EP3714030A1 - Milieu de culture anti-adhésion - Google Patents

Milieu de culture anti-adhésion

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Publication number
EP3714030A1
EP3714030A1 EP18804013.3A EP18804013A EP3714030A1 EP 3714030 A1 EP3714030 A1 EP 3714030A1 EP 18804013 A EP18804013 A EP 18804013A EP 3714030 A1 EP3714030 A1 EP 3714030A1
Authority
EP
European Patent Office
Prior art keywords
varieties
microorganisms
culture
microalgae
culture medium
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.)
Pending
Application number
EP18804013.3A
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German (de)
English (en)
French (fr)
Inventor
Philippe BOUDIER
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.)
Fermentalg SA
Original Assignee
Fermentalg SA
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Filing date
Publication date
Application filed by Fermentalg SA filed Critical Fermentalg SA
Publication of EP3714030A1 publication Critical patent/EP3714030A1/fr
Pending 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
    • C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms
    • 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
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0018—Culture media for cell or tissue culture
    • 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
    • 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
    • C12N1/10—Protozoa; Culture media therefor
    • 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
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C12N2500/22—Zinc; Zn chelators
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C12N2500/24—Iron; Fe chelators; Transferrin
    • 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
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/30—Synthetic polymers

Definitions

  • the present invention relates to a method for culturing microorganisms, in particular protists, designed to prevent the adhesion of microorganisms to the walls of the reactors in which the culture is carried out.
  • Microorganisms tend to adhere to the walls during long-term cultures (from days to months). This phenomenon known as "fouling" is well known. It has very harmful consequences both on crop productivity and on the longevity of the material that must be regularly cleaned.
  • the cells that adhere to the walls of bioreactors reduce the amount of light penetrating inside, they can die for lack of resources (nutrients, oxygen ... etc) and thus cause the development of necrophagous organisms.
  • active surface work methods such as mechanical scraper-type systems (Wet Labs / Sea-Bird BioWiper), pressurized water jet, ultrasound, biocides such as chlorine or bromine (Alconox), anionic detergents for manual washing, UV radiation.
  • C-Spray YSI This can be a surface composed of a biocide that gradually releases the neutralizing element of the biofilm into the medium.
  • biocides are usually composed of copper salts, it releases Cu2 + which interferes with enzymes on membranes preventing their division (YSI 6-Series Anti-Fouling Kits).
  • the invention also relates to a method for culturing microorganisms in a culture medium suitable for their growth, characterized in that the culture medium comprises an appropriate amount of a flocculating agent to prevent the adhesion of microorganisms on the walls of the reactors. in which the culture is carried out.
  • Figure 3 shows the viability of microalgae with different concentrations of starch
  • Figure 5 shows the points on the wall of the transparent column used as a reactor for measuring the light passing through the column.
  • the microorganisms grown are protists.
  • protists all unicellular eukaryotic microorganisms.
  • Microalgae Chlorophytes such as Chlorella, Senedesmus, Tetraselmis, Haematococcus, Charophytes, chrysophytes including diatoms, Nannochloropsis, Euglenophytes such as Euglena, Phacus; Rodophytes including Galdieria ... etc), unicellular fungi (Thrautochytrids such as Schizochytrium, Aurantiochytrium ...
  • cyanobacteria Arabaena, Nostoc, Microcistis, Arthrospira, Spirulina Certainly or cyanobacteria are part of the group of protists.
  • microalgae When the microalgae are of the Chlorella genus, they may be selected from C. acuminata, C. angustoellipsoidea, C. anitrata, C. antarctica, C. aureoviridis, C. autotrophica, C. botryoides, C. caldaria, C. candida , C. capsulata, C. chlorelloides, C. cladoniae, C. coelastroides, C. colonialis, C. com munis, C. conductrix, C. conglomerata, C. desiccata, C. ellipsoidea, C. elongata, C. emersonii , C. faginea, C.
  • the algae of the genus Chlorella may be algae selected from the species C. sorokiniana or C. vulgaris.
  • microalgae When the microalgae are of the Euglena genus, they may be chosen among others from the species E. viridis, E. gracilis, E. limosa, E. globosa, E. prowsei, E. polomorpha.
