The document discusses bioreactors and various aspects related to their use and design. It describes bioreactors as vessels that provide a controlled environment for optimal growth and product formation of cell cultures. Various types of bioreactors are discussed based on factors like oxygen need, mode of use, operation, and type of microbial growth. Key components like vessels, agitation, aeration, and their functions are summarized. Applications in secondary metabolite production and downstream processing are also mentioned.

1. BIOREACTOR
GLAXY EZEKEL.V
ROLL NO: 2
M.PHIL BOTANY
ST.TERESAS COLLEGE. ERNAKULAM

2. • Bioreactor: scaling up and cost reduction
• Production of useful compounds via
biotransformation
• Secondary metabolite production
• Suspension cultures
• Immobilization
• Examples of chemicals being produced for use in
pharmacy, medicine and industry
• Management and marketing

3. BIOREACTOR
SIDE VIEW FRONT VIEW

4. • It is a cylindrical vessel
• It is a vessel in which chemical
process which involves
organisms or biochemically
active substances derived
from such 9rganisms are
carried out.
• under sterile condition &
control of environmental
conditions
• This process can either be
aerobic or anaerobic.
• The bioreactors are commonly
cylindrical, ranging in size
from litres to cubic metres,
• are often made of stainless
steel.

5. AIM OF BIORERACTOR
• To provide a controlled environment to achieve
optimal growth and/or product formation in the
particular cell system employed.
• Bioreactors are used in the biotechnological
production of substances such as pharmaceuticals,
antibodies, or vaccines, or for the bioconversion of
organic waste.

6. • The function of the bioreactor is to provide a suitable
environment in which an organism can efficiently
produce a target product
• the target product might be
Cell biomass
Metabolite
Transformed Product
• The performance of any bioreactor depends on the key
factors such as:
Agitation rate
Oxygen transfer
Temperature
Foam production
pH

7. BIOREACTOR SHOULD HAVE FOLLOWING
QUALITIES
• The vessel – capable of being operated aseptically
for a number of days.
• Adequate aeration and agitation
• Power consumption - should be as low as possible.
• Temperature control and pH should be provided.
• Sampling facilities should be provided.
• Evaporation losses from fermenter should not be
excessive.

8. • Minimal use of labor in operation, harvesting,
cleaning and maintenance.
• Should have internal smooth surfaces .
• Containment involves prevention of escape of
viable cells from a fermenter or downstream
equipment.
• Aseptic operation involves protection against
contamination.

9. PARTS OF A BIOREACTOR

10. BIOREACTOR

12. VESSEL
• Glass vessel with a round or flat bottom and a top
flanged carrying plate.
• The large glass containers originally used were
borosilicate battery jars.
• They are sterilized by autoclaving.
• Glass is useful because it gives smooth surfaces, is
nontoxic, corrosion proof and it is easy to examine
the interior of the vessel
• More expensive.
• Pilot-scale and industrial scale – stainless steel

13. AERATION AND AGITATION
• Aeration provide microorganisms in submerged
culture with sufficient oxygen for metabolic
requirements.
• Agitation is the mixing or uniform suspension of
microbial cells in homogeneous nutrient medium.
• Structural components involved in aeration and
agitation are…
1. Agitator (impeller)
2. Baffles
3. Aeration system (sparger)

14. AGITATOR
• Also known as impeller
 bulk fluid and gas-phase mixing
air dispersion
oxygen transfer
 heat transfer
 suspension of solid particles
maintaining uniform environment throughout
vessel contents.

15. BAFFLES
• Baffles incorporated into agitated vessels of all sizes
to prevent vortex and to improve aeration
efficiency.
• Metal strips roughly one-tenth of vessel diameter
and attached radially to the wall.
• Usually, four baffles are used, but larger bioreactor
may have 6 or 8 baffles.
• Extra cooling coils may be attached to baffles to
improve cooling.

