US3607659A

US3607659A - Fermentation apparatus

Info

Publication number: US3607659A
Application number: US786570A
Authority: US
Inventors: James L Bloomer
Original assignee: Temple University of Commonwealth System of Higher Education
Current assignee: Temple University of Commonwealth System of Higher Education
Priority date: 1968-12-24
Filing date: 1968-12-24
Publication date: 1971-09-21
Anticipated expiration: 1988-09-21

Abstract

Apparatus is provided in the form of fermentation vessels which are provided with caps for substantially sealing the same against contamination, and with tube means for communicating between the inside and outside of the vessels. Stirring is effected by a magnetic element disposed within the vessels which is actuated by an external electromagnetic source.

Description

United States Patent 1 3,607,659

[72] Inventor James Bloomel' 2,432,132 12/1947 Allen 215/56 Philadelphia, Pa. 2,990,339 6/1961 Frank et al.. 195/139 [21] Appl. No. 786,570 3,172,235 3/1965 Bjorklund... 7 195/143 [22] Filed Dec. 24,1968 3,275,528 9/1966 Ainis 195/143 [73] Assignee Temple University of the Commonwealth system of Higher Education 605,467 7/1948 Great Britain 23/292 Philadelphia, Pa. Primary ExaminerAlvin E. Tanenholtz AtlorneyPau1 and Paul [54] FERMENTATION APPARATUS 12 Claims, 5 Drawing Figs.

[52] US. Cl 195/142, 195/143 [51 Int. Cl ClZh 1/00 T T; Apparatus i provided in the form of fermenta- [SOI Field of Search l95/139,

tiun vessels which are provided with caps for substantially 41,21 LE; 23/292; 25/56 sealing the same against contamination, and with tuhu means for communicating between the inside and outside of the ves- [56] References Cited sels. Stirring is effected by a magnetic element disposed within UNITED STATES PATENTS the vessels which is actuated by an external electromagnetic 2,287,746 6/1942 Morton 195/139 (LE) source.

PMENTED SEPZI 197i SHEET 1 BF 2 INVENTOR. James L. Bloomer ATTORNEYS.

PATENIEU SEPZI c971 SHEET 2 BF 2 INVENTOR. James L. Bloomer mwwa ATTORNEYS.

F ERMENTATION APPARATUS BACKGROUND OF THE INVENTION In fermentation vessels (and other vessels used for organism growth) of the prior art, several apparatus have been developed, principally for use in analyzing organic mixtures while the mixtures are in motion, as a continuous process. Generally, such apparatus have been rather complicated, and have required the use of rotary seals, principally for providing stirring from an external source, such seals often sticking when the vessels are used for many solutions, such as sugar solutions.

Furthermore, there has developed a need for apparatus which is sufficiently inexpensive to be used in quantity, as for example, for laboratory use by students and the like, but which will provide sufficient safeguards against contamination when used for such studies as culture growths, fungal metabolic studies and the like. In such studies, it is often desirable to inoculate the culture or growth, and it is additionally desirable to often take samples of the growth at various stages of its development. A primary consideration at all times is that contamination of the apparatus must be prevented, and in this regard, along with the other above-discussed parameters, it has been necessary to develop vessels which would satisfy all of the requirements mentioned above, but which would still be economical, as well as readily capable of being assembled.

SUMMARY OF THE INVENTION The present invention seeks to provide an apparatus which may be economically manufactured and assembled, to provide contamination-free apparatus for various studies, wherein inoculation and sampling facilities are provided for the apparatus, by using a vessel and a cap fitted over its upper end, sufficiently snug with the top of the vessel, but wherein means are provided for delivering a gas, such as sterile air to the vessel, the outflow of such gas from the vessel providing a seal against contamination from the ambient air. Novel means are also provided whereby gas inlet, inoculation and sampling tubes communicate between the inside and outside of the vessel.

Accordingly, it is a primary object of this invention to provide a novel fermentation apparatus, whereby a gas seal is provided between a vessel and its cap, but whereby the apparatus remains free from contamination during use, and also whereby access is provided between the interior of the vessel and externally thereof.

