Abstrict
The invention relates to apparatus for cultivating microbiological
substances. The fermenter comprises a container (1) accommodating
an aerator (4) and a stirring unit (5) having the form of a circulation
tube (6) and membrane (7). A partition (9) having a through hole
(10) offset relative to the center thereof is disposed between the
membrane (7) and the end of the tube (6) to define with the membrane
(7) a closed volume (11). The aerator (4) and the stirring unit
(5) are integrated into a single assembly secured on the partition
(9) and having a chamber (12) for distributing liquid and gas flows.
The fermenter is intended for intermittent cultivation of various
substances of biological origin feeding on liquid, solid or mixed
nutrient substrates.
Claims
We claim:
1. A fermenter comprising a container with an inlet having secured
therein a bacterial air filter and provided with a heat-transfer
jacket, the container having accommodated therein an aerator and
a stirring unit in the form of a vertical circulation tube and a
membrane underlying its lower end and connected to a control drive,
characterized in that a rigid partition wall (9) is disposed between
the membrane (7) and the lower end of the circulation tube (6),
this rigid partition wall (9) being provided with a through hole
(10) offset relative to the center thereof and defining with the
membrane (7) a closed volume (11), the aerator (4) and the stirring
unit (5) being made in the form of an integrated assembly secured
on the partition wall (9) and having a chamber (12) for distribution
of liquid and gas flows and separated in terms of height by two
non-return valves (13, 14) into three cavities (15, 16, 17) of which
the cavity (16) lying between the valves communicates with the hole
(10) made in the rigid partition wall (9), the cavity (15) underlying
the valves communicating with a liquid zone of the container (1),
and the cavity (17) overlying the valves communicating with the
interior of the circulation tube (6) the upper end of which is extended
toward a gas zone of the container (1).
2. A fermenter as claimed in claim 1, characterized in that a headpiece
(aerator 4) for forming a flow of liquid ejected from the tube (6)
is secured on the upper end of this tube (6).
3. A fermenter as claimed in claim 1, characterized in that the
container (1) accommodates a horizontal platform (19) having holes
(20) and serving to support a solid nutrient substrate.
4. A fermenter as claimed in claim 2, characterized in that the
container (1) accommodates a horizontal platform (19) having holes
(20) and serving to support a solid nutrient substrate.
Description BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to apparatus for periodic cultication of
various microbodies of biological origin fed on solid, liquid or
combination nutrient substrates, and more particularly to fermenters.
2. Description of Related Art
Any fermentation process involves a complex of interconnected biochemical,
physicochemical and diffusive transformations dependent in their
intensity mostly on the conditions of operation of the fermenter
and its hydrodynamic characteristics.
For cultivating microorganisms, fungi, algae or separate tissues
the microbiological practice has widely used flasks stopped by cotton
filters and accommodating microbodies to be grown in a temperature-controlled
rocker. Such devices are sufficiently simple and allow the growth
of a range of various microbodies. However, despite their multipurpose
application, their use is inherently confined in laboratory practice
due to insufficient mass transfer. In addition, the flasks are not
adaptable for cultivating microbodies of biological origin fed on
solid nutritive substrates.
There are known different structural arrangements of fermenters;
each such arrangement posessing hydrodynamic characteristics providing
for the active growth of this or that type of microbody.
In particular, there is known a fermenter comprising a container
with a stirring unit in the form of a multistage mixer (cf., e.g.,
E. U. Viestur, et al "Kultivirovanie mikroorganizmov"--Cultivation
of Microorganisms, in Russian, Moscow, 1980, pp. 141 to 171).
This fermenter can be used for growing bacteria of yeast on a liquid
nutritive medium and is not adapted for operation on solid or combination-type
substrates.
The stirring units used in these fermenters tend to produce excessive
foam in the cultural liquid, whereas for damping or supressing the
foam use is made of mechanical, chemical or combined foam suppression,
which affects the process parameters. In addition, the above apparatus
provide only for cultivation of one or two types of microorganisms,
such as yeast or bacteria.
There is also known a fermenter comprising a container provided
with a heat-transfer jacket, a bacterial filter disposed in its
inlet, an aerator, and a stirring unit in the form of a vertical
circulation tube having a membrane accommodated under its lower
end and connected with a drive (cf., a pamphlet of the Bioengineering
AG Company, Switzerland).
The fermenter of its construction is adapted for use with bacteria
or yeast fed on liquid nutritive substrates. The saturation of the
cultural liquid with the oxygen of the air and mixing thereof in
such a fermenter is assured by sucking in the air through the aerator
to the vertical circulation tube accompanied by the formation therein
of a gas-liquid mixture.
