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WO1994007993A1 - Method of producing microalgae - Google Patents

Method of producing microalgae Download PDF

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Publication number
WO1994007993A1
WO1994007993A1 PCT/FR1993/000989 FR9300989W WO9407993A1 WO 1994007993 A1 WO1994007993 A1 WO 1994007993A1 FR 9300989 W FR9300989 W FR 9300989W WO 9407993 A1 WO9407993 A1 WO 9407993A1
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Prior art keywords
microalgae
culture
biomass
forcing
metabolites
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PCT/FR1993/000989
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French (fr)
Inventor
Catherine Dupre
Dominique Grizeau
Jean-Claude Guary
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Conservatoire National Des Arts Et Metiers
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Publication of WO1994007993A1 publication Critical patent/WO1994007993A1/en

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    • 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

Definitions

  • microalgae culture lies a • *> share large quantities of biomass production, subsequently exploited, particularly in the food industry, ticks cosmé ⁇ or agriculture, and on the other hand in the production of metabolites of economic interest.
  • any product in natural production will be considered in terms of industrial economic balance, more profitable than its production by synthesis.
  • pigments such as phycoerythrin, phycocyanin and beta-carotene
  • lipids such as eicosapentaenoic acid, phosphoric and sulphate-lipids, amino acids such as proline, and molecules with therapeutic activity such as certain derivatives of sterols.
  • microalgae cultures can be obtained in either extensive or intensive mode.
  • the biomasses obtained at the end of the culture range from 0.1 to a few kg / m 2 ⁇ 5 dry matter, relatively low values in comparison with other cultures of microorganisms such as bacteria and hetero trophic fungi.
  • microalgae are harvested according to various methods involving flocculation / decantation, flocculation / flotation and / or centrifugation operations, etc.
  • the known methods and the installations for implementing them are distinguished by their respective economic returns.
  • solid / liquid separations by mechanical means involve the use of oversized material, - ⁇ reducing the profitability of this type of production all the more.
  • the accumulation of molecules with high added value is practiced during the culture of microalgae by playing on environmental factors. The methods used differ according to the molecules sought and the strains cultivated.
  • the object of this invention is therefore to optimize the implementation of the conditions causing the accumulation of molecules of economic interest while optimizing the harvest of microalgae.
  • the process according to the present invention is characterized by the following steps: a) cultivating the microalgae in a liquid medium under appropriate conditions essentially ensuring the formation of a large biomass, b) concentrating the biomass obtained, c) placing this concentrated biomass in conditions essentially ensuring the accumulation of metabolite, and d) harvesting the biomass containing the metabolite and / or the medium containing the metabolite.
  • Step a) of biomass production implements all of the techniques known and developed in the prior art for the optimal production of algal biomass.
  • the culture of microalgae can be carried out in continuous or discontinuous mode, either in semi-control mode (extensive or semi-intensive cultures), or in controlled mode (intensive cultures).
  • the culture medium conventionally used for the production of algal biomass, under photoautotrophic conditions can be enriched with the aid of solid or liquid fertilizer formulations, marketed for horticultural uses, provided that they are used in optimal doses.
  • the concentrated biomass obtained in step b) represents less than 1/5 of the volume of the culture in step a).
  • this concentrated biomass represents between 1/10 and 1/20 of the volume of the culture in step a).
  • the concentration operated in step b) will be advantageously carried out by flocculation of microalgae.
  • the alkalinity of the medium is preferably varied after stopping - 0 carbonation for 2 to -f hours in light period.
  • Another preferred means of flocculation in the process according to the present invention consists in using self-flocculating microalgae, and / or in adding chemical, biological flocculating agents and their mixtures.
  • Such agents are known from the prior art, such as, for example, polysaccharides such as chitosan or metal salts such as iron perchloride.
  • the 0 flocculated algae are then concentrated by appropriate means of static or mechanical decan ⁇ tion or by appropriate means of flotation.
  • the accumulation of metabolites in step c) is obtained by physiological forcing.
