Bioreactors for Solid-State

Cultivation

David Mitchell
Department of Biochemistry and Molecular Biology
Federal University of Paraná
Curitiba, Brazil

davidmitchell@ufpr.br

1
The basic features of the system – aerobic SSC
Aerobic SSC involves the growth of microorganisms
(usually filamentous fungi, but not always) on a bed of
particles of a moist solid substrate, with the minimum of
free water in the spaces between the particles, with these
spaces being filled a continuous gas phase
microbial biomass
concentrated at the
moist solid particle surface
particles
Bioreactor a continuous inter-
containing
nutrients particle gas phase
(e.g. grain,
meal, flour) liquid water within the
particles, almost none
between particles

2
Topics
The importance of transport phenomena
Bioreactor types
• Tray bioreactors
• Packed-bed bioreactors
• Rotating drums and horizontal stirred-drum bioreactors
• Forcefully aerated agitated bioreactors
Continuous SSC bioreactors

3
 The importance of
transport phenomena in
solid-state cultivation
bioreactors

4
 Growth of a microorganism in a bioreactor depends on
the local conditions

bed of • growth will depend on the local

particles concentrations of the nutrients
and void (sources of C, N, etc)
spaces • the carbon source is typically
present within the substrate in
Air the form of non-diffusible
polymers, needing to be
liberated by hydrolytic
pores enzymes
Particle
(mm) • for aerobic processes, growth will
depend on the local concentration
of O2
• the local pH , T and water activity
mycelium will affect growth

5
The local conditions experienced by the microorganism depend
on the balance between microbial activities and a complex
network of “microscale” and “macroscale” transport phenomena

Liquid film at the MICROSCALE MACROSCALE

Wall
particle surface Stagnant gas
translocation

Surroundings
Particle
interior uptake
agitation
uptake
diffusion diffusion
oxygen convection
uptake
evaporation
starch
water
rxn release of vapor
glucose metabolic
conduction
gluco- heat conduction conduction
amylase diffusion

entry of air flow of cooling

at the inlet water 6
 These transport phenomena determine our ability to
control the local environment

macroscale microscale

microorganism

mass transfer
within the
solid particle
heat and mass transfer
across the bed of particles
and exchange between the
air and solids phases

the growth kinetics of the

microorganism depend on the
operating variables conditions in its local environment:
e.g. air flow rate and temperature [nutrients], pH, T, aw & [O2]...
agitation regime 7
 Our ability to control conditions within the bed is limited

macroscale microscale
If we want to control the temperature
and moisture content of the bed, we
microorganism
are limited to manipulating operating
variables outside the bed
mass transfer
The combination of metabolic heat
within the
production by the microorganism and
solid particle
heat and mass transfer
transport phenomena affect the
across the bed of particles
efficiency with which the operating
and exchange between the
variables outside the bed affect
air and solids phases
conditions within the bed
the growth kinetics of the
microorganism depend on the
operating variables conditions in its local environment:
e.g. air flow rate and temperature [nutrients], pH, T, aw & [O2]...
agitation regime 8
 The design and operating conditions that we can manipulate are limited

for optimum growth The operational and design variables that can
the microbe needs be manipulated in an attempt to maintain the
optimum values of conditions at their optimal values are:
[nutrients], pH, T,
aw & [O2]... agitation system or not, additions such as
if so, frequency, water, nutrient
intensity and duration solutions, dilute
...but the microbe is of agitation events acid or base
• consuming nutrients
• altering the local pH bioreactor
• releasing metabolic heat geometry
• consuming oxygen
and probably evaporation cooling
is occurring. jacket/plates or not
if so, water flow rate
Therefore the local and temperature,
conditions of the microbe number and
will tend to deviate from aeration system or not spacing of plates
the optimum conditions. temperature, humidity
and flow rate of air 9
 Our ability to control conditions within individual substrate
particles is severely limited

Between
macroscale
(i) the operating variables that we
microscale
manipulate outside the bed and
(ii) the inside of the particle, microorganism

