Classifications

• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D11/00—Solvent extraction
  • B01D11/02—Solvent extraction of solids
  • B01D11/0261—Solvent extraction of solids comprising vibrating mechanisms, e.g. mechanical, acoustical
  • B01D11/0265—Applying ultrasound
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
  • A23L5/40—Colouring or decolouring of foods
  • A23L5/42—Addition of dyes or pigments, e.g. in combination with optical brighteners
  • A23L5/46—Addition of dyes or pigments, e.g. in combination with optical brighteners using dyes or pigments of microbial or algal origin
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
  • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
  • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
  • A23L33/17—Amino acids, peptides or proteins
  • A23L33/195—Proteins from microorganisms
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/66—Microorganisms or materials therefrom
  • A61K35/74—Bacteria
  • A61K35/748—Cyanobacteria, i.e. blue-green bacteria or blue-green algae, e.g. spirulina
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
  • A61K36/02—Algae
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D11/00—Solvent extraction
  • B01D11/04—Solvent extraction of solutions which are liquid
  • B01D11/0484—Controlling means
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D21/00—Separation of suspended solid particles from liquids by sedimentation
  • B01D21/26—Separation of sediment aided by centrifugal force or centripetal force
  • B01D21/262—Separation of sediment aided by centrifugal force or centripetal force by using a centrifuge
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
  • A61K2236/30—Extraction of the material
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
  • A61K2236/30—Extraction of the material
  • A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D2259/00—Type of treatment
  • B01D2259/80—Employing electric, magnetic, electromagnetic or wave energy, or particle radiation
  • B01D2259/816—Sonic or ultrasonic vibration

Definitions

• the present invention relates to the field of valorization of biomass, in particular of algal biomass, more precisely the subject of the present invention is a process for extracting water-soluble compounds from microalgae and / or cyanobacteria, as well as the product obtained. by this method and its applications especially in the food field or as food supplements.
• microalgae In order to cope with the increase in global food demand, many manufacturers are turning to alternative resources such as microalgae. These microorganisms, of great biodiversity, represent a unique source of biomass. Their production differs from that of market garden plants and cereals, grown on arable land, by their very high surface yield.
• microalgae are a food product in its own right.
• the first consists of targeting a consumption of the whole microalga
• the second concerns the extraction, transformation and packaging of bioactive molecules derived from them.
• spirulina or Arthrospira platensis the green microalgae Chlorella and the diatom Odontella aurita.
• These microorganisms are emerging as major elements in the challenge of global food, thanks to their unique protein content and low environmental impact of their production: low consumption of water and energy.
• spirulina has in recent years attracted the attention of a large number of researchers and industrialists. It is already widespread on the market in the unprocessed dried form for its consumption as a dietary supplement.
• Supercritical fluids are mainly used to extract the hydrophobic, liposoluble compounds such as carotenoids, tocopherols, fatty acids ... (Supercritical Carbon Dioxide and Microwave-Assisted Extraction of Functional Lipophilic Compounds from Arthrospira platensis, Esquivel-Hernandez et al, 2016).
• Chlorella which is made of cellulose and has a high rigidity resulting in a high resistance of the algae to mechanical stresses, these processes would not allow effective extraction of the water-soluble compounds such as chlorophyll.
• the Applicants have today developed a method that can modulate the cavitation bubbles generated by ultrasound, the origin of the extraction efficiency, by other parameters applied during the treatment of microalgae. ultrasonic, such as pressure, temperature, and flow rate, thereby enabling intensification and modulation of the water-soluble compound extraction process, depending on the nature and amount of the desired water-soluble compounds in the extract.
• the process according to the invention makes it possible to generate in an optimized manner, on an industrial scale, from a biomass of starting microalgae, an aqueous solution and a process various extracts of high nutritional value, rich in water-soluble compounds, in particular proteins, water-soluble vitamins, chlorophyll, phycocyanin and minerals, intended for human consumption, said extracts having compositions that can be modulated into said water-soluble compounds.
• said extracts According to the parameters of ultrasonic power (Pus), temperature, pressure and flow chosen initially, it is thus possible to vary the nutritional composition of the extract but also its color.
• a first object of the present invention relates to a process for obtaining water-soluble compounds from eukaryotic microalgae (microalgae) or prokaryotes (cyanobacteria), characterized in that .
• said method comprises at least one step of extracting said water-soluble compounds by ultrasonic treatment with application of an ultrasonic power (Pus) of between 10 and 1000 W / L applied to the microalgae or cyanobacteria eh mixture with an aqueous solution to which is applied, preferably concomitantly a pressure (P) of between 1 and 2 bar, a temperature (T) between 5 and 70 ° C, and a flow (D) between 1 and 1000 L / hour.
• Pus ultrasonic power
• T temperature
• D flow
• microalgae By microalgae is meant according to the present invention eukaryotic microalgae, which are characterized by a cell wall and a nucleus, including chlorophytes, chrysophytes and pyrrophytes, said eukaryotic microalgae commonly known as "microalgae", and prokaryotic microalgae, which do not have a nucleus and a cell wall, including cyanophytes, hereinafter specifically referred to as "cyanobacteria”.