  • inducing compound any chemical or biological compound which, when added to the culture medium
  • the inducing compounds are, for example nitrogen sources such as nitrate, ammonium or urea, sources of trace elements such as sodium moiybdate, sources of hormones such as certain auxins or sources of acids and / or bases for regulate the pH to a ** -> optimal value.
  • the duration of physiological forcing is preferably between 2 and 15 hours.
  • the biomass and / or the medium containing the metabolites will be harvested by centrifugation and / or filtration.
  • a tangential filtration type filtration will preferably be used.
  • the algal paste obtained then comprises between 20 and 25% of dry matter.
  • the method according to the present invention applies equally to microalgae of marine, fresh or terrestrial water, prokaryotic or eukaryotic origin. 5
  • the examples below describe preferential embodiments of the method according to the present invention, without however seeking to limit its scope.
  • Example 1 Production of phycoerythrin by culture of Porphyridum cruetum, Rhodosorus marinus or Rhodella violacea.
  • the algae are cultivated in a conventional manner and pre-harvested by flocculation / decantation. They are transferred to a physiological forcing tank allowing optimal transfers of matter and light.
  • the induction of phycoerythrin synthesis is favored by an illumination of the order of 50 ⁇ E.cm 2 .s ⁇ and an enrichment in nitrate (2gJ ⁇ ) and sodium heptamolybdate (25 gJ " ) for biomass of a few gl " (dry weights).
  • the final harvest takes place after 3 hours of incubation.

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  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Botany (AREA)
  • Biomedical Technology (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
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Abstract

A method of producing microalgae giving metabolites comprises the steps of cultivating the microalgae, concentrating them, invoking accumulation of metabolites, and harvesting the biomass and/or the enriched medium. The method of physiological forcing of microalgae or for the over-accumulation of metabolites is characterized in that forcing is carried out in culture volumes smaller than 1/5 of the normal production volumes for algal biomass.

Description

" Procédé de Production de Micro-algues" "Microalgae Production Process"
La présente invention concerne un procédé de production de microalgues produisant, en particulier, des metabolites d'intérêt écono¬ mique.The present invention relates to a process for the production of microalgae producing, in particular, metabolites of economic interest.
Le principal intérêt de la culture de microalgues réside d'une •*> part dans la production de biomasse en grande quantité, exploitée ultérieurement, notamment dans l'industrie agro-alimentaire, les cosmé¬ tiques ou pour l'agriculture, et d'autre part dans la production de metabolites d'intérêt économique.The main interest of microalgae culture lies a • *> share large quantities of biomass production, subsequently exploited, particularly in the food industry, ticks cosmé¬ or agriculture, and on the other hand in the production of metabolites of economic interest.
Par metabolites d'intérêt économique, on entend en particulier l O des molécules à haute valeur ajoutée produites naturellement et dont la synthèse est délicate du fait de leur complexité.The term metabolites of economic interest means in particular the O of molecules with high added value produced naturally and the synthesis of which is difficult due to their complexity.
Entrera également dans cette catégorie, tout produit ddnt la production naturelle sera considérée en terme de bilan économique industriel, plus rentable que sa production par voie de synthèse. l*-5 Parmi les metabolites d'intérêt économique, on trouvera, par exemples, des pigments tels que phycoérythrine, phycocyanine et béta-carotène, des lipides tels que l'acide éicosapentaénoique, des phospho- et sulfo-lipides, des acides aminés tels que la proline, et des molécules à activité thérapeutique tels que certains dérivés de stérols.Also falling into this category, any product in natural production will be considered in terms of industrial economic balance, more profitable than its production by synthesis. l * - 5 Among the metabolites of economic interest, we find, for example, pigments such as phycoerythrin, phycocyanin and beta-carotene, lipids such as eicosapentaenoic acid, phosphoric and sulphate-lipids, amino acids such as proline, and molecules with therapeutic activity such as certain derivatives of sterols.