It is another object of this invention to provide a novel apparatus comprising a vessel and a cap therefore, whereby tubes extend from the inside of the vessel to the outside thereof, such tubes being closely fitted with the vessel cap for purposes of preventing contamination of the vessel.

It is a further object of this invention to provide a novel vessel having a cap, wherein the vessel is provided with inoculation, sampling and gas inlet tubes, as separate communication routes between the inside and the outside of the vessel.

It is a further object of this invention to accomplish the above object, wherein magnetic stirring means are utilized interiorly of the vessel.

Other objects and advantages of the present invention will become readily apparent to those skilled in the art from a reading of the following brief description of the drawing figures, detailed description of the preferred embodiments and the appended claims.

IN THE DRAWINGS FIG. 1 is a vertical sectional view, taken through the apparatus of this invention, generally along the line I-I of FIG. 2.

FIG. 2 is a transverse sectional view taken through the apparatus of FIG. 1, generally along the line IIII, wherein several of the components of the apparatus are illustrated in plan view.

FIG. 3 is a vertical sectional view of an alternative form of the apparatus of this invention.

FIG. 4 is a transverse sectional view of the apparatus of FIG. 3, taken generally along the lines IV-IV of FIG. 3.

FIG. 5 is a fragmentary vertical sectional view of a modified form of the apparatus of FIG. 3.

Referring now to the drawings in detail, reference is first made to FIGS. 1 and 2, wherein there is illustrated a laboratory growth apparatus of this invention, generally designated by numeral 10.

The apparatus 10 includes a vessel 11, of glass construction, which may be in the form of a twenty liter carboy, having a bottom wall 12, a main body or sidewall portion 13, an upper portion 14 of reduced diameter relative to the body 13, and an upper rim 15 defining an opening therein, the vessel 11 being otherwise imperforate. A glass cap 16, in the form of an inverted beaker, having an end wall 17 and an annular sidewall 18, terminating at its lower end in a rim 20 defining a generally circularly configured hole or opening is provided, in inverted position disposed about the neck portion 14 of the vessel 11.

An inoculation tube 21, also of glass construction is provided, having a generally perpendicularly disposed major portion 22, and a lower portion 23 angularly bent toward the lower corner of the vessel 11 defined by the junction of the body wall 13 and bottom wall 12, with the tube 21 having an open lower end 24 The upper end of the tube 21 is reversely bent at 25, back over the vessel rim 15, to be disposed downwardly between the vessel neck 14 and the cap sidewall 18, in disposition close to the sidewall 18, with the tube 21 being upwardly bent in the vicinity of the rim 20 of the cap 16, to again have a vertically upwardly extending portion 27 which is fitted with a glass closure in the form of a serum cap 28. Reversely bent portion 26 of the glass tube 21 extends through a slot 30 in the sidewall 18 of the cap 16, the slot 30 being cut upwardly from the rim 20 of the cap 16, and being flared slightly outwardly to be closely fitting about the tube portion 26, to permit minimum air passage therethrough about the tube portion 26. The tube 21 thus provides a means for inoculation of a culture 31 within the vessel 11. An additional glass tube 32 is provided, virtually identical to the inoculation tube 21, but being used for sampling, for example by extraction of a portion of the culture 31 by means of application ofa vacuum to an upper portion 33 of the glass tube 32, after removal of the serum cap 34 therefrom.