This fermenter is not capable of growing fungi, algae and separate
tissue, and is not adapted for carrying out the processes of microbiological
transformation of biologically active substances by using immobilized
cells; it is also not adapted to operate on solid or combination
substrates.
This is accounted for by the fact that aeration in said fermenter
takes place by virtue of suction of air by a jet of liquid with
the escape velocity thereof depending on its viscosity. Thus, during
cultivation of mycelium the speed of flow of the cultural liquid
is sharply reduced, and aeration terminates completely.
The free access of the substrate to the membrane prevents the fermentation
process using solid or combination substrates, because such insoluble
substrates (e.g., straw, wood saw dust, coal, etc.) tend to block
the inlet of the circulation tube to result in plugging thereof
and termination of the fermentation process.
For similar reasons the use of the above fermenter for carrying
out processes involving a separate tissue or immobilized cells is
impossible.
SUMMARY OF THE INVENTION
The invention is directed toward the provision of a fermenter having
a construction as to expand its functional capabilities and make
it applicable for growing microorganisms, algae or separate tissues
on solid, liquid or combination substrates.
The aim of the invention is attained by a fermenter comprising
a container with an inlet having secured therein a bacterial air
filter and provided with a heat-transfer jacket. The container accommodates
therein an aerator and a stirring unit in the form of a vertical
circulation tube and a membrane underlying its lower end and connected
to a control drive. According to the invention, a rigid partition
wall is disposed between the membrane and the lower end of the circulation
tube, this rigid partition wall being provided with a through hole
off-set relative to the center thereof and defining with the membrane
a closed volume, whereas the aerator and the stirring unit are made
in the form of an integrated assembly secured on the partition wall
and having a chamber for distribution of liquid and gas flows and
separated in terms of height by two non-return valves into three
cavities of which one cavity lies between the valves and communicates
with the hole made in the rigid partition wall, the other cavity
underlies the valves and communicates with a liquid zone of the
container, and the third cavity overlies the valves and communicates
with the interior of the circulation tube the upper end of which
is extended toward a gas zone of the container.
Preferably, a headpiece for forming a flow of liquid ejected from
the tube is secured on the upper end of this tube.
Advisably, the container accommodates a horizontal platform having
holes therein and serving to support a solid nutrient substrate.
The fermenter embodying the features of the present invention can
be used for cultivating microorganisms, algae, or separate tissues
on liquid, solid or combination substrates.
The use in the proposed fermenter of pulsewise stirring in combination
with the structural peculiarities of the stirring unit made integral
with the aerator provides in the fermenter jet or bubble mixing
of the cultural liquid, which is especially advantageous for selecting
proper hydrodynamic conditions for growing particular types of microorganisms.
The provision of the horizontal perforated platform and replaceable
headpieces on the aerator makes it possible to grow a variety of
aerobic microorganisms, mycelium, separate tissues, and photoautotrophic
organisms on liquid, solid or combination substrates, as well as
to carry out processes involving immobilized ferments or cells.
BRIEF DESCRIPTION OF THE DRAWINGS
The invention will now be described in greater detail with reference
to various specific embodiments thereof taken in conjunction with
the accompanying drawings, in which:
FIG. 1 is a longitudinal sectional view of the fermenter according
to the invention;
FIG. 2 is a longitudinal sectional view of a headpiece for forming
a flow of liquid;
FIG. 3 illustrates substantially the same as shown in FIG. 1; non-return
valves for distributing liquid and gas flows being in an upside
down position;
FIG. 4 is a modified form of the fermenter according to the invention
comprised of seven containers and a common membrane movable from
one drive; and
FIG. 5 is a modification of a multifunctional fermenter adapted
for industrial application.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
A fermenter illustrated in FIG. 1 comprises a container 1 provided
with a heat transfer jacket 2 and having a bacterial filter 3 disposed
at an inlet of the container 1. The container 1 also accommodates
an aerator 4 and a stirring unit 5 in the form of a vertical circulation
tube 6 and a membrane 7 underlying its lower end and connected to
a controlling pneumatic drive (not shown) by means of a pipe 8.
Interposed between the membrane 7 and the circulation tube 6 is
a rigid partition wall 9 having a through hole 10 offset relative
to the center of the partition wall 9 to form with the membrane
7 a closed volume 11. The aerator 4 and the stirring unit 5 are
integrated into a single assembly secured on the partition wall
9 and having a chamber 12 for distributing liquid or gas flows.
The chamber 12 is separated in terms of its height by two non-return
valves 13 and 14 into three cavities 15, 16 and 17, of which the
intervalve cavity 16 communicates with the hole 10 made in the rigid
partition wall 9, the undervalve cavity 15 communicating through
holes 18 with a liquid zone of the container 1, and the overvalve
cavity 17 communicates with the interior of the circulation tube
6 the upper end of which is extended toward a gas zone of the container
1.