20 Toutefois, l'exploitation des microalgues implique pour être rentable la maîtrise des conditions de culture et de récolte, tout en préservant la qualité biochimique de la matière algale. Les cultures de microalgues peuvent être obtenues en mode soit extensif, soit intensif. Les biomasses obtenues en fin de culture vont de 0,1 à quelques kg/m de 2*5 matière sèche, valeurs relativement faibles par comparaison avec d'autres cultures de microorganismes tels que bactéries et champignons hétéro- trophes.20 However, the exploitation of microalgae implies to be profitable the control of the culture and harvest conditions, while preserving the biochemical quality of the algal matter. Microalgae cultures can be obtained in either extensive or intensive mode. The biomasses obtained at the end of the culture range from 0.1 to a few kg / m 2 × 5 dry matter, relatively low values in comparison with other cultures of microorganisms such as bacteria and hetero trophic fungi.
Classiquement, les microalgues sont récoltées selon divers procédés impliquant des opérations de floculation/décantation, flo- 30 cula.tion/flottation et/ou centrifugation, etc. Les procédés connus et les installations pour les mettre en oeuvre se distinguent par leurs rentabilités économiques respectives. Lorsqu'il s'agit de cultures cellulaires photoauto- trophes, donc relativement diluées, les séparations solides/liquides par voie mécanique impliquent la mise en oeuvre de matériel surdimensionné, -^ réduisant d'autant la rentabilité de ce type de production. Selon l'art antérieur, l'accumulation de molécules à forte valeur ajoutée, est pratiquée durant la culture des microalgues en jouant sur les facteurs environnementaux. Les méthodes utilisées diffèrent selon les molécules recherchées et les souches cultivées. La maîtrise des conditions optimales pour ces biosynthèses est loin d'être assurée dès qu'il s'agit de volumes de cultures supérieurs à quelques m . En effet à ces échelles de production apparaissent des limitations liées à des problèmes de transfert de matière et/ou de lumière. Dans ces conditions, la maîtrise des facteurs environnementaux permettant l'accumulation des metabolites recherchés implique, de manière générale des coûts de production très élevés.Conventionally, microalgae are harvested according to various methods involving flocculation / decantation, flocculation / flotation and / or centrifugation operations, etc. The known methods and the installations for implementing them are distinguished by their respective economic returns. When it comes to photoautotrophic cell cultures, therefore relatively diluted, solid / liquid separations by mechanical means involve the use of oversized material, - ^ reducing the profitability of this type of production all the more. According to the prior art, the accumulation of molecules with high added value is practiced during the culture of microalgae by playing on environmental factors. The methods used differ according to the molecules sought and the strains cultivated. Controlling the optimal conditions for these biosyntheses is far from being guaranteed when it comes to crop volumes greater than a few m. Indeed on these production scales appear limitations linked to problems of transfer of matter and / or light. Under these conditions, the control of the environmental factors allowing the accumulation of the metabolites sought implies, in general, very high production costs.
L'objet de cette invention est donc d'optimiser la mise en oeuvre des conditions provoquant l'accumulation de molécules d'intérêts économi¬ ques tout en optimisant la récolte des microalgues.The object of this invention is therefore to optimize the implementation of the conditions causing the accumulation of molecules of economic interest while optimizing the harvest of microalgae.
La présente invention concerne donc un procédé de récoite de microalgues permettant de dissocier les conditions optimales de production de biomasse de celles relatives à la production des metabolites recherchés.The present invention therefore relates to a process for receiving microalgae making it possible to dissociate the optimal conditions for the production of biomass from those relating to the production of the desired metabolites.
Le procédé selon la présente invention se caractérise par les étapes suivantes : a) cultiver les microalgues en milieu liquide dans des conditions appropriées assurant essentiellement la formation d'une biomasse importante, b) concentrer la biomasse obtenue, c) placer cette biomasse concentrée dans des conditions assurant essentiellement l'accumulation de métabolite, et d) récolter la biomasse contenant le métabolite et/ou le milieu contenant le metabolite.The process according to the present invention is characterized by the following steps: a) cultivating the microalgae in a liquid medium under appropriate conditions essentially ensuring the formation of a large biomass, b) concentrating the biomass obtained, c) placing this concentrated biomass in conditions essentially ensuring the accumulation of metabolite, and d) harvesting the biomass containing the metabolite and / or the medium containing the metabolite.