  • microalgae When the microalgae are of the Scenedesmus genus, they may be selected from the species S. abundans, S. aciculatus, S. aculeolatus, S. aculeotatus, S. acuminatus, S. acutiformis, S. acutus, S. aldavei, S. alternans , S. ambuehlii, S. anhuiensis, S. anomalus, S. antennatus, S. antillarum, S. apicaudatus, S. apiculatus, S. arcuatus, S. aristatus, S. armatus, S. arthrodesmiformis, S. arvernensis, S.
  • ornatus S. ovalternus, S. pannonicus, S. papillosum, S. parisiensis, S. parvus, S. pecsensis, S. pectinatus, S. perforatus, S. planctonicus, S. plarydiscus, S. platydiscus, S. pleiomorphus, S. polessicus, S. polydenticulatus, S. polyglobulus, S. polyspinosus, S. praetervisus, S. prismaticus, S. producto-capitatus, S. protuberans, S. pseudoarmatus, S. pseudobernardii, S. pseudodenticulatus, S. pseudogranulatus, S.
  • pseudohystrix S. pyrus, S. quadrialatus, S. quadricauda, S. quadricaudata, S. quadricaudus, S. quadrispina, S. raciborskii, S. ralfsii, S. reginae, S. regularis, S. reniformis, S. rostrato-spinosus, S. rotundus, S. rubescens, S. scenedesmoides, S. schnepfii, S. schroeteri, S. securiformis, S. semicristatus, S. semipulcher, S. sempervirens, S. senilis, S. serrato-perforatus, S.
  • serratus S. serrulatus, S. setiferus, S. sihensis, S. smithii, S. soli, S. sooi, S. spicatus, S. spinoso-aculeolatus, S. spinosus, S. spinulatus, S. striatus., S. subspicatus, S. tenuispina, S. terrestris, S. tetradesmiformis, S. transilvanicus, S. Tricostatus, S. tropicus, S. tschudyi, S. vacuolatus, S. variabilis, S. velitaris, S. verrucosus, S.
  • microalgae When microalgae are diatoms, they can be chosen from the following genera: Nitzschia, Navicula, Gyrosigma, Phaeodactylum, Thalassiosira ... etc.
  • microalgae When the microalgae are of the genus Nitzschia, they may be selected from the species N. abbreviata, N. abonuensis, N. abridia, N. accedens, N. accommodata, N. aciculariformis, N. acicularioides, N. acicularis (including all varieties), N. acidoclinata, N. actinastroides, N. actydrophila, N. acula, N. acuminata (including all these varieties), N. acuta, N. adamata, N. adamatoides, N. adapta, N. adducta, N. adductoids, N. admissa, N.
  • N. aequalis N. aequatorialis, N. aequora, N. aequorea
  • N. aerophila N. aerophiloides
  • N. aestuari N. affinis, N. africana, N. agnewii , N. agnita, N. alba, N. albicostalis, N. alexandrina, N. alicae, N. allanssonii, N. alpina, N. alpinobacillum, N. amabilis, N. ambigua, N. americana, N. amisaensis, N. amphibia, N. amphibia (including all these varieties), N. amphibioides, N.
  • N. ardua N. aremonica
  • N. arenosa N. areolata
  • N. armoricana N. asperula, N. astridiae, N. atomus, N. attenuata, N. aurantiaca, N. aurariae, N. aurica, N. auricula, N. australis, N. austriaca, N. bacata (including all these varieties), N. bacillariaeformis, N. bacilliformis, N. bacillum, N. balatonis, N. balcanica, N. baltica, N. barbieri (including all these varieties), N. barkleyi, N. barronii, N.
  • N. brevirostris N. brevissima (including all these varieties), N. brevistriata, N. brightwellii, N. brittonii, N. brunoi, N. bryophila, N. buceros, N. bukensis, N. bulnheimiana, N. buschbeckii, N. calcicola, N. caledonensis, N. calida (including all these varieties), N. californica, N. campechiana, N. capensis, N. capitata, N. capitellata (including all these varieties), N. capsuleuspalae, N. carnicobarica, N. carnico-barica, N. challengeri, N. chalonii, N.
  • congolensis N. constricta (including all these varieties), N. consummata, N. diverenta, N. costei, N. coutei, N. creticola, N. cucumis, N. cursoria, N. curta , N. curvata, N. curvilineata, N. curvipunctata, N. curvirostris (including all these varieties), N. curvula (including all these varieties), N. cuspidata, N. cylindriformis, N. cylindrus, N. dakariensis, N. davidsonii, N. dealpina, N. debilis, N. decipiens, N. delauneyi, N. americanissima, N.
