WO2014085869A1 - Bioréacteur et procédé d'utilisation - Google Patents

Bioréacteur et procédé d'utilisation Download PDF

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Publication number
WO2014085869A1
WO2014085869A1 PCT/AU2013/001424 AU2013001424W WO2014085869A1 WO 2014085869 A1 WO2014085869 A1 WO 2014085869A1 AU 2013001424 W AU2013001424 W AU 2013001424W WO 2014085869 A1 WO2014085869 A1 WO 2014085869A1
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WO
WIPO (PCT)
Prior art keywords
liquid permeable
bioreactor
microorganisms
permeable layer
fluid
Prior art date
Application number
PCT/AU2013/001424
Other languages
English (en)
Inventor
Florian BERNER
Kirsten Heimann
Madoc SHEEHAN
Original Assignee
James Cook University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from AU2012905377A external-priority patent/AU2012905377A0/en
Application filed by James Cook University filed Critical James Cook University
Publication of WO2014085869A1 publication Critical patent/WO2014085869A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/02—Membranes; Filters
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/02—Photobioreactors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/04—Flat or tray type, drawers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/50—Means for positioning or orientating the apparatus
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • Y—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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W10/00—Technologies for wastewater treatment
    • Y02W10/30—Wastewater or sewage treatment systems using renewable energies
    • Y02W10/37—Wastewater or sewage treatment systems using renewable energies using solar energy

Definitions

  • FIELD OF THE INVENTION relates generally to a bioreactor for cultivating microorganisms.
  • the invention relates to a bioreactor, method and system for cultivating photosynthetic microorganisms.
  • Culture systems for microorganisms are generally classified according to their engineering and hydraulic characteristics in open systems, examples of which include ponds, deep channel and shallow circulating units and closed or fully hydraulic systems commonly called bioreactors or photo-bioreactors (if cultured organisms are capable of using light as an energy source).
  • bioreactors or photo-bioreactors (if cultured organisms are capable of using light as an energy source).
  • photosynthetic microorganisms are capable of growing both in suspension cultures and as a biofilm attached to a surface (examples of which include benthic microorganisms such as algae, microalgae and cyanobacteria).
  • algae and other benthic microorganisms are grown as a biofilm attached to a cultivating surface, the biomass is naturally concentrated (containing less water) and more easily harvested, leading to more direct removal of the algal biomass and reduced processing.
  • the cultivation of algae is currently being considered for a number of different applications, including: removal of C0 2 or other gases from industrial flue gases by algae bio-fixation; the reduction of Green House Gas (GHG) emissions from a company or process while producing biodiesel; wastewater treatment through the removal of unwanted compounds; the production of biomass for processing into ethanol and methane, the production of hydrogen gas, livestock feed, use as organic fertilizer due to its high N:P ratio, energy cogeneration (electricity and heat); and the extraction of compounds including a large range of fine chemicals and bulk products, such as fats, polyunsaturated fatty acids, oil, natural dyes, sugars, pigments, antioxidants and high-value bioactive compounds.
  • cultivatable microorganisms include but are not limited to methanotrophic bacteria for the remediation of methane, a GHG with a warming potential 23 times that of C0 2 over a 100 year period and saprophytic fungi (for the treatment of problematic waste waters with a high load of complex and often chemically inert organic materials).
  • the biomass of these organisms can be utilised as described above for algae and microalgae.
  • microorganisms in particular phototrophic microorganisms, could potentially revolutionise a large number of biotechnology areas including biofuels, cosmetics, pharmaceuticals, nutrition and food additives, aquaculture, and pollution prevention.
  • an improved bioreactor, method and system for cultivating microorganisms in particular photosynthetic microorganisms, including algae and microalgae, is required.
  • an aim of the present invention to provide an improved bioreactor, method and/or system for cultivating microorganisms, in particular photosynthetic microorganisms.
  • the present invention has arisen, after the inventors discovered a novel bioreactor system for the cultivation of microorganisms.