16. CLASSIFICATION OF BIOREACTOR
Based on the need
of oxygen
Based on the mode
of use
Based on the mode
of operations
Based on the mode
of growing
microbes
Based on the mode
of type of
bioreactors used in
the industries

17. BASED ON THE NEED OF OXYGEN
•Need adequate
mixing and aerationAEROBIC
•No need of sparging
and agitationANAEROBIC

18. BASED ON THE MODE OF USE
SINGLE USE
• Also known as disposable
bioreactor
• For small scale purposes
• Low contamination
• Low cost
• Labour charge will be less
MULTIPLE USE
• For the industrial purposes
• Used by the large
investment companies for
large production
• High cost
• Labours are required for
cleaning processes
• The contaminations will be
high

19. BASED ON THE MODE OF OPERATION
BATCH
FED-BATCH
CONTINOUS

20. Based on the mode of growing microbes
suspended
• Petridish is the simplest
immobilized bioreactor
• the large scale immobilized
bioreactors are used for
commecial manufacturingof
metabolites.
• Moving bed, fibrous bed,
packed bed, membrane
immobilized

21. BASED ON THE TYPES OF BIOREACTORS
USED IN INDUSTRIES
• Stirred tank bioreactor
• Continuous stirred tank bioreactor
• Bubble column bioreactor
• Air lift bioreactor
• Tower bioreactor
• Jet-loop bioreactor
• Fixed bed bioreactor
• Fluidised bed bioreactor
• Propeller bioreactor
• Flocullated bioreactor
• Gaseous phase bioreactor
• photobioreactor

22. Stirred tank bioreactor

23. ADVANTAGES
• Continous operation
• Good temperature control
• Simplicity of construction
• Low operating cost
• Easy to clean
DISADVANTAGES
• The need for shaft seals and
bearings
• Size limitation by motor
size, shaft length and weight

24. CONTINUOUS STIRRED TANK BIOREACTOR

25. Advantages
• Continuous operation
• Good temperature control
• Easily adapts to two phase
runs
• Good control
• Simplicity of construction
• Low operating (labor) cost
• Easy to clean
Disadvantages
• The need for shaft seals and
bearings.
• Size limitation by motor
size, shaft length and
weight.

26. BUBBLE COLUMN BIOREACTOR

27. ADVANTAGES
• They have excellent heat
and mass transfer
charectristics
• Little maintenance
• Low operating cost
• Lack of moving parts and
compactness
DISADVANTAGES
• The durabilitry of the
catalyst or other packing
material is high
• Oriejnted only on one
phase, i.e either liquid or
gas

28. AIR LIFT BIOREACTOR

29. ADVANTAGES
• Simple design
• Easier steriliusation
• Low energy requirement vs
stirred tank
• Greater heat removal vs
stirred tank
• Very low cost
DISADVANTAGES
• Greater air throughput and
higher pressures needed
• Inefficient break the foam
when foaming occurs
• No bubbles breaker

30. TOWER BIOREACTOR

31. • Non agitated type of bioreactor
• Used in both continuous and batch process
• It has long tube with closed ends
• The presence of baffles increase the turbulent
and residual time of waves inside the
bioreactor and also helps in aeration
• This type of reactor is used in the production
of beer, citric acid, bakers yeast

32. FLUIDISED BED BIOREACTOR

33. ADVANTAGES
• Uniform particle mixing
• Uniform temperature
gradients
• Ability to operate reactor in
continous state
DISADVANTAGES
• Increased reactor vessel size
• Pumping requirements and
pressure drop
• Particle entrainment
• Lack of current
understanding
• Erosion of internal
components
• Pressure loss scenarios

34. Propeller bioreactor
• Propeller is present
• Loop circulation is
promoted by a
propeller which acts as
a pump to force fluid
either up or down
through a central draft
tube

