Device for Producing Metabolites
20220372416 · 2022-11-24
Inventors
Cpc classification
C12M29/04
CHEMISTRY; METALLURGY
Y02P20/133
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
C12M25/12
CHEMISTRY; METALLURGY
International classification
C12M1/12
CHEMISTRY; METALLURGY
Abstract
A bioreactor for producing hydrogen gas and other metabolites. The bioreactor utilizes light, fermentation, and other metabolic processes for the production of metabolites, derived from various microorganisms contained within the bioreactor through respective metabolic pathways. The bioreactor comprises a main reactor chamber, a semipermeable membrane, a sleeve, a power supply, a substrate medium, a heating member, a plurality of tubing members, a collection reservoir, a pressure-sealed connecter member, and an agitator.
Claims
1. A bioreactor comprising: a main reactor chamber, including a substrate containment space; a semipermeable membrane with a hollow inner lumen side and an external surface side; a sleeve perimetrically enveloped around the main reactor chamber; an electric power supply; a substrate medium; a heating member; a plurality of tubing members; a collection reservoir; a pressure-sealed connecter member; a metabolite purifying filter member; an agitator; the pressure-sealed connecter member hermetically sealed to the main reactor chamber, opposite the heating member and the agitator; the substrate medium fills the substrate containment space by traveling through an inlet tubing member into the substrate containment space; the substrate medium envelopes the external surface side of the semipermeable membrane and permeates through the semipermeable membrane to fill the inner lumen side; the heating member maintains a predefined temperature to maintain a predefined substrate temperature; and the agitator maintains a predefined cyclical rate.
2. The bioreactor of claim 1, wherein the semipermeable member is housed within the main reactor chamber within the substrate containment space.
3. The bioreactor of claim 1, wherein the plurality of tubing members includes an inlet tubing member, an outlet tubing member, and a collection tubing member; the plurality of tubing members are hermetically sealed to the pressure-sealed connecter member; the inlet tubing member and outlet tubing member traverse through the pressure-sealed connecter member to enter the substrate containment space; and the collection tubing member traverses through the pressure-sealed connecter member to enter the hollow inner lumen space of the semipermeable membrane.
4. The bioreactor of claim 3, further comprising: a light source; the light source contained within the sleeve; and the light source having a variable wavelength, wherein the wavelength is varied to promote metabolite production by a plurality of microorganisms contained within the hollow lumen space of the semipermeable membrane.
5. The bioreactor of claim 3, wherein the substrate medium is supplied to the substrate containment space at a predefined flow rate to permeate through the semipermeable membrane and into the hollow inner lumen space without disrupting or removing the contents therein.
6. The bioreactor of claim 5, wherein the semipermeable membrane is wrapped cylindrically around the outlet tubing member, forming multiple layers with space between the layers.
7. The bioreactor of claim 6, wherein the semipermeable membrane further comprises, an inert scrim; the inert scrim positioned between each layer of the semipermeable membrane to prevent the surfaces of the semipermeable membrane in the rolled-up configuration from being juxtaposed; and the inert scrim being of a net configuration so gases or nutrients flow freely through the fenestrations and allow the full surface of the membranes to be separated from each other.
8. The bioreactor of claim 5, wherein a dial increases or decreases the temperature of the heating member and cyclical rate of the agitator.