L'étape a) de production de biomasse met en oeuvre l'ensemble des techniques connues et développées dans l'art antérieur pour la production optimale de biomasse alguale.Step a) of biomass production implements all of the techniques known and developed in the prior art for the optimal production of algal biomass.
Ainsi, la culture des microalgues pourra être réalisée en mode continu ou discontinu, soit en mode semi-contrôle (cultures extensives ou semi-intensives), soit en mode contrôlé (cultures intensives). Le milieu de culture classiquement utilisé pour la production de biomasse algale, en conditions de photoautotrophie, pourra être enrichi à l'aide de formulations d'engrais solides ou liquides, commercialisées pour des usages horticoles, à condition d'être utilisées aux doses optimales. D'une manière avantageuse, la biomasse concentrée obtenue dans l'étape b) représente moins de 1/5 du volume de la culture dans l'étape a).Thus, the culture of microalgae can be carried out in continuous or discontinuous mode, either in semi-control mode (extensive or semi-intensive cultures), or in controlled mode (intensive cultures). The culture medium conventionally used for the production of algal biomass, under photoautotrophic conditions, can be enriched with the aid of solid or liquid fertilizer formulations, marketed for horticultural uses, provided that they are used in optimal doses. Advantageously, the concentrated biomass obtained in step b) represents less than 1/5 of the volume of the culture in step a).
Préférentiellement, cette biomasse concentrée représente entre 1 /10 et 1 /20 du volume de la culture dans l'étape a). •-> Selon la présente invention, la concentration opérée dans l'étape b) sera effectuée avantageusement par floculation des microalgues.Preferably, this concentrated biomass represents between 1/10 and 1/20 of the volume of the culture in step a). • -> According to the present invention, the concentration operated in step b) will be advantageously carried out by flocculation of microalgae.
De préférence, la floculation est effectuée par augmentation de l'alcalinité du milieu de culture de l'étape a).Preferably, the flocculation is carried out by increasing the alkalinity of the culture medium from step a).
On fait de préférence varier l'alcalinité du milieu après arrêt de -- 0 la carbonatation durant 2 à -f heures en période lumineuse.The alkalinity of the medium is preferably varied after stopping - 0 carbonation for 2 to -f hours in light period.
Cette augmentation de l'alcalinité pourra être obtenue par l'addition de soude et/ou de phosphate de sodium.This increase in alkalinity can be obtained by the addition of soda and / or sodium phosphate.
Une attention particulière devra être apportée à cette étape afin que la floculation par modification de l'alcalinité n'entraîne pas une -* -5 dégradation des microalgues à des pH trop élevés.Particular attention should be paid to this stage so that the flocculation by changing the alkalinity does not cause - * - 5 degradation of microalgae at too high pH.
On pourra toutefois employer comme soude ou comme phosphate de sodium des produits destinés à l'usage agricole, particulièrement économiques pour la mise en oeuvre du procédé selon l'invention.However, it is possible to use as soda or as sodium phosphate products intended for agricultural use, which are particularly economical for implementing the process according to the invention.
Un autre moyen préférentiel de floculation dans le procédé selon 0 la présente invention consiste à employer des microalgues autofloculantes, et/ou à ajouter des agents floculants chimiques, biologiques et leur mélanges.Another preferred means of flocculation in the process according to the present invention consists in using self-flocculating microalgae, and / or in adding chemical, biological flocculating agents and their mixtures.
De tels agents sont connus de l'art antérieur, comme par exemple des polysaccharides tels que le chitosan ou des sels métalliques tels que le *5 perchlorure de fer.Such agents are known from the prior art, such as, for example, polysaccharides such as chitosan or metal salts such as iron perchloride.
Bien entendu, l'homme du métier saura optimiser les conditions de floculation, en particulier par combinaison des différents moyens décrits ci-dessus.Of course, those skilled in the art will be able to optimize the flocculation conditions, in particular by combining the various means described above.
