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1 Module 1 – Basics of Bioprocess Technology

Bioprocess versus Chemical Process
Bioprocess Chemical Process
 Bioprocess is the production of commercially useful  A Chemical Process involves the conversion of a
component by manipulation of natural metabolic substrate or component A into component B or product
processes such as fermentation, degradation, etc. via a chemical reaction.

 It requires the intervention of biological entities either  It only requires the substrate chemical and in some
as intermediates or as substrates, such as plant cells, cases catalysts also.
enzymes, whole plant/animal tissues, etc

 The process is mostly organic  The process can be organic or inorganic

 The product, being produced from a biological source,  The product, being a sole production of a chemical
is more preferred for human consumption or shows entity, may not be beneficial for human as the SAR
more pharmacological activity due to its SAR may not be the same or pharmacological activity may
be less or lost.

 The process requires the intervention of enzyme or  Catalysts may be required for the better and efficient
group of enzymes for the conversion of the substrate to conversion of substrate to product. Catalysts are
the product. Enzymes are biocatalyst, biological entities inorganic/organic compounds which may or may not
/proteinaceous compounds which require special require any special conditions.
conditions to function.

 The process requires the critical control over the  The process mostly works at higher temperatures, pH,
optimal conditions like optimal temperature, optimal osmotic pressure etc and mostly does not require much
pH, osmotic pressure, water/fluid content, agitation and critical control.
aeration, etc.

 The mixture obtained from the main process  The mixture obtained contains maximum amount of the
(fermentation, degradation, etc) is required to be product in free suspended form. There is no
processed further i.e. purified so as to obtain the purification required as such to obtain the product.
product. This is also called Downstream Processing,
and the part before it is called upstream processing.

 Presence of impurities is very high but can be dealt  Presence of impurities is very low but is more potent to
with using proper downstream purification processes be a toxic content which can be hazardous in nature.
like filtration, centrifugation, etc.

 The specificity is very high because of the presence of  Specificity is very low compared to the bioprocess.
enzymes in the process.

 The compendia and regulatory authorities consider  It may or may not be a greener approach and
bioprocess as a better and greener approach and commercially viable.
commercially viable option.
Basic Principles in Bioprocess Technology
Bioprocess engineering emphasizes the engineering and
sciences of industrial processes that are bio based:
1) Biomass feedstock conversion for a sustainable
society or bio refinery;
2) Bio catalysis-based processing; and
3) Manipulation of microorganisms for a
sustainable and socially desirable goal.
Bioprocess engineering is neither product-based nor is
substrate based. Therefore, bioprocess engineering deals
with biological and chemical processes involved in all
areas, not just for a particular substrate or species (of
feedstock or intermediate), outcome, or product.
Thus, bioprocess engineering intercepts chemical, mechanical, electrical, environmental, medical, and industrial
engineering fields, applying the principles to designing and analysis of processes based on using living cells or
subcomponents of such cells, as well as nonliving matters. Bioprocess engineering deals with both microscale
(cellular/molecular) and large-scale (system wide/industrial) designs and analyses. Science and engineering of processes
converting biomass materials to chemicals, materials, and energy are therefore part of bioprocess engineering by
extension. Predicting and modeling system behaviors, detailed equipment and process design, sensor development,
control algorithms, and manufacturing or operating strategies are just some of the challenges facing bioprocess
engineers. At the heart of bioprocess engineering lays the process kinetics, reactor design, and analysis for Biosystems.
The basic questions which need to be asked for the process development and design are as follows:
 What change can be expected to occur?
To answer this question, one must have an
understanding of the basic sciences for the
process involved. These are microbiology,
biochemistry, molecular biology, genetics, and
so on. Biochemical engineers need to study
these areas to a certain extent. It is also true that
the contribution of biochemical engineers in
selecting and developing the best biological
catalyst is quite limited unless the engineer
receives specialized training. However, it is
important for biochemical engineers to get
involved in this stage, so that the biological
catalyst may be selected or genetically modified
with a consideration of the large-scale operation.
 How fast will the process take place?
If a certain process can produce a product, it is
important to know how fast the process can take
place. Kinetics deals with rate of a reaction and
how it is affected by various chemical and
physical conditions. This is where the expertise
of chemical engineers familiar with chemical
kinetics and reactor design plays a major role.
Similar techniques can be employed to deal with
enzyme or cell kinetics. To design an effective
bioreactor for the biological catalyst to perform,
it is also important to know how the rate of the
reaction is influenced by various operating
conditions. This involves the study of
thermodynamics, transport phenomena, biological interactions, clonal stability, and so on.
  How can the system be operated and controlled for the maximum yield?
For the optimum operation and control, reliable on-line sensing devices need to be developed. On-line
optimization algorithms need to be developed and used to enhance the operability of bioprocess and to ensure
that these processes are operated at the most economical points.
 How can the products be separated with maximum purity and minimum costs?
For this step, the downstream processing (or bio separation), a biochemical engineer can utilize various
separation techniques developed in chemical processes such as distillation, absorption, extraction, adsorption,
drying, filtration, precipitation, and leaching. In addition to these standard separation techniques, the
biochemical engineer needs to develop novel techniques which are suitable to separate the biological materials.
Many techniques have been developed to separate or to analyze biological materials on a small laboratory scale,
such as chromatography, electrophoresis, and dialysis. These techniques need to be further developed so that
they may be operated on a large industrial scale.