  • the system provides a bioreactor and method for generating high density cultivated biomass with an increased productivity and reduced harvesting time when compared to existing technologies.
  • the present invention is broadly directed to a bioreactor and/or a method and system for cultivating microorganisms, preferably photosynthetic microorganisms.
  • a bioreactor comprising:
  • At least one or a plurality of liquid permeable layers comprising a first surface and a second surface
  • microorganisms are cultivatable on the first surface of the liquid permeable layer and the second surface is connectable in fluid communication with a fluid source; and wherein the liquid permeable layer is rotatable.
  • a method for cultivating microorganisms comprising:
  • a system for cultivating microorganisms comprising:
  • liquid permeable layers comprising a first surface comprising microorganisms and a second surface in fluid communication with a fluid source;
  • a rotation member for rotating the liquid permeable layer, to facilitate microorganism growth.
  • the liquid permeable layer is rotated or rotatable to provide the microorganisms being cultured with sufficient conditions for maximum growth, such as for example, light and C0 2 .
  • the bioreactor further comprises at least one rotation member.
  • the rotation of the at least one liquid permeable layer is facilitated by said rotation member.
  • the rotation member may be manually operated and/or may be automated.
  • the rotation member may be manually operated by a handle or may be automated by a motor, or by wind, solar and/or water powered apparatus, or a combination of the above.
  • the at least one liquid permeable layer may be rotated or rotatable by a handle.
  • the at least one liquid permeable layer may be rotated or rotatable by a motor.
  • the at least one liquid permeable layer may be rotated or rotatable by wind, solar and/or water powered apparatus.
  • the at least one liquid permeable layer is rotated or rotatable continuously.
  • the at least one liquid permeable layer is rotated or rotatable intermittently.
  • the rotation member rotates a platform comprising said at least one liquid permeable layer.
  • said at least one liquid permeable layer is rotated or rotatable about an axis. More preferably said at least one liquid permeable layer is rotated or rotatable about a central axis of the liquid permeable layer or the bioreactor.
  • rotation provides light exposure to a substantial portion of the first surface of the at least one liquid permeable layer.
  • the bioreactor comprises a single liquid permeable layer.
  • the single liquid permeable layer may be cultivated with microorganisms.
  • the single liquid permeable layer may be cultivated with photosynthetic microorganisms.
  • the bioreactor comprises a plurality of liquid permeable layers.
  • the plurality of liquid permeable layers may be cultivated with microorganisms.
  • the plurality of liquid permeable layers may be cultivated with photosynthetic microorganisms.
  • the liquid permeable layers may be provided in a double layer, between which a fluid source is located.
  • the liquid permeable layers may comprise a first surface upon which microorganisms may be cultured to form a biofilm and a second surface connectable in fluid communication with the fluid source.
  • the same fluid source supplies fluid and nutrients to both liquid permeable layers.
  • a separate fluid source for each liquid permeable layer may be provided.
  • the liquid permeable layer is configured as a sheet.
  • the sheet may be of any size or shape, according to the culturing requirements of the microorganisms. Shapes contemplated by the invention, include without limitation, cylindrical, octahedral, tetrahedral etc.
  • the liquid permeable layer is substantially cylindrical. The liquid permeable layer acts like a membrane allowing fluid to pass from the second surface of the liquid permeable layer through to the first surface of the liquid permeable layer, upon which microorganisms are capable of being cultured.
  • the liquid permeable layer does not allow the passage of microorganisms from the first surface of the liquid permeable layer to the second surface of the liquid permeable layer, thereby reducing the risk of contamination.
  • the liquid permeable layer may be formed from porous and/or perforated material.
  • the liquid permeable layer is formed from porous and/or perforated material that allows the selective passing of fluid.
