about bioreactors (collecting)

1. BIOREACTORS
THE STATE OF THE ART
PROF.DR/SAID SAAD SOLIMAN
By / Ahmed Shehata Ali
Gail

2. DEFINITIONS
1- bioreactor is a vessel in which a chemical process is carried out which
involves organisms (mainly microbes-viruses or bacteria, fungi and yeasts
–traditionally designated as „fermenters“) or biochemically active
substances (enzymes, e.g.) derived from such organisms –in opposite to
fermenters frequently considered as „true“ bioreactors. this process can
either be aerobic or anaerobic.
2- an apparatus, such as a large fermentation chamber, for growing
organisms such as bacteria or yeast that are used in the biotechnological
production of substances such as pharmaceuticals, antibodies, or
vaccines, or for the bioconversion of organic waste.

3. DEFINITIONS
a bioreactor may also refer to a device or system meant to grow cells or
tissues in the context of cell culture. cell culture is the process by which
cells are grown under cultivated conditions (animal cells, plant cells, algae).

4. INTRODUCTION
•the bioreactor´s environmental condition like gas (oxygen, nitrogen, carbon
dioxide) and liquid flow rates, temperature, ph, concentration of substrate and
products, cells number and their composition (proteins and nucleic acids),
dissolved oxygen levels,, and agitation speed (or circulation rate) need to be
closely and continuously monitored and controlled. in many cases, strictly
aseptic conditions have to be maintained.

5. INTRODUCTION
•in an aerobic process, optimal oxygen transfer is perhaps the most difficult task
to accomplish.
•there are, limits to the speed of agitation, due both to high power consumption
and to the damage to organisms caused by excessive tip speed.

6. CLASSIFICATION
ON the basis of mode of operation
1- batch
2- fed batch
3- continuous

7. CLASSIFICATION
on the basis of mode of flow of fluids
1- cstr bioreactor (continuous flow stirred reactor-the content of the bioreactor
is ideally mixed)
2- bioreactor with piston flow and bioreactors with non- ideal flow of fluids
(cascade of ideal mixtures
3- dispersed flow of fluids).
-the quality of flow of fluids significantly influences the rate of grow of
cells and the degree of conversion of substrate

8. CLASSIFICATION
On the basis of a number of phases treated
1- Homogeneous bioreactors (e.G. One phase tubular bioreactor with enzyme
diluted in the liquid substrate)
(e.G. Two phase solid-liquid bioreactor like the column type bioreactor2-
heterogeneous bioreactors with immobilized enzyme and liquid substrate and/or
three phase bioreactors with submersed culture: gas (air bubbles)-liquid
(substrate)-solids (cells).

9. SUMMARY OF BIOREACTOR SYSTEMS
ProductsCell Systems usedBioreactor product design
SCP, Enzymes, Secondary
metabolites, Surfactants
Bacteria, Yeast and other fungiAir-Lift Bioreactor
Ethanol, Secondary ,
metabolites, Wastewater
treatment
Immobilized bacteria, yeast and
other fungi, Activated sludge
Fluidized-Bed Bioreactor
Interferons, Growth factors,
Blood factors, Monoclonal
antibodies, Vaccines, Proteases,
Hormones
Immobilized (anchored)
mammalian cells on solid
particles
Microcarrier Bioreactor

10. SUMMARY OF BIOREACTOR SYSTEMS
ProductsCell Systems usedBioreactor product design
Interferons, Growth factors,
Blood factors , Monoclonal
antibodies, Vaccines,
Proteases, Hormones
mammalian, tissue growth on
solid surface, tissue
engineering
Surface Tissue Propagator
Ethanol, Monoclonal anti-
bodies, Interferons, Growth
factors
Bacteria, Yeasts, Mammalian
cells, Plant cells
Membrane Bioreactors,
Hollow fibers and membranes
used, products
Rotorfermentor
Ethanol, Monoclonal anti-
bodies, Interferons, Growth
factors
Immobilized Bacteria, Yeast,
Plant cells
Modified Stirred Tank
Bioreactor