A third glass tube 35 is provided, generally configured similar to the

tubes

21 and 32, but having at its lower most end 36, inside the vessel 11, a portion 37 in the form of a course porosity glass frit 37, which provides a screening effect, as well as a diffusing effect for air passing through the tube 35. An outer portion 38 of the tube 35 is not bent upwardly as is, for example, the portion 27 of the tube 21, but is downwardly bent and is connected to an enlarged glass universal joint 40, or ball joint, which contains a cotton plug 41, several inches long, with the lower end of the universal joint 40 also containing a cotton plug 42, to further facilitate the filtering of particles from the air supply. A source of a gas, such as sterile air is provided through the inlet 43 to the ball joint 40, to enter into the culture 31 within the vessel 11 through the frit 37, to provide an atmosphere effective for facilitating culture growth and the like. As this air bubbles up through the culture 31, and into the upper portion of the interior of the vessel 11, it continually escapes from the vessel 11, by passing up over the rim 15 of the vessel 11, and downwardly between the neck 14 of the vessel 11 and the sidewall 18 of the cap 16, to pass outwardly of the apparatus 10 about the point of contact of the rim 20 with the tapering or expanding portion of the body wall 13 of the vessel 11, as well as outwardly through the slots 30 in the cap sidewall 18. Thus, this continuous discharge of air from the apparatus 10 provides an effective seal preventing contamination of the vessel 11 from an external source, at the ambient pressure, in that the escaping pressure of the air from the vessel 11 is at a greater pressure than the ambient pressure.

At the upper most end of the glass tube 32, just below the serum cap 34, a small cotton plug (not shown) is disposed within the tube 32, held in place by small indentations in the glass, to provide a means for removal or sampling through the tube 32. Similarly, the tube 21 may be used for sampling, if such use is more desirable than its use for inoculation, depending upon the particular culture being studied, in which case, the tube 21 would also be provided with a cotton plug 44 held in place by indentations in the glass tube 21.

It is to be noted, that, while two

such tubes

21 and 32 are provided, either or both may be used for sampling and/or inoculation, as desired, depending upon the particular experiment.

The apparatus, for example the vessel 11 an the beaker 16 are generally sterilized in an autoclave prior to use, to guard against contamination, the tubes and other apparatus also being similarly sterilized.

Stirring is provided by using a large diameter (for example 2 inch) magnet 45 which has a coating 46 of glass, nylon or the like, to render the same inert to the culture being studied, the magnet 45 being disposed approximately centrally of the vessel 11, at the bottom thereof, and with the vessel 11 resting upon a magnet actuating apparatus 47.

The magnetic stirring apparatus 47 includes a housing 48, a motor 50, which may be connected to the usual AC source, and a rotating ferromagnet 51 driven by the motor 50, which, in turn, rotates the magnetic element 45 within the vessel 1 I, for stirring the culture 31.

It is to be noted, in the apparatus mentioned above, the use of ring seals is avoided, which works towards minimizing the cost of the apparatus. Also, the use of seals on the vessel 11 is avoided.

Referring now to FIGS. 3 and 4, an alternative apparatus 60 is illustrated, for studies involving organisms which prefer a high degree of aeration, such as lichen fungi. Because lichen fungi are particularly notorious as being slow growers, the apparatus must provide protection against contamination for long periods of time. Also, provision must also be made for inoculation and removal of solution and/or cell material, also with maximum protection against contamination, as is required with the apparatus mentioned above. The apparatus 60 utilizes a flask 61 which may be constructed in the configuration of a one liter Erlenmeyer flask which has a pair of

tubes

62 and 63 in the form of sidearms millimeter diameter medium wall glass tubing) heat sealed thereto at 64 and 65, respectfully, to communicate the interior bottom of the flask 61 with the outside. Suitable serum caps 66 and 66' are also provided. In the tube 63, provision is made for removal of the solution with filtration through a cotton plug 67, which is held in place by glass indentations 68. The tube 63 would thus ordinarily be used for removal of solution, and the tube 62 would be used for inoculation.

A cap 70 is provided, having an annular sidewall portion 71 and an end wall 72, for covering the open end 69 of the flask 61, seating on the rim 74 thereof.

Sterile air is provided through the air inlet 75, which is filtered through

cotton plugs

76 and 77, in the ball joint 78, which then passes into a glass tube 80 through an opening 81 in the end 72 of the cap 70, with the tube 80 being connected in heat sealed relation within the tube 72. v

The tube 80 is angularly bent at 82 toward the periphery of the bottom portion of the flask 61, and is also provided with a course porosity frit 83.