...we have transport phenomena mass transfer

• in the air phase across the bed within the
• from the air phase to the particle solid particle
heat and mass transfer
• within the particle
across the bed of particles
and exchange between the
We have very little ability
air andtosolids
controlphases
the
O2 and nutrient concentrations in the
local environment of the microorganism the growth kinetics of the
microorganism depend on the
conditions in its local environment:
[nutrients], pH, T, aw & [O2]...
10
The growth conditions that we have most control over in an
SSC bioreactor are the temperature and moisture content
• Our difficulties in controlling these growth conditions increase as scale
increases (in other words, the transport phenomena become ever
more important with increase in scale)
• as a result, temporal and spatial profiles are almost unavoidable in
large-scale SSC bioreactors – for example, for temperature

wall wall
given position in the

temperature at a
temperature at a

given time
bioreactor

Toptimum Toptimum

time position
The objective of a bioreactor is to minimize spatial and temporal variations,
to keep the conditions as close as possible to the optimum for growth and
product formation
11
In other words...
• for as much of the bed and for as much of the time as possible
– the temperature should be maintained as near as possible to the
optimum temperature for growth of the microorganism
– the water activity of the substrate should be maintained as near
as possible to the optimum water activity for growth of the
microorganism
growth rate

growth rate

temperature water activity

12
Due to the production of metabolic heat, the control of
the bed temperature is a key issue in bioreactor
design, especially for large scale bioreactors

Before we start talking about the types of bioreactors

let us consider the types of heat removal mechanisms
• conduction
• convection
• evaporation

13
 Conduction
Transfer of heat due to transfer of vibrations of
molecules – with no associated mass flow
It is a relatively slow method of heat removal

Particles and void spaces

within a substrate bed

At the molecular level, for both the air phase and

the solids phase (but with different efficiencies)

transmission of
energy through
molecular
vibrations
higher temperature lower temperature
direction of conduction
14
Convection

Transfer of heat due to a moving phase that transports

the thermal energy away

Heat transfer from the solids

to the air because Tsolids > Tair

The arriving The air that

air is cooler leaves is hotter

a particular
region in a bed

15
Evaporation

Transfer of heat from the solids due to evaporation of

liquid water in the solids, with the vapor being swept away
a particular Evaporation occurs because
region in a bed awsolids > aw(air)

The arriving The air that

air is less leaves is more
humid humid

• Evaporation decreases the temperature of the solids phase

• Evaporation does not increase the temperature of the gas phase
(the energy removed from the solids is transformed into the latent
heat of vaporization of the vapor)
16
 What types of
bioreactors are used in
solid-state cultivation?

17
 The scenario

• You are studying an SSC process that is performing

well at small scale
• You are interested in establishing a large-scale process
• what kind of bioreactor will you use at large scale?
• to what degree will you be able to control the temperature
and moisture content of the bed?

I will focus on
• bioreactors for processes involving aerobic growth
• situations in which it is not interesting for the temperature to
rise during the process (i.e. I will not talk about bioreactors for
composting)
18
What types of bioreactors are used in SSC?

Based on our discussion of transport phenomena

and our limitations in controlling bed conditions, the
main functions of an aerobic SSC bioreactor are to
• bring O2 to the particle surface
• allow control of the temperature of the bed
• allow control of the water activity of the bed

Although particular SSC bioreactors may have

specific features that might make them seem to be
unique, it is useful to classify them into four groups
based on mixing and aeration strategies...

19
Mixing →
No mixing Continuous mixing
 Aeration
(or very infrequent) (or frequent intermittent mixing)

No
I III
forced
aeration
(passes
around
the bed) Rotating drum Stirred drum
Tray chamber
II IV

Forced
aeration
(air
forced
through
the bed)

Gas-solid
Packed bed Stirred bed
fluidized bed 20
Mixing →
No mixing Continuous mixing
 Aeration
(or very infrequent) (or frequent intermittent mixing)

No
I III
forced There are not necessarily sharp
aeration distinctions between the different
(passes classifications
around
the bed) Often bioreactorsRotating
Tray chamber
fall between
drum two Stirred drum
groups...
II IV
What is the dividing line between
Forced infrequent and frequent mixing?
aeration
Bioreactors might be forcefully aerated,
(air
the air distribution may not be uniform
forced
through
the bed)

Gas-solid
Packed bed Stirred bed
fluidized bed 21
 One possible guide to bioreactor selection
Does the process involve batches of only a few kilograms?
no yes
Use other bioreactor types Can use a tray bioreactor