Suivant le procédé selon la présente invention, les algues 0 floculées sont ensuite concentrées par des moyens appropriés de décan¬ tation statique ou mécanique ou par des moyens appropriés de flottation.According to the method according to the present invention, the 0 flocculated algae are then concentrated by appropriate means of static or mechanical decan¬ tion or by appropriate means of flotation.
D'une manière préférentielle, on emploiera un décanteur cylindro-conique à flux vertical dont la pente de la partie conique estPreferably, a cylindrical-conical settling tank with vertical flow will be used, the slope of the conical part of which is
5 comprise entre 50° et 70° pour une vitesse ascentionnelle moyenne comprise entre 1,5 et 2,5 m/h.5 between 50 ° and 70 ° for an average ascending speed between 1.5 and 2.5 m / h.
D'une manière avantageuse, l'accumulation de metabolites dans l'étape c) est obtenue par forçage physiologique.Advantageously, the accumulation of metabolites in step c) is obtained by physiological forcing.
En fonction du métabolite recherché, le forçage physiologique sera induit par différents paramètres, notamment la température de culture, l'exposition à la lumière, ou l'enrichissement du milieu de culture en différents composés inducteurs. Par composé inducteur, on entend tout composé chimique ou biologique qui, lorsqu'additionné au milieu de cultureDepending on the metabolite sought, physiological forcing will be induced by different parameters, in particular the culture temperature, exposure to light, or the enrichment of the culture medium with different inducing compounds. By inducing compound is meant any chemical or biological compound which, when added to the culture medium
10 modifie le métabolisme des microalgues favorisant la production de metabolites. Les composés inducteurs sont, par exemplaes des sources azotées telles que nitrate, ammonium ou urée, des sources d'oligoéiéments tel le moiybdate de sodium, des sources d'hormones telles que certaines auxines ou des sources d'acides et/ou de bases pour réguler le pH à une ** -> valeur optimale.10 modifies the metabolism of microalgae promoting the production of metabolites. The inducing compounds are, for example nitrogen sources such as nitrate, ammonium or urea, sources of trace elements such as sodium moiybdate, sources of hormones such as certain auxins or sources of acids and / or bases for regulate the pH to a ** -> optimal value.
La durée du forçage physiologique, fonction de la nature des metabolites recherchés et de l'espèce de microaigue cultivée, est de préférence comprise entre 2 et 15 heures.The duration of physiological forcing, depending on the nature of the metabolites sought and the species of microalgae cultivated, is preferably between 2 and 15 hours.
Enfin, après avoir assuré l'accumulation de metabolites, la 0 biomasse et/ou le milieu contenant les metabolites seront récoltés par centrifugation et/ou filtration.Finally, after ensuring the accumulation of metabolites, the biomass and / or the medium containing the metabolites will be harvested by centrifugation and / or filtration.
Dans ce dernier cas, on emploiera préférentiellement une filtration de type filtration tangentielle.In the latter case, a tangential filtration type filtration will preferably be used.
On constatera donc que le procédé selon la présente invention s'applique aussi bien à la production de metabolites sécrétés dans les microalgues qu'à celle de metabolites excrétés dans le milieu de culture.It will therefore be seen that the process according to the present invention applies both to the production of metabolites secreted in microalgae and to that of metabolites excreted in the culture medium.
La pâte alguale obtenue comprend alors entre 20 et 25% de matière sèche.The algal paste obtained then comprises between 20 and 25% of dry matter.
Après sa récolte, la biomasse enrichie peut éventuellement être 0 conditionnée, en particulier par congélation, cette dernière pouvant être effectuée sous vide ou sous atmosphère contrôlée.After its harvest, the enriched biomass can optionally be packaged, in particular by freezing, the latter being able to be carried out under vacuum or under a controlled atmosphere.
Le procédé selon la présente invention s'applique indifféremment aux microalgues d'origine marine, d'eau douce ou terrestre, procaryotes ou eucaryotes. 5 Les exemples ci-après décrivent des modes de réalisation préférentiels du procédé selon la présente invention, sans toutefois chercher à en limiter la portée.The method according to the present invention applies equally to microalgae of marine, fresh or terrestrial water, prokaryotic or eukaryotic origin. 5 The examples below describe preferential embodiments of the method according to the present invention, without however seeking to limit its scope.