35. FLOCULLATED BIOREACTOR
• They are egg shaped
• The cells are trapped in
the reactor due to an
induced or natural
flocculation
• in flocculation, cells tends
to group together
causing them to came out
of solution and to sink
towards the base of the
reactor
• They are mainly used in
the anaerobic waste
water treatment

36. GASEOUS PHASE BIOREACTOR
• This type of bioreactor is equipped with filters on which
the culture is supported and with a shower nozzle for
spraying on the medium.
• Seed cultures are inoculated on the filters and the
medium is supplied to the culture by spraying from a
shower nozzle.
• The drained medium is collected on the bottom of the
bioreactor. This type of bioreactor is excellent for plant
cell, tissue, and organ cultures because there is no
mechanical agitation (e.g., driven impeller, aerator)
and, therefore, the growth rate and the secondary
metabolite production are enhanced.

38. PHOTOBIOREACTOR

39. ADVANTAGES
• Higher productivity
• Large surface to volume
ratio
• Better control of gas
transfer
• Reduction in evaporation of
growth medium
• More uniform temperature
DISADVANTAGES
• Capital cost is very high
• The productivity and
production cost in some
enclosed photobioreactor
systems are not much
better than those
acheivable in open pond
cultures
• The technical difficulty in
sterilizing

40. PACKED BED BIOREACTOR

41. ADVANTAGES
• Higher conversion per unit
mass of catalyst than other
catalytic reactor
• Low operating cost
• Continous operation
• No moving parts to wear out
• Catalyst stays in the reactor
• Reaction mixture/catalyst
seperation is easy
• Effective at high temperatures
and pressures
DISADVANTAGES
• Undesired heat gradients
• Poor temperature control
• Difficult to clean
• Difficult to replace catalyst
• Undesirable side reactions

42. ROTARY DRUM REACTOR.

43. ADVANTAGES
• In this, the drum is filled
with 40% of its volume and
rotated by means of rollers.
It is particularly suitable for
the cultivation of the plant
cell cultures.
• High oxygen transfer.
• Good mixing Facilitated
better growth and impart
less hydrodynamic stress
DISADVANTAGES
• Difficult to scale up.

44. OPERATIONAL STAGES IN A BIO-PROCESS

45. • Products need to be produced at large volume
• This requires scaling up
• We do scaling up studies to ensure that the
fermentation process is technically and
economically viable to be produced in the end
at a large scale.
• Preliminary work is carried out at the level of
petridishes

46. SCALING UP

47. • After laboratory scale fermenters have been
used to work out the optimum growth
conditions, the process is next scaled upto a
pilot plant fermenter.
• The pilot plant fermenter is used to trial the
industrial process and production
• It is also using to confirm that the growth
conditions that were optimal in the laboratory
scale fermenter are the same when the process
is scaled up.
• Also ensuring that the products are not at all
harmful to the workers and those who
eventually buy it.

48. • After upstream processing step, the resulting feed
is transferred to one or more Bioreaction stages.
• The Biochemical reactors or bioreactors form the
base of the Bioreaction step.
• This step is mainly consists of three operations
namely,
i. production of biomass
ii. metabolize biosynthesis
iii. biotransformation.
• Finally, the material produced in the bioreactor
must be further processed in the downstream
section to convert it into more useful form.

49. • The downstream process is mainly consists of
physical separation operations which includes,
solid liquid separation, adsorption, liquid-liquid
extraction, distillation, drying etc.
• The raw material can be of biological or non-
biological origin.
• It is first converted to more suitable form for
processing.
• This is done in upstream processing step which
involves chemical hydrolysis, preparation of liquid
medium, separation of particulate, air purification
and many other preparatory operations

50. DOWNSTREAM PROCESSING
• Separation of particles
• Disintegration of cells
• Extraction
• Concentration
• Purification
• Drying

51. BIOTRANSFORMATION
• It is the chemical modification (or modifications)
made by an organism on a chemical compound.
• If this modification ends in mineral compounds like
CO2, NH4
+, or H2O, the biotransformation is called
mineralisation.
• Biotransformation means chemical alteration of
chemicals such as nutrients, amino acids, toxins,
and drugs in the body.
• It is also needed to render non-polar compounds
polar so that they are not reabsorbed in renal
tubules and are excreted.