A stirring element 84 which comprises a magnet having a Teflon, glass or other suitable coating 85 is also provided in the bottom of the flask 61, and is magnetically rotated by means of a rotating ferromagnet 86 driven similarly the the magnet 51 of FIG. 1. The device of FIG. 3 also operates in a manner similar to that of FIG. 1, in that it is the air passing through the culture 87, upwardly outwardly around the rim 74 of the flask 61 and downwardly outwardly of the cap 70 which provides a seal by means of the pressure differential between the escaping air and the ambient air pressure.

In the apparatus of FIGS. 3 and 4, the

cotton plugs

76 and 77 also provide a means for filtering particulate matter from the incoming air. It will be noted, with respect to the air seal mentioned above with respect to both apparatus, that the principal of the ordinary petri dish is utilized, such that the outflow of air from the apparatus tends to expel any foreign spores.

The apparatus has been used for the growth of lichen fungi, fungi, and yeast, without visible evidence of contamination. These apparatus have been used without contamination for periods as much as several months. In some instances, as with fast growing fungi such as those of the penicillin species, the inoculation may result in such a heavy growth that the magnetic stirring becomes ineffective in a matter of 2 or 3 days.

It will further be noted that, when the apparatus is used with media having foaming tendencies, antifoams must be added to the

solutions

31 or 87, as necessary. However, in some instances, the use of antifoams would not be desired, and accordingly, the apparatus, particularly that of FIGS. 3 and 4 may be modified as illustrated in FIG. 5, such that the tube would extend downwardly in a perpendicular manner, to a position directly above the magnet 84, without being bent in the manner of FIG. 3, such that the end of the tube 80 would be in fairly close proximity to the magnet 84. By close control of this spacing and the size of the magnet, as well as the rate of rotation of the magnet, a vortex effect may be obtained which will create sufficient dispersion of the incoming air into the solution 87.

It will be noted, that while two basic forms of the apparatus of this invention have been disclosed in detail herein, various modifications may be made in the structural details of the apparatus of this invention, as well as in the use thereof, without departing from the spirit and scope of the invention as defined in the appended claims.

I claim:

1. An apparatus for use in organism growth comprising an open-topped vessel having a body portion and having a neck portion of reduced diameter, an open ended cap having side and end walls and being disposed in inverted relation over said vessel open end, with lower sidewall portions of said cap being in sufficiently close fitted relation to an adjacent outer portion of said vessel that gaseous outflow therebetween from inside the vessel will maintain sterility inside the vessel, including tube means for providing gas inflow to the interior of the vessel; said tube means comprising a duct extending from outside the vessel, passing between said close fitted cap and vessel portions, extending between said cap and vessel to the upper end of said vessel, being reversely bent beneath the cap endwall and over the vessel open upper end, and extending downwardly into a lower interior portion of said vessel, with at least one of an innoculation and sampling tube means also being provided and comprising an additional duct extending from outside the vessel, passing from outside said cap between said close fitted cap and vessel portions, extending between said cap and vessel to the upper end of said vessel, being reversely bent beneath the cap endwall andover the vessel open upper end, and extending downwardly into a lower interior portion of said vessel, wherein said gas inflow tube means includes gas filter means and means for connection to an external gas source, and wherein removable cover means for each said innoculation and sampling tube means is provided.

2. The apparatus of claim I, including a magnetic stirring element disposed within the vessel, and adapted for use with an external magnetic actuator for rotating the same.

3, The apparatus of claim 1, wherein a diffusing frit is provided on the lowermost end of said inflow tube means inside said vessel.

4. The apparatus of claim I, wherein said additional duct has a second reverse bend around the lower end of the inverted cap and terminates in an upwardly extending free end.

5. The apparatus of claim 1, wherein both said innoculation and sampling tube means are provided.

6. The apparatus of claim I, wherein each said tube means is angularly outwardly bent inside said vessel near the lower end thereof.