Does the organism tolerate frequent or continuous mixing?

not very well yes
Mixing should be minimized. Use Consider agitated types
static or infrequently-mixed beds
Does the organism
Will the water activity of the substrate grow quickly?
decrease significantly? no yes
Will the bed pull away from the walls?
Rotating or Forced
no to both yes to one or both
stirred aeration
Completely static Infrequent drums might will be
packed bed mixing be sufficient necessary
22
 Tray bioreactors
Bioreactors without agitation
(or with very infrequent agitation)
and without forced aeration

23
 Tray-type bioreactor systems are typically used to
produce fungal spores for use as biopesticides
– the substrate bed needs to be maintained static in
order not to damage reproductive hyphae
For example

Naeimi et al. (2020) Agronomy https://doi.org/10.3390/agronomy10091258 24

 A flask is like a tray!

Solid state cultivation of Trichoderma harzianum on broom sorghum grains

Naeimi et al. (2020) Agronomy https://doi.org/10.3390/agronomy10091258 25

A tray bioreactor is not simply “a tray”

• It consists of many trays in a chamber with control

of the temperature and humidity of the atmosphere
chamber = bioreactor

individual tray

headspace

air of controlled
air
temperature
out
and humidity

26
The “tray” itself can have different forms
An individual tray
Air circulated around the tray
a thin layer of the sides and
substrate, either bottoms can
completely static be perforated
or agitated by to aid in gas
hand one or two exchange
times per day

Microporous plastic bags Erlenmeyer

have also been used flasks represent
this type of
system

27
The chamber may be a whole room or a smaller incubator

A whole room

https://www.takabio.com/en/la-fermentation-solide-
et-la-fermentation-liquide/
28
An incubator

https://www.aquaculturealliance.org/ https://link.springer.com/article/
advocate/solid-state-fermentation- 10.1007/s00253-018-9157-4
novel-process-improving-nutritional-
value-plant-feedstuffs/

29
The scale of production and degree of sophistication of
the production process can vary greatly

30
31
32
A plastic bag system

http://www.agr.gc.ca/eng/science-and-innovation/technology-
transfer-and-licensing/licensing-opportunities/agriculture-and-agri-
food-canada-biopesticide-phoma-macrostoma/?id=1407525855663 33
 In this case, a large room in which the trays have lids

http://www.visualphotos.com/image/ http://www.sciencephoto.com/image/
1x6057538/solid_state_fermentation 211803/530wm/G2520124-
_of_gm_microbes_india Microbe_fermentation_unit-SPL.jpg

34
At large scale, you may need efficient systems for
handling the trays

Automated systems for

tray washing and filling

https://www.feedstrategy.com/animal
-feed-manufacturing/the-quest-for-
improved-fiber-utilisation/

35
 Tray bioreactors can have a high level of sophistication
“Platotex” (Institut de Chimie des Substances Naturelles)

Trays containing Trays containing

maize grains agar medium

Hydrazides from
Streptomyces sp.

In these cases, the

tray chamber is a
stainless-steel vessel

http://www.icsn.cnrs-gif.fr/spip.php?article927&lang=en 36
What questions arise if you want to establish a
commercial process based on the use of trays?
• How many trays will you need?
• How much space will be required?
• Is the production scale sufficiently large to warrant
automated tray-handling processes?
• How will the air in the room be conditioned? What
temperature and humidity are required?
• How to position the trays to ensure good air circulation
and no stagnant regions in the room?
• Will the air be forced to move past the tray surface with a
good velocity in order to improve heat removal?

37
More questions – with respect to design aspects of the
individual trays
• Will you use trays or plastic bags?
• Will the tray have a lid?
• Will the base and sides of the tray be perforated?
• What material to use for the trays?
• metal
• plastic
• wood/bamboo
• How thick will the bed in the tray be?