• the eukaryotic microalgae are chosen from chlorophytes, preferably from Chlorella, Nannocloropsis, Dunaliella and Euglena.
• the cyanobacteria are chosen from spirulina (Arthrospira platensis or Spirulina maxima) and AFA (Aphanizomenon Floes-aquae).
• microalgae according to the invention is spirulina
• water-soluble compounds is intended to mean, according to the present invention, compounds which are soluble in water and contained in microalgae and / or cyanobacteria, preferably chosen from proteins and peptides, water-soluble vitamins, preferably vitamins of group B (in particular Thiamin (B1), Riboflavin (B2), Niacin (B3), Pantothenic acid (B5), Pyridoxine (B6), Biotin (B8), Folate (B9), Cobalamin (B12)), minerals, preferably sodium, calcium, potassium, magnesium and iron, C-phycocyanin (or phycocyanin), and chlorophyll.
• vitamins of group B in particular Thiamin (B1), Riboflavin (B2), Niacin (B3), Pantothenic acid (B5), Pyridoxine (B6), Biotin (B8), Folate (B9), Cobalamin (B12)
• minerals preferably sodium, calcium, potassium, magnesium and iron, C-phycocyanin (or
• the microalgae according to the invention is spirulina and the water-soluble compound is C-phycocyanin.
• ultrasound treatment with application of ultrasonic power is meant according to the present invention a physical treatment implementing an ultrasonic reactor.
• the ultrasonic power is between 10 and 1000 W / L of aqueous solution mixed with microalgae or cyanobacteria, preferably between 30 and 100 W / L, more preferably between 50 and 80 W / L.
• the energy deployed by the ultrasonic reactor varies between 500 and 5000 J
• the feed rate of the ultrasonic reactor varies from 1 L / hour to 100 OL / hour, preferably 1 L / hour
• the treatment ultrasound is performed for a time varying from 30 seconds to 90 minutes.
• aqueous solution is meant according to the present invention an aqueous solution compatible for food use.
• a buffered water preferably phosphate buffer at pH 7, the buffer used being sodium phosphate.
• the aqueous solution according to the invention does not include chemical preservatives, or organic solvents of petrochemical origin (methanol, chloroform, hexane, ethanol) which have a toxicity that would make them incompatible for use. of the water-soluble compounds obtained by the process according to the invention.
• an acidic solvent since certain water-soluble compounds such as phycocyanin are very sensitive to pH and any significant variation in pH below pH 5 and above pH 7.5 causes its precipitation.
• An alcoholic solvent is not preferred because certain water-soluble compounds such as phycocyanin then lose much of their antioxidant properties.
• the ratio of microalgae or cyanobacteria / aqueous solution is between 1/5 and 1/50 .
• P pressure
• T temperature
• the temperature is kept constant between 20 and 30 ° C throughout the duration of the process for extracting the water-soluble compounds.
• the microalgae / aqueous solution mixture is received in a double-walled tank.
• Said tank can be thermostated by means of a circulation of water and a thermostat making it possible to regulate the desired temperature.
• the mixture is then sent via a pump and a valve to an ultrasonic reactor comprising an ultrasonic probe connected to an ultrasound generator.
• Said ultrasonic reactor may also be preferably provided with a double wall to be thermostated by a circulation of water and a thermostat to adjust the desired temperature.
• Valve allows to impose a resistance to the circulation of the mixture and thus to increase the pressure experienced by the mixture.
• a pressure gauge makes it possible to follow, upstream of the ultrasonic reactor, the pressure exerted on the mixture, and to modulate the opening of the valve to apply the desired pressure to the mixture.
• the temperature (T) is controlled by means of a thermostat, preferably via a circulation of thermostatically controlled water in the double wall of a reactor comprising the microalgae / aqueous solution mixture, and the mixture is maintained at the desired temperature.
• a thermostat preferably via a circulation of thermostatically controlled water in the double wall of a reactor comprising the microalgae / aqueous solution mixture, and the mixture is maintained at the desired temperature.
• the Applicant has shown that when the temperature exceeds 40 ° C some molecules of interest such as phycoyanine can be masked (by chlorophyll for example).
• the optimum temperature for the extraction of water-soluble compounds of interest is around 25 ° C.
• the modulation of the temperature makes it possible to extract different types of compounds: at low temperature the extraction of thermolabile compounds is favored whereas at high temperature the heat-sensitive compounds are masked but other molecules that are more difficult to extract cold are found in solution like chlorophyll (of apolar nature, soluble in organic solvents in normal times).
• the pressure (P) is applied by means of a valve to the system making it possible to supply the reactor to which Pus will be applied and makes it possible to impose a resistance to the circulation of the microalgae mixture. aqueous solution in the system.
• the ultrasonic power, pressure, temperature and flow rate are modulated to vary the water-soluble compounds extracted from said eukaryotic or cyanobacterial microalgae.