  • N. draveillensis, N. droebakensis, N. dubia including all these varieties
  • N. dubiformis N. dubioides
  • N. ebroicensis N. eglei, N. elegans, N. elegantula, N. elegens, N. elliptica, N. elongata, N. entomon, N. epiphytica, N. epiphyticoides, N. epithemiformis, N. epithemioides, N. epithemoides (including all these varieties), N. epsilon, N. erlandssonii, N. erosa, N. etoshensis, N. examanda, N.
  • eximia N. famelica, N. fasciculata, N. febigeri, N. ferox, N. ferrazae, N. fibula-fissa, N. filiformis (including all these varieties), N. flexa, N. flexoides, N. fiuminensis, N. fluorescens, N. fluvialis, N. fogedii, N. fonticola (including all these varieties), N. fonticoloides, N. fonticula, N. fontifuga, N. forfica, N. formosa, N. fossalis, N. fossilis, N. fragilariiformis, N. franconica, N. fraudulenta, N. frauenfeldii, N.
  • N. goetzeana including all these varieties
  • N. gotlandica N. graciliformis
  • N. gracilis including all these varieties
  • N. gracillima N. graciloides
  • N. granulata including all these varieties
  • N. granulosa, N. groenlandica N. grossestriata, N. grovei, N. gruendleri, N. gru nowii, N. guadalupensis, N. guineensis, N. guttula, N. gyrosigma, N.
  • N. homburgiensis N. hudsonii, N. hummii, N. hungarica (including all these varieties), N. hustedti, N. hustedtiana, N. hyalina, N. hybrida (including all these varieties), N. hybridaeformis, N. ignorata (including all these varieties), N. iltisii, N. impressa, N. improvisa, N. incerta, N. incognita, N. inconspicua, N. incrustans, N. incurva (including all these varieties), N. indica, N. indistincta, N. inducta, N. inflatula, N. ingenua, N.rysta, N.
  • innominata N. insecta, N. insignis (N. including all varieties), N. intermedia (including all these varieties), N. intermissa, N. interrupta, N. interruptstriata, N. invicta (including all these varieties), N. invisa, N. invisitata, N. iranica, N. irregularis, N. irremissa, N. irrepta, N. irresoluta, N. irritans, N. italica, N. janischii, N. jelineckii, N. johnmartinii, N. juba, N. jucunda, N. jugata (including all these varieties ), N. jugiformis, N.
  • lorenziana (including all these varieties), N. lucisensibilis, N. lunaris, N. lunata, N. lurida, N. luzonensis, N. macaronesica, N. macedonica, N. macera, N. machardyae, N. macilenta (including all these varieties), N. magnacarina, N. mahihaensis, N. mahoodii, N. maillardii, N. major, N. majuscula (including all these varieties) ), N. makarovae, N. manca, N. mancoides, N. manguini, N. marginata, N. marginulata (including all these varieties), N. marina, N. martiana, N.
  • N. nathorsti N. navicularis, N. navis-varingica, N. navrongensis, N. neglecta, N. nelsonii, N. neocaledonica, N. neoconstricta, N. neofrigida, N. neogena, N. neotropica, N. nereidis, N. nicobarica, N. nienhuisii, N. normannii, N. notabilis, N. nova, N. novae-guineaensis, N. novae-guineensis, N. novaehollandiae, N. nova-zealandia, N.
  • N. nyassensis N. operheimiana, N. obesa, N. obliquecostata, N. obscura, N. obscurepunctata, N. obsidialis, N. obsoleta, N. obsoletiformis, N. obtusa (including all these varieties), N. obtusangula, N. oceanica, N. ocellata, N. oiiffi, N. omega, N. osmophila, N. ossiformis, N. ostenfeldii, N. ovalis, N. paaschei, N. pacifica, N. palacea, N. palea (including all these varieties), N. paleacea, N.
  • N. Perversa N. petitiana, N. philippinarum, N. pilum, N. pinguescens, N. foundedarum, N. plana (including all these varieties), N. planctonica, N. plicatula, N. plioveterana, N. polaris, N. polymorpha, N. ponciensis, N. praecurta, N. praefossilis, N. praereinholdii, N. princeps, N. procera, N. prolongata (including all these varieties), N. prolongatoides, N. promare, N. propinqua, N. pseudepiphytica, N.
