.
Tissue engineering of replacement organs.
Gene therapy.

Applications
Agriculture
Plant breeding to improve resistance to pests, diseases, drought and salt
conditions.
Bioinsecticide development.
Modification of plants to improve nutritional and processing characteristics.

Veterinary Practice
Vaccine production.
Fertility control.
Livestock breeding.

Environment
Biological recovery of heavy metals from mine tailings and other industrial
sources.
Bioremediation of soil and water polluted with toxic chemicals.
Sewage and other organic waste treatment.

The Birth of the Concept of

Bioprocess Engineering

Penicillin

Alexander Flemings contaminated plate, 1928

[Link]
[Link]

Contaminated Plate
Bacteria culture
dishes contaminated
with a fungus.
Bacteria growth
inhibited by the
presence of the
fungus!

Mold was releasing a substance (Penicillium chrysogenum)

that was inhibiting bacterial growth !

Penicillin was then exploited

for treatment of bacterial infections

A Milestone in the History

of Human Medicine
by Alexander Fleming in 1928
An accidental discovery of an antibiotic
produced by a common mold of Penicillium
notatum.
High demand of antibiotics during World War II.

US became the major player: Merck, Pfizer,

Squibb, and USDA Northern Regional
Research Lab (USDA-NRRL).

Large-Scale Antibiotic Fermenters

How Was this Achieved?

Mercks model: partnership of biologists
and chemical engineers.
0.001 g/L (1939) ----- 50 g/L (1990).
The birth of the concept of

Bioprocess Engineering!

Introduction to Bioprocess
Factors Affecting Microorganism Growth

Concentration and time

Temperature

pH

Availability of nutrients

Source of carbon

Source of water

Electron Acceptor

 Engineer must provide

optimum environment for
desired microbial cell culture.
Seeding and special cultures
needed in special
circumstances.

Enzyme-Catalyzed Reactions

Enzymes
They are produced only by living organisms and
commercial enzymes are generally produced by
bacteria.
Most of enzymes are high-molecular weight proteins or
protein-like substances that acts on a substrate (reactant
molecule) to transform it chemically at a greatly
accelerated rate, usually 103 to 1017 times faster than the
uncatalyzed rate.
Enzymes are specific: one enzyme can usually catalyze
only one type of reaction.

Enzymes
They are present in small quantities in the reaction and are
not consumed during the course of the reaction nor do they
affect the chemical reaction equilibrium.

Uncatalyzed reaction:

SP

Catalyzed reaction: S + E

ES E + P

Catalytic activity is revealed by studying enzyme kinetics!

Activation Energy
They provide an alternate pathway for the reaction to occur
thereby requiring lower activation energy.

Because
enzymatic
pathways have
lower activation
energies,
enhancements in
reaction rates
can be
enormous!

Enzyme-Substrate Complex
Substrate binds with a specific site of the enzyme to form
the ES complex.

Two models for substrate-enzyme interactions:

Lock and key model

Induced Fit Model

Michaelis-Menten Kinetics
When the enzyme concentration is the limiting
factor, the kinetics of enzymes reactions are well
represented by the Michaelis-Menten equation:
dS
-

Vmax (S)
= - rS =

dt

Km+ (S)

- rS is the volumetric rate of reaction.

S is the concentration of substrate.
Vmax is the maximum rate of reaction at infinite
substrate concentration.
Km is the Michaelis constant for the substrate.

Michaelis-Menten Plot
Zero-order region

Michaelis-Menten region

rS

Cs
First-order region

This means that low values of Km imply

the enzyme achieves maximal catalytic
efficiency at low S.

Michaelis-Menten Kinetics
Vmax S
- rS=
Km+ S
High Substrate
Concentration

Low Substrate
Concentration

S KM

KM S

- rS

Vmax S
First-order

Km

Zero-order - rS Vmax

Michaelis-Menten Kinetics
Consider the case when the substrate concentration is
such that the reaction rate is equal to one-half the
maximum rate:

Vmax
- rS =
2

rS

Cs

Vmax
Vmax S (1/2)
=
2
Km + S (1/2)

Km = S (1/2)

-rS

Problems

Estimate
Vmax?
K m?

Problems
1) The enzyme-catalyzed conversion of a substrate at 25oC has a Km of 0.035 M.
The rate of the reaction is 1.15 x 10-3 M s-1 when the substrate concentration is
0.110 M. What is the maximum velocity of this reaction?

Solution : Vmax= 1.52 x 10-3 M s-1

An enzyme with a Km of 1x10-3 M was assayed using an initial substrate

concentration of 3x10-5 M. After 2 min, 5 percent of the substrate was converted.
What is the maximum velocity of this reaction?
2)

Solution : Vmax = 2.71 x 10-5 M min-1

Most Imp!!!

Discuss about Example 7-3 (Page No: 401)

Enzyme Inhibition (Mechanism)

I

Competitive

Non-competitive

Equation and Description

Cartoon Guide

Substrate

E
I

Compete for
Inhibitor active site
E + S
ES E + P
+
I

EI

I
S

Uncompetitive
E
I

Different site
E + S
ES E + P
+
+
I
I

EI + S EIS

E + S
ES E + P
+
I

EIS

[I] binds to free [E] only,

[I] binds to [ES] complex
[I] binds to free [E] or [ES]
and competes with [S];
complex; Increasing [S] can only, increasing [S] favors
increasing [S] overcomes
not overcome [I] inhibition. the inhibition by [I].
Inhibition by [I].
Juang RH (2004) BCbasics

Enzyme Inhibition (Plots)

I

Competitive

Non-competitive

Vmax

Double Reciprocal

Direct Plots

vo
I

[S], mM

Uncompetitive

Vmax

vo

Km Km

Km = Km

Vmax

Vmax

[S], mM

Km Km

Vmax

[S], mM

Vmax unchanged
Km increased

Vmax decreased
Km unchanged

Both Vmax & Km decreased

1/vo

1/vo

1/vo

Intersect
at Y axis

1/Km

Two parallel
lines

1/ Vmax
1/[S]

Intersect
at X axis

1/Km

1/ Vmax
1/[S]

1/ Vmax
1/Km

1/[S]

Juang RH (2004) BCbasics

Batch or Plug Fermenter

Where
CM=KM
k3CE0= Vmax

Mixed Flow Fermenter

Alternate Method for Evaluating

the M-M Parameters

Cell Growth
Natural proses mitosis - cells multiply their number to survive
4 phases
Phase 1 - lag phase little increase of cells number
Phase 2 exponential growth phase- cell dividing at the
maximum rate
Phase 3 stationary phase reach zero growth rate due to
lack of nutrient and space
Phase 4 death phase- decrease of live cells due to toxic
by-product, harsh environment and depletion of nutrient
Phase
Phase
No. of
Phase
Cells
1

Phase
4
Time

Rate Law
Cells + Substate More cells + Product
Monod equation

rg

m axCs Cc
K s Cs

Can use Moser or Tessier growth laws

Affected by temperature

Stoichiometry
Yield coefficients

YC S

CC
Mass of new cells formed

Mass of substrate consumed

CS

Yield coefficients of product

YP S

Mass of products formed

C P

Mass of substrate consumed

CS

Cell maintenance

Mass of substates consumed for ma int enance

m
Mass of cells T

Look at Table 7-5

Mass Balance
During Growth phase

dCs
YS C (rg ) mCc
dt
During Stationary Phase

dCs
V
mCcV YS P (rp )V
dt
Batch Stationary Growth rate

dCP
V
rpV Yp s (rs )V
dt
Look Example 7-6

The End