52. DRUG METABOLISM
• The metabolism of a drug or toxin in a body is an
example of a biotransformation.
• The body typically deals with a foreign compound by
making it more water-soluble, to increase the rate of its
excretion through the urine.
• There are many different processes that can occur; the
pathways of drug metabolism can be divided into:
1. phase І
2. phase II
• Drugs can undergo one of four potential
biotransformations: Active Drug to Inactive Metabolite,
Active Drug to Active Metabolite, Inactive Drug to
Active Metabolite, Active Drug to Toxic Metabolite
(biotoxification).

53. PHASE І REACTION
• Includes oxidative, reductive, and hydrolytic
reactions.
• In these types of reactions, a polar group is either
introduced or unmasked, so the drug molecule
becomes more water-soluble and can be excreted.
• Reactions are non-synthetic in nature and in
general produce a more water-soluble and less
active metabolites.
• The majority of metabolites are generated by a
common hydroxylating enzyme system known as
Cytochrome P450.

54. PHASE II REACTION
• These reactions involve covalent attachment of
small hydrophilic endogenous molecule such as
glucuronic acid, sulfate, or glycine to form water-
soluble compounds, that are more hydrophilic.
• This is also known as a conjugation reaction.
• The final compounds have a larger molecular
weight.

55. SECONDARY METABOLITE PRODUCTION
• Plant secondary metabolism produces products that
aid in the growth and development of plants but
are not required for the plant to survive
• A common role of secondary metabolites in plants
is defence mechanism
• It is believed that production of secondary
metabolites is linked to the induction of
morphological differentiation-

56. WHY SECONDARY METABOLITES
• Chemically warfare to protect plants from the
attacks by predators, pathogens or competitors
• Attract pollinators are seed dispersal agent
• Important for abiotic stress
• Medicine
• Industrial additives

57. Seven production types of secondry
metabolite production in bioreactors
 shikonin production
 Berberine production
 rosmarinic acid production
 Indole alkaloid production
 Organ culture
 Hairy root culture
 commercialization

58. SUSPENSION CULTURES
• Suspension culture is a type of culture in
which single cells or small aggregates of cells
multiply while suspended in agitated liquid
medium.
• It is also referred to as cell culture or cell
suspension culture.

59. PRINCIPLE
• Callus proliferates as an unorganised mass of cells.
• So it is very difficult to follow many cellular events
during its growth and developmental phases.
• To overcome such limitations of callus culture, the
cultivation of free cells as well as small cell
aggregates in a chemically defined liquid medium as
a suspension was initiated to study the
morphological and biochemical changes during
their growth and developmental phases.

60. • To achieve an ideal cell suspension, most
commonly a friable callus is transferred to agitated
liquid medium where it breaks up and readily
disperses.
• After eliminating the large callus pieces, only
single cells and small cell aggregates are again
transferred to fresh medium and after two or
three weeks a suspension of actively growing cells
is produced.

61. • This suspension can then be propagated by regular sub-
culture of an aliquot to fresh medium.
• Ideally suspension culture should consist of only single cells
which are physiologically and biochemically uniform.
• Although this ideal culture has yet to be achieved, but it can
be achieved if it is possible to synchronize the process of cell
division, enlargement and differentiation within the cell
population.
• The culture of single cells and cell aggregates in moving liquid
medium can be handled as the culture of microbes.
• The suspension culture eliminates many of the disadvantages
ascribed to the callus culture on agar medium. Movement of
the cells in relation to nutrient medium facilitates gaseous
exchange, removes any polarity of the cells due to gravity and
eliminates the nutrient gradients within the medium and at
the surface of the cells.

