Slides have details about various types of Fermentors, Bioreactors and Downstream processing of Bioreactors

1. DR. HARISINGH GOUR CENTRAL UNIVERSITY, SAGAR
Presented by:-
Sunny Rathee
Subject – Industrial Biotechnology
DEPARTMENT OF PHARMACEUTICAL SCIENCES
1

2. 1. Bioreactor (Introduction).
2. Classification of Bioreactor process.
3. Types of Bioreactors.
4. Downstream processing of Bioreactors.
5. Immobilized cell Bioreactors.
6. Applications of Bioreactors.
7. Modelling of Bioreactors.
8. Single cell proteins (Introduction).
9. Production of single cell proteins.
10. Applications of single cell proteins .
CONTENTS
2

3. BIOREACTORS
3
Bioreactor : A Bioreactor is basically
a device in which the organism (cells) are
cultivated to form the desired product(s).
 It is a containment system designed to
give right environment for optimal growth
and metabolic activity of the organism.
 A Fermenter usually refers to the
containment system for the cultivation of
prokaryotic cells (bacteria, fungi), while a
bioreactor used to grow the eukaryotic
cells (mammalian, insect).
FIGURE1: SCHEMATIC
REPRESENTATION OF AN
CONTINUOUS STIRRED
TANK BIOREACTOR

4. FIGURE 2 :DIAGRAM OF A TYPICAL BIOREACTOR

5. CLASSIFICATION OF BIOREACTOR PROCESSES
FOR SUSPENSION CULTURE
5
Types of
Bioreactors
(In terms of
process
requirements)
2)Anaerobic
1)Aerobic
3)Immobilized
cell Bioreactor

6. On the basis of mode of operation bioreactors may
be classified as :
6
Types of
Bioreactors on
basis of Mode
of Operation
2) Fed Batch
Bioreactor e.g. –
Fluidized Bed
Bioreactor
1) Batch Type
Bioreactor e.g. –
Stirred tank
Bioreactor
3) Continous Type
Bioreactor e.g. –
Chemostat and
Turbidostat

7. Types of Bioreactors
 Stirred types of bioreactor ,
 Air lift bioreactor ,
 Fluidized bed bioreactor ,
 Tower bioreactor ,
 Continuous bioreactor ,
 Gaseous phase bioreactor . 7

8. Air Lift Bioreactor
The bioreactor consists of two concentric cylinders.
The bottom of the inner chamber carries a sintered steel ring
through which 5%Carbon dioxide or Oxygen-8% in air is
bubbled.
The bubbles rise, carrying the cell suspension with them. O2/CO2
is vented from the top and displacement ensures the return of the
cell suspension down the outer chamber.
8

9. 9
FIGURE 3 – Schematic Representation of Air lift Bioreactor

10.  Advantages :
1. Suited for aerobic culture.
2. Low energy consumption.
3. No agitator shaft is needed.
4. Greater heat removal vs stirred tank.
Disadvantages
1. Greater air through put and higher pressure needed.
2. No bubble breaker. 10
ADVANTAGES AND DISADVANTAGES OF
AIR LIFT BIOREACTORS

11. Continuous Bioreactor
 Chemostat-(Chemical environment is constant) cell grow at max
density when some nutrient like vitamin , is growth limiting.
A continuous stirred tank bioreactor consists of a cylindrical vessel with
motor driven central shaft that supports one or more agitators (impellers).
Turbidostat - Cells growto achieve higher density.
The shaft is fitted at the bottom of the bioreactor.
The number of impellers is variable and depends on the size of the
bioreactor i.e. height to diameter ratio, referred to as aspect ratio.
The aspect ratio of a stirred tank bioreactor is usually between 3-5.
11

12. Advantages
1. Generally maintain cells in log phase for longer period.
2. Process maintain at steady state.
3. Large amount of production can be done.
Disadvantage
• Difficult to maintain pH.
12
ADVANTAGES AND DISADVANTAGES OF
CONTINUOUS TYPE BIOREACTORS

13. TOWER BIOREACTOR
 Elongated non - mechanically stirred
fermenters.
 Aspect ratio 6:1.
 Unidirectional flow of gases.
Used for continues production.
 E.g. Used for singled cell protein
production.
 Cells are used as biocatalyst in three
phase system (solid ,liquid and gas).
FIGURE 4 – TOWER
BIOREACTOR
13

14.  Basically particles used in FBB can be of three types-
 Inert core in which cell can attached.
 Porous particles in which biocatalyst is entrapped.
 Cell aggregates or flocs.
 Because of the higher density of the microcarriers they can be perfused
slowly from below, at such a rate that their sedimentation rate matches
the flow rate.
 Gas exchange is external to the reactor, and no mechanical mixing is
required.
 Fluidized bed bioreactor is comparable to bubble column bioreactor
except the top position is expanded to reduce the velocity of the fluid.
 These bioreactors are suitable for use to carry out reactions involving
fluid suspended biocatalysts , such as immobilized enzymes , cells.
FLUIDIZED BED BIOREACTOR
14

