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CHAPTER 10:
INTEGRATED BIOSEPARATION
SCHEME FOR PRODUCT
ISOLATION, PURIFICATION
AND FORMULATIONS UNITS
FOR BIOPROCESS
MISS. RAHIMAH BINTI OTHMAN
(Email: rahimah@unimap.edu.my)
 COURSE OUTCOME 1 CO1)
CHAPTER 7 : Upstream Processing In Bioprocess Plant.
CHAPTER 8 : Bioreaction Design In Bioprocess Plant.
CHAPTER 9 : Downstream Processing In Bioprocess Plant.

CHAPTER 10: Integrated Bioseparation Scheme for Product

Isolation, Purification and Formulation Units
For Bioprocess.
CHOOSE the appropriate sequencing of processes;
SYNTHESIZE and EVALUATE integrated bio-separation
schemes that could be used to achieved the required purification
of important biological molecules (product).
1. What is the “Integration Process”?
2. What is “Product Isolation, Purification and
Formulation Units”.
3. How to CHOOSE the appropriate sequencing
of processes.
4. SYNTHESIZE and EVALUATE integrated
bio-separation schemes.
- [Propose: to achieve the required purification of
important biological molecules (product)].
GENERALIZED VIEW OF BIOPROCESS
RAW MATERIALS

UPSTREAM PROCESSES

Media Formulation
Inoculum Equipment
and
Preparation Sterilization
Sterilization

BIOREACTOR - FERMENTER

Reaction Kinetics Transport Phenomena Instrumentation

and Bioactivity and Fluid Properties and Control

DOWNSTREAM PROCESSES

Recovery and Waste Recovery,

Separation
Purification Reuse and Treatment

THE BOTTOM LINE

REGULATION ECONOMICS HEALTH AND SAFETY

WHAT IS INTEGRATION PROCESS?

INTEGRATION
In engineering, system
integration is the bringing
together of the component
subsystems into one system and
ensuring that the subsystems
function together as a system.
In information technology,
systems integration is the
process of linking together
different computing systems and
software applications physically
or functionally.
 EXAMPLE: The research
roadmap of Process Integration
study in Yeast & Ethyl Alcohol
(Y&EA) plant.
Physical model of Y&EA plant.
Fig. 5. Scheme of the integrated bioprocess. Limonene-saturated air was
supplied to the bioreactor by leading it through a washing flask filled with
pure limonene (A).Glycerol (B) and limonene (C) were continuously fed via
peristaltic pumps. Using another peristaltic pump (D) the culture broth was
continuously recirculated through a recovery loop containing a column with
a fluidized bed of anion exchanger Amberlite IRA 410 Cl (E).
CHEMICAL MOLECULE STRUCTURE
INTEGRATION
 What is “Product Isolation,
Purification, and Formulation Units?
Removal of insolubles is the first step and involves the capture of the
product as a solute in a particulate-free liquid, for example the separation
of cells, cell debris or other particulate matter from fermentation broth
containing an antibiotic.
- Typical operations to achieve this are filtration, centrifugation,
sedimentation, flocculation, electro-precipitation, and gravity settling.

- Additional operations such as grinding, homogenization, or leaching,

required to recover products from solid sources such as plant and
animalIsolation
Product tissues, are
is usually included
the removal in this
of those group. whose properties
components
vary markedly from that of the desired product.
- For most products, water is the chief impurity and isolation steps are
designed to remove most of it, reducing the volume of material to be
handled and concentrating the product.
- Solvent extraction, adsorption, ultrafiltration, and precipitation are
some of the unit operations involved.
 What is “Product Isolation,
Purification, and Formulation Units?
Product Purification is done to separate those contaminants that resemble
the product very closely in physical and chemical properties.
- Consequently steps in this stage are expensive to carry out and
require sensitive and sophisticated equipment.
- This stage contributes a significant fraction of the entire downstream
processing expenditure.
- Examples of operations include affinity, size exclusion, reversed
phase chromatography, crystallization and fractional precipitation.