7. An apparatus for use in organism growth, comprising an open-topped vessel having body portion and having an open upper end of reduced diameter relative to the vessel bottom, with the vessel body diameter progressing to a greatest diameter at the bottom thereof, and with a cap being provided having an open lower end, and being disposed over the vessel open upper end in inverted close-fitting relation thereto, with the cap sidewalls extending downwardly along, but outside the upper ends of the vessel sidewalls, and with the cap upper end being closed except for the passage of a gas inlet tube means therethrough in heat sealed relation thereto, with the gas inlet tube means comprising a duet extending through the upper end wall of the cap, and down into the lower interior portion of the vessel from outside the cap, comprising means for providing gas to the interior of said vessel, and with at least one of an innoculation and sampling tube means permanently connected to the vessel at the bottom thereof, in integrally sealed relation therewith, and extending vertically upwardly therefrom, and terminating in an upwardly opening free end having a removable cover therefor, to comprise means for providing access to the interior bottom portion of said vessel, and wherein said gas inflow tube means includes means for connection to an external gas source, and wherein filtering means are provided within said gas inlet tube means.

8. The apparatus of claim 7, wherein a magnetic stirring element is disposed within the vessel, and is adapted for use with an external device adapted for rotating the magnetic stirring element from outside the vessel.

9. The device of claim 7, wherein both said innoculation and sampling tube means are provided.

10. The device of claim 7, wherein said one of said innoculation and sampling tube means is provided with removable filtration means disposed therein.

11. The device of claim 7, wherein said gas inlet tube means is provided with a diffusing frit at its lowermost portion inside the vessel, for distribution of gas passing through the tube means into a liquid contained within the vessel.

12. The apparatus of claim 7, wherein said gas inlet tube means is angularly bent in an outward direction, at the lowermost interior end thereof, within the vessel.

Claims

2. The apparatus of claim 1, including a magnetic stirring element disposed within the vessel, and adapted for use with an external magnetic actuator for rotating the same.
3. The apparatus of claim 1, wherein a diffusing frit is provided on the lowermost end of said inflow tube means inside said vessel.
4. The apparatus of claim 1, wherein said additional duct has a second reverse bend around the lower end of the inverted cap and terminates in an upwardly extending free end.
5. The apparatus of claim 1, wherein both said innoculation and sampling tube means are provided.
6. The apparatus of claim 1, wherein each said tube means is angularly outwardly bent inside said vessel near the lower end thereof.
7. An apparatus for use in organism growth, comprising an open-topped vessel having body portion and having an open upper end of reduced diameter relative to the vessel bottom, with the vessel body diameter progressing to a greatest diameter at the bottom thereof, and with a cap being provided having an open lower end, and being disposed over the vessel open upper end in inverted close-fitting relation thereto, with the cap sidewalls extending downwardly along, but outside the upper ends of the vessel sidewalls, and with the cap upper end being closed except for the passage of a gas inlet tube means therethrough in heat sealed relation thereto, with the gas inlet tube means comprising a duct extending through the upper end wall of the cap, and down into the lower interior portion of the vessel from outside the cap, comprising means for providing gas to the interior of said vessel, and with at least one of an innoculation and sampling tube means permanently connected to the vessel at the bottom thereof, in integrally sealed relation therewith, and extending vertically upwardly therefrom, and terminating in an upwardly opening free end having a removable cover therefor, to comprise means for providing access to the interior bottom portion of said vessel, and wherein said gas inflow tube means includes means for connection to an external gas source, and wherein filtering means are provided within said gas inlet tube means.
8. The apparatus of claim 7, wherein a magnetic stirring element is disposed within the vessel, and is adapted for use with an external device adapted for rotating the magnetic stirring element from outside the vessel.
9. The device of claim 7, wherein both said innoculation and sampling tube means are provided.
10. The device of claim 7, wherein said one of said innoculation and sampling tube means is provided with removable filtration means disposed therein.
11. The device of claim 7, wherein said gas inlet tube means is provided with a diffusing frit at its lowermost portion inside the vessel, for distribution of gas passing through the tube means into a liquid contained within the vessel.
12. The apparatus of claim 7, wherein said gas inlet tube means is angularly bent in an outward direction, at the lowermost interior end thereof, within the vessel.