9. A bioreactor comprising: a main reactor chamber, including a substrate containment space; a semipermeable membrane with a hollow inner lumen side and an external surface side; a sleeve perimetrically enveloped around the main reactor chamber; an electric power supply; a substrate medium; a heating member; a plurality of tubing members; a collection reservoir; a pressure-sealed connecter member; a metabolite purifying filter member; an agitator; the pressure-sealed connecter member hermetically sealed to the main reactor chamber, opposite the heating member and the agitator; the semipermeable member is housed within the main reactor chamber within the substrate containment; the plurality of tubing members includes an inlet tubing member, an outlet tubing member, and a collection tubing member; the plurality of tubing members are hermetically sealed to the pressure-sealed connecter member; the inlet tubing member and outlet tubing member traverse through the pressure-sealed connecter member to enter the substrate containment space; and the collection tubing member traverses through the pressure-sealed connecter member to enter the hollow inner lumen space of the semipermeable membrane; the substrate medium fills the substrate containment space by traveling through the inlet tubing member into the substrate containment space; the substrate medium envelopes the external surface side of the semipermeable membrane and permeates through the semipermeable membrane to fill the inner lumen side; the heating member maintains a predefined temperature to maintain a predefined substrate temperature; and the agitator maintains a predefined cyclical rate.
10. The bioreactor of claim 9, further comprising: a light source; the light source contained within the sleeve; and the light source having a variable wavelength, wherein the wavelength is varied to promote metabolite production by a plurality of microorganisms contained within the hollow lumen space of the semipermeable membrane.
11. The bioreactor of claim 9, wherein the substrate medium is supplied to the substrate containment space at a predefined flow rate to permeate through the semipermeable membrane and into the hollow inner lumen space without disrupting or removing the contents therein.
12. The bioreactor of claim 11, wherein the semipermeable membrane is wrapped cylindrically around the outlet tubing member forming multiple layers with space between the layers.
13. The bioreactor of claim 12, wherein the semipermeable membrane further comprises, an inert scrim; the inert scrim positioned between each layer of the semipermeable membrane; and the inert scrim being of a net configuration.
14. The bioreactor of claim 13, wherein a dial increases or decreases the temperature of the heating member and cyclical rate of the agitator.
15. The bioreactor of claim 9, further comprising: at least two capillary membranes; at least one capillary channel; connecting segments; a capillary bed; wherein the connecting segments adjoin the at least two capillary membranes and enclose the at least one capillary channel within the at least two capillary membranes; the at least one capillary membranes are housed within the main reactor chamber between the inlet tubing member and outlet tubing member; the at least one capillary membranes are fashioned into parallel rows; the substrate medium fills the at least one capillary channel by traversing through the inlet tubing member; and the substrate medium envelopes the at least two capillary membranes and permeates through the channels of capillaries.
16. The bioreactor of claim 15, further comprising: a pressure source; a pulsation source; an inflow port; an exit port; the pulsation source adjacent to the inflow or outflow; the inlet tubing member traverses through the capillary bed and exits through the exit port to the collection reservoir; the plurality of tubing members are hermetically sealed and fashioned at opposite ends of the main reactor chamber; and the pulsation source is attached adjacent the inflow tubing member, opposite the at least two capillary membranes.
17. The bioreactor of claim 16, wherein the semipermeable membrane comprises, an inert scrim; the inert scrim oriented between each layer of the semipermeable membrane; and the inert scrim being of a net configuration.
18. The bioreactor of claim 17, further comprising: a canister; a cap member; the canister made of a rigid inert material or a more flexible but semi rigid material; the canister housing all the components of the bioreactor device; the canister having an inflow and an outflow; the inflow and outflow regulated by the pressure source which adds predefined pressure values to the at least two capillary membranes to include it or open it for flow; and the cap member is hermetically sealed to the canister.