  • pseudoamphioxoides N. pseudoamphioxys, N. pseudoamphyoxys, N. pseudoatomus, N. pseudobacata, N. pseudocapitata, N. pseudocarinata, N. pseudocommunis, N. pseudocylindrica, N. pseudodelicat issima, N. pseudofonticola, N. pseudohungarica, N. pseudohybrida, N. pseudonana, N. pseudoseriata, N. pseudosigma, N. pseudosinuata, N. pseudostagnorum, N. pubens, N. pulcherrima, N. pumila, N. punctata (including all these varieties), N. pismes (including all these varieties), N. pungiformis, N.
  • N. puriformis N. pusilla (including all these varieties), N. putrida, N. quadrangula, N. quickiana, N. rabenhorstii, N. radicula (including all these varieties), N. rautenbachiae, N. recta (including all these varieties), N. rectiformis, N. rectilonga, N. rectirobusta, N. rectissima, N. regula, N. reimeri, N. reimerii, N. reimersenii, N. retusa, N. reversa, N. rhombica, N. rhombiformis, N. rhopalodioides, N.
  • N. rigida richterae, N. rigida (including all these varieties), N. ritscheri, N. robusta, N. rochensis, N. rolandii, N. romana, N. romanoides, N. romanowiana, N. rorida, N. rosenstockii, N. rostellata, N. rostrata, N. ruda, N. rugosa, N. rupestris, N. rusingae, N. ruttneri, N. salinarum, N. salinicola, N. salpaespinosae, N. salvadoriana, N. sansimoni, N. sarcophagum, N. scabra, N.
  • N. scalaris N. scaligera, N. scalpelliformis, N. schoenfeldii, N. schwabei, N. schweikertii, N. scutellum, N. sellingii, N. semicostata, N. semirobusta, N. separanda, N. seriata (including all these varieties ), N. serpenticola, N. serpentiraphe, N. serrata, N. sibula (including all these varieties), N. sigma (including all these varieties), N. sigmaformis, N. sigmatella, N. sigmoidea (including all these varieties), N. silica, N. silicula (including all these varieties), N. siliqua, N.
  • N. streinikovae N. stricta, N. strigillata, N. striolata, N. subaccommodata, N. subacicularis, N. subacuta, N. subamphioxioides, N. subapiculata, N. subbacata, N. subcapitata, N. subcapitellata, N. su bcohaerens (including all these varieties), N. subcommunis, N. subconstricta, N. subcurvata, N. subdenticula, N. subfalcata, N. subfraudulenta, N.
  • subfrequens N. subfrustulum, N. subgraciloides, N. subinflata, N. subinvicta, N. sublaevis, N. sublanceolata, N. sublica, N. subirinearis, N. sublongirostris, N. submarina, N. submediocris, N. subodiosa, N. subpacifica, N. subpunctata, N. subromana, N. subrostrata, N. subbrostratoides, N. subrostroides, N. subsalsa, N. subtilioides, N. subtilis (including all these varieties), N. subtubicola, N. subvitrea, N. suchlandtii, N.
  • N. sulcata N. sundaensis, N. supralitorea, N. tabellaria, N. taenia, N. taeniiformis, N. tantata, N. tarda, N. taylorii, N. temperei, N. tenella, N. tenerifa, N. tenuiarcuata, N. tenuirostris, N. tenuis (including all these varieties), N. tenuissima, N. tergestina, N. terrestris, N. terricola, N. thermalis (including all these varieties), N. thermaloides, N. tibetana, N. tirstrupensis, N. tonoensis, N.
  • towutensi s N. translucida, N. tropica, N. tryblionella (including all these varieties), N. tsarenkoi, N. tubicola, N. tumida, N. turgidula, N. turgiduloides, N. umaoiensis, N. umbilicata, N. umbonata, N. vacillata, N. vacua, N. valdecostata, N. valdestriata, N. valens, N. valga, N. valida (including all these varieties), N. vanheurckii, N. vanoyei, N. vasta, N. ventricosa, N.