Secured on the upper end of the circulation tube 6 is a replaceable
headpiece or aerator 4 in the embodiment described in the form of
a jet injector.
The container 1 also accommodates a horizontal platform 19 having
holes 20 and serving to support a solid nutrient substrate.
The fermenter shown in FIG. 1 is a multifunction inoculator-type
fermenter intended for cultivation of microorganisms in laboratories,
particularly for growing such living cells as bacteria, yeast and
other anaerobic microorganisms fed on liquid or mixed nutrient substrates.
The fermenter operates in the following manner.
The container 1 is filled with a culture liquid with a level thereof
not in excess of the level at which the jet injector (aerator 4)
is disposed to be then stopped by the bacterial filter 3. Compressed
air is supplied from a generator of pneumatic pulses (not shown)
at a predetermined pulse frequency through the pipe 8 to a chamber
21 underlying the membrane 7.
The membrane 7 is caused to oscillate and during its downward travel
acts to suck the cultural liquid through the holes 18, valve 14,
cavity 15 and hole 10 into the closed volume 11; the non-return
valve 13 being closed.
During the upward movement of the membrane 7 the cultural liquid
occupying the volume 11 is forced therefrom through the hole 10,
cavity 16, non-return valve 13, circulation tube 6 and aerator 4
(jet injector) in the form of jets back to the container 1; the
non-return valve 14 being closed. The jets of cultural liquid formed
in the injector (aerator 4) penetrate through the body of cultural
liquid occupying the container 1 to disperse oxygen of the air therein
entrained by the jets from the gas zone of the container 1.
During oscillations of the membrane 7 the level of the cultural
liquid in the container 1 also tends to oscillate to follow the
movements executed by the membrane 7 and thereby carry out the exchange
of air occupying the gas zone of the container 1 with the atmospheric
air through the bacterial filter 3.
In the case of using combination nutrient substrates, use is made
of the horizontal perforated platform 19 secured on the circulation
tube 6 and walls of the container 1 by means of elastic rings 22.
When microorganisms are grown on such nutrient substrates, solid
insoluble components of such nutrient media are retained on the
platform 19 and fail to contaminate the stirring unit 5. The pulsewise
operation of the membrane 7 and the oscillations of the entire body
of the cultural liquid prevents obstruction of the holes 20 of the
platform 19 by the insoluble solids contained in the substrate.
When the cultural liquid generates too much foam, the foam covers
the injector or aerator 4 and the jets of the cultural liquid act
to reduce the dispersion of air in the cultural liquid and thereby
stabilize froth formation at this level.
In such conditions of operation of the fermenter mass transfer
of the cultural liquid is effected through vigorous mixing and recirculation
of the foam; such a mass transfer is no less efficient than during
jet dispersion of the oxygen of the air in the cultural liquid.
With reference to FIG. 2, there is shown one more embodiment of
the headpiece-aerator in the form of a semisphere 23 acting to generate
a substantially flat annular flow of liquid.
The use of the semisphere 23 instead of the jet injector represented
in FIG. 1 allows production of a very thin film of cultural liquid
flowing down on the transparent walls of the container 1 for the
fermenter according to the invention to be capable of growing photoautotrophic
microorganisms and algae.
A modified form of the fermenter according to the invention illustrated
in FIG. 3 is different from the fermenter shown in FIG. 1 only in
positioning of the valves 13 and 14. The valves 13 and 14 in the
fermenter of FIG. 3 are turned upside down as compared to the modification
of the fermenter represented in FIG. 1 to provide for pulsewise
bubbling operation of the fermenter.
This modification is also a multifunctional inoculator-type fermenter
intended for laboratory cultivation of aerobic microorganisms or
fungus mycelium grown on solid, liquid or mixed substrates, as well
as for carrying out the processes of transformation of steroids
with immobilized cells or ferments, or for growing a separate living
tissue.
The modified form of the fermenter shown in FIG. 2 operates in
the following manner.
The container 1 is filled with a cultural liquid to a level not
exceeding the level at which the aerator 4 is disposed, the container
is then closed by the bacterial filter 3. Compressed air is fed
from a generator of pneumatic pulses (not shown) at a predetermined
pulse frequency through the pipe 8 to the chamber 21 under the membrane
7.
The membrane 7 is therefore caused to reciprocate or oscillate,
and during an upward travel to force air from the closed volume
11 through the hole 10, cavity 16, valve 14 and holes 18 to the
part of the container 1 occupied by the liquid, the thus forced
air tending to cause bubbling and vigorous stirring of the body
of cultivated liquid to saturate it with the oxygen of the air.