38
Some considerations about trays
• the bed is limited to a height of 2-10 cm (depending on the process)
• if the bed is too thick, the center will suffer from O2 depletion and
high temperatures
air circulated around the tray

heat removal by heat removal by

gas exchange
convection evaporation

heat transfer in the gas transfer in the (there may

bed essentially bed essentially be some
limited to conduction limited to diffusion natural
convection)

O2 CO2
air circulation improves heat transfer/evaporation/gas exchange between
the bed surface and the air, but does little to help transfer within the bed
39
 Heat and mass transfer within the tray can be improved by
placing the trays in a pressure vessel and cycling the pressure

https://link.springer.com/article/
10.1007/s11947-012-0956-9

repeated
cycling of
pressure
during the
process

http://dx.doi.org/10.1016/j.procbio.2014.11.007
40
 Packed-bed bioreactors
Bioreactors without agitation
(or with very infrequent agitation)
and with forced aeration

41
Packed-bed bioreactors can be divided into two main
types

“Traditional packed-bed bioreactors”

• There are no heat transfer plates within the bed

Zymotis-type packed-bed bioreactor

• The bed contains heat transfer plates

First I will talk about traditional packed-bed bioreactors

42
Basic design aspects of traditional packed-bed bioreactors

static substrate
bed (particles
and void spaces)

perforated
base plate

Forced aeration
• controlled T
• typically the air
is saturated 43
Axial temperature gradients are an intrinsic
feature of traditional packed-bed bioreactors

convective
cooling of the
bed leads to
In wide traditional packed- an axial
bed bioreactors, conduction temperature
in the horizontal direction gradient
makes a relatively small
contribution to heat removal

44
 Why does convective cooling unavoidably lead to
axial temperature gradients?

Cool Warm Hot

air air air

a region nearer a region further

to the air inlet from the air inlet

45
The steepness of the axial temperature gradients
depend on several factors

• The rate of growth of the organism, which affects heat

production rates
• The porosity of the bed – which affects the Watts of
metabolic heat per cubic meter
• The air flow rate – which affects the rate of convective
heat removal

• Temperature differences between the inlet and outlet

have been reported
• as being as low as 1 °C
• as being around 20 °C
in the absence of
other problems, this
is a sign that the air
velocity is too low!

46
 Axial temperature gradients mean that evaporation is
unavoidable, even if saturated air is supplied to the bed

...with the increase

in the temperature
even if the of the air, its vapor-
air enters air Tsolid air carrying capacity
saturated... Tair
increases, so it
heat and mass transfer to the air removes water from
the bed

• The bed can dry out to low water activities that restrict
growth
• Water replenishment is needed
• How to replenish water without agitating the bed?
47
There can be wall effects

This can lead to

non-uniform growth
lower porosity within the bed
further from
the wall

the presence
of the wall
causes a
higher porosity
near the wall

Caroline Lopes Perez (2017) Avaliação da viabilidade

técnica da ampliação de escala da produção de enzimas
celulolíticas e hemicelulolíticas por FES em biorreatores
de leito empacotado. Master’s Dissertation,
Universidade Estadual Paulista, Brasil
48
Fungal growth fills up the space between particles,
leading to an increased pressure drop across the bed

The fungal mycelium

grows into the spaces
between the particles,
increasing greatly the
resistance to air flow
http://www.tub-
Much higher pressures are collection.com/pic.html
needed to force the air
through the bed

This can
exacerbate
non-uniform air
flow and promote
channeling

49
Channeling can occur due to shrinking of the bed

The fungus consumes matter of the particle, which shrinks

The fungus binds the particles into an agglomerate, which shrinks as a
whole
The bed can then decrease in height and also pull away from the walls...
50
An example of the agglomeration of wheat bran during the cultivation
of Aspergillus niger

51
 Channeling = flow of air through the gaps, not through the
bed, reducing the supply of O2 and removal of heat

Particle
agglomeration decrease in
bed height
and shrinkage
gap
between
bed and
wall

Although bed shrinkage and channeling are well known qualitatively,

they are poorly characterized quantitatively.
We need to understand not only particle shrinkage, but also the
interparticle forces exerted by the intertwining hyphae
52
Immediately after After 24 h of
inoculation cultivation
53
The gap is
about 1 cm
wide

54
With flow around the bed rather than through it, the bed will overheat

55
It is also possible for the bed to crack in the middle

Again, air fill flow preferentially

through gaps and cracks, meaning
the bed is not properly aerated

56
Non-monotonic temperature profiles indicate that channeling
is occurring
If we have uniform air flow... ...and if we have thermocouples at
different heights, then we expect...

