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WO2013048543A1 - Systèmes photobioréacteurs et procédés de culture d'organismes photosynthétiques - Google Patents

Systèmes photobioréacteurs et procédés de culture d'organismes photosynthétiques Download PDF

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Publication number
WO2013048543A1
WO2013048543A1 PCT/US2011/064599 US2011064599W WO2013048543A1 WO 2013048543 A1 WO2013048543 A1 WO 2013048543A1 US 2011064599 W US2011064599 W US 2011064599W WO 2013048543 A1 WO2013048543 A1 WO 2013048543A1
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WIPO (PCT)
Prior art keywords
photobioreactor
plate
organism
culture
photosynthetic
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Ceased
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PCT/US2011/064599
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English (en)
Inventor
C. Mark TANG
Fan Lu
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CHLOR BIOENERGY Inc
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CHLOR BIOENERGY Inc
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Publication of WO2013048543A1 publication Critical patent/WO2013048543A1/fr
Anticipated expiration legal-status Critical
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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G33/00—Cultivation of seaweed or algae
    • 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
    • C12M23/22—Transparent or translucent parts
    • 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
    • C12M23/58—Reaction vessels connected in series or in parallel
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6463—Glycerides obtained from glyceride producing microorganisms, e.g. single cell oil

Definitions

  • This invention relates to photobioreactors for cultivation of oil or lipid-enriched photosynthetic organisms and methods of cultivating the organisms, in particular algae, useful as feedstock for biodiesel production and for manufacture of, among others, valuable fine chemicals, nutraceuticals, and animal feed byproducts.
  • Microalgal culture systems can be roughly classified into two categories: open raceways (or open ponds) and enclosed photobioreactors (Richmond, J. Appl. PhycoL, 4: 281-286 (1992); Chaumont, J. Appl. PhycoL, 5: 593-604 (1993); Tredici, Handbook of Microalgal Culture: Biotechnology and Applied Phycology, pp. 178-214 (2004)).
  • Open raceways are made of shallow circular channel of 1,000 to 5,000 m either with clayed surface or lined with plastic sheets. Culture mixing is often provided by paddle wheels to affect circulation of the culture suspension around the raceway.
  • One of the major advantages of open raceways is that they are relatively cheap to build and easy to maintain.
  • this cultivation mode suffers from low yield of algae (5-10 g m - " 2 d - " 1 of dry weight) and unstable quality of algal biomass, due mainly to lack of temperature control, long light paths (ca. 15 - 30 cm in depth) and poor mixing. Exposure of the culture suspension directly to the atmosphere also makes the culture system highly vulnerable to airborne contamination.
  • closed photobioreactors can have considerably reduced culture depth or increased ratios of illuminated culture surface to the culture volume.
  • closed photobioreactors can be subdivided into four major groups: 1) Horizontal tubular photobioreactors; 2) Vertical column photobioreactors; 3) "Bio-coil” tubular photobioreactors and 4) Vertical or inclined flat-plate photobioreactors. (Borowitzka, J. BiotechnoL, 70: 313-321 (1999); Tredichi, Handbook of Microalgal Culture: Biotechnology and Applied Phycology, pp. 178-214 (2004)).
  • Photobioreactor system design and optimization are key R&D issues that often determine the success of the mass cultivation of microalgae.
  • an inherent drawback of tubular photobioreactors is that they can cause cell damage or mortality due to hydrodynamic stress, or shearing force, created by lipid pumps.
  • Shilva et al. conducted a thorough study of Dunaliella mortality affected by pump-related hydrodynamic stress (J. Chem. Tech. BiotechnoL, 40: 253-264(1987)).
  • Gudin and Chaumont also observed that significant cell fragility occurred in Haematococcus culture maintained in a large-scale tubular photobioreactor ⁇ Bioresour. TechnoL, 38, 145-151 (1991)).
  • Culture mixing and circulation in a closed photobioreactor is usually provided by either mechanical liquid pumps or aeration.
  • Doucha et al. (1999) reported an inclined thin-layer cultivation system on which algal suspension form a thin layer of 5-18 mm flowing through the inclined cultivation area. The algal suspension is re-circulated by a continuously operating water pump.
  • Our preliminary study indicated that nearly 20% of the cell wall-deficient mutant cells were damaged by a volumetric pump used in a 40-liter bench-top tubular photobioreactor. It can be particularly difficult to achieve a consistent high yield of cultured biomass when operating under space and/or financial constraints. Therefore, more efficient photobioreactors and methods of using them to cultivate biomass feedstock for biofuel production are still in high demand.
  • the current invention is intended to meet the foregoing demand, providing an innovative photobioreactor system and method of using the system for cultivating photosynthetic organisms, including but not limited to algae, cyanobacteria, plants, and plant cells, to enrich oil and lipids.
  • the present invention provides a flat-plate photobioreactor system.
  • the photobioreactor unit contains front and rear sides, which are made of transparent materials (such as glass, plastics, or other light transmitting materials).
  • the front and rear sides are mounted in a "U"-Shape frame, which is made of supporting materials, such as fiberglass, stainless steel, or other metal materials.
  • the individual flat-plate photobioreactor units can be connected to each other in a cascade forming a photobioreactor system, which can be used for large- scale cultivation of algae of microalgae and other photosynthetic organisms.
  • Each individual photobioreactor unit contains a circulation pipe mounted in the bottom, so that the compressed gas (such as air, carbon dioxide, nitrogen) can be diffused from the pores of the circulation pipe to provide sufficient turbulence for keeping the photosynthetic organisms in liquid phase and preventing sedimentation.
  • the present invention provides a stepwise photobioreactor containing a series of units, with two adjacent units overlapping at the edges, each unit containing at least one plate held in a frame and having a substantially flat surface, a collection tank for holding photosynthetic culture materials, a tunnel positioned at one end of the plate and arranged to conduct culture materials from the plate to the collection tank, and a pump configured to transfer culture materials from the collection tank back to the surface of the plate.
  • the plate is supported in a substantially horizontal orientation.
  • the surface of the plate is water resistant.
  • the present invention provides a method of culturing photosynthetic materials using the flat-plate photobioreactor sytem, the stepwise photobioreactor system, or combination thereof.
  • the present invention provides a method of cultivating photosynthetic organisms using these photobioreactors in combination with other systems, including but not limited to agar plates, regular flask reactors, fermentation reactors, and open systems to achieve efficient production of oil-enriched organisms.
  • the present invention provides methods of manufacturing biodiesel, valuable fine chemicals, nutraceuticals, and animal feed byproducts using the lipids and oils extracted from the oil-enriched microorganisms produced in the photobioreactor system as the starting material.
  • FIG. 1 is a drawing of one embodiment of a photobioreactor unit according to the present invention.
  • FIG. 2 is an overview of the cascade containing interconnected photobioreactor units of the photobioreactor system.
  • FIG. 3 depicts an alternate embodiment of the photobioreactor unit of the present invention.
  • FIG. 4 is a perspective view of a stepwise photobioreactor.
  • FIG. 5 is a front view of the stepwise photobioreactor of FIG. 4.
  • FIG. 6 is a side view of the stepwise photobioreactor of FIGs. 4 and 5.
  • FIG. 7 is a top schematic view of a stepwise photobioreactor.
  • FIG. 8 is a front schematic view of the stepwise photobioreactor of FIG. 7.
  • FIG. 9 is a side schematic view of the stepwise photobioreactor of FIGs. 7 and 8.
  • FIG. 10 illustrates an overview of the cultivation system.
  • the present invention provides new methods for producing oil-enriched photosynthetic organisms, in particular microalgae, using two unique photobioreactor systems, which enable efficient photosynthesis and rapid growth and oil-enrichment in the organisms.
  • the phototsynthetic organisms suitable for the cultivation methods disclosed herein include, but are not limited to, algae, cyanobacteria, plants, and plant cells.
  • the first type of photobioreactor of the present invention is a flat-plate photobioreactor as illustrated in FIG. 1.
  • the flat-plate photobioreactor includes a U-shape frame, which affords a strong configuration, flexible dimensions, convenient construction and operation, and potential to form a cascade of many individual photobioreactors.
  • the U-Shape frame can be made of any strong durable material, for example, stainless steel (or other supporting materials), making the photobioreactor strong and; the dimension of the photobioreactor can be made in a wider range (very thin or very long reactor units are possible with the U-shaped configuration);
  • each photobioreactor unit makes it possible to form a cascade of many individual photobioreactor units, so that efficiency of operation can be greatly improved.
  • the photobioreactor system may be illuminated with artificial light (such as fluorescent light) from both front and rear sides.
  • artificial light such as fluorescent light
  • An appropriate supporting frame is required to support the light bulbs so that the illumination is provided evenly from the top to the bottom of the photobioreactor system.
  • the photobioreactor system is illuminated with natural sunlight.
  • an appropriate frame is required to support the photobioreactor system, so that each photobioreactor has an angle of 45° ⁇ 60° towards the ground. This angular arrangement assures the maximum absorption of sunlight.
  • the temperature of the photobioreactor system is controlled using a water spray system in the summer for cooling and electric heating rods in winter to heat the photobioreactor system.
  • various cultivation media is used in the photobioreactor system.
  • the height of the culture media is between about 70% and about 90% of the height of the photobioreactor.
  • Culture mixing is provided by releasing air through air tubing running along the bottom of the photobiorector at a rate of about 0.1 to about 0.5 liter of air per liter of culture suspension per minute.
  • Compressed fine air bubbles enriched with about 0.5% to about 1 % (v/v) C0 2 not only enhances photosynthesis but also maintains a favorable culture pH (preferably from about 7.5 to about 8.0).
  • the photobioreactor system is equipped with a monitoring system (not illustrated) that measures dissolved oxygen, pH, conductivity, and temperature. Each monitor is internet-connected via TCP/IP communication port and is set to send browser-based HTML text.
  • the second type of photobioreactor sytem of the present invention is a stepwise photobioreactor as disclosed in U.S. Provisional Application No. 61/334,120, filed on May 12, 2010, and PCT Application No. PCT/US2011/036273, filed on May 12, 2011, both of which are hereby incorporated by reference in their entirety.
  • the photobioreactor contains at least one plate held in a frame and has a substantially flat surface, a collection tank for holding photosynthetic culture materials, a tunnel positioned at one end of the plate and arranged to conduct culture materials from the plate to the collection tank, and a pump configured to transfer culture materials from the collection tank back to the surface of the plate.
  • the plate is supported in a substantially horizontal orientation.
  • the surface of the plate is water resistant.
  • the photobioreactor includes a first plate and a second plate, the second plate being positioned at a higher elevation than the first plate. In embodiments, an edge of the second plate partially overlaps the surface of the first plate. Further, in some embodiments, the pump is configured to transfer the photosynthetic culture materials from the collection tank back to a substantially flat surface of the second plate.
  • the photobioreactor further includes a cover positioned over a portion of the surface of the plate. Further, in some embodiments, a portion of the cover is substantially transparent.
  • the plate includes a raised edge configured to maintain a predetermined depth of culture suspension on the surface of the plate. In some embodiments, the raised edge includes a first portion and a second portion, the first portion having a greater height than the second portion. Further in some embodiments, the height of the second portion is between about 2 cm and 3 cm.
  • the photobioreactor includes a series of plates arranged in a cascading sequence, such that culture materials fall from an edge of an upper one of the plates onto a lower one of the plates, the tunnel being positioned at one of end of a lowest plate in the series of plates, and the pump being configured to transfer culture materials from the holding tank back to the surface of a highest plate of the series of plates.
  • the present invention provides a method of cultivating photosynthetic organisms, the method including introducing a selected amount of photosynthetic culture material to the surface of a plate of a photobioreactor, and circulating the culture material by collecting the culture material from the plate in a collection tank and operating a pump to transfer the culture material from the collection tank back to the surface of the plate.
  • the present invention provides a method of producing an oil- enriched photosynthetic organism, including cultivating the organism in a flat-plate photobioreactor followed by cultivating it in a stepwise photobioreactor.
  • the method of producing an oil-enriched photosynthetic organism includes cultivating the organism in a flat-plate photobioreactor followed by fermentation in a container under anaerobic conditions in the presence of nutrients, such as sugar or other organic compounds. Fermentation can be conducted under heterotrophic or mixotrophic conditions as disclosed in U.S. Provisional Application No. 61/470,820, filed on April 1, 2011, which is hereby incorporated by reference in their entirety.
  • the organism thus fermented may be advantageously transferred to and cultivated in the stepwise photobioreator in the presence of light and carbon dioxide.
  • the method of producing an oil-enriched photosynthetic organism includes cultivating the organism in a reactor containing a growth medium followed by cultivating it in the stepwise photobioreactor directly.
  • the method of producing an oil-enriched photosynthetic organism includes further cultivating it in an open system, such as an open pond or an open race way.
  • an open system such as an open pond or an open race way.
  • Such a step can be subsequent to cultivating in the stepwise photobioreactor, or directly from cultivating in a flat-plate photobioreactor.
  • the method of producing an oil-enriched photosynthetic organism includes cultivating the organism in agar plates followed by cultivating in a flat- plate photobioreactor, or by fermentation under anaerobic conditions feeding with sugar, or directly by cultivating in a stepwise photobioreactor in the presence of light and carbon dioxide.
  • the present invention provides a process for producing an oil- enriched photosynthetic organism, the process including:
  • photobioreactor system contains at least one photobioreactor unit containing:
  • the process further includes:
  • the maintaining step contains supplying nutrition for the organism cells to continue to grow and proliferate.
  • the nutrition contains carbon dioxide.
  • the open system is a pond or an open raceway.
  • the maintaining step is performed until after the organism biomass reaches approximately 80% to 95% of the capacity of the open system, or until the biomass stops growing due to limited supply of carbon dioxide, or until the biomass stops growing in the presence of additional carbon dioxide and/or other nutrition.
  • the culture of the organism species is circulated through the photobioreactor system.
  • the culture of the organism species is illuminated with an artificial light or sunlight while being circulated through the photobioreactor system.
  • the high content of lipid or oil is at least 40%, at least 45%, at least 50%, at least 55%, or at least 60% based on dry weight of the biomass.
  • the pre-determined amount of lipid or oil is at least 20%, at least 30%, at least 40%, at least 50%, or at least 60% based on dry weight of the biomass.
  • the photosynthetic organism is an oil- containing algae selected from the group consisting of diatoms (bacillariophytes), green algae (chlorophytes), blue-green algae (cyanophytes), golden-brown algae (chrysophytes), haptophytes, and combinations thereof.
  • oil-containing algae is selected from
  • the organism species is microalgae
  • the process further includes pre-cultivating the organism species in a first container containing a growth medium prior to cultivating it in the photobioreactor system.
  • the pre-cultivating includes: a) purifying a sample of the organism species; b) maintaining the sample in agar plates containing a growth medium; and c) cultivating the organism in the first container.
  • the first container in the pre-cultivating step, contains a growth medium in the range of about 50 mL to about 5000 mL.
  • the photobioreactor unit has a capacity of about 1 L to about 1000 m 3 .
  • the photobioreactor unit is between about 0.3 and about 100 meters in length; between about 0.3 and 10 meters high; and between about 0.01 and about 1 meter deep.
  • the photobioreactor unit further contains horizontal or vertical supporting grids in the front and rear sides of the frame.
  • the photobioreactor unit further contains holes having a diameter ranging between about 0.01 and about 0.5 meters in the bottom and sides of the U-shape frame.
  • the photobioreactor unit contains one or more pipes passing through the holes in the bottom and/or sides of the U-shape frames for connecting with an adjacent photobioreactor unit.
  • the photobioreactor unit further contains one or more porous circulation pipes mounted in the bottom of the unit to release compressed gas from the pores of the circulation pipes to provide sufficient turbulence for keeping photosynthetic organisms in liquid phase and to prevent sedimentation.
  • the photobioreactor unit contains at least one connection hole in each side of the U-shape frame.
  • the photobioreactor unit contains at least one hole in the bottom of the U-shape frame.
  • the parallel transparent plates are sealed to the frame with a biologically acceptable sealing material.
  • the sealing material is an acrylic resin.
  • the present invention provides a method of producing an oil- enriched photosynthetic organism, including cultivating the photosynthetic organism in a liquid medium in a photobioreactor cascade, the photobioreactor cascade containing a plurality of photobioreactor units interconnected with one another through a pipe passing through connection holes in the side of each photobioreactor unit, wherein the photobioreactor unit contains:
  • the culture medium can be circulated throughout the photobioreactor through a collection tank and a pump, and wherein the photobioreactor system can be illuminated by an artificial light source or sunlight.
  • the maintaining step further contains supplying nutrition for the organism cells to continue to grow and proliferate.
  • the nutrition contains carbon dioxide.
  • the open system is a pond or an open raceway.
  • the present invention provides a method of producing an oil- enriched photosynthetic organism as feedstock for biodiesel production, including:
  • a tunnel positioned at one end of the plate and arranged to conduct culture materials from the plate to the collection tank; and (iv) a pump configured to transfer culture materials from the collection tank back to the surface of the plate.
  • the first photobioreactor contains at least one connector hole in the side of the u-shaped frame and a drainage hole in the bottom of the u- shaped frame.
  • the first photobioreactor contains a cover for sealing the reactor contents from the atmosphere.
  • the first photobioreactor further contains an artificial light source.
  • the cover includes at least one aperture for admitting or withdrawing culture medium from the reactor unit.
  • the at least one plate of the second photobioreactor contains a first plate and a second plate, the second plate being positioned at a higher elevation than the first plate.
  • an edge of the second plate partially overlaps the surface of the first plate.
  • the pump of the second photobioreactor is configured to transfer photosynthetic culture materials from the collection tank back to the surface of the second plate.
  • the at least one plate of the second photobioreactor is supported in a substantially horizontal orientation.
  • the second photobioreactor further contains a cover positioned over a portion of the surface of the at least one plate.
  • a portion of the cover is substantially transparent.
  • the at least one plate of the second photobioreactor contains a raised edge around a portion of its periphery, and wherein the raised edge is configured to maintain a predetermined depth of culture suspension on the surface of the at least one plate.
  • the raised edge contains a first portion and a second portion, the first portion having a greater height than the second portion.
  • the height of the second portion is between about 1 cm to 5 cm, preferably about 2 cm and 3 cm.
  • the at least one plate of the second photobioreactor contains a series of plates arranged in a cascading sequence, such that culture materials fall from an edge of an upper one of the plates onto a lower one of the plates, wherein the tunnel is positioned at one of end of a lowest plate in the series of plates, and the pump is configured to transfer culture materials from the holding tank back to the surface of a highest plate of the series of plates.
  • the surface of the at least one plate of the second photobioreactor contains a water resistant surface.
  • the cultivating in the second photobioreactor including:
  • the circulation system contains a collection tank to collect the cultural material flowed from the photobioreactor plates and a pump to transfer the culture material back to the photobioreactor plate(s).
  • the method further contains a fermentation step following the cultivation in the first photobioreactor, the fermentation step including maintaining the organisms under a hetertrophic or mixotrophic condition in a liquid medium containing sugar and/or other organic matter.
  • the fermentation step is conducted under an anaerobic condition.
  • the sugar or organic matter is selected from the group consisting of glucose, fructose, sodium acetate, and combinations thereof.
  • the method further contains steps of: (d) transferring the cultured organism material to an open pond or open raceway suitable for the organism to grow; and (e) maintaining the organism in the open system for the organism to continue to grow into a biomass enriched with oil or lipids.
  • the present invention provides a process for producing an oil- enriched photosynthetic organism as feedstock for biodiesel production, including:
  • a pump configured to transfer culture materials from the collection tank back to the surface of the plate
  • the at least one plate contains a first plate and a second plate, the second plate being positioned at a higher elevation than the first plate. In another embodiment of this aspect, an edge of the second plate partially overlaps the surface of the first plate.
  • the pump is configured to transfer photosynthetic culture materials from the collection tank back to the surface of the second plate.
  • the at least one plate is supported in a substantially horizontal orientation.
  • the photobioreactor further contains a cover positioned over a portion of the surface of the at least one plate.
  • a portion of the cover is substantially transparent.
  • the at least one plate contains a raised edge around a portion of its periphery, and wherein the raised edge is configured to maintain a predetermined depth of culture suspension on the surface of the at least one plate.
  • the raised edge contains a first portion and a second portion, the first portion having a greater height than the second portion.
  • the height of the second portion is between about 1 cm and about 5 cm, preferably between about 2 cm and about 3 cm.
  • the at least one plate contains a series of plates arranged in a cascading sequence, such that culture materials fall from an edge of an upper one of the plates onto a lower one of the plates, wherein the tunnel is positioned at one of end of a lowest plate in the series of plates, and the pump is configured to transfer culture materials from the holding tank back to the surface of a highest plate of the series of plates.
  • the surface of the at least one plate contains a water resistant surface.
  • the cultivating in the photobioreactor includes: introducing a selected amount of photosynthetic culture material to the photobioreactor system;
  • the circulation system contains a collection tank to collect the cultural material flowed from the photobioreactor plates and a pump to transfer the culture material back to the photobioreactor plate(s).
  • the present invention provides a process for making synthetic biodiesel, including: a) cultivating a photosynthetic organism species enriched with lipid or oil according to any of claims 1, 27, 32, and 53; b) extracting lipids and oils from the oil- enriched organism; and c) converting the extracted algal oil or lipids to biodiesel through a chemical or enzymatic transformation.
  • the chemical or enzymatic transformation includes a process selected from hydrogenation, esterification, transesterification, and combinations thereof.
  • the present invention provides methods of manufacturing valuable fine chemicals, nutraceuticals, and animal feed byproducts, among others, using the lipids and oils extracted from the oil-enriched microorganisms produced in the photobioreactor system as the starting material through chemical or chemical engineering transformations, including but not limited to hydrogenation, esterification, transesterification, and combinations thereof.
  • FIG. 1 is a schematic drawing of one photobioreactor unit illustrating one embodiment of the present invention.
  • the front side 11 and rear side 12 of the unit are made of clear transparent materials, such as glass, polycarbonate or acrylic sheets.
  • the front side 11 and rear side 12 of the photobioreactor unit are mounted on a "U"-Shape frame 13, that is made of supporting materials such as fiberglass, stainless steel, or other rust resistant metal materials.
  • the "U"-Shape frame 13 forms the lateral sides and the bottom of the photobioreactor unit.
  • the dimensions of the "U"-Shape frame 13 are: length 0.3-100 meters; height 0.3-10 meters; depth 0.01-1 meter.
  • the glass plates are sealed to the frame 13 using conventional biologically acceptable sealing materials such as Polytetrafluoroethyline (PTFE) or acrylic resin.
  • PTFE Polytetrafluoroethyline
  • the top of the "U"-Shape frame 13 is connected by two horizontal supporting grids 14. There are several vertical supporting grids 14 in the front side 11 and rear side 12 of the photobioreactor, respectively. The interval between the vertical supporting grids is between 0.5-1.0 meter.
  • the vertical and horizontal supporting grids may be made of the same material as the "U"-Shape frame 13.
  • connection holes 15 are in each lateral side, and one harvesting hole 16 in the bottom of the photobioreactor unit.
  • the connection holes 15 can be used to connect the photobioreactors of the invention together in a cascade. Depending on the orientation of the photobioreactor unit, the connection holes 15 may also be used for harvesting the microalgae from the culture medium.
  • the diameter of the holes at the bottom and sides of the U-shape frame is between about 0.01 to 0.5 meter.
  • the number of outlets 16 in the bottom can be adjusted depending on the length of the photobioreactor units. Longer photobioreactor units have 2 or more outlet holes 16 on the bottom of the reactor unit.
  • the photobioreactor system consists of a series of photobioreactors (21) connected to each other as a cascade.
  • the connection between two photobioreactor units (21) is made by connecting pipes (22) (which can be made of non-transparent materials) through the connection holes (15) on the bottom or the lateral sides of the "U"-Shape frame (13). Therefore the photobioreactor system can vary its total volume by increasing or decreasing the number of photobioreactors units in the system (23).
  • the level of the culture media in the reactor is kept below the level of grids (14).
  • FIG. 3 An alternate embodiment of the photobioreactor is depicted in FIG. 3.
  • the reactor unit has a cover (24).
  • An aperture (25) in the cover can be used for adding culture medium to the reactor.
  • each individual photobioreactor unit contains a circulation pipe (17) mounted in the bottom.
  • the pipe is preferably porous so that the compressed gas (such as air, carbon dioxide, nitrogen) can be diffused from the pores of the circulation pipe to provide sufficient turbulence for keeping photosynthetic organisms in liquid phase and preventing sedimentation.
  • the photobioreactor system consists of a series of photobioreactors 21 connected to each other as a cascade.
  • the connection between two photobioreactor units 21 is made by connecting pipes 22 through the connection holes 15 on the bottom or the lateral sides of the "U"-Shape frame 13. Therefore the photobioreactor system can vary its total volume by increasing or decreasing the number of photobioreactors units in the system 23.
  • the level of the culture media in the reactor is kept below the level of grids 14.
  • the reactor unit has a cover 24. An aperture 25 in the cover can be used for adding culture medium to the reactor.
  • Each individual photobioreactor unit can contain a circulation pipe 17 mounted in the bottom.
  • the pipe is preferably porous so that the compressed gas (such as air, carbon dioxide, nitrogen) can be diffused from the pores of the circulation pipe to provide sufficient turbulence for keeping photosynthetic organisms in liquid phase and preventing sedimentation.
  • FIG. 4 is a perspective view of a stepwise photobioreactor.
  • the photobioreactor includes a series of horizontally positioned flat plates (1) organized in cascade fashion.
  • the number of flat plates (1) in the cascade can be one, or more than one, depending on the culture requirements and available space.
  • Each flat plate (1) is at a lower position than the preceding flat plate.
  • an aqueous culture of photosynthetic organisms can be deposited on the highest (first) plate in the cascade, flow across the surface of the plate, and then, by gravity, flow down to the next (lower) plate in the cascade.
  • the culture may be collected at the last flat plate and subsequently be transferred back to the first flat plate by pumping, aeration or other available methods.
  • Such an apparatus can be used for cultivation of any suitable photosynthetic organism (such as algae, cyanobacteria, and plants).
  • each individual plate (1) can be square, rectangle, triangle, circle, and alike.
  • the length can range from 30 cm to 1000 m; the width can range from 30 cm to 1000 m.
  • the flat plate (1) can be made of any suitable water resistant materials, such as glass, plastic, cement, brick, metal.
  • Each flat plate is fixed in a supporting frame (2).
  • FIG. 5. illustrates a front view of the photobioreactor.
  • each flat plate is placed horizontally, and is at a lower elevation than the preceding flat plate in the cascade, so that the culture can flow from the upper flat plate to the lower one by gravity.
  • each plate (1) has a raised edge on three of its sides to inhibit or prevent water overflow (e.g., a 10 cm high raised edge).
  • the height of the raised edge is lower than the other sides (e.g., between about 1 and 3 cm), so that the depth of the culture suspension on the plate will be kept at a desirable level.
  • the depth of the culture can be adjusted to about 1cm; conversely, if the cell concentration is low, 2 cm or 3 cm culture depth may be desirable to provide more self shading for cells to prevent photoinhibition.
  • FIG. 6 is a side view of the photobioreactor.
  • the flat plate cascade can be covered by transparent materials (3), such as glass or plastic sheets, to prevent dust and water from contacting the culture.
  • the culture can be collected at the tank (5) and transferred to the first flat plate (1) in the cascade by a water pump (6).
  • a selected amount of photosynthetic culture material is introduced to the surface of the first plate in the cascading series of plates.
  • the raised edges (9) surrounding the periphery of the plates (1) inhibit overflow of the culture.
  • the culture material flows across the surface of each plate and falls to the surface of the next plate in the series.
  • the culture is collected by the tunnel (4) having an orifice (8), through which the culture is conducted to the collection tank (5).
  • the culture is transferred from the collection tank (5) to the first flat plate (1) by a pump (6) and a conveying conduit (7).
  • the transfer can be made by means of any suitable transfer device, such as aeration devices, or mechanical lifting devices.
  • the continuous or semi-continuous transfer of the culture from the collection tank (5) to the first flat plate (1) will keep the culture moving though the flat plate cascade system. In this way, the cultivated cells will be prevented from sedimentation and have an equal opportunity for exposure to sunlight.
  • the circulating time of the culture suspension can be controlled by the operation of the water pump (6) at different time intervals, such as 5/15; 5/25; 5/35; 5/45, 5/55; etc. (minutes for on/off), depending on the total volume of the culture suspension. In such way, much less electricity will be required to operate the water pump (6).
  • cultivation of photosynthetic organisms can be conducted in a number of alternative embodiments by combining use of the photobioreactors and/or conditions of the present invention with use of other types of culturing/cultivating systems, including but not limited to agar plates, small or large culturing reactors, fermentation equipments, and/or open systems.
  • the cultivation can start in a flat-plate photobioreactor and then transfer to a stepwise photobioreactor (a), followed by harvesting of the cells and extraction of oil/lipids from them (m); the cultivation can also start in a fermentation container under anaerobic conditions, and then transfer to a stepwise photobioreactor (c).
  • the cultivation can start in a flat-plate photobioreactor, then transfer to a fermentation container in which fermentation is conducted (b), followed by transfer to the stepwise photobioreactor for large scale cultivation (c).
  • Culturing of cells can also start in agar plates and then transfer directly to a stepwise photobioreactor (e), or alternatively going through a culturing reactor (h), fermentation (i, j), and/or a flat-plate photobioreactor (g, h), depending on the needs and conditions of the cell cultures.
  • a person skilled in the art would be able to determine whether and how to combine these reactors in order to obtain optimum results.
  • the bioreactors are illuminated with a manmade light.
  • a manmade light For smaller reactors, such as the culturing reactor, the fermentation reactor or single flat-plate reactors, strong incandescent lights may be used to speed up cultivation or promote photosynthesis, especially in the night or when daylight is not sufficient.
  • large reactors such as cascades of the flat-plate reactors or the step-wise reactors, although a manmade light can also be used, it is economically preferable to make good use of daylight, especially sunlight. Therefore, the bioreactor system of the present invention is preferably built in an area having plenty of sunshine throughout the year, for example, in an open area such as desert.
  • C0 2 carbon dioxide
  • the carbon dioxide can be provided by any conventional means from any conventional sources.
  • C0 2 may be transported from a C0 2 storage, or directly from a power plant through a transportation pipe.
  • the culture system from the stepwise photobioreactor may be further transferred to an open system, for example, an open pond or open raceway, for further cultivation (k).
  • the oil/lipid-enriched organism will then be subject to isolation and extraction processes (m, n) to obtain algae oil (o) as feedstock for production of biodiesel (p) or manufacture of valuable fine chemicals, nutraceuticals, and animal feed byproducts, etc (q).
  • photosynthetic organisms including but not limited to Chlorococcum pamirum
  • a growth medium containing the following components: about 0.1-2 g/L NaN0 3 , about 0.05-1.75 g/L MgS0 4 , about 0.5-3.6 g/L Na 2 C0 3 , about 0.05-0.2 g/L CaCl 2 , about 0.001 g/L EDTA, about 0.02- 1.2 g/L K 2 HP0 4 , about 0.006 g/L Citric Acid, about 0.006 g/L Fe(NH 4 ) 2 Citric, about 0.2- 1.0 mL/L A5 micronutrients, and about 1.0-100 g/L glucose.
  • Heterotrophic cultivation of the species Chlorococcum pamirum can contain the following steps:
  • All culture devices have to be axenic, no light is required, and temperature is between about 5 °C and about 40 °C; sterilized compressed air is provided to the cell culture as a source of oxygen and as a way for mixing the culture.
  • the mixotrophic cultivation of the species Chlorococcum pamirum consists of the following steps:
  • Steps la through Id The steps are the same as described in Steps la through Id, except that culture device must be transparent to permit light penetration to the cells within the device.
  • the light intensity can range from 10-5000 ⁇ m - " 2 s - " 1.
  • All culture devices have to be axenic, and temperature must be maintained between 5 °C and 40 °C; sterilized compressed air is provided to the cell culture as a source of oxygen and as a way for mixing the culture.
  • photosynthetic organisms including but not limited to Chlorococcum pamirum
  • heterotrophic conditions as follows:
  • Oil production in the microalgae can be significantly enhanced from about 20-30% to about 40%-50% of dried weight after about 5 to about 20 days.
  • 2b. Transfer the cells of Chlorococcum pamirum to a phosphate-deficient medium, containing the composition described in Section 2a with nitrogen added; Oil production in the microalgae can be significantly enhanced from about 20-30% to about 40%-50% of dried weight after about 10-24 days.
  • Transfer the cells of the microalgae to a low temperature (1-15 °C) with the same growth medium as in Section 2a can also enhance oil production to about 40% -45% of dried weight of the microalgae after about 10-30 days.
  • photosynthetic organisms including but not limited to
  • Chlorococcum pamirum can then be cultivated under mixotrophic conditions as follows:
  • Transfer cells of the microalgae to a low temperature (about 1-15 °C) under light intensity of about 10-5000 ⁇ m - " 2 s - " 1 with the same growth medium as set forth in Section 2a can also enhance oil production to about 40%-45% of dried weight of the microalgae after 7-20 days.
  • Ranges can be expressed herein as from “about” one particular value, and/or to "about” another particular value. When such a range is expressed, another aspect includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent "about,” it will be understood that the particular value forms another aspect. When the term “about” appears in front of a number, it means that the value can vary by at least +10%, preferably by within ⁇ 5%.

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Abstract

Cette invention concerne un système photobioréacteur à plaque plate, un système photobioréacteur par étape et des procédés de culture d'organismes photosynthétiques, à l'aide de ces photobioréacteurs, dans des conditions de photosynthèse aérobie, facultativement en combinaison avec d'autres systèmes de culture, par exemple des plaques d'agar, des réacteurs classiques de petite ou moyenne dimension et des systèmes ouverts, tels que des bassins ouverts ou des circuits ouverts. Les procédés comprennent également une combinaison de la photosynthèse dans ces photobioréacteurs avec une fermentation dans des conditions hétérotrophes ou mixotrophes, anaérobie ou aérobie. Les procédés sont particulièrement utiles pour la production de microalgues enrichies en huile comme source d'alimentation dans la production d'un biodiesel, de produits chimiques fins valables, de produits nutraceutiques et de sous-produits d'aliments pour animaux, etc.
PCT/US2011/064599 2011-09-29 2011-12-13 Systèmes photobioréacteurs et procédés de culture d'organismes photosynthétiques Ceased WO2013048543A1 (fr)

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US20150250113A1 (en) * 2014-03-04 2015-09-10 Greenonyx Ltd Systems and methods for cultivating and distributing aquatic organisms
BE1021386B1 (fr) * 2013-05-07 2015-11-12 Agc Glass Europe Dispositif pour cultiver des organismes phototrophes.
WO2016060892A1 (fr) * 2014-10-16 2016-04-21 University Of South Florida Systèmes et procédés pour cultiver des algues
CN106399105A (zh) * 2015-07-23 2017-02-15 中国石油化工股份有限公司 一种单针藻的养殖方法
WO2017148893A1 (fr) * 2016-02-29 2017-09-08 Aveston Grifford Ltd. Procédé de récolte de biomasse d'un photobioréacteur
CN111164197A (zh) * 2017-10-10 2020-05-15 智康工程顾问有限公司 用于微藻的异养和混合营养培养的方法和系统
WO2023073454A1 (fr) * 2021-10-29 2023-05-04 Bluemater, S.A. Photobioréacteur pour la culture de macro ou de micro-organismes, l'évaporation liquide ou la fermentation liquide
EP4183862A1 (fr) * 2021-11-17 2023-05-24 Agfa Nv Biomatériau pour la production d'hydrogène

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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
BE1021386B1 (fr) * 2013-05-07 2015-11-12 Agc Glass Europe Dispositif pour cultiver des organismes phototrophes.
US20150250113A1 (en) * 2014-03-04 2015-09-10 Greenonyx Ltd Systems and methods for cultivating and distributing aquatic organisms
US10039244B2 (en) * 2014-03-04 2018-08-07 Greenonyx Ltd Systems and methods for cultivating and distributing aquatic organisms
WO2016060892A1 (fr) * 2014-10-16 2016-04-21 University Of South Florida Systèmes et procédés pour cultiver des algues
CN106399105A (zh) * 2015-07-23 2017-02-15 中国石油化工股份有限公司 一种单针藻的养殖方法
WO2017148893A1 (fr) * 2016-02-29 2017-09-08 Aveston Grifford Ltd. Procédé de récolte de biomasse d'un photobioréacteur
CN111164197A (zh) * 2017-10-10 2020-05-15 智康工程顾问有限公司 用于微藻的异养和混合营养培养的方法和系统
US20200231923A1 (en) * 2017-10-10 2020-07-23 Gicon Grossmann Ingenieur Consult Gmbh Method and System for Heterotrophic and Mixotrophic Cultivation of Microalgae
CN111164197B (zh) * 2017-10-10 2024-03-22 智康工程顾问有限公司 用于微藻的异养和混合营养培养的方法和系统
US12012581B2 (en) * 2017-10-10 2024-06-18 Gicon Grossmann Ingenieur Consult Gmbh Method and system for heterotrophic and mixotrophic cultivation of microalgae
WO2023073454A1 (fr) * 2021-10-29 2023-05-04 Bluemater, S.A. Photobioréacteur pour la culture de macro ou de micro-organismes, l'évaporation liquide ou la fermentation liquide
EP4183862A1 (fr) * 2021-11-17 2023-05-24 Agfa Nv Biomatériau pour la production d'hydrogène

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