  • materials may include but are not limited to any woven, knitted, pleated, printed (e.g., by a 3D printer), felted or otherwise cross-linked synthetic or natural material (e.g., cotton, wool, silk, tree fibres, celluloses, including differing grades of paper (e.g., -acetate, -sulfonate, nitro- ), chitins (e.g., sponges)) or synthetic polymers (e.g., polyesters, artificial sponges, polyacrylates, polyesters, polyamines, polysulfone, polyamides) or specifically nanotechnologically engineered 3-D nano-fibres of either natural or synthetic origin, with or without surface modifications, (e.g., bonded enzymes, adhesive proteins etc.); polystyrene, silicon, nitrocellulose, cellulose acetate, glass fibre, polycarbonate, polyethylene, ceramics, glass
  • the fluid source may comprise any one or more of the following, without limitation: water; nutrients; water and nutrients; growth media for microorganism cultivation; waste waters and/or secondary treated sewage.
  • the waste water and/or secondary treated sewage may be treated and/or filtered by the microorganisms.
  • the fluid source is provided in the form of a fluid conducting layer.
  • the fluid conducting layer is in fluid communication with the second surface of the liquid permeable layer.
  • the fluid conducting layer is configured to provide fluid to a substantial portion of the liquid permeable layer.
  • the fluid conducting layer comprises a fluid conducting material, examples of which include but are not limited to: fabric; foam; glass fibres; synthetic polymers (e.g., polyesters, polyacrylate, polyamine, polyamide, artificial sponge etc); natural fibres (e.g., cotton, wool, sponge, hemp, tree fibre etc); 3D materials (e.g., meshed nano-fibres, spun or printed 3D matrixes) or a combination thereof.
  • the fluid conducting material comprises fabric.
  • the fabric maybe woven, knitted, a felt, mesh, cross-linked, or a combination thereof. More preferably, the fluid conducting material comprises capillary matting.
  • the fluid conducting material provides at least fluid and/or nutrients and/or support to the growing microorganisms.
  • the bioreactor further comprises a fluid reservoir.
  • the fluid reservoir supplies the fluid source with fluid and optionally nutrients.
  • the fluid source may be transported from the fluid reservoir to the second surface of the liquid permeable layer.
  • the bioreactor further comprises a pump.
  • fluid may be pumped from the fluid reservoir to the second surface of the liquid permeable layer. More preferably, fluid may be pumped from the fluid reservoir to the fluid conducting layer.
  • fluid transport include without limitation, capillary action, gravity, rotational forces, etc.
  • the bioreactor further comprises a light source.
  • the light source provides light to a substantial portion of the first surface of the liquid permeable layer.
  • the light source provides continuous light.
  • the light source provides intermittent light.
  • the light source may provide a combination of both continuous and intermittent light.
  • the light requirements may depend on the requirements of the microorganisms.
  • the light source may be provided by natural light (e.g., sunlight) or may be provided by an artificial source such as a lamp.
  • the bioreactor is contained within a housing.
  • the housing allows light to penetrate through to the culturing microorganisms. More preferably, the housing is transparent.
  • the bioreactor further comprises a gas source for the supply of C0 2 or other gases to the liquid permeable layer (e.g., methane).
  • a gas source for the supply of C0 2 or other gases to the liquid permeable layer (e.g., methane).
  • the gas requirements may depend on the requirements of the microorganisms for facilitating growth.
  • the present invention is applicable to all microorganisms.
  • the microorganisms are photosynthetic microorganisms. More preferably, the microorganisms are algae, microalgae or cyanobacteria.
  • the microorganisms are capable of forming a biofilm, examples of which include, without limitation: photosynthetic microorganisms (e.g., algae, microalgae and blue algae); methanotrophic microorganisms; yeast; benthic microorganisms and combinations thereof.
  • photosynthetic microorganisms e.g., algae, microalgae and blue algae
  • methanotrophic microorganisms e.g., yeast, benthic microorganisms and combinations thereof.
  • the photosynthetic microorganisms include cyanobacteria, the Rhodophyta (red algae), the Chlorophyta (green algae), Dinophyta, Chrysophyta (golden-brown algae), Prymnesiophyta (haptophyta), Bacillariophyta (diatoms), Xanthophyta, Eustigatophya, Rhaphidophyta and Phaeophyta (brown algae) or a combination thereof.