Exemple 1 : Production de phycoérythrine par culture de Porphyridum cruetum, Rhodosorus marinus ou Rhodella violacea.Example 1: Production of phycoerythrin by culture of Porphyridum cruetum, Rhodosorus marinus or Rhodella violacea.
Les algues sont cultivées de manière classique et pré-récoltées par floculation/décantation. Elles sont transférées dans un bac de forçage physiologique permettant des transferts optimaux de matière et de lumière. L'induction de la synthèse de phycoérythrine est favorisée par un éclairement de l'ordre de 50 μE.cm2.s~ et un enrichissement en nitrate (2gJ~ ) et heptamolybdate de sodium (25 gJ" ) pour des biomasses de quelques g.l" (poids secs). La récolte finale a lieu après 3 heures d'incubation.The algae are cultivated in a conventional manner and pre-harvested by flocculation / decantation. They are transferred to a physiological forcing tank allowing optimal transfers of matter and light. The induction of phycoerythrin synthesis is favored by an illumination of the order of 50 μE.cm 2 .s ~ and an enrichment in nitrate (2gJ ~ ) and sodium heptamolybdate (25 gJ " ) for biomass of a few gl " (dry weights). The final harvest takes place after 3 hours of incubation.
Exemple 2 : Production de -carotène par culture de Dunallella salina.Example 2: Production of -carotene by culture of Dunallella salina.
La production de biomasse de cette algue est optimale à une salinité de 60%o. Les algues sont pré-récoltées par floculation (en accroissant l'alcalinité du milieu) et décantation. Elles sont transférées dans un bac de forçage physiologique permettant des transferts optimaux de matière et de lumière.The biomass production of this alga is optimal at a salinity of 60% o . The algae are pre-harvested by flocculation (by increasing the alkalinity of the medium) and decantation. They are transferred to a physiological forcing tank allowing optimal transfers of matter and light.
La carotenogenèse est induite par augmentation de la salinité du milieu ( 100%o) et un éclairement supérieur à 2000 μE. cm -2.s -1. Après 1 2 heures d'incubation les algues sont récoltées et congelées sous vide ax ant extraction.Carotenogenesis is induced by an increase in the salinity of the medium (100% o ) and an illumination greater than 2000 μE. cm -2.s -1. After 1 2 hours of incubation the algae are harvested and frozen under vacuum for extraction.
Exemple 3 : Production d'acide eicosapentaénoique par culture de Phaeodactylum tricornutum.Example 3: Production of eicosapentaenoic acid by culture of Phaeodactylum tricornutum.
Les algues sont cultivées dans un milieu de culture constitué d'eau de mer enrichie, en particulier, en silice. Les diatomées sont pré-récoltées par floculation (induite par accroissement de l'alcalinité du milieu) et décantation. Elles sont transférées dans un bac de forçage physiologique où elles sont soumises à une température inférieure de 10°C à celle des cultures initiales. Après 6 heures d'incubation sous agitation, les microalgues sont récoltées sous forme de pâte alguale. En comparaison avec la technique antérieure, la présente invention permet de produire des microalgues en conditions semi-contrôlées tout en assurant, à moindre frais, leur qualité biochimique. D'une part, la mise en oeuvre d'un procédé de récolte impliquant une concentration par floculation-décantation, floculation-flottation ou floculation-filtration permet de réduire les coûts liés à cette opération, en réduisant d'un facteur 10 à 20 les volumes à traiter, donc la dimension et la durée de fonctionnement des appareils. D'autre part, la pratique du forçage physiolo¬ gique sur des volumes réduits de cultures concentrées et pour des durées plus courtes permet également de réduire les coûts de sa mise en oeuvre. La présente invention concerne donc également un procédé de forçage physiologique de microalgues pour la suraccumulation de metabolites, caractérisé en ce que le forçage est effectuée à un volume de culture inférieur à 1 /5, de préférence compris entre 1 /10 et 1 /20 du volume usuel de culture des microalgues. The algae are cultivated in a culture medium consisting of seawater enriched, in particular, with silica. Diatoms are pre-harvested by flocculation (induced by increasing the alkalinity of the middle) and decantation. They are transferred to a physiological forcing tank where they are subjected to a temperature 10 ° C lower than that of the initial cultures. After 6 hours of incubation with shaking, the microalgae are harvested in the form of algal paste. In comparison with the prior art, the present invention makes it possible to produce microalgae under semi-controlled conditions while ensuring, at low cost, their biochemical quality. On the one hand, the implementation of a harvesting process involving concentration by flocculation-settling, flocculation-flotation or flocculation-filtration makes it possible to reduce the costs associated with this operation, by reducing by a factor of 10 to 20 the volumes to be treated, therefore the size and duration of operation of the devices. On the other hand, the practice of physiological forcing on reduced volumes of concentrated cultures and for shorter durations also makes it possible to reduce the costs of its implementation. The present invention therefore also relates to a method of physiological forcing of microalgae for the overaccumulation of metabolites, characterized in that the forcing is carried out at a culture volume of less than 1/5, preferably between 1/10 and 1/20 of the usual volume of microalgae culture.