  • N. vermicularioides N. vermicularis (including all these varieties), N. vermicularoides, N. vexans, N. victoriae, N. vidovichii, N. vildaryana, N. villarealii, N. virgata, N. visurgis, N. vitrea (including all these varieties), N. vivax (including all these varieties). ), N. vixnegligenda, N. vonhauseniae, N. vulga, N. weaveri, N. weissflogii, N. westii, N. williamsiii, N. wipplingeri, N. witkowskii, N. wodensis, N. woltereckii, N. woltereckoides, N. wuellerstorfii, N. wunsamiae, N. yunchengensis, N. zebuana, N. zululandica.
  • the algae of the genus Nitzschia may be algae selected from the species N. sp.
  • microalgae When the microalgae are of the genus Haematococcus, they may be selected from the species H. allmanii, H. buetschlii, H. capensis, H. carocellus, H. droebakensis, H. grevilei, H. insignis, H. lacustris, H. murorum , H. pluvialis, H. salinus, H, sanguineis, H. thermalis, H. zimbabwiensis.
  • microalgae When microalgae are of the genus Aurantiochytrium, they may be chosen from among the species: A. limacinum, A. mangrovei
  • microalgae When the microalgae are of the genus Schizochytrium, they may be chosen from among the species: S. aggregatum, S. limacinum, S. mangrovei, S. minutum, S. octosporum.
  • microalgae When the microalgae are of the genus Crypthecodinium, they may be chosen from among the species: C. cohnii, C. setense.
  • the microalgae When the microalgae are of the Tetraselmis genus, they may be chosen from among the species: T. alacris, T. apiculata, T. arnoldii, T. ascus, T. astigmatica, T. bichlora, T. bilobata, T. bolosiana, T. chui, T. contracta, T. convolutae, T. cordiformis, T. desikacharyi, T. elliptica, T. fontiana, T. gracilis, T. hazenii, T. helgolandica, T. impellucida, T. incisa, T. inconspicua, T. indica, T. levis, T.
  • the methods of culturing microorganisms are also well known to those skilled in the art, whether in the auxotrophic, heterotrophic or mixotrophic mode (references).
  • the culture media used for these different culture methods adapted to the various microorganisms mentioned above are also well known to those skilled in the art (reference).
  • the invention is particularly suitable for so-called long-term cultures (from several days to several months), subject to the phenomenon of adhesion of microorganisms to the walls of the reactors in which they are grown.
  • reactors employed for these different modes of cultivation are also known to those skilled in the art, such as in bubble columns, airlifts (Lesson: Industrial). Applications of Microbes, Dr. Parvinder Kaur, page 11-12), fermenters (Tryton TM), tubular photobioreactors (Schott, Synoxis Algae), raceways and bioreactors.
  • the walls of the bioreactors are made of materials known to those skilled in the art, in particular flexible plastic, stainless steel, concrete or brick.
  • the invention is particularly suitable for cultures carried out in reactors comprising at least one transparent wall to let light, be it natural light or artificial light, in autotrophic mode or in mixotrophic mode.
  • the light-transparent walls may be made of glass, borosilicate, plastic of the polymethyl methacrylate (PMMA), polyvinyl chloride (PVC), polyethylene (PE) type, in particular low polyethylene type. density (PE-LD), polycarbonate, polystyrene (PS).
  • PMMA polymethyl methacrylate
  • PVC polyvinyl chloride
  • PE polyethylene
  • PE-LD polyethylene
  • PS polystyrene
  • the transparent walls are made of PMMA, PVC, PE or polycarbonate.
  • the invention is particularly suitable for transparent wall type reactors with autotrophic bubble column type for the cultivation of microalgae, such as carbon sinks.
  • These carbon sinks are bubble columns or "air lifts" for growing microalgae in auxotrophic mode and in which the ambient air is passed through which carbon dioxide is absorbed by the microorganisms as carbon source. By multiplying, they clean up the atmosphere by absorbing carbon dioxide.
  • Such systems are described in particular in WO 2014/063229 or WO 2017/077061.
  • microalga To divide the microalga needs light.
  • the availability of light in the environment is essential for the development of microalgae.
  • the adhesion on the walls of the reactor decreases the available light and growth capacity, and therefore carbon absorption of microalgae.
  • the carbon sink corresponds to a variable volume column containing microalgae that will be able to capture between 1 and 10,000 tons of CO2 per year.
  • the light does not come from outside the bioreactor through transparent walls, but light is brought into the bioreactor by plunging sources of sealed lights into the vessel.