  • Non-limiting examples of photosynthetic microorganisms that form biofilms include: Scenedesmus obliquus (Chlorophyta), Isocrysis glabana (haptophyta), Nannochloropsis sp. (Eustigatophya), Tetraselmis Suecica (Chlorophyta), Phaeodactylum tricomutum (Bacillariophyta), Botryococcus braunii (Chlorophyta) and Spirulina (Cyanobacteria).
  • the microorganisms may be harvested from the liquid permeable layer.
  • Harvesting may occur through loosening the microorganisms from the liquid permeable layer with a variety of means, examples of which include, although are not limited to physical scraping, fluid washing, chemical treatment, agitation, mechanical manipulation and/or drying.
  • Harvesting may be undertaken with the liquid permeable layer in place, or after removal of the liquid permeable layer.
  • the microorganisms are harvested from the liquid permeable layer when the desired level of biomass is cultivated.
  • the bioreactor, method and system for cultivating microorganisms of the invention may be used for the production of biomass, biofuels, animal feed, waste water remediation, the production of high value compounds (e.g., oils), C0 2 remediation and/or the production of fertilizer or a combination thereof.
  • high value compounds e.g., oils
  • indefinite articles “a” and “an” may refer to one entity or a plurality of entities and are not to be read or understood as being limited to a single entity.
  • FIG. 1 shows a perspective view of the bioreactor according to an embodiment.
  • FIG. 2 shows a cross section view of a portion of the bioreactor according to an embodiment.
  • FIG. 3 shows a cross-sectional view of the bioreactor, according to an embodiment.
  • FIG. 4 shows a cross-sectional view of the bioreactor according to an embodiment.
  • FIG. 5 shows a perspective view of the bioreactor according to two different embodiments.
  • FIG. 6 shows both a side view and top view of six alternate embodiments of the bioreactor (labeled I- VI).
  • FIG. 7 shows a perspective view of the bioreactor as shown in embodiment V of FIG. 6.
  • FIG. 8 shows an example of a bioreactor arrangement as shown in embodiment IV of FIG. 6.
  • FIG. 9 shows a dense bio film of algae growing on a first surface of a vertical liquid permeable layer.
  • FIG. 10 shows the harvesting process of three different microalgal cultivation systems, including an algal culture grown on a liquid permeable layer; a conventional laboratory algal liquid suspension culture; and a further algal liquid suspension culture sourced from a different location, as discussed in the Examples.
  • the present invention resides in a method and/or system for cultivating microorganisms, preferably photosynthetic and other benthic microorganisms and a bioreactor comprising at least one liquid permeable layer, comprising a first surface and a second surface, wherein photosynthetic microorganisms are cultivatable on the first surface of the liquid permeable layer and the second surface is connectable in fluid communication with a fluid source, and wherein the liquid permeable layer(s) is rotatable.
  • microorganisms preferably photosynthetic and other benthic microorganisms and a bioreactor comprising at least one liquid permeable layer, comprising a first surface and a second surface, wherein photosynthetic microorganisms are cultivatable on the first surface of the liquid permeable layer and the second surface is connectable in fluid communication with a fluid source, and wherein the liquid permeable layer(s) is rotatable.
  • the bioreactor may further comprise a rotation member, the rotation member operating to rotate the liquid permeable layer(s).
  • bioreactor 100 comprises a liquid permeable layer 110 comprising a first surface 1 1 1 and a second surface 1 12, which in the embodiment shown in FIG. 1, the liquid permeable layer 110 is cylindrical and in a vertical orientation.
  • the first surface 1 1 1 of the liquid permeable layer 1 10 is shown to cultivate photosynthetic microorganisms 150, such as algae inclusive of micro algae.
  • the bioreactor 100 of FIG. 1 is mounted on a stand 140 with a rotation member 130, which when in use, in the embodiment shown facilitates the rotation of the liquid permeable layer 110 vertically about a central axis 180.