63. PROTOCOL
• Take 150/250 ml conical flask containing autoclaved
40/60 ml liquid medium (Fig 4.1).
• 2. Transfer 3-4 pieces of pre-established callus tissue
(approx. wt. 1 gm. each) from the culture tube using
the spoon headed spatula to conical flasks.
• 3. Flame the neck of conical flask, close the mouth of
the flask with a piece of alluminium foil or a cotton
plug. Cover the closure with a piece of brown paper.
• 4. Place the flasks within the clamps of a rotary shaker
moving at the 80-120 rpm (revolution per minute)
• 5. After 7 days, pour the contents of each flask through
the sterilized sieve pore diameter -60µ- 100µ and
collect the filtrate in a big sterilized container. The
filtrate contains only free cells and cell aggregates.

64. • 6. Allow the filtrate to settle for 10-15 min. or centrifuge the
filtrate at 500 to 1,000 rpm and finally pour off the supernatant.
• 7. Re-suspend the residue cells in a requisite volume of fresh liquid
medium and dispense the cell suspension equally in several ster-
ilized flasks (150/250 ml). Place the flasks on shaker and allow the
free cells and cell aggregates to grow.
• 8. At the next subculture, repeat the previous steps but take only
one-fifth of the residual cells as the inoculum and dispense equally
in flasks and again place them on shaker.
• 9. After 3-4 subcultures, transfer 10 ml of cell suspension from
each flask into new flask containing 30 ml fresh liquid medium.
• 10. To prepare a growth curve of cells in suspension, transfer a
definite number of cells measured accurately by a
haemocytometer to a definite volume of liquid medium and
incubates on shaker. Pipette out very little aliquot of cell
suspension at short intervals of time (1 or 2 days interval) and
count the cell number. Plot the cell count data of a passage on a
graph paper and the curve will indicate the growth pattern of
suspension culture.

65. USE OF CELL SUSPENSION CULTURE
• Understanding of biosynthetic pathway
• Mutant selection
• Secondary metabolite production
• In plant propagation

66. IMMOBILIZATION
• Enzyme immobilization may be defined as
a process of confining the enzyme
molecules to a solid support over which a
substrate is passed and converted to
products.

67. TECHNIQUE OF ENZYME IMMOBILIZATION:-
1.Carrier binding
· Physical adsorption
· Covalent bonding
· Ionic bonding
2. Cross linking
3. Entrapment
· Occlusion within a cross linked gel
· Microencapsulation

68. High Stability.
Reusable.
Products are enzyme free.
Controls of enzyme function is easy.
Suitable for industrial & medical use.
Minimize effluent disposal problems.
ADVANTAGES OPF IMMOBILISED
ENZYMES

69. Possible loss of biological activity of an enzyme
during immobilization or while it is in use
Immobilization is expensive technique requires
sophisticated equipment
DISADVANTAGES

70. Immobilized glucosidase & glucose isomerase
are used in production of fructose from
starch.
Immobilized L-Aminoacylase which resolves a
mixture of D- and L- amino acids.
Immobilization of Microbial cells.

71. APPLICATIONS
• These devices are being developed for use in tissue
engineering
• The bioreactor is modular in nature and carry out all
the processes of fermentation in a single contained
environment.
• Bioreactor plays a core role in bioprocess.
• Stirred tank bioreactors are commonly used in
fermentation

72. • Due to simple technology and higher yield solid
state bioreactors are widely used in industries.
• Ethanol fermentation is done by saccharomyces
cerevisiae in bioreactor.
• Organic acids e.g. acetic acid and butyric acid are
formed in bioreactor by the Eubacterium
limosum.
• Thienamycine an antibiotic also produced in
bioreactor.
• Glucomylase is produced by Auerobasidium
pullulans

73. • The use of immobilized enzymes, antibodies
or reactive proteins was recently considered
as a promising approach for detoxification
purposes.