• the higher the ultrasonic power, the pressure and the temperature delivered to the medium the more the extract will be rich in proteins and chlorophyll.
• the lower the ultrasonic power and the temperature the less the extract will be rich in water-soluble compounds.
• the duration of the ultrasonic treatment step is between 1 to 90 minutes.
• the modulation of the duration makes it possible to vary the composition in water-soluble compounds, and therefore also the concentration. This is explained by the fact that the more the microalgae / aqueous solution mixture frequently passes into the ultrasonic reactor, the more it will be subjected to ultrasound and release of water-soluble compounds.
• the ultrasonic power, pressure, temperature and flow rate are modulated to vary the color of the extract obtained by said process.
• the higher the ultrasonic power, the pressure and the temperature delivered to the medium the more the extract will be rich in proteins and chlorophyll and therefore of intense green color.
• less the ultrasonic power and the temperature is high the less the extract will be rich in water-soluble compounds and the color of the extract generated will be blue (due to the absence of chlorophyll).
• the steps are applied to the mixture of eukaryotic or cyanobacterial microalgae and aqueous solution continuously, preferably with the aid of a pump ensuring the circulation of the mixture in a closed or open circuit.
• the method for obtaining water-soluble compounds according to the invention further comprises an additional step of solid-liquid separation, preferably by centrifugation, of the extract obtained in step d 'extraction.
• the process for obtaining water-soluble compounds according to the invention further comprises an additional step of filtering the liquid part obtained by separating the solid part (cell debris) from the extract.
• the filtration is carried out frontally, advantageously on a food filter, for example polyamide, especially nylon, with a fineness of 2 to 50 microns, preferably less than 25 microns.
• a food filter for example polyamide, especially nylon, with a fineness of 2 to 50 microns, preferably less than 25 microns.
• the invention relates to products as obtained by the process according to the invention containing water-soluble compounds present in microalgae or cyanobacteria.
• said method makes it possible to extract microalgae and cyanobacteria from the water-soluble compounds separated from the liposoluble fraction of the microalga or cyanobacteria.
• the extract produced will have organoleptic qualities as well as an optimal nutritional composition.
• the invention relates to the use of the products as obtained by the method according to the invention in chemical, food, cosmetic or pharmaceutical compositions.
• the invention also aims as new products retentates obtained at the end of the filtration step. These retentates can also be used in the food, medical or cosmetic field or as food supplements.
• FIG. 1 is a diagram of the mano-thermo-sonication mounting device (MTS) for setting up use of the method according to the invention, comprising a probe and an ultrasonic reactor for generating Pus, a thermostat for setting T, a valve and a pressure gauge for adjusting P and a pump for setting D.
• MTS mano-thermo-sonication mounting device
• Said method is in the form of a closed or open circuit (depending on the number of cycles chosen) allowing the treatment of a solution of microalgae (5% in MS) continuously.
• the tank can accommodate the microalgae mixture and aqueous solution to be treated.
• This is a double-walled tank that can be thermostatically controlled by water circulation.
• a pump regulates the circulation of the mixture in the constant flow system.
• the ultrasonic probe is connected to a generator to set the desired ultrasonic power.
• the probe is immersed in the liquid to be treated at 2/3 and emits a fixed frequency of its own (about 20 kHz).
• the ultrasonic reactor is provided with a double wall for thermostating the mixture via a thermostatically controlled circulation of water.
• the ultrasonic probe penetrates the reactor so as to deliver ultrasonic waves into the medium.
• the valve makes it possible to impose a resistance to the circulation of the mixture and thus to increase the pressure in the system. It is possible to visually follow this pressure variation on the manometer present upstream of the ultrasonic reactor.
• the system is drained in order to bring the mixture into a centrifugation module allowing separation.
• a filtration step makes it possible to obtain a clear and clear extract.
• Example 1 US Low Power, No Pressure - Blue Extract
• the biomass ⁇ Arthrospira platensis is mixed with an aqueous extraction solution (phosphate buffer pH 7) according to the ratio 1/20; 1 g of biomass per 20 g of aqueous solution.
• aqueous extraction solution phosphate buffer pH 7
• the mixture is sonicated at low ultrasonic power (30 W) for 10 min, with thermoregulation at 20 ° C. No pressure is imposed on the system.
• the extracts are then centrifuged for 10 min at 8000 rpm and then filtered by frontal filtration on Buchner (porosity 8 ⁇ ) in order to remove any particles in suspension.
• the extract obtained is blue in color because of the presence of phycocyanin (8.34 g / 100 g dry matter (MS)) and contains 33.36 g / 100 g MS of proteins.
• the biomass ⁇ Arthrospira platensis is mixed with the aqueous extraction solution (phosphate buffer pH 7) at the ratio 1/20; 1 g of biomass per 20 g of aqueous solution.
• aqueous extraction solution phosphate buffer pH 7
• the mixture is subjected to ultrasonic power ultrasound (100