15. ADVANTAGES OF FLUIDIZED BED
BIOREACTOR
Uniform temperature gradient.
 Ability to operate reactor in continuous
state.
 Uniform mixing of particles.
 Recycling of liquid is important to
maintain continuous contact between the
constants and biocatalysts. This enable
good efficiency of bioprocessing.
 Fluidized bed bioreactors are well
suited to exothermic reactions.
FIGURE 5 : FLUIDIZED BED
BIOREACTOR
15

16. Downstream processing of Bioreactor
Filtration –Surface filtration , Depth filtration, Cross flow
filtration ,Membrane filtration etc.
Centrifugation- Difficulty arise due to small differences in the
density of particle and medium.
Ion – Exchange Resin – Dextrose , Hemicellulose .
Chromatography- Affinity chromatography ,
Adsorption chromatography.
16

17. NASA TISSUE CLONING BIOREACTORS
The bioreactors in which cells or tissues grow for experimental or
therapeutic purposes, the design is significantly different from
industrial bioreactors.
Many cells and tissues, especially mammalian ones, must have a
surface or other structural support in order to grow, and agitated
environments are often destructive to these cell types and tissues.
NASA has developed a new type of bioreactor that artificially
grows tissue in cell cultures.
NASA's tissue bioreactor can grow heart tissue, skeletal tissue,
ligaments, cancer tissue for study, and other types of tissue.
17

18. IMMOBILIZED CELL BIOREACTORS
 These are based on immobilized cells.
 Advantages –
1. Useful for manufacture of intracellular enzymes.
2. Can be used when extracted enzymes are unavailable.
3. For preparing low molecular weight products which are released into
the medium.
4. Suitable for production of amino acids , organic acids , etc.
 Commonly Fluidized bed bioreactors and hollow fiber membrane
bioreactors are used.
 Hollow fiber membrane bioreactors have hollow fibers made from
cellulose acetate ,acrylic polymers , poly –sulphones.
18

19. Pharmaceutical applications of bioreactors
1. Production of industrially valuable products such as enzymes,
proteins, antibiotics, etc.
1. Bioreactors are used for biomass production (cells or embryogenic
propagules, shoots, or roots as a final product).
2. Bioreactors are used for biotransformation of exogenously added
metabolites.
3. Stirred tank bioreactors are used for the production of alpha –
interferon that have a clinical use in cancer and viral infection.
4. Bioreactors are extensively used for the ethanol production and
distillation, commercial production of citric acid. 19

20. 6. Bioreactors are commonly used for the production of penicillin by
fermentation process.
7. Packed bed bioreactors have been used widely for perfusion culture of
immobilized mammalian cells.
8. Stirred tank bioreactors are used for free and immobilized cells and
enzymes.
 Other applications of Bioreactors :
1. Bioreactors can be used to produce Biodiesal from algae with help of
tubular algae bioreactors.
2. The most common application of bioreactors is in wastewater
treatment, where bioreactor supports desired bacteria in flocculated
forms known as activated sludge flocs. Such flocs allow for
microorganisms to convert biodegradable organic substances into
carbon dioxide and new biomass. This generated biomass is easily
removed with membranes before distribution of the treated water. 20

21. MODELLING OF BIO-REACTOR
 Mathematical models act as an important tool in various bio-reactor
applications including wastewater treatment.
 These models are useful for planning efficient process control
strategies and predicting the future plant performance.
 Moreover, these models are beneficial in education and research areas.
 Bioreactors are generally used in those industries which are concerned
with food, beverages and pharmaceuticals.
 The emergence of Biochemical engineering is of recent origin.
 Processing of biological materials using biological agents such as
cells, enzymes or antibodies are the major pillars of biochemical
engineering.
 Applications of biochemical engineering cover major fields of
civilization such as agriculture, food and healthcare, resource recovery
and fine chemicals.

22. PRESENTED BY:
SUNNY
RATHEE
22

23. DEFINITION:
 It refers to edible micro-organisms.
 The biomass or total protein extract from pure or mixed cultures of
algae, fungi, bacteria.
 Yeast used as an ingredient or substitutes for protein rich foods.
 Dietary protein supplement.
 Scp can be used as a protein supplement for humans &animals.
 Food grade for humans.
 Feed grade for animals. 23
SINGLE CELL PROTEIN

24. 24
Several micro-organisms are used for the production of SCP:
 BACTERIA – Pseudomonas fluroescens , Lactobacillus.
 YEAST – Saccharomyces cerevisiae , Candida tropicalis.
 FUNGI – Aspergillus fumigatus , Aspergillus Niger.
 ALGAE – Spirulina species , Chlorella pyrenoidosa.
 ACTINOMYCETES.
SCP PRODUCTION IN INDIA -
 National Botanical Research Institute (NBRI).
 Central Food Technological Research Institute (CFTRI).
SOURCES OF SINGLE CELL PROTEIN
PRODUCTION

