WO2010103154A2 - Procédé de culture de microorganismes et photobioréacteur utilisé dans ce procédé - Google Patents

Procédé de culture de microorganismes et photobioréacteur utilisé dans ce procédé Download PDF

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Publication number
WO2010103154A2
WO2010103154A2 PCT/ES2010/070132 ES2010070132W WO2010103154A2 WO 2010103154 A2 WO2010103154 A2 WO 2010103154A2 ES 2010070132 W ES2010070132 W ES 2010070132W WO 2010103154 A2 WO2010103154 A2 WO 2010103154A2
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WO
WIPO (PCT)
Prior art keywords
duct
microorganisms
photobioreactor
culture
reactor
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/ES2010/070132
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English (en)
Spanish (es)
Other versions
WO2010103154A3 (fr
Inventor
Carlos DÍAZ GARCÍA
Eduardo Romero Palazón
Guillermo GARCÍA-BLAIRSY REINA
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Repsol SA
Original Assignee
Repsol YPF SA
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
Application filed by Repsol YPF SA filed Critical Repsol YPF SA
Publication of WO2010103154A2 publication Critical patent/WO2010103154A2/fr
Publication of WO2010103154A3 publication Critical patent/WO2010103154A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/002—Photo bio reactors
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/14—Pressurized fluid
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate

Definitions

  • the present invention can be included within the field of cultivation of microorganisms in reactors, in particular it describes a method for obtaining biomass from algae and a high performance vertical reactor employed in said method.
  • Microalgae are unicellular beings very varied in size and shape that exist in almost all known habitats. Most of them are aquatic habitats, both marine and freshwater, although some live on land.
  • the seas and oceans contain huge amounts of planktonic algae, with an estimated 90% of the Earth's total photosynthesis being performed by these aquatic plants.
  • microalgal biomass can be used for applications as biofertilizers, in the purification of wastewater, as soil conditioners and as food.
  • microalgae for the production of a wide variety of substances, such as fatty acids, pigments, vitamins, antibiotics, pharmaceuticals and other chemical products of interest, as well as hydrogen, hydrocarbons and other biological fuels has been revealed.
  • the so-called photobioreactors can be used. It is essential, for said production, to properly select the design of the reactor to be used, as well as a series of parameters depending on the microorganism to be used, such as optimal growth conditions and resistance to environmental variations. In this way, the design of reactors that allow to achieve high productivity at minimum costs is one of the most important lines of work. important in the field of biotechnology, and without whose progress a worldwide expansion of this industry will be impossible.
  • the present invention provides a method for the cultivation of oil-generating microalgae for use as fuels, as well as a reactor employed in said method.
  • an aqueous solution is available with microalgae that grow by photosynthesis due to the combined effect of sunlight and injection of both air and eventually CO2.
  • the cultivation of algae requires, therefore, these two basic conditions; The entry of light that allows the simultaneous existence of illuminated and shaded areas to provide the cycle of photosynthesis and agitation of the culture liquid that promotes the exchange of algae between both areas.
  • the present invention provides a high efficiency advanced vertical photobioreactor (hereinafter FBR) for algae culture, in a cylindrical shape, with a bottom that can be conical or flat, of transparent material and with an external recirculation duct, in which a vortex effect is generated in the upper part of the aqueous medium that allows a greater entry of light energy into the system and a more efficient agitation of the medium, which allows to increase the production of biomass cultivated by unit of volume and surface.
  • Said vortex or swirl is generated by the effect of the tangential entry into the system of the culture medium itself, which is driven from the lower part of the reactor to the upper part by a gas injection system located in the recirculation duct.
  • a first aspect of the present invention refers to a reactor for the cultivation of microorganisms, or FBR photobioreactor, which comprises a transparent cylindrical body, a bottom located next to the cylindrical body and an outlet conduit connected to an outlet located at the bottom, characterized in that it additionally comprises: a recirculation duct whose inlet is located next to the outlet duct and which flows into the upper part of the cylindrical body, and injection means, located in the vicinity of the inlet of the duct inlet. recirculation, which introduce gases.
  • the reactor of the invention is a cylindrical reactor, made of a transparent material, which may be included, but not limited to, the group comprising glass fiber, low density polyethylene (LDPE), polycarbonate (PC), polymethylmethacrylate (PMMA) or any other material that meets the optimal light transmission conditions PAR (photosynthetically active radiation, from English Photosynthetically Active Radiation).
  • LDPE low density polyethylene
  • PC polycarbonate
  • PMMA polymethylmethacrylate
  • PAR photosynthetically active radiation, from English Photosynthetically Active Radiation
  • the bottom of the reactor can be conical or flat, preferably flat.
  • the height of the reactor can be between 1 and 5 meters and the diameter can vary between 15 and 100 cm.
  • the reactor of the invention contains an aqueous solution that in turn contains microalgae that grow by photosynthesis by combined effect of sunlight and air, more preferably CO2, injected.
  • a second aspect of the present invention relates to a process for producing biomass from a culture of microorganisms using the reactor of the invention.
  • the conditions that this algae culture procedure requires, in addition to the use of a reactor like the one described above, are: light input with illuminated areas and in shaded areas to allow photosynthesis; and a stirring of the culture liquid that promotes the exchange of algae between the lighted areas and the shaded areas.
  • Said reactor is subjected to adequate lighting to provide illuminated areas and shaded areas, and to agitation of the culture liquid by means of a biphasic current self-propelled by the injection of gases.
  • the agitation in the reactor of the invention is achieved by means of a biphasic liquid-gas stream self-propelled by means of gas injection and its subsequent rise in buoyancy bubbles.
  • Said current is ejected tangentially in the plane formed by the free surface of the culture liquid with the aim of obtaining a swirl sufficiently intense to generate a conical free surface whose vertex is located in the vicinity of the outlet duct.
  • the additional effect of free surface increase It increases the area of the illuminated area and can increase the capacity of the system to capture light from the environment and transform it into plant matter (biomass).
  • the eddy is obtained by introducing gas into the recirculation duct ("riser"), in the vicinity of the outlet duct.
  • the air rises bubbling through the recirculation duct to the height of the free surface of the culture liquid.
  • the entrainment of liquid due to the continuous injection of gas called the "holdup" effect, must provide the necessary impulse to generate rotation.
  • the microorganisms are microalgae that can be selected from the list comprising, but not limited to, the following genera: Spirulina, Chlorella, Chlorococcum, Neochloris Isochrysis ,, Tetraselmis, Oocytis Ettlia, Porphyridium , Nannochloris Synechocystis, Crucigenia, Muriellopsis, Haematococcus, Clamidomonas, Synechococcus, Phaeodactylum,
  • Platymonas Amphora, Auxenochlorella, Ankistrademus, Nanochloropsis, Nav ⁇ cula, Boekelovia, Scenedesmus or Rhodopseudomonas.
  • the microalgae can be selected from the following species: Phaeodactylum trichornutum, Platymonas sp., Amphora sp., Boekelovia sp., Swedish Tetraselmis, Nav ⁇ cula sp. Synechococcus sp., Scenedesmus quadricuada, Rhodopseudomonas palustres, Muriellopsis sp.