11. SUMMARY OF BIOREACTOR SYSTEMS
ProductsCell Systems usedBioreactor product design
Ethanol, Enzymes, Medicinal
products
Immobilized Bacteria, Yeasts
and other fungi
Modified Packed Bed
Bioreactor
Single Cell Protein (SCP)Bacteria, YeastsTower and Loop Bioreactors
Ethanol, Volatile productsBacteria, Yeasts, FungiVacuum Bioreactors

12. SUMMARY OF BIOREACTOR SYSTEMS
ProductsBioreactor product design
Commodity products,SCPBacteria, Yeasts, FungiCyclone Bioreactors
SCP, Algae, Medicinal plant
products Monoclonal
antibodies,
Vaccines, Interferons
Photosynthetic bacteria
Algae, Cyanobacteria, Plant
Cell culture, r-DNA plant
cells
Photochemical Bioreactors
Cell Systems used

13. BASIC BIOREACTOR DESIGN CRITERIA
1- Microbiological and biochemical characteristics of the cell system (microbial, mammalian,
plant)
2- Hydrodynamic characteristics of the bioreactor
3- Aeration and oxygen, mass and heat transfer characteristics of the bioreactor
4- Kinetics of the cell growth and product formation
5- Genetic stability characteristics of the cell system

14. BASIC BIOREACTOR DESIGN CRITERIA
6- Aseptic equipment design
7- Control of bioreactor environment (both macro-and micro-environment)
8- Implications of bioreactor design on downstream products separation
9- Capital and operating costs of the bioreactor
10- Potential for bioreactor scale-up

15. BIOREACTOR MAIN DESIGNS
1- Stirred tank
2- air lift reactors
3- bubble column
4- packed bed reactors
5- trickle bed reactors
6- fluidized bed reactor

16. STIRRED TANK REACTOR
mixing method: mechanical agitation
high input required
baffles are constructed within the
built-in.
applications include production of
antibiotics and free/immobilized
enzymes
draw back is that high shear forces
may break the cells

17. AIR LIFT REACTORS
Mixing method: airlift
Central draft tube
Up-flowing stream and down flowing
stream
Homogenization of all components
present
Applications include bacterial, animal,
plant, fungi and yeast cells.

18. BUBBLE COLUMN REACTOR
Mixing method: gas sparging
Simple design
Good heat and mass transfer rates
Low energy input
Gas-liquid mass transfer
coefficients depend largely on
bubble diameter and gas hold-up

19. PACKED BED REACTOR
Column with attached biofilm
Biocatalysts
Pump is required to make fluid
move through the packed bed
Applications include waste
water treatment

20. FLUIDIZED BED REACTOR
When the packed beds are
operated in up-flow mode, the bed
expands at high liquid flow rates
due to upward motion of the
particles.
Energy is required
Waste water treatment

21. TRICKLE BED REACTORS
Liquid is sprayed onto the top of the
packing and trickles down through the
bed in small rivulets.
In the process, the gaseous pollutants
on the surface of the carriers is
adsorbed and immediately biologically
mineralized (degraded) by the
microorganisms.

23. - A bioreactor is a device or vessels which are designed to obtain an effective
environment for conversion of one material into some product by appropriate
biochemical reactions
- conversion is carried out by …… enzymes, microorganisms, cells of animals
and plants, or sub cellular structures such as chloroplasts and
mitochondria.
- Plants can be used as cheap chemical factories that require only water,
minerals, sun light and carbon dioxide to produce thousands of chemical
molecules with different structures.

24. Design gene for
high level
expression
Plant
transformation
Regeneration of
Cell
Selection of
transgenic
Growth of plants
in field
Harvesting of
plant materials
Purification of
product
Biosafety &
Functionality test

25. Where they are
produced?
Shown are various intracellular
organelles or extracellular spaces
(ES) that can be used to store the
recombinant proteins expressed
in a plant bioreactor.
G - Golgi;
PSV - Protein
storage
vacuole;
OB - Oil body;
C -
Chloroplast;
ES -
Extracellular

26. TYPES OF PLANT REACTORS
- seed-based plant bioreactors
- plant suspension cultures
- hairy root system bioreactor
- chloroplast bioreactor

27. SEED-BASED PLANT BIOREACTORS
An example is the successful expression of the human lysosomal enzyme alpha-
l-iduronidase in arabidopsis thaliana seeds.
The advantage of these systems is that, proteins do not degrade at ambient
temperature and are stable for long term storage.