Claims

REVENDICATIONS
1. Procédé de production de microalgues produisant en particulier des metabolites, caractérisé en ce qu'il consiste à effectuer les étapes1. Method for producing microalgae producing in particular metabolites, characterized in that it consists in carrying out the steps
5 suivantes : a) cultiver les microalgues en milieu liquide dans des conditions appropriées assurant essentiellement la formation d'une biomasse importante, b) concentrer la biomasse obtenue, c) placer cette biomasse concentrée dans des conditions assurant 0 essentiellement l'accumulation de metabolites, et d) récolter la biomasse contenant le métabolite et/ou le milieu contenant le métabolite.5 below: a) cultivating the microalgae in a liquid medium under appropriate conditions essentially ensuring the formation of a large biomass, b) concentrating the biomass obtained, c) placing this concentrated biomass under conditions ensuring essentially the accumulation of metabolites, and d) harvesting the biomass containing the metabolite and / or the medium containing the metabolite.
2. Procédé selon la revendication 1, caractérisé en ce que dans l'étape b), la biomasse concentrée obtenue représente moins de 1/5 du *5 volume de la culture dans l'étape a).2. Method according to claim 1, characterized in that in step b), the concentrated biomass obtained represents less than 1/5 of the * 5 volume of the culture in step a).
3. Procédé selon la revendication 2, caractérisé en ce que la biomasse concentrée représente entre 1/10 et 1 /20 du volume de la culture dans l'étape a) . Procédé selon l'une des revendications 1 à 3, caractérisé en ce 0 que la concentration est effectuée par floculation.3. Method according to claim 2, characterized in that the concentrated biomass represents between 1/10 and 1/20 of the volume of the culture in step a). Method according to one of claims 1 to 3, characterized in that the concentration is carried out by flocculation.
5. Procédé selon la revendication -+, caractérisé en ce que la floculation est effectuée par augmentation de l'alcalinité du milieu de culture de l'étape a).5. Method according to claim - +, characterized in that the flocculation is carried out by increasing the alkalinity of the culture medium from step a).
6. Procédé selon la revendication 5, caractérisé en ce que l'on *5 fait varier l'alcalinité du milieu après arrêt de la carbonatation durant 2 à heures en période lumineuse.6. Method according to claim 5, characterized in that * 5 varies the alkalinity of the medium after stopping carbonation for 2 to hours in the light period.
7. Procédé selon l'une des revendications 5 ou 6, caractérisé en ce que l'on augmente l'alcalinité du milieu par l'addition de soude et/ou de phosphatre de sodium. 0 s. Procédé selon l'une des revendications à 7, caractérisé en ce que la floculation est obtenue en employant des microalgues naturellement floculantes, et/ou en ajoutant des agents floculants chimiques, biologiques ou leurs mélanges.7. Method according to one of claims 5 or 6, characterized in that the alkalinity of the medium is increased by the addition of soda and / or sodium phosphate. 0 sec. Method according to one of claims to 7, characterized in that the flocculation is obtained by using naturally flocculating microalgae, and / or by adding chemical, biological flocculating agents or their mixtures.