The valve 13 is closed during this stirring action. During the reverse
travel of the membrane 7 the valve 14 is closed, and the air is
again passed to the closed volume 11 from the gas zone of the container
1 through the aerator 4, circulation tube 6, valve 13, cavity 16
and hole 10. Exchange of air occupying the gas zone of the container
1 with the atmospheric air is effected through the bacterial filter
3 synchronously with the movements of the membrane 7 similarly to
what has been described with reference to the modification shown
in FIG. 1.
In the case of excessive foaming of the cultural liquid, the foam
tends to envelope the aerator 4 to be delivered instead of air to
the closed volume 11, whereby aeration of the cultural liquid will
be less vigorous and the level of foam in the container 1 will be
stabilized.
Mass transfer in the fermenter in this case will take place due
to a highly intensive circulation of the foam; this mass transfer
being as efficient as when bubbling takes place in the fermenter.
When growing microorganisms on solid substrates or during carrying
out the processes of transformation of steroid compounds with an
immobilized biocatalyst, these compounds are placed on the horizontal
platform 19 to be aerated by air fed in a pulsewise manner through
the holes 20 in response to the movement of the membrane 7.
During growing fungus mycelium on such solid substrates as hay,
the air conveyed for aeration can be moistened by accommodating
in the space under the horizontal platform 19 a liquid nutrient
medium or water. The mycelium growth process in this case is carried
out similarly to what has been described with reference to bubbling
aeration of the cultural liquid.
The fermenter shown in FIG. 4 is a multifunctional cultivator type
intended for cultivation of aerobic microorganisms by using various
nutrient substrates, separate tissue cultures, and photoautotrophic
microorganisms and algae, as well as for carrying out the processes
of transformation of biological compounds making use of immobilized
ferments or cells.
This modified form of the proposed fermenter is a set of seven
fermenters generally similar to the one described with reference
to FIG. 1 and accommodated in a single heat transfer jacket 2. Cavities
21 of all of the fermenters are integrated into a single common
cavity by a conduit 24 communicated by the pipe 8 with a controlling
pneumatic drive (not shown).
This fermenter is intended predominantly for laboratory use. It
can find a wide application in research and development, for microbiological
and medicinal practice, as well as in sanitation control laboratories
for checking the level of bacterial contamination of environments,
etc. It can further be used in research practice for routine analytical
and inspection operations where for these purposes bulky and energy-consuming
flasks are used. The apparatus has a provision for a self-contained
or independent temperature control, and has no need for temperature-adjusted
rooms.
This modification of the fermenter is especially advantageous for
selecting optimized dynamic conditions for growing a chosen microorganism.
The fermenter illustrated in FIG. 4 operates substantially in the
same manner as those represented in FIGS. 1 and 3, although a feature
which distinguishes it from the previously described modifications
resides in the use of a common drive providing for a synchronous
operation of all the seven fermenters in both equal and different
stirring conditions therein. This provides for growing microorganisms
simultaneously in seven reproductions, which is especially important
for research practice.
As distinct from the aforedescribed modified forms of the fermenters
according to the invention, the fermenter shown in FIG. 5 comprises
several, in the case described three, stirring units 5 in one container
1, these stirring units 5 being mounted on a partition wall indicated
by the same numeral 9 and adaptable for different rates of aeration
and stirring in a range of various ratios.
For example, one unit 5 intended for producing bubbles may be disposed
in the central part of the container 1 with two of jet stream operated
units 5 arranged at the periphery.
Therefore, the fermenter thus constructed can provide for combination
conditions of stirring assuring counter-directed flows of gas and
liquid promoting a more vigorous saturation of the cultural liquid
with gas, which expands the functional capabilities of the fermenter,
especially with a foamy substrate, when it is necessary to increase
the rate of mass transfer thanks to more active circulation of the
foam without resorting to an increase in its volume.
In addition, such an arrangement of the units 5 promotes additional
removal from the walls of the container 1 of particles of the substrate
of microorganisms tending to adhere thereto.
The fermenter operates from a single drive through the pipe 8 and
conduit 24 communicable with the cavities 21 arranged under the
membranes 7.
The fermenter embodying the present invention can perform a multitude
of functions and can find application:
in the microbiological practice, as a fermenter for carrying out
the processes of microbiological synthesis of protein products,
bioconversion of vegetable materials, utilization of wastes, etc.;
in the food industry as a mixer or an apparatus for stirring liquid
media, for conducting fermentation processes, etc.;
in the pharmaceutical industry, as a reactor for making medicinal
preparations, such as during transformation of steroids, etc.; and
in the chemical industry, as a reactor for mixing liquid media
and carrying out the reactions involving the use of solid catalysts.
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