19. A bioreactor comprising: a main reactor chamber, including a substrate containment space; a semipermeable membrane with a hollow inner lumen side and an external surface side; a sleeve perimetrically enveloped around the main reactor chamber; an electric power supply; a substrate medium; a heating member; a plurality of tubing members; a collection reservoir; a pressure-sealed connecter member; a metabolite purifying filter member; a light source; an agitator; an inert scrim; a pulsation source; the pulsation source alternately expanding and contracting the capillary bed; the semipermeable member is housed within the main reactor chamber within the substrate containment space; the semipermeable membrane is positioned cylindrically around the outlet tubing member in multiple layers with space between the layers; the inert scrim positioned between each layer of the semipermeable membrane; the inert scrim being of a net configuration; the pressure-sealed connecter member hermetically sealed to the main reactor chamber, opposite the heating member and the agitator; the plurality of tubing members includes an inlet tubing member, an outlet tubing member, and a collection tubing member; the plurality of tubing members are hermetically sealed to the pressure-sealed connecter member; the inlet tubing member and outlet tubing member traverse through the pressure-sealed connecter member to enter the substrate containment space; the collection tubing member traverses through the pressure-sealed connecter member to enter the hollow inner lumen space of the semipermeable membrane; the substrate medium fills the substrate containment space by traveling through the inlet tubing member into the substrate containment space; the substrate medium envelopes the external surface side of the semipermeable membrane and permeates through the semipermeable membrane to fill the inner lumen side; the sleeve contains the light source; the light source having a variable wavelength, wherein the wavelength is varied to promote metabolite production by a plurality of microorganisms contained within the hollow lumen space of the semipermeable membrane; the heating member maintains a predefined temperature to maintain a predefined substrate temperature; and the agitator maintains a predefined cyclical rate.
20. The bioreactor of claim 19, further comprising: a canister; a cap member; the canister made of a rigid inert material or a more flexible but semi rigid material; the canister housing all the components of the bioreactor device; the canister having an inflow and an outflow; the inflow and outflow regulated by the pressure source which adds predefined pressure values to the at least two capillary membranes to include it or open it for flow; and the cap member is hermetically sealed to the canister.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0003]
[0004]
[0005]
[0006]
[0007]
[0008]
[0009]
DETAIL DESCRIPTIONS OF THE INVENTION
[0010] All illustrations of the drawings are for the purpose of describing selected versions of the present invention and are not intended to limit the scope of the present invention.
[0011]
[0012] In an alternative embodiment of the present invention a metabolite purifying filter member 21, such as a calcium bicarbonate filter, is attached adjacent the collection tubing member 18. When the gaseous metabolites 8 traverse through the collection tubing member 18 then will pass through the purifying filter member 21 and the desired metabolites will be separated from the precipitate.
[0013] In an alternative embodiment of the present invention a light source 22, such as light-emitting diodes (LED) at an optimal wavelength, is affixed within the sleeve 10. When activated, the light source 22 emits light into the substrate containment space 3 and penetrates the hollow inner lumen side 5 of the semipermeable membrane 9a, 9b, thereby, facilitating metabolite 8 production.
[0014] In another embodiment of the present invention a dial 26 is affixed to the heating member 14 and agitator 23. When the dial 26 is manipulated, it controls the speed of the agitator 23 which operates to mix the microorganisms 11 and substrate medium 13 within the hollow inner lumen side 5 of the semipermeable membrane 9a, 9b.
[0015] The substrate medium 13 is supplied to the main reactor chamber 2 of the bioreactor 1 at a predefined frequency—dependent on various factors such as the type of organism, the size of the bioreactor, volume of substrate medium desired, and pressure in the system—to permeate through the semipermeable membrane 9a, 9b and provide the microorganisms 11 with a consistent influx of necessary substrate medium 13 to facilitate the production of desired metabolites 8 without disrupting or removing the microorganisms 11 from the hollow inner lumen space 5 of the semipermeable membrane 9a, 9b. Further, the heating member 14, positioned opposite the pressure-sealed connecter member 20 and within the hollow inner lumen side 5, maintains a consistent, predefined temperature to maintain a predefined substrate medium temperature. A preferred temperature for the system is 40 F-140 F but can vary dependent on external and internal factors. Further, the agitator 23, positioned opposite the pressure-sealed connecter member 20 and within the hollow inner lumen side 5, maintains a consistent, predefined cyclical rate to maintain a homogenous mixture of substrate medium 13 and microorganism 11 within the hollow inner lumen side 5 of the semipermeable membrane 9a, 9b.