  • a light backlight system may be used (WO2014 / 174182). The invention then makes it possible to reduce the adhesion of the cells to the light source.
  • the culture medium used includes "BG-11 growth media", described and marketed by UTEX (UTEX Culture Collection of Algae, 205 W. 24th St., Biological Labs 218, The University of Texas at Austin (F0402), Austin, TX 78712 USA).
  • UTEX UTEX Culture Collection of Algae, 205 W. 24th St., Biological Labs 218, The University of Texas at Austin (F0402), Austin, TX 78712 USA.
  • the culture medium does not include a carbon source other than the carbon dioxide contained in the air that passes into the culture medium.
  • a carbon source other than carbon dioxide such as glucose.
  • the ambient air flow rate in the culture medium corresponds to a flow rate ranging from 0.3 to 0.5 vvm.
  • Outside air containing CO2 passes through a bubbler to be injected into the column.
  • the injected air can also come from factory smoke (eg incineration plants) which contain between 1 and 20% CO2.
  • the carbon sink is drained. Finally, the well is again filled with medium to be inoculated with microalgae.
  • Flocculating agents are well known to those skilled in the art. They are usually used at the end of the microorganism culture, added to the fermentation must when the culture is complete to facilitate the separation of the biomass from the culture medium (Hanotu J. & al., 2012). These additions once the crop is complete are generally not detrimental to the use of recovered microalgae, for example for feed for aquaculture (US 2013/205850).
  • flocculants are organic or inorganic, such as chitosan, arboxymethylcelluloses, starches or starch derivatives such as rice starch, corn starch, tapioca starch, yellow dextrin, starch potato, pregelatinized starch or cationic starch for organic flocculants, or aluminum sulphate, calcium chloride, zinc chloride or iron chloride for inorganic flocculants.
  • organic or inorganic such as chitosan, arboxymethylcelluloses, starches or starch derivatives such as rice starch, corn starch, tapioca starch, yellow dextrin, starch potato, pregelatinized starch or cationic starch for organic flocculants, or aluminum sulphate, calcium chloride, zinc chloride or iron chloride for inorganic flocculants.
  • the use of flocculants according to the invention is during the culture, in the culture medium, not to promote the separation of the biomass from the culture medium, but to favor the maintenance of this biomass in the culture medium by preventing its adhesion on the walls of the reactors.
  • the flocculating agents according to the invention must be able to be used without affecting the growth capacities and the biological activity of the microorganisms cultivated. Indeed the viability tests have revealed that flocculants and iron chloride in particular do not disturb the growth of microalgae. ( Figures 1, 2 and 3)
  • the skilled person will be able to select by simple laboratory tests the agents flocculants best suited to cultured microorganisms and their growing conditions.
  • the amount of flocculating agent in the culture medium ranges from 50 to 450 mg / l, more particularly from 150 to 350 mg / l.
  • the addition of flocculating agents in the culture medium can lead to the formation of suspended cell aggregates.
  • the culture will be done under usual conditions of agitation, such as mechanical or passive agitation such as in a bubble column or an air lift reactor.
  • the invention also relates to a culture medium suitable for the culture of microorganisms, characterized in that it comprises from 50 to 450 mg / l, more particularly from 150 to 350 mg / l of flocculating agent as defined above, in particular of iron chloride.
  • the columns were inoculated at 0.5 g / L of microalgae.
  • the temperature remained constant at 30 ° C.
  • the brightness was set at 500 pEinstein / m 2 / s.
  • the pH is stabilized at 6.3 up to 160 hours (control) 189 hours (Fecl3) and is no longer regulated, the pH will rise to 8 to stabilize at this level until the end of the experiment. It was chosen not to regulate the pH because a high pH makes it possible to increase the bonding capacities of microalgae to the walls.
  • the biomass is recovered with a pump, one obtains the biomass in suspension as well as that remained at the bottom of the column. Then a volume of water is added to the column in order to take off the microalgae remaining on the walls. A dry mass measurement is performed on each batch.
  • the total biomass suspended in the medium represents 18 g and the biomass remained at the bottom of the column amounted to 1.11 g.
  • the biomass bonded to the walls is 0.043 g, which represents 0.22% of the total biomass.
  • the light measurements were carried out on 24 points as shown in the schematic representation of the column in Figure 5.
  • the light was measured on a clean column without microalgae, on a control column with microalgae and a column containing FeCl3 with microalgae.

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