highest
temperature

mid
temperature

lowest
temperature

57
 These experimental results show that channeling is occurring

This is wrong! If the air is

flowing through the bed,

Pectinase activity at top of bed (U g-1)

then the air leaving the
bed must be hotter
18 cm bed height
than the bed!
Bed temperature (°C)


5 cm
bed 
height
air leaving Due to the gaps, the
the bed air is NOT flowing
through the bed, but
rather around it!
Time (h)

58
 These experimental results show that channeling is NOT occurring

This is correct! If the

air is flowing through

))
(Ugg-1-1
the bed, then the air

bed(U
leaving the bed must
Bed temperature (°C)

top of bed
air leaving be hotter than the
the bed bed!

attop
18 cm

activityat
5 cm

activity


Pectinase


Pectinase


In this case, there are

no gaps and the air is
Time (h)
flowing uniformly!
59
Intermittent mixing may be required to “reseat” the bed
or to replenish water to prevent the bed from drying out
• We are talking about a few mixing events (we will
deal with more frequent intermittent mixing later)
• It may be difficult to break up agglomerates!
• In fact, it may be the case to mix early in order to
prevent agglomerates from forming in the first place!
agglomeration
during static
operation agitation

hyphae crushed
onto the particle ripped
surface, forming a hyphae
moist biofilm 60
It may be possible to avoid the need for intermittent mixing
by adding a “bed porosity modifier”

With 100% wheat bran...

The fungus agglomerated

The bed shrank.
the substrate
Gaps appeared between the
bed and the wall

61
With 90% wheat bran +10% sugarcane bagasse...

The substrate retained The bed did not shrink.

its porous, loose Gaps did not appear between
structure throughout the bed and the wall

62
With 50% citrus pulp +50% sugarcane bagasse...

The bed remained loose and fluffy

Sugarcane bagasse

Citrus pulp The bed did not shrink.

(which tends to agglomerate Gaps did not appear between
by itself when wet!) the bed and the wall
63
Several alternative designs have been suggested
for packed-bed bioreactors...

64
An intermittently stirred drum with air being introduced
through a perforated base

Side view End view

air

Problems
• The bed depth is not constant across a cross section
• During static operation, this will lead to preferential flow where
the bed thickness is smaller
• Therefore, continuous agitation will be necessary
• This is not a good design for an intermittently mixed packed bed!
65
A perforated tube within the bed

However, the air flow through the perforated

tube is unlikely to be well distributed

How would it be done for wide packed beds?

What if you want to agitate the bed intermittently?

66
A “radial flow” bed
air out
top view

air in air in
Problems
• How to scale-up such a design?
• It cannot be made ever wider
• It will need to be made taller
67
Multi-layered beds

This is a potentially interesting design

Care must be taken to avoid channeling

Cooling plates can be inserted between

the layers

heat
exchanger
plate

cooling air cooling

water in water out
68
 The use of water jackets has been proposed

cooling Problems
water in • How to scale-up such a design?
• It cannot be made ever wider
• It will need to be made taller
cooling
water out

in a wide
packed bed, a
water jacket will
only cool a few
cm near the wall

69
Water jacketing can be useful at small scale...
“Raimbault columns” can be used for metabolic studies at small scale

The temperature can be well controlled in columns 2-4 cm in diameter

blower

cotton wool
filter

analyzer substrate bed

rotameters

filter
cotton wool
rubber stopper
sparger

water

secondary
Waterbath humidifier
70
 The “Zymotis” packed-bed bioreactor has heat transfer
plates inserted within the bed
internal heat
transfer plates
cooling
water

Heat transfer plates in the bed help to

minimize axial temperature gradients

Evaporation is therefore minimized Air

(if saturated air is used)

Conduction in the horizontal direction to the heat transfer

plates contributes significantly to heat removal...
...but is only effective over distances of 5 – 10 cm
71
A small pilot-scale Zymotis packed-bed bioreactor was built

Durand A (2003). Bioreactor designs for solid state fermentation.

Biochemical Engineering Journal 13, 113–125 72
However, there are few reports of the use of the Zymotis bioreactor

cooling
Possibly there are water
problems with channeling?

If gaps appear, it will be

difficult to reseat the bed!

