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 enzyme-
catalyzed 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 bio-
manufacture 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

http://fig.cox.miami.edu/~cmallery/255/255enz/penicillin.gif
http://www.time.com/time/time100/scientist/profile/fleming.html
 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 S
This means that low values of Km imply
First-order region the enzyme achieves maximal catalytic
efficiency at low S.
 Michaelis-Menten Kinetics
Vmax S
- rS=
Km+ S

Low Substrate High Substrate

Concentration
Concentration
S KM
KM S

Vmax S
- rS First-order Zero-order - rS Vmax
Km
 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
- rS =
2
Cs S

Vmax Vmax S (1/2)

= Km = S (1/2)
2 Km + S (1/2)
 Problems
-rS

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

2)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?

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 I Non-competitive I Uncompetitive

Substrate E
S S E I
S
Cartoon Guide

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

Equation and Description

EI EI + S EIS EIS

[I] binds to free [E] only, [I] binds to free [E] or [ES] [I] binds to [ES] complex
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 I Non-competitive I Uncompetitive

Vmax Vmax Vmax
vo vo
Vmax Vmax
I I I
Direct Plots

Km Km [S], mM Km = Km [S], mM Km Km [S], mM

Vmax unchanged Vmax decreased
Km increased Km unchanged Both Vmax & Km decreased

1/vo I 1/vo I 1/vo I

Double Reciprocal

Two parallel
Intersect lines
at Y axis 1/ Vmax Intersect 1/ Vmax 1/ Vmax
at X axis

1/Km 1/[S] 1/Km 1/[S] 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 3 Phase
No. of
Phase 2 4
Cells
1
Time
 Rate Law
Cells + Substate More cells + Product

Monod equation

max Cs Cc
rg
K s Cs

Can use Moser or Tessier growth laws

Affected by temperature
 Stoichiometry
Yield coefficients
Mass of new cells formed CC
YC S
Mass of substrate consumed CS

Yield coefficients of product

Mass of products formed C P
YP S
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

dC P
V rpV Yp s (rs )V
dt
Look Example 7-6
The End