  • the liquid permeable layer 110 of the bioreactor 100 of FIG. 1 is supplied with water and nutrients from a fluid reservoir 160.
  • Liquid permeable layer 1 10 may be constructed from any materials, either naturally porous or engineered to be porous, and/or perforated, for allowing fluid to permeate and facilitating adherence of the photosynthetic microorganisms 150 to be cultivated.
  • the bioreactor 100 shown in FIG. 1, in one embodiment is situated in a housing 200, which allows light to penetrate through to the culturing photosynthetic microorganisms 150.
  • FIG. 2 shows a cross section view of a portion of a bioreactor.
  • the liquid permeable layer 110 is shown to comprise a first surface 111 upon which photosynthetic microorganisms 150 are cultivated as a biofilm 151 and a second surface 112.
  • the second surface 112 of the liquid permeable layer 110 is shown to be in fluid communication with a fluid source 120.
  • the fluid source 120 comprises a fluid conducting layer 121. It will be appreciated that fluid conducting material may be used to provide fluid and nutrients to the photosynthetic microorganisms 150.
  • the photosynthetic microorganisms 150 are constantly supplied with water and nutrients via the liquid permeable layer 110 while also being provided with light and C0 2 .
  • FIG. 3 shows a cross sectional view of one embodiment of the bioreactor 100.
  • the inventors as described in the examples, cultivated photosynthetic microorganisms on two static liquid permeable layers either in full light or limited light to determine whether the level of light had an effect on the level of cultivated biomass.
  • the liquid permeable layers were static during the experiments, although are capable of being rotated.
  • the bioreactor 100 may be static as described above or may be rotated.
  • the bioreactor 100 is arranged vertically on a stand 140, and comprises two liquid permeable layers 110, between which a fluid source 120 is located and wherein the liquid permeable layers 110 comprise a first surface 1 1 1 wherein photosynthetic microorganisms 150 are cultured to form a biofilm 151 and a second surface 1 12 in fluid communication with the fluid source 120.
  • the same fluid source 120 supplies fluid and nutrients to both liquid permeable layers 110 and the fluid source is shown to comprise a fluid conducting layer 121.
  • the fluid source can be seen in FIG. 3 to be supplied with water and nutrients from a fluid reservoir 160, wherein, when in use, the water and nutrients are pumped via pump 170 to the fluid conducting layer 121.
  • a separate fluid source 120 for each liquid permeable layer 110 is also contemplated. It will also be appreciated that varying the growth conditions of photosynthetic microorganisms 150, and in particular algae, will vary the rate of culturing and the final biomass concentration. As further shown in FIG. 3, photosynthetic microorganisms 150 provided with limited light grown on a liquid permeable layer 1 10 of the bioreactor 100 results in a reduced culturing rate and a reduced overall biomass concentration when compared with photosynthetic microorganisms 150 provided with full light grown on a liquid permeable layer 1 10 of the bioreactor 100.
  • the light source 190 as shown in FIG. 3 may be natural light (e.g., the sun) or may be provided by an artificial source.
  • FIG. 4 shows a cross section view of a bioreactor 100, in use, wherein the photosynthetic microorganisms 150, are shown as a uniform layer cultured on the first surface 111 of liquid permeable layer 1 10, wherein the liquid permeable layer 1 10 has been configured as a cylinder and mounted on a stand 140 in a vertical orientation.
  • the stand 140 comprises a rotation member 130 which in the embodiment shown in FIG. 4 is releasably coupled or connected to a motor 210 which facilitates rotation.
  • the stand 140 upon which the bioreactor 100 shown in FIG. 4 is positioned, is constructed such that the substantially cylindrical liquid permeable layer 110 rotates about a central axis 180.
  • the second surface 1 12 of the liquid permeable layer 110 as shown in FIG.
  • a fluid source 120 which is shown to comprise a fluid conducting layer 121.
  • the fluid conducting layer 121 can be seen in FIG. 4 to be supplied with water and nutrients from a fluid reservoir 160, wherein, when in use, the water and nutrients are pumped via a pump 170 to the fluid conducting layer 120.