25. 25
SCP PRODUCTION
 Selection of suitable strain
 Fermentation
 Harvesting
 Post harvest treatment
 SCP processing for food

26. 1. RAW MATERIALS
Production of SCP requires micro-organisms that serve as the
protein source and the substrate that is biomass on which they
grow.
There is a variety of both the sources that can be used for the
production of SCP.
The biomass used can be plant biomass or organic biomass.
The micro-organisms used belongs to the group of algae
,fungi and bacteria.
26

27. • Micro-organisms used are fungi , yeast, algae & bacteria .
The following table shows average different compositions of
main groups of micro-organisms (% dry wt.).
COMPOSITON FUNGI ALGAE YEAST BACTERIA
PROTEIN 30- 40 % 40- 60 % 45- 55 % 50- 65 %
FAT 9-14 % 8-10 % 5-10 % 3-7 %
NUCLEIC ACID 7-10 % 3-8 % 6-12 % 8-12 %
27
2 . MICRO-ORGANISMS

28. 3. Selection of strain
 It a very critical step as the quality of protein depends totally on the
microbe that is used for the production.
 Thus careful selection of the strain should be done.
 Care should be taken that the selected strain should not produce any
toxic or undesirable effects in the consumer.
 It can be carried out in the fermenter which is equipped with aerator,
thermostat, pH, etc. or in the trenches or ponds.
Microbes are cultured in fed- batch culture.
 Engineers have developed deep lift fermenter & air lift fermenter .28
4. FERMENTATION

29. 5 .Harvesting
 When the colonies of microbes are fully developed, they are then
harvested.
 The bulk of cells are removed from the fermenter by decantation.
 After harvesting, the cells are subjected to a variety of processes.
 Post-harvesting treatments includes steps like washing , drying ,etc.
29
6 .Post harvest treatment

30. 7 . PROCESSING FOR FOOD
It includes -
1. Liberation of cell proteins by destruction of indigestible cell wall.
A. MECHANICALMETHODS
 Crushing, crumbling, grinding, pressure homogenization, etc.
B. CHEMICALMETHODS
 Enzymes & salts are used to digest or disrupt the cell wall.
 Salts like NaCl, sodium dodecyl sulfate, etc. whereas nuclease enzymes
are used.
C. PHYSICALMETHODS
 Freeze- thaw, osmotic shock, heating & drying.
30

31.  Chemical & enzymatic treatments are preferred.
 Chemicals which are used includes acidified alcohol, salts, acids
& alkalies.
Use of such chemicals leads to formation of lygino-alanine which causes
hypersensitivity skin reactions.
Enzymes which are used include ribonuclease & nuclease enzymes.
 These enzymes can be used exogenously or can be induced
endogenously.
31
2. Reduction of nucleic acid content

32. APPLICATIONS OF SCP
 Also source of vitamins, amino acids, minerals, crude
fibers, etc.
 Supplemented food for under - nourished children.
 Controls obesity.
 Provides instant energy .
 Example- Spirulina- part of diet of US Olympic team.
1. As protein supplemented food-
2. As health food-
32

33. 3. In therapeutic and natural medicines-
Reduce body weight, cholesterol, stress.
Lowers blood sugar level in diabetis (due to presence of B -linolenic
acid).
Prevents accumulation of cholesterol in body.
Healthy eyes and skin (beta carotene).
Beta carotene ( Anti - cancer substance - UN National Cancer Research
Institute).
Increase lactation.

33

34. 4. In cosmetics-
 Important role in maintaining healthy hair (vitamin A andB).
 Many herbal beauty products.
 Biolipstics and herbal face cream (Phycocyanin).
 Capable of replacing coal tar dye based cosmetics.
 Excellent, convenient source of protein.
 Used to feed cattle, fishes etc.
5 . Poultry and Cattle Field
34

35. References :
1. Satyanarayana .U , “Textbook of biotechnology” ,Reprint edition
Third revised edition 2009, Books and allied (P) Ltd. 239-269.
2. Sharma R, Rohilla A, Arya V. International Journal of
Pharmaceutical Sciences Review and Research, 2011; 9(2): 20-21.
3. Stanbury .P.f., Whitaker .A , “Principles of fermentation technology
” Second edition, 2012, Elsevier publications, 234-246.
4. Reddy .S.M . “Basic fermentation technology” Second revised
edition ,2010, New age publications , 125-132.
5. Mandenius.C. “Bioreactors, design, operation, and novel
applications, First edition 2009, Wiley – VCH publications, 54,78,
92,12.
35

36. 36