Basic Principles of Engineering Analysis

It is said that any good engineering calculation ends with a number bearing a dollar sign. The purpose of engineering
analysis, therefore, is to determine “how much” and “how fast.” These questions are answered in the form of equations
that can be solved for product capacity, processing rate, and possibly product purity. To obtain values for such
variables, a set of governing equations derived from the three principal ingredients of engineering analysis is required:
equilibria, material balance, and flux (or transport phenomena).
1. Material balance
Operating curves (for each unit operation) are built on the basis of material balances and used in consort with
equilibrium curves. The material balance equation states
Accumulation=Inflow – Outflow + Amount produced− Amount consumed
Examples of Equilibria
Traditionally, a chemical reaction that is at equilibrium,
A+ B C
Can be characterized by an equilibrium constant
[C ]
K eq =
[ A ] [B]
Where all concentrations are in mole fractions, molarity, or molality. Likewise, in an extraction process that has gone to
equilibrium,
Y
K=
X
Where, Y = Concentration of a separand in the extract phase or Extractant phase
X = Concentration of the same separand in the Raffinate (usually heavy) phase or Raffinate Phase
K = partition coefficient or distribution coefficient.
In the case of adsorption, the equilibrium constant relates the concentration in the adsorbent phase, [CS], to the
concentration in the liquid phase, [C], at equilibrium, where C is a chemical species and S is an adsorption site. When
the adsorption equilibrium is linear, for example, the following relation is valid at low concentrations:
[CS ]
K=
[C ]
Flux Relationships (Transport Phenomena)
Flux relationships abound in chemistry and physics, and they all state that
Flux=Coefficient × Driving Force
Where flux is in units flowing per unit area per unit time, driving force is a gradient down which units flow, and the
“coefficient” is a permeability or the inverse of a resistance, derivable from properties of the medium. Thus, we have
Ohm’s law
J e =CE
Where Je is current density, C is electrical conductivity (property of the medium), and E is electrical potential gradient.
Fick’s first law applies for diffusive flux due to a concentration gradient dc/dx in one dimension
dc
J D=−D
dx
Where D, the diffusion coefficient, is also a property of the medium and in some cases calculable from the Stokes-
Einstein equation for spheres
kT
D=
6 πμa
Where k is the Boltzmann constant, T is absolute temperature, μ is viscosity, and a is particle radius.
Of great interest in downstream processing is the empirical law (Darcy’s law) of flow through a porous medium or the
transport of permeate through the membrane follows the basic principles of flow through porous media. For membrane
filtration, Darcy’s law is written as follows:
J W =LP ∆ p (1)
Where Jw is the fluid flux/permeate flux, Lp is Darcy’s law permeability or hydraulic permeability, and Δp is pressure
drop across the porous medium, such as a filter or a column of adsorbent resin—systems very frequently used in
bioprocessing. The hydraulic permeability is an important characteristic of the membrane, because it affects strongly the
filtration capacity of the system.
What are the physical properties that determine the hydraulic permeability of membranes? If the membrane is
considered as a medium perforated by straight, parallel capillaries of radius r, then the hydraulic permeability, based on
Poiseuille law, is given by:

ε r2
Lp = (2)
8 μz
Where, ε = membrane porosity, dimensionless
z = membrane thickness, m
µ = viscosity of the permeate, Pa.sec

2. Process and Product Quality

The measures of product quality due to processing are purity, fold purification, specific activity, and yield. Purity is
defined as follows:
Amount of Product Purity at Stage B
Purity= And Fold Purity=
Amount of Product +Total Impurities Purity At Stage A
Fold purification is the ratio of the purity at any stage in the process to the purity at the start of the purification process.
Another measure of purity is
Units of Biological Activity( IU )
Specific activity=
Mass(g)
Where units of biological activity are assayed by means of a biological test, such as moles of substrate converted per
second per liter or fraction of bacterial cells killed. For proteins, the mass is usually total protein; on this basis, the
specific activity reaches a constant value when the protein is pure.
Yield is given by
Amount of Product produced
Yield=
Amount of Product ∈feed
Purity is a strictly quantitative measure and not always an expression of the quality of the product. A therapeutic protein
can be 99.99% pure but still unacceptable if any pyrogen (a substance that produces a fever) is present. On the other
hand, if the product is not a therapeutic protein but an industrial enzyme, then practically any impurities that do not
inhibit the activity of the product or endanger the user are allowed. In short, two kinds of measurements of purity are
required: activity, composition, and structure on the product itself; and host-cell materials, degraded product, and
excipients (additives) on the impurities.