  • the bioreactor 100 shown in FIG. 4 is rotated about a central axis 180 and the first surface 1 11 of the liquid permeable layer 1 10 upon which the photosynthetic microorganisms 150 are cultured is exposed to full light and limited light intermittently as shown by the light sources 190.
  • FIG. 5 shows a perspective view of the bioreactor 100 according to two different embodiments. It will be appreciated that the positioning of the liquid permeable layer 1 10, upon which the photosynthetic microorganisms 150 are cultured relative to the light source 190 and/or any further required growth conditions may be varied to enhance the culturing of the specific photosynthetic microorganisms 150.
  • the liquid permeable layer 110 is cylindrical and mounted on a stand 140 in a vertical orientation.
  • the stand 140 comprises a rotation member 130 which in the embodiment shown in FIG. 5 is releasably coupled or connected to a motor 210 which facilitates rotation.
  • the stand 140 pon which the bioreactor 100 shown in FIG.
  • the liquid permeable layer 110 in FIG. 5A is in fluid communication with a fluid source.
  • the fluid source in FIG. 5A is supplied with water and nutrients from a fluid reservoir 160, wherein, when in use, the water and nutrients are pumped via a pump 170 to the fluid source.
  • the cylindrical liquid permeable layer 110 is positioned at an angle to the light source 190 to ensure the greatest exposure of the photosynthetic microorganisms 150 cultured on the liquid permeable later 1 10 to the light source 190.
  • the liquid permeable layer 110 is in fluid communication with a fluid source.
  • the fluid source in FIG. 5B is supplied with water and nutrients from a fluid reservoir 160, wherein, when in use, the water and nutrients are pumped via a pump 170 to the liquid permeable layer 110.
  • the bioreactor 100 is situated in a housing 200, which allows light to penetrate through to the cultivating photosynthetic microorganisms 150.
  • FIG. 6 shows both a side view and top view of six alternate embodiments of the bioreactor 100 (labeled I- VI), wherein the liquid permeable layer 110 is configured to comprise different structures arranged around a central axis 180 and when in use, rotated about that axis 180.
  • Embodiment I of FIG.6 comprises at least one liquid permeable layer 1 10 as a panel, wherein, either one or both sides of the panel comprise a first surface of the liquid penneable layer 1 10 for culturing photosynthetic microorganisms.
  • Embodiment II of FIG. 6 comprises one liquid permeable layer 110 as a cylinder.
  • Embodiment III of FIG. 6 comprises at least one liquid permeable layer 110 as an octagon.
  • Embodiment IV of FIG. 6 comprises at least one liquid permeable layer 1 10 as two panels forming a cross.
  • Embodiment V of FIG.6 comprises at least one liquid permeable layer 110 as three intersecting panels forming a stellate structure and
  • embodiment VI of FIG. 6 comprises at least one liquid permeable layer 110 configured as four intersecting panels forming a stellate structure.
  • Each of the six different embodiments in use would be rotated around a central axis 180 to ensure maximum culturing conditions.
  • FIG. 7 shows a perspective view of an embodiment of the bioreactor 100 as shown in embodiment V of FIG. 6.
  • there are two liquid permeable layers 110 comprising a first surface 11 1 wherein photosynthetic microorganisms 150 are cultured to form a biofilm 151 and a second surface 112 in fluid communication with a fluid source 120 which is shown to comprise a fluid conducting layer 121 provided on each separately spaced panel 220.
  • a separate fluid conducting layer 121 supplies fluid and nutrients to the liquid permeable layers 1 10.
  • the same fluid conducting layer 121 for each liquid permeable layer 1 10 is also contemplated.
  • Fluid source 120 is supplied with water and nutrients via a pump 170 from fluid reservoir 160 and situated on stand 140.
  • FIG. 8 shows an example of a bioreactor 100 as shown in embodiment IV of FIG. 6, which rotates about a central axis, via rotation member 130, which in this particular embodiment is releasably coupled or connected to a motor 210, which facilitates rotation and acts as a stand 140.