Product Polishing describes the final processing steps which end with
packaging of the product in a form that is stable, easily transportable and
convenient.
- Crystallization, desiccation, lyophilization and spray drying are
typical unit operations.
How to CHOOSE the appropriate sequencing of processes?

Process Integration (PI), is primary focused to system

oriented process design for energy efficient and cost
effective mapping of the plant.

This approach can be summarised in one sentence:

‘‘PI is probably the best approach that can be used to

obtain significant energy and water savings as well as
pollution reductions for different kind of industries’’.
How to CHOOSE the appropriate sequencing of processes?

Integrated bioprocesses have been developed to optimise yield and

cost-effectiveness of production of low and high molecular weight
molecules.
Most of the recent research describes the integration of
processes, especially the ISPR (in situ product removal) technique,
during the production of low molecular weight compounds.
To achieve a continuous product recovery (i.e. ISPR), techniques
based on extraction (i.e. the most commonly used method),
adsorption, electrodialysis and crystallisation are used.

Other integrated processes include integrated product formation in

which the breakdown product of a polymer is converted to a low
molecular compound in a one-pot process, and integrated
extraction and enzymatic conversion.
How to CHOOSE the appropriate sequencing of processes?

In situ product removal (ISPR)

I n situ extraction of other systems
 In situ adsorption and ion exchange
 In situ electrodialysis
 In situ crystallisation

Integrated product formation

EXAMPLE: The decomposition of a polymer (starch, cellulose,
hemicellulose, protein) and the conversion of the monomer are
performed sometimes in a one-pot process. Such a process is the
simultaneous saccharification of corncobs by the enzyme of
Trichoderma reesei and the conversion of the product by L.rhamnosus
to lactic acid .
How to CHOOSE the appropriate sequencing of processes?

 EXAMPLE: Membrane-based separations

SYNTHESIZE and EVALUATE integrated bio-separation schemes.

Integrated stack-monitoring emission system.

Detailed flowsheet of Yeast & Ethyl Alcohol Plant.
 References and recommended reading
1. Freeman A, Woodley J, Lilly MD: In situ product removal as a tool for bioprocessing.
Biotechnology 1993, 11:1007-1012.
2. Schuegerl K: Solvent Extraction in Biotechnology. Springer Verlag; 1994.
3. Schuegerl K, Kretzmer G, Freitag R, Scheper T: Integrierte biotechnologische produktionsprozesse.
Chem Ing Technik 1994, 66:1585-1592.
[Title translation: Integrated biotechnological processes.]
4. Schuegerl K: Integrated processing of biotechnology products. Biotechnol Adv 2000, 18:581-599.
5. Fernandes P, Prazeres DM, Cabral JMS: Membrane assisted extraction bioprocesses. Adv Biochem
Eng Biotechnol 2003, 80:115-148. This review covers all of the integrated membrane assisted
extractive processes.
6. Stark D, von Stockar U: In situ product removal (ISPR) in whole cell biotechnology during the last
twenty years. Adv Bioche Eng/Biotechnol 2003, 80:149-175.
[This review covers all types of in situ product removal techniques.]
7. Maas D, Gerigk MR, Kreutzer A, Weuster-Botz D, Wubbolts M, Takors R: Integrated L-
phenylalanine separation in an E. Coli fed-batch process from laboratory to pilot scale. Bioproc
Biosynth Eng 2002, 25:85-96.
8. Gerigk MR, Maas D, Kreutzer A, Sprenger G, Bonaerts J, Wubbolts M, Takors R: Enhanced pilot-
scale fed-batch Lphenylalanine production with recombinant Escherichia coli by fully integrated
reactive extraction. Bioprocess Biosyst Eng 2002, 25:43-52.
9. Ru¨ ffer N, Heidersdorf U, Kretzers I, Sprenger GA, Raeven L, Takors R: Fully integrated L-
phenylalanine separation and concentration using reactive-extraction with liquid centrifuges in a
fed-batch process with E. coli. Bioprocess Biosyst Eng 2004, 26:239-248.
 Thank
Thank you
you
Prepared by,
MISS RAHIMAH OTHMAN