Air

73
 Rotating drums and
horizontal stirred-drum
bioreactors
Bioreactors with frequent or
continuous agitation, with air being
blown through the headspace and
not forcefully through the bed
74
Basic features of horizontal stirred drums

Horizontal cylinder

paddles or scrapers or other

agitator designs, mounted on a
central axis
air inlet air
(controlled outlet
T, %RH
and flow rate)

75
Basic features of rotating drums
direction air outlet
of rotation

it is possible to use lifters

in order to improve the
agitation of the bed

air inlet
(controlled
T, %RH & substrate particles
flowrate) tumble down the
upper surface of
the bed

76
Some example of rotating drums and horizontal-stirred drums

An industrial-scale bioreactor with a total volume of 800 L

77
 A quite large rotating drum used for composting

http://xactsystemscomposting.com/tag/rotating-drum/ 78
 A laboratory-scale “roller bottle” system

https://link.springer.com/protocol/10.1007%2F978-1-62703-122-6_5 79
 A 35-L stirred drum for laboratory studies

Nagel (2002) PhD Thesis, University of Wageningen 80

 Some considerations about rotating drums
For drums without lifters, the flow regime of the solids
depends on the rotational speed
increasing rotational speed

slumping tumbling cataracting centrifuging

• the bed tends to • the bed is static

move as whole • operating costs
• it is not well one would need to operate in are high
mixed these regimes
81
It is better to install lifters in the drum... you can operate
at low speeds while still ensuring agitation of the bed

the curtain of falling

substrate particles has good
contact with the air flowing
through the headspace

82
 What are the heat and mass transfer processes in rotating
and stirred drums?

Convection to
surroundings
HEADSPACE

Models suggest that in Conduction Convection to

to wall headspace
large-scale bioreactors
bed-to-headspace heat
transfer and evaporation
Metabolic Evaporation to
will be the major route for
heat headspace
heat removal from the bed
Bulk flow
with
conduction
agitation BED
across wall

conduction within wall 83

 The air flow through the headspace will remove heat through
convection and evaporation – from the bed surface (and falling
curtain of substrate particles, if lifters are used) to the air flowing
through the drum

heat

heat
moist moist
air air
vapor
heat

water held
Metabolic within solids
heat
heat

84
However, there is no guarantee that there will be good contact between
the air flowing through the headspace and the bed
There may be dead spaces
Design of the air inlet and outlet are important
Residence-time distribution studies can be done

Dead region in headspace

air
air Plug flow region
inlet outlet

Dead region in the voids within the bed

85
Several variations of design and operation have been
proposed for rotating drums and horizontal stirred
drums...

86
Cooling the outside surfaces of rotating drums

A water jacket is unlikely to be

appropriate for a rotating drum
• Increased weight that needs
to be rotated
• The need to have a rotating
seal

It may be useful to spray cooling water

on the outside surface of the drum
However, this is probably only likely to
be appropriate at small scale

87
 We can use inclined drums and curved baffles to improve the mixing of
the bed itself
The drum axis would be inclined at an angle less than the dynamic
angle of response of the solids
The curved baffles would move the substrate to the higher end, and it
would tumble back down to the lower end

direction direction
of rotation of rotation

lifters
gravity dynamic
angle of
repose
inclination of drum
axis to the horizontal
Schutyser et al. (2002) https://doi.org/10.1002/bit.10277 88
Some authors have suggested using rotating drums
with intermittent agitation and long static periods

During static periods, even with air flow through the headspace,
the bed will overheat!

air

This will not be a useful strategy at large-scale

If you want to agitate infrequently, then use a packed-bed!

89
 Forcefully aerated
agitated bioreactors
Bioreactors with frequent or
continuous agitation, with air being
blown forcefully through the bed

90
Many different designs have been proposed

A 50-L cylindrical bioreactor with a planetary mixer

air outlet

air inlet (controlled T,

%RH & flowrate)
Chamielec et al. (1994) https://doi.org/10.1007/BF00155415
Bandelier et al. (1997) https://doi.org/10.1016/S0032-9592(96)00063-5 91
 A conical solids mixer aerated from the top
Capacity for 20 kg of cooked wheat

air in
motor

spiral
mixing
blade

water
jacket
perforated
air out
base plate

Schutyser et al. (2003) https://doi.org/10.1002/bit.10739 92

A rectangular bioreactor with a travelling screw mixer
developed at INRA, Dijon, France
Bed heights up to 1 m, capacity for ~1 tonne of substrate

agitation can be continuous or intermittent

(with relatively frequent mixing events)

air inlet
(controlled
T, %RH &
flowrate
http://www2.dijon.inra.fr
/biotec/ppban.htm