  • FIG. 9 shows a dense biofilm of photosynthetic microorganisms 150, specifically a proprietary mix of freshwater algae, growing on a first surface 111 of a vertical liquid permeable layer 110, as cultivated in the experiments undertaken by the inventors.
  • Example 1 is a non-limiting example of a bioreactor.
  • Proprietary mixed freshwater algae culture was successfully grown on two vertical 50cm x 13cm liquid permeable layers (FIG. 3 and FIG. 10), A and B, constantly supplied with nutrients and water from a fluid source through the liquid permeable layers.
  • the vertical liquid permeable layers (A and B) did not rotate.
  • the first surface of liquid permeable layer A was exposed to artificial light 12 hours a day and the first surface of liquid permeable layer B was shaded.
  • a conventional suspended culture in a 2 Litre Schott-bottle was grown in parallel to allow for direct comparison.
  • liquid permeable layers were formed from unprinted newspaper and the fluid source was fibre capillary matting fed with water and nutrients from a fluid reservoir.
  • the algae culture was cultivated for 28 days, using LI medium. Nitrate and phosphate consumption were monitored daily and both nutrients replenished as necessary. In addition, the culture medium for the algae biofilm culture was completely replaced after 7 and 14 days.
  • the biofilm culture grown on liquid permeable layer A grew the most during the experiment on the top half of the first surface and less well on the bottom half and on the first side of liquid permeable layer B, due to shading.
  • both biofilms from the first and second liquid permeable layers A and B, in full light and shade were harvested and analysed for ash-free biomass, Fatty Acid Methyl Ester content (FAME) profile and carbohydrate content.
  • FAME Fatty Acid Methyl Ester content
  • the biofilms from liquid permeable layers A and B were harvested by scraping the algae off the liquid permeable layers with pre-weighted glass microscope slides.
  • Ash-free dry weight was measured directly on the slides and for biochemical profiling, the biofilm was washed off the slides with deionising water.
  • the suspended algal culture was harvested by centrifugation of 40 ml of culture in a laboratory centrifuge and then transferred into glass beakers for dry weight determination.
  • 400 ml of culture was centrifuged and collected in several steps. Both cultures were then freeze-dried and analyzed.
  • the biomass density/total solid content of the cultures could be calculated: 13.60% for the light exposed first surface of liquid permeable layer A, 14.49% for the shaded first surface of liquid permeable layer B and 0.048% for the suspended culture.
  • the biofilm was 283 and 302 times more concentrated, depending on the cultivation side. On average, adjusted for total biomass produced on each surface, the biofilm has a total solid content of 13.64% equivalent to a concentration factor of 284 compared to the suspended culture.
  • the biochemical analysis showed that the biofilm had a higher lipid content than the suspended culture: 21.09% for the biofilm cultured on the light exposed first surface of liquid permeable layer A and 19.54% for the biofilm cultured on the shaded first surface of liquid permeable layer B, compared to 12.54% for the suspended culture.
  • FAME content and carbohydrate content was only measured for the biofilm on the light exposed first surface of liquid permeable layer A and the suspended culture, as there was not enough biomass on the shaded first surface of liquid permeable layer B.
  • FAME content was 9.45% (44.9% of total lipid) for the biofilm vs. 5.30% (42.3% of total lipid) for the suspended culture.
  • Carbohydrate content was 46.01% for the biofilm versus 41.96% for the suspended culture.
  • biomass productivity and quality were equal to or better than that of a suspended algae culture under the same conditions.
  • the biomass was about 300 times more concentrated during cultivation on a liquid permeable layer and could be easily harvested by scraping the algae off the surface. As determined, productivity clearly differed when the algae growing on the first surface of the liquid permeable layer was provided with differing amounts of light.
  • the present invention provides an improved bioreactor, method and/or system for culturing microorganisms, preferably photosynthetic microorganisms.
  • the bioreactor, method and system disclosed herein advantageously exhibits increased and uniform microorganism growth and productivity for generating high density cultivated biomass, specifically photosynthetic microorganisms, such as algae, as well as reduced contamination of the microorganisms and reduced harvesting time.

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  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
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Abstract

La présente invention concerne un bioréacteur amélioré, un procédé et un système de culture de microorganismes, en particulier de microorganismes photosynthétiques. Ledit bioréacteur comprend au moins une ou une pluralité de couches perméables au liquide comportant une première surface et une seconde surface. Les microorganismes peuvent être cultivés sur la première surface de la couche perméable au liquide, la seconde surface peut être en communication fluidique avec une source de fluide, et la couche perméable au liquide peut pivoter.
PCT/AU2013/001424 2012-12-07 2013-12-06 Bioréacteur et procédé d'utilisation WO2014085869A1 (fr)

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AU2012905377 2012-12-07
AU2012905377A AU2012905377A0 (en) 2012-12-07 Bioreactor and method of use

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WO2014085869A1 true WO2014085869A1 (fr) 2014-06-12

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107502545A (zh) * 2017-10-12 2017-12-22 重庆工商大学 一种玻璃纤维固定化光合细菌连续产氢方法及装置
WO2020161519A1 (fr) 2019-02-05 2020-08-13 Solaga Ug Bioréacteur pour des biofilms stationnaires composés de micro-organismes photosynthétiquement actifs
US20220259533A1 (en) * 2019-07-22 2022-08-18 Auburn University Culture systems and methods of using same
DE202019005811U1 (de) 2019-02-05 2022-08-18 Solaga UG (haftungsbeschränkt) Bioreaktor für stationäre Biofilme aus photosynthetisch aktiven Mikroorganismen

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4861725A (en) * 1988-03-29 1989-08-29 Liau Ming Y Mammalian cell culture apparatus
US5376548A (en) * 1990-06-29 1994-12-27 Fuji Photo Film Co., Ltd. Bioreactor apparatus
US5403742A (en) * 1993-09-01 1995-04-04 Ramot University Authority Ltd. Bioreactor for production of products with immobilized biofilm
US20090017541A1 (en) * 2004-08-17 2009-01-15 Kyushu Institute Of Technology Porous Sheet-Form Material For Cell Culture, And Bioreactor And Culturing Method Utilizing Same
CN101735948A (zh) * 2010-01-11 2010-06-16 新奥科技发展有限公司 一种微藻培养方法及装置

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4861725A (en) * 1988-03-29 1989-08-29 Liau Ming Y Mammalian cell culture apparatus
US5376548A (en) * 1990-06-29 1994-12-27 Fuji Photo Film Co., Ltd. Bioreactor apparatus
US5403742A (en) * 1993-09-01 1995-04-04 Ramot University Authority Ltd. Bioreactor for production of products with immobilized biofilm
US20090017541A1 (en) * 2004-08-17 2009-01-15 Kyushu Institute Of Technology Porous Sheet-Form Material For Cell Culture, And Bioreactor And Culturing Method Utilizing Same
CN101735948A (zh) * 2010-01-11 2010-06-16 新奥科技发展有限公司 一种微藻培养方法及装置

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107502545A (zh) * 2017-10-12 2017-12-22 重庆工商大学 一种玻璃纤维固定化光合细菌连续产氢方法及装置
WO2020161519A1 (fr) 2019-02-05 2020-08-13 Solaga Ug Bioréacteur pour des biofilms stationnaires composés de micro-organismes photosynthétiquement actifs
DE202019005811U1 (de) 2019-02-05 2022-08-18 Solaga UG (haftungsbeschränkt) Bioreaktor für stationäre Biofilme aus photosynthetisch aktiven Mikroorganismen
US20220259533A1 (en) * 2019-07-22 2022-08-18 Auburn University Culture systems and methods of using same
US12024695B2 (en) * 2019-07-22 2024-07-02 Auburn University Culture systems and methods of using same

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