28. Plant Suspension
Cultures
express recombinant proteins, secondary metabolites and
antibodies transported to subcellular organelles.
for example, is the expression of 80-kda human
lysosomal protein
Hairy Root System Bioreactor
it offers extreme biosynthetic stability and is suitable for
making biopharmaceuticals as for example scopolamine
in hyoscyamus muticus l. hairy root culture.

29. CHLOROPLAST BIOREACTOR
- insulin, interferon and other biopharmaceutical proteins can
be made using chloroplast bioreactor.
- foreign genes are inserted into nuclear chromosomes and
with peptides target expressed proteins into chloroplast.
- an example is the high yield in the expression of human
serum albumin protein in chloroplast.

31. PLANTS GENETICALLY ENGINEERED TO MAKE PRODUCTS THAT
ARE NOT OF PLANT ORIGIN
PRODUCTS:
vaccines antigens
therapeutics products
nutritional components
industrial products
bio plastics

32. Vaccine antigens:
antigens like insulin, rotavirus enterotoxin, anthrax lethal factor, hiv
antigen, foot and mouth disease virus antigen, heat stable toxin have been
produced in plants.
Therapeutic products:
the first successful production of a functional antibody, namely a mouse
immunoglobulin iggi in plants, was reported in 1989.
In 1992, C.J. Amtzen and co-workers expressed hepatitis B surface antigen
in tobacco to produce immunologically active ingredients via genetic
engineering of plants

33. Nutritional components:
β-carotene (naqvi et al.,
2009),
lycopene (fraser et al.,
2002),
flavonoid (butelli et al.,
2008),
nutraceuticals (kang et al.,
2009),
fatty acid (hoffmann et al.,
2008),
vitamins (nunes et

34. Biodegradable plastics:
Polyhydroxyalkanoates: biodegradable polymers which occur
naturally in plants.
Plant was engineered to produce PHAs or PHBs in the various
plant cell compartments.
When PHB expression targeted to cytoplasm, accumulation level
was low.
Expression was increased by targeting plastids, (40% of dry
weight was obtained).

35. • Industrial products:
Most expensive Drug – Hgc
hST (Human somatotropin)
rHLF (Recombinant human lactoferrin)
Synthetic fiber: Produced from Potato and
tobacco.

36.  Low cost source.
 Simple & Cost effective.
 Plant pathogens do not infect humans or animals.
 Produce large biomass.
 Easy storage for long time.
 Plant proteins have different sugar residues from human
or animal proteins.

37. Problems need to be
addressed
- Storage issues related to transgenic fruits or leaves.
- Most inserted genes are expressed at very low level in plants.
- Enhancing the stability of products obtained.
- Standardization of dosage in case of edible vaccine.
- Examining issues related to commercialization.
- Issues relating to the ethical, social, biosafety and environmental
impact.
- Some plants produce allergenic compounds.

38. Photobioreactors

39. - micro-algae are source of unique metabolites that can
be used to produce novel high-added value bioactive
compounds with industrial potential in medical
technologies or as food, feed or cosmetic ingredients or
as potential source of biofuels.
- among these substances play important role
polyunsaturated fatty acids (pufas). production of novel
pufas by micro algae is highly challenging as current
production processes from fish oil threatens natural
marine organism’s populations.

40. algae can accumulate large amounts of polysaccharides,
lipids and proteins with potential as nutrients/energy or
biofuel source

41. Photobioreactors - experimental
Algae does not need expensive reactors to be
cultivated

42. Photobioreactors – plastic bags

43. Photobioreactors - experimental

44. PHOTOBIOREACTORS
Advantages
- controlled, optimized conditions
- contamination can be minimized
- high rates of production
Disadvantages
- expensive

45. Flat panel photobioreactors

47. Tubular photobioreactors

49. Open ponds

50. Czech“ way
Open inclined Thin Layer Flat Plate
Photobioreactor with high rates of
algae suspension on the top of the
plates.
Harvest concentration 10-30 g/l.