A version with a capacity of 25 tonnes has been constructed in China

93
The idea can be adapted to a circular bioreactor

air

94
 The instrumented and controlled bioreactor at PUC, Santiago, Chile
(50 kg capacity)
measurement
response

differential motor water

pressure tank
sensor
static CO2 & O2
mixing analyzers
blades

rotating basket PC
containing the thermo-
substrate bed couples steam
generator
relative
humidity
probe blower
heater cooler
air
box

95
The 200-kg capacity bioreactor at PUC, Santiago, Chile

motor for
turning the air out
agitators
upper cover
(stationary)
mechanical seal
the middle thermocouples are
section with raised out of the bed
the bed rotates during mixing periods
during a mixing and lowered into it
event during static operation
helical mixing
blades motor for
thermocouples
turning
toothed skirt perforated base plate
the
attached to middle
middle section section
bottom “air box”
water seal (stationary) air in

96
 Commercial bioreactors produced for the soy sauce koji industry

http://image.tradett.com/images/products/FA20101112163129961ulynn2359068/
solid-state-fermentation-equipment.jpg
97
Soy sauce koji bioreactor with 15-tonne capacity used by Nagata
Brewing Industry Co Ltd., Takarazuka, Japan

after 1 day, it
is transferred screw conveyer substrate held
to the lower within a 12 m

air preparation system

disk diameter disk

screw conveyer agitator

The agitator rotates in place

while the circular bed moves
Adapted from Sato K, Sudo S (1999) Small-scale solid-state fermentations.
In: Demain AL, Davies JE (eds) Manual of industrial microbiology and
biotechnology, 2nd edn. ASM Press, Washington DC, pp 61–79 98
There are some reports of the use of fluidized beds
air outlet

Space for bed expansion

and disengagement (larger
diameter decreases the air
velocity, causing the
particles to fall)
agitator for
breaking of The soy sauce company
agglomerates Kikkoman built an 8 m3
fluidized bed bioreactor in
the 1970s
air inlet (controlled T,
%RH & flowrate)
The air is blown at sufficient velocity
to fluidize the substrate particles
99
Continuous solid-state
cultivation bioreactors

100
Is continuous operation appropriate in solid-state
cultivation?
When continuous culture is done in submerged liquid culture with a
CSTR, the added substrate is mixed uniformly into the broth in the
bioreactor

https://en.wikipedia.org/wiki/Chemostat

101
What if you were to try something similar in SSC?
Would the added substrate be inoculated?
Or would it need to be colonized from the other particles?

Freshly inoculated The product would

substrate particles contain particles at
different stages of
the growth cycle

102
 It seems as though the plug-flow type of continuous operation
would be much more adequate

Freshly inoculated
substrate particles

plug flow

The product will be

much more uniform

103
A possible design for a continuous bioreactor with plug
flow

solids in
air out
solids flow

air in

solid flow recycle (with screw conveyor)

solids out

Recycled substrate used to inoculate fresh substrate

The screw can be rotated continuously or intermittently
Uniform aeration of the bed is difficult to achieve
104
 A continuous multilayer packed-bed bioreactor

10

9
new layer
8 layers
Each layer is like a separate batch with moved
7
a cultivation time equal to the residence

airflow
downwards
time in the bioreactor 6
at regular
So, what is the advantage?
5
intervals
4
• Upstream and downstream
3
processes can work continuously harvested
with a lower capacity 2 layer
• Once a steady-state is established, 1

all layers have the same

fermentation profile, which should
promote product uniformity
105
 Take home messages
There are many possible designs

There are many considerations in selecting a

bioreactor and substrate, including...
• to what degree does the organism tolerate mixing?
• what are the properties of the substrate bed and how do
they change during the process?

Should you build and test pilot-scale versions of

each design?

Mathematical models are useful tools for helping

to optimize bioreactor design and operation
106