  • Chlorella sorokiniana Spirulina platensis, Chlorella sp., Neochloris oleoabundans, Scenedesmus sp., Auxenochlorella protothecoides, Synechocystis sp., Chlorococcum sp., Isochrysis galbana, Oocytis sp., Ettlia carotinosa, Porphyridnois ouent.
  • Crucigenia tetrapedia Haematococcus pluviales, Clamidomonas sp., Ankistrademus, sp., Nanochloropsis occulata or Nanochloropsis gaditana.
  • the main advantages of the reactor of the present invention over others already known are: a) Compared to conventional vertical systems. It improves the dissolution of nutrients and ensures the light-dark periods of the microalgae in crops with high densities. b) Against open systems (Race-Ways): better agitation of the culture medium and better control of the culture conditions (pH, Temperature, dissolution of nutrients in a homogeneous way) as well as avoiding the entry into the environment of contaminants that may affect to the yield of biomass production. c) Against helical tubular systems with small tubes: ease of operation (control of crop conditions such as pH,
  • Another aspect of the present invention relates to the biomass obtainable by the process of the invention.
  • Another aspect of the present invention relates to the use of the biomass of the invention for the manufacture of fuel, preferably this fuel is biodiesel although it can also be used for other energy uses, such as gasification.
  • Figure 1. Represents a diagram of an FBR reactor of the invention.
  • Figure 2.- Represents an operating scheme of an FBR reactor of the invention with the non-return valve closed.
  • Figure 3. Represents an operating scheme of an FBR reactor of the invention with the non-return valve open.
  • the FBR reactor of the invention is composed of the following elements: a) A vertical body (1) of cylindrical shape, with a height between 1 and 5 meters, and a diameter between 15 and 100 cm, built in a transparent material, which can be both rigid and flexible, such as the group comprising fiberglass, low density polyethylene (LDPE), polycarbonate (PC), polymethylmethacrylate (PMMA) or any other material that meets the optimal conditions of PAR light transmission.
  • the upper part of the body (1) is provided with a cover that is not necessarily airtight that allows the aeration of said reactor and the placement of different crop control systems (thermocouples, pH meters, salinity, O2 concentration, etc.).
  • the lower part of the body rests on a bottom (2) described below in point b).
  • a bottom (2) on which the body (1) rests which can be conical or flat, of material that can be opaque, such as for example steel, aluminum, polyvinyl chloride (PVC), or transparent according to materials similar to those described for the body (1).
  • the body (1) rests by means of a hermetic seal that ensures the tightness of the union between the body (1) and the bottom (2).
  • the outlet duct (3) In the lower part of the bottom (2) there is an outlet, connected to an outlet duct (3), with a diameter between 2 cm and 15 cm, which in turn connects with a recirculation duct (4) described below in the next point.
  • the bottom (2) is provided with a stable, strong and safe system to support the entire weight of the reactor on the ground.
  • An external recirculation conduit (4) which connects the lower part and the upper part of the reactor by means of upper (6) and lower (not shown) sealed connections, respectively.
  • the material of the circulation duct (4) can be opaque, such as steel, aluminum, polyvinyl chloride (PVC), or transparent, according to materials similar to those described for the body (1).
  • the upper connection (6) is located so that the culture medium accesses the reactor tangentially to the surface of said reactor.
  • the recirculation duct (4) incorporates, in the vicinity of the outlet duct (3), a third connection (8) to which an injector (5) is connected, which injects air or air + CO2 mixture into the reactor.
  • the reactor of the invention comprises a non-return valve (12) located in the outlet duct (3).
  • a non-return valve (12) located in the outlet duct (3).
  • the recirculation duct (4) in its lower section is provided with a purge duct (9) to facilitate the emptying of the reactor.
  • the recirculation system is provided with a diverting duct (10) that allows the recirculation of the culture medium to be diverted to any other element such as a second reactor, a harvesting tank or a drain.
  • the reactor must be filled to a pre-established maximum level with an aqueous solution that will act as a culture medium for the microalgae.
  • a sufficient inoculum of individual individuals will be incorporated into this medium to initiate the growth phase.
  • the start-up stage of the recirculation begins. For this, air is injected through the injection means (5), with a flow rate of 20-150 l / min. The "hold up" effect of the bubbles created will propel the culture medium upwards through the recirculation system to the upper entrance (6) of the body (1), generating a circular movement on the surface that will eventually become a vortex (11).
  • the reactor of the invention has been started. After the period of growth set according to the species of microalgae that is cultivated, the harvesting process will be carried out using either the purge duct (9) or the diversion duct (10), located in the recirculation duct ( 4).
  • PSSC max is the maximum dry weight without ashes
  • PSSC Range is the optimum harvest range
  • g is the generation time in days
  • ⁇ is the growth rate
  • P vol. It is the average volumetric production for the different species tested abroad (E) and in the greenhouse (I) in FFBR.
  • BNA are the codes of microalgae deposited in the National Algae Bank located in Las Palmas de Gran Canaria (Spain).
  • the PSSC values were calculated from the relationship between the optical density of the culture at 680 nm and the dry weight without ashes for each species / strain.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Sustainable Development (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un photobioréacteur comprenant un corps (1) vertical transparent de forme cylindrique, un fond (2) sur lequel repose ledit corps (1), et une sortie située dans la partie inférieure du fond (2) et reliée à un conduit de sortie (3), qui débouche sur un conduit de recirculation (4) relié à une entrée supérieure (6) située dans la partie supérieure du corps (1). Des moyens d'injection (5) injectent de l'air et éventuellement du CO2 dans le conduit de recirculation (4), produisant ainsi un tourbillon (11) à l'intérieur du corps (1). En outre, l'invention concerne un procédé d'obtention d'une biomasse à partir de micro-algues au moyen dudit photobioréacteur.
PCT/ES2010/070132 2009-03-09 2010-03-09 Procédé de culture de microorganismes et photobioréacteur utilisé dans ce procédé Ceased WO2010103154A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ES200900652A ES2351566B1 (es) 2009-03-09 2009-03-09 Método de cultivo de microorganismos y fotobiorreactor empleado en dicho método.
ESP200900652 2009-03-09