5 9. Procédé selon l'une des revendications k à 8, caractérisé en ce que les algues floculées sont concentrées par des moyens appropriés de décantation statique ou mécanique ou par des moyens appropriés de flottation. 10. Procédé selon la revendication 9, caractérisé en ce que le moyen approprié de décantation est un décanteur cylindro-conique à flux vertical, dont la partie conique est comprise entre 50° et 70°, pour une vitesse ascentionnelle comprise entre 1,5 et 2,5 m/h.5 9. Method according to one of claims k to 8, characterized in that the flocculated algae are concentrated by suitable means of static or mechanical settling or by appropriate means of flotation. 10. Method according to claim 9, characterized in that the appropriate means of decantation is a cylindro-conical decanter with vertical flow, the conical part of which is between 50 ° and 70 °, for an ascent rate between 1.5 and 2.5 m / h.
1 1. Procédé selon l'une des revendications l à 10, caractérisé en •- 0 ce que l'accumulation de metabolites dans l'étape c) est obtenue par forçage physiologique.1 1. Method according to one of claims l to 10, characterized in • - 0 that the accumulation of metabolites in step c) is obtained by physiological forcing.
12. Procédé selon la revendication 1 1, caractérisé en ce que le forçage physiologique est induit par la température de culture et/ou l'exposition à la lumière et/ou l'enrichissement du milieu de culture en12. The method of claim 1 1, characterized in that the physiological forcing is induced by the culture temperature and / or exposure to light and / or the enrichment of the culture medium in
1 composés inducteurs.1 inducing compounds.
13. Procédé selon l'une des revendications 1 1 ou 12, caractérisé en ce que la durée du forçage physiologique est comprise entre 2 et 15 heures.13. Method according to one of claims 1 1 or 12, characterized in that the duration of the physiological forcing is between 2 and 15 hours.
1 *+. Procédé selon l'une des revendications 1 à 13, caractérisé en 0 ce qUe ja biomasse et/ou le milieu contenant le métabolite est récolté par centrifugation et/ou filtration.1 * +. Method according to one of claims 1 to 13, characterized in 0 that q Ue ja biomass and / or the medium containing the metabolite is harvested by centrifugation and / or filtration.
15. Procédé selon l'une des revendications 1 à 14, caractérisé en ce que les microalgues sont d'origine marine, d'eau douce ou terrestre, procaryotes ou eucaryotes. -5 16. Procédé de forçage physiologique de microalgues pour la suraccumulation de métabolite, caractérisé en ce que le forçage est effectuée à un volume de culture inférieur à 1/5, de préférence compris entre 1/10 et 1/20 du volume usuel de culture des microalgues.15. Method according to one of claims 1 to 14, characterized in that the microalgae are of marine, freshwater or terrestrial, prokaryotic or eukaryotic origin. -5 16. A method of physiological forcing of microalgae for the overaccumulation of metabolite, characterized in that the forcing is carried out at a culture volume of less than 1/5, preferably between 1/10 and 1/20 of the usual volume of culture of microalgae.
00
5 5
PCT/FR1993/000989 1992-10-07 1993-10-06 Method of producing microalgae WO1994007993A1 (en)

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FR9211877A FR2696475B1 (en) 1992-10-07 1992-10-07 PROCESS FOR PRODUCING MICROALGAE.
FR92/11877 1992-10-07

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WO2007046685A1 (en) * 2005-10-17 2007-04-26 Universiti Putra Malaysia Starter kit for the production of pure and high quality microalgae
CN101292023B (en) * 2005-10-17 2011-08-10 马来西亚博特拉大学 Starter kit for the production of pure and high quality microalgae
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RU2753766C1 (en) * 2020-02-27 2021-08-23 Федеральное государственное бюджетное образовательное учреждение высшего образования "Государственный университет по землеустройству" Biocomplex for production of chlorella
CN112760343A (en) * 2020-11-23 2021-05-07 惠州卫生职业技术学院 Novel biological flocculant and preparation method thereof

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