B.Tech.

IV (CH), Semester VII L T P C

ES-1: Fundamentals of Biochemical Engineering 3 0 0 3
(Elective Subject from Department)
INTRODUCTION (06 Hours)
Scope and possibilities; Characteristics and classification of biological matter; Basics of microbial growth;
OVERVIEW OF BIOSEPARATIONS (02 Hours)
An Overview of bioseparations; Cell disruptions; Genetically modified organism
SEPARATION METHODS (25 Hours)
Filtration; Centrifugation; Adsorption; Extraction; Membrane separation processes; Concepts of precipitation,
Chromatography Basic concepts; Gel filtration Ion exchange chromatography; Hydrophobic chromatography;
Affinity chromatography; Suitable examples; Electrokinetic methods of separations; Finishing operations and
formulations
INDUSTRIAL APPLICATIONS (12 Hours)
Biomass to Biofuels; Bioremediation; Biocatalysts; Biofouling; Microbial Polymer and plastics; Natural resources
recovery
Total Contact Hours: 45)
BOOKS RECOMMENDED:
1.Shuichi Aiba, Arthur E. Humphrey, Nancy F. Millis, Biochemical Engineering, 2nd Ed., Academic Press, New
York, 1973.
2. James E. Bailey, David F. Ollis, Biochemical Engineering Fundamentals, 2nd Ed., McGraw hill, 1986.
3. L. Weatherley, Engineering Processes for Bioseparations, Butterworth-Heinemann Ltd., Oxford, 1995.
4. D.L. Pyle, Separation for biotechnology, Royal Society of Chemistry, Cambridge, 1994
5. A. Scragg, Environmental Biotechnology, 2nd Ed., Oxford University Press, 2005.
 Fundamentals of Biochemical
Engineering
Topics already Discussed
The importance of the need of the day of the subject
Similarity and difference between the Chemical and
Biochemical Engineering
Advantages of bioprocesses (ambient temperature,
high product specificity, relatively clean technology)
Application areas like Pharmaceuticals, drug
intermediates, food chemicals, beverages, organic fine
chemicals and solvents, industrial enzymes, dairy
products etc.
Characteristics of Bioproducts
 Need for Downstream Processing
Downstream processing (bioseparation)is
essential part of bioprocess Technology
The products are manufactured using a variety
of equipment like Fermenters or bioreactors,
other reactors such as airlift, membrane and
immobilized cell reactors are also used.
The products formed are usually in low
concentrations and for separation unit
operations are involved
Characteristics of Fermentation Broths
The characteristics of fermentation broths that
influence the downstream processing of
biomolecules include:

The type of microorganisms and their

morphological features (size and shape)

Concentration of cells, products and byproducts

Physical and rheological characteristics

Morphology of Cells
Cells size ranges from 1 micro m
to 4000 micro m

The cells and the cell

agglomerates exhibit a variety of
shapes.

Bacterial and yeast cells occur

mostly as homogeneously
suspended particles in the
fermentation broth

Fungi form a network of hyphi

thereby increasing the viscosity
of the broth

Cell size decreases capacity of

Figure 1: Morphology of Cell the separation process decreases
and cost increases
Concentration of cells, products and
byproducts
The concentrations of the biomass and that of the products in the fermentation
broth are important in deciding on the choice of the separation process
 Physical and Rheological
Characteristics
The density of the dry biomass would be about 1400 kg/m3;
however the density of the fermentation broth is lower , around
1100 kg/m3 as the cells have a high water content of about 70-80%.

The rheological property of fermentation broth is of importance for

downstream processing in the case of centrifugation and
membrane separations.

Newtonian and Non-newtonian models can describe most of the

fermentation broths.

The viscosity of the broth is strongly influenced by the cell

concentration as well as cell shape and to a minor extent by the
changes in the concentration of the nutrients and other
metabolites.
 The stages of bioseparations
The bioseparation stages are:
1. Product release and pre-treatment: Involves cell disruption
and release of intracellular products
2. Removal of insoluble's or particulates that is solid- liquid
separation: Filtration or centrifugation achieves the removal
or separation of particulates
3. Product concentration and recovery: Involves adsorption,
extraction, precipitation or membrane separation
4.Purification: Involves high resolution techniques like
chromatography and affinity separations
5. Finishing operations and formulations: Includes drying,
crystallization etc.
Note: This is purely for academic purpose and there is no rigid
stages for bioseparation
The separation Techniques
Typical bioprocesses
Typical bioprocesses
Cell wall of microorganisms
 Cell wall
The murein layer is about 10-80 nm thick, made of
petidoglycan exists in one form or other in almost all the
species.
The space below the murien layer called periplasmic space, is
about 8 nm thick and often contains enzymes
The inner layer called the plasma or cytoplasmic membrane is
about 8 nm is a double layer made of phospholipids and some
proteins and metal ions
The cell interior, called the cytoplasm, is an aqueous solution
of salts, amino acids and biopolymers including proteins,
enzymes, RNA and DNA.
It is necessary to rupture the cell to release the protein in the
cytoplasm
 Cell Disruption

Cells for disruptions

Chemical
Physical and Mechanical
Methods Enzymatic Disruptions
methods

Detergent Cell wall

Osmotic Ultrasonicati Alkali Enzyme Bead Mill High prressure
Thermolysis treatment
solubilizatio permeabiliz disruption homogenizer
shock on digestion
n ation

Lytic enzyme
to the cell
Ultrasound suspension
Cells in pure
For waves of Lipid
water (double
products of frequency solubilization by
amt) , cells
stable of .20kHz ruptures organic solvents
swells and
heat shock the cells
disrupts
Disruption of microbial cell
Bead mill disruption