[0016] This device and system allows the microorganisms 11 to be supplied with an enriched substrate medium 13, without disturbing or removing the microorganisms 11 from the main reactor chamber 1, thereby facilitating continuous substrate medium 13 processing and metabolite 8 synthesis by the microorganisms 11 contained in the semipermeable membrane 9a, 9b. The type of microorganisms can vary from light-dependent bacteria like cyanobacteria to various algae. Alternatively, a direct metabolic pathway, such as fermentation, can be used in this system as well.
[0017] Commercial production of hydrogen gas and other metabolites by microorganisms in the device and system described herein is possible. Thus, it is important to note that changing small factors like adding modular fitting in order to chain them in a series does not change or improve upon the spirit of the invention described herein.
[0018]
[0019]
[0020] In this alternative embodiment the plurality of tubing members 15 are hermetically sealed and fashioned at opposite ends of the main reactor chamber 2. The at least two capillary membranes 29 are housed within the capillary bed 36 between the inlet tubing member 16 and outlet tubing member 17. The at least two capillary membranes 29 are fashioned into parallel rows and the substrate medium then fills the at least one capillary channel 30 by traveling through the inlet tubing member 16 via forces exerted from the pulsation source 32, which is hermetically sealed and attached adjacent the inlet tubing member 16, opposite the at least two capillary membranes 29. The substrate medium 13 envelopes the at least two capillary membranes 29 and permeates through the at least one capillary channel 30. The microorganisms 11 within the system are positioned within the substrate medium 13 and the at least two capillary membranes 29 in a homogenous mixture. The unfiltered metabolites 8 produced from the microorganisms traverse through the outlet tubing member and are collected in a collection reservoir 19.
[0021]
[0022]
[0023] The inlet tubing member 16 passes through the capillary bed 36 and exits through an exit port 39 to the collection reservoir 19. The canister 37 is semi-ridged and houses the entire system. The canister has an inflow controlled by a pressure source 40, such as an air pump or vacuum, so that inflow and outflow of the homogenous mixture of microorganism 11 and metabolites 8 can be regulated to add sufficient pressure to the membrane by either including it or opening it for flow.
Unexpected Results
[0024] For the experiment depicted in
[0025] Control:
[0026] In the control group 0.7 g of instant yeast was mixed with 35 ml water in the reactor bottle then placed it on a hotplate set to 60 degrees Celsius. Once the mixture was at temperature, the 3 g of sugar was poured in, and the sensor cap was placed on the bottle with skewers creating a 3 mm space between the cap and bottle so the CO2 could vent out of the bioreactor. Once assembled, logger software was started, and CO2 readings were taken every 4 seconds in units of parts per million. After CO2 production fell to an inactive level, the reaction was stopped.
[0027] Control Group 2 (No Membrane):
[0028] For the second control group, a test was conducted to determine what would happen if the same 0.7 g of yeast was mixed with 30 g of sugar and 350 ml of water, just as in the first experimental group, but without a membrane. This was testing whether mixing the same amount of yeast with more nutrient mix would yield comparable levels of production to the two trials in experimental group 1. 0.7 grams of instant yeast was mixed with 350 ml water in the bioreactor bottle then placed on a hotplate set to 60 degrees Celsius. Once the mixture was at temperature, the 30 g of sugar was poured in, and the sensor cap was placed on the bottle with skewers. Once assembled, the logger software was started, and CO2 readings were taken every 4 seconds in units of parts per million. After the CO2 production fell to an inactive level, the reaction was stopped.
[0029] Group 1:
[0030] In the first experimental group, the membrane was fixed in the reactor with a spacer to raise the reactor opening, so the membrane could hang down without touching the bottom. Another spacer was then used to secure the sensor above the membrane opening. The membrane was fixed in between these two spacers. The membrane tubing was sealed at one end and secured to a 1″ piece of tubing at its opening, which was then passed through 6″×6″ square of layered cellophane wrap so that the membrane could hang in between the two spacers like an eardrum or diaphragm. Once the assembly was made, the reactor bottle was filled with 350 ml of water and mixed in 30 g of table sugar, then placed on a hotplate set to 60 degrees Celsius for the substrate medium to heat up. In the meantime, the membrane was prepared and filled with 35 ml water (like the control), to which the 0.7 g of yeast was mixed in. Once the membrane mixture was prepared, the bioreactor bottle was assembled with spacers and a membrane, and the CO2 sensor was placed at the top. After assembly the logging software was started, which took CO2 readings every 4 seconds, in parts per million. After the CO2 production fell to an inactive level, the reaction was stopped. This experimental group was run twice and is identifiable on
[0031] Findings and Discussion:
[0032] As
[0033] The second control group was prepared like the first control group but with 350 ml of water and 30 g of sugar instead. The second control group was intended to demonstrate that even with more nutrient mix, the 0.7 g of yeast would be no more productive than the control. Ultimately, it was determined that the second control group could not compete with the first control group. The trial never reached a production level higher than 3050 ppm and was recorded to have stopped at just over 20,000 seconds, though it probably could have been stopped sooner.
[0034] The experimental group had two trials. In the first trial, the production curve started out flat because, when mixed, the yeast and water in the membrane did not have any added sugars. For the reaction to take place, enough sugars would need to enter the inner membrane space to cross the reaction's activation threshold, meaning that any significant CO2 production would clearly indicate the membrane allowed sugars to permeate and mix in with the yeast. After the activation threshold was crossed at approximately 4,500 seconds, there was a swift climb up to nearly 23,000 ppm. The production levels stayed in a range of 22,000 ppm to 25,000 ppm during the peak of the reaction, which was nearly 50,000 seconds. Slowly, the productivity began to decrease, and the reaction was recorded to have ended at 68,000 seconds. The experiment was repeated for the second trial, however the same membrane was used for the second trial that was used in the first trial, to show system resiliency. Surprisingly, the second trial did not have the same initial period of low to no productivity and ran for a longer duration than the first trial. There was an almost immediate rise to nearly 40,000 ppm from where it dropped to a constant production level right above 22,000 ppm that it carried for about 76,000 seconds, at which point the productivity fell off and the reaction was recorded to have ended at just under 85,000 seconds. The two trials both ran for about four times the duration as the control group and demonstrated how the bioreactor with a membrane can create an environment for microorganisms that allows for continuous metabolism.
[0035] One question raised by these experimental results was: why are the first experimental group curves so much lower than the first control group? Right away the area under the curves can make it appear that the control group may have a greater or comparable total CO2 production to the first experimental group. To investigate this, the fluid left over in the container from the second trial of the first experimental group was collected and distilled to separate the water and ethanol from the remaining sugars and any other particulates. After distilling, 330 ml ethanol-water solution was left. This distilled liquid was then taken and tested using an ethanol by volume meter. With this meter, it was determined that the second trial of the first experimental group produced a solution that was 3.5% ethanol by volume, meaning approximately 11.55 ml of pure ethanol was produced. Now that the amount of ethanol produced was determined, some metabolic calculations were performed to extrapolate how much sugar was consumed. The calculations revealed that the second trial was able to consume nearly 19 g of sugar, which is over six times the maximum amount of sugar the 0.7 g of yeast could have consumed in the control, and many times greater than the sugar consumed in the second experimental group. These findings were unexpected and showed that a bioreactor with a membrane facilitated increased nutrient consumption by yeast as opposed to a bioreactor without the membrane.
CONCLUSION
[0036] As shown by both the production curves and the metabolic calculations in
[0037] Although the invention has been explained in relation to its preferred embodiment, it is to be understood that many other possible modifications and variations obvious to anyone familiar with the art can be made without departing from the spirit and scope of the invention.