Publications (2)

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WO2010103154A2 true WO2010103154A2 (fr) 2010-09-16
WO2010103154A3 WO2010103154A3 (fr) 2010-10-21

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102086437A (zh) * 2010-12-02 2011-06-08 中国海洋大学 薄膜挂袋式单胞藻光生物反应器及其制作方法
WO2014006232A1 (fr) * 2012-07-03 2014-01-09 Acciona Energía, S. A. Système de fixation de co2 pour la culture de microalgues
WO2014044883A1 (fr) * 2012-09-19 2014-03-27 Universidad De Alicante Photobioréacteur combiné de type "air-lift" pour la production de biomasse
CN114752502A (zh) * 2022-04-29 2022-07-15 云南绿A生物产业园有限公司 一种综合利用碳源提高螺旋藻生长速率的养殖方法及系统
US12264304B2 (en) 2021-04-08 2025-04-01 Premium Oceanic Inc. Systems and methods for deepwater photobioreactor
US12410388B2 (en) 2021-04-08 2025-09-09 Blue Evolution Inc. Photobioreactor systems and methods

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR3073860B1 (fr) * 2017-11-23 2024-07-12 Fermentalg Milieu de culture anti-adhesion

Family Cites Families (10)

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Publication number Priority date Publication date Assignee Title
ES2071572B1 (es) * 1993-07-13 1996-01-16 Univ Granada Dispositivo para el cultivo de microorganismos fotosinteticos y la produccion de biomasa rica en acido e icosapentaenoico.
JP3950526B2 (ja) * 1997-10-17 2007-08-01 次郎 近藤 光合成培養装置及び集合光合成培養装置
ITFI20030047A1 (it) * 2003-02-24 2004-08-25 Univ Firenze Reattore per la coltura industriale di microrganismi fotosintetici
KR100490641B1 (ko) * 2003-12-16 2005-05-19 인하대학교 산학협력단 다중 광생물반응기 및 이를 이용한 광합성 미생물 배양방법
DE102004019234B3 (de) * 2004-04-16 2005-11-24 Sartorius Ag Bioreaktor zur Kultivierung von Mikroorganismen
MXPA05007801A (es) * 2005-07-22 2006-01-30 Univ Autonoma Metropolitana Cultivo de haematococcus pluvialis y produccion de astaxantina en un fotobiorreactor tipo quimiostato.
US20070048848A1 (en) * 2005-08-25 2007-03-01 Sunsource Industries Method, apparatus and system for biodiesel production from algae
US9637714B2 (en) * 2006-12-28 2017-05-02 Colorado State University Research Foundation Diffuse light extended surface area water-supported photobioreactor
US8404004B2 (en) * 2006-12-29 2013-03-26 Genifuel Corporation Process of producing oil from algae using biological rupturing
EP3546588A3 (fr) * 2007-06-01 2019-12-18 Corbion Biotech, Inc. Cellule d'algue recombinante et son utilisation pour la production de lipides

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102086437A (zh) * 2010-12-02 2011-06-08 中国海洋大学 薄膜挂袋式单胞藻光生物反应器及其制作方法
WO2014006232A1 (fr) * 2012-07-03 2014-01-09 Acciona Energía, S. A. Système de fixation de co2 pour la culture de microalgues
WO2014044883A1 (fr) * 2012-09-19 2014-03-27 Universidad De Alicante Photobioréacteur combiné de type "air-lift" pour la production de biomasse
ES2482015A1 (es) * 2012-09-19 2014-07-31 Universidad De Alicante Fotobiorreactor combinado tipo air-lift para la producción de biomasa
US12264304B2 (en) 2021-04-08 2025-04-01 Premium Oceanic Inc. Systems and methods for deepwater photobioreactor
US12410388B2 (en) 2021-04-08 2025-09-09 Blue Evolution Inc. Photobioreactor systems and methods
CN114752502A (zh) * 2022-04-29 2022-07-15 云南绿A生物产业园有限公司 一种综合利用碳源提高螺旋藻生长速率的养殖方法及系统

Also Published As

Publication number Publication date
ES2351566A1 (es) 2011-02-08
WO2010103154A3 (fr) 2010-10-21
ES2351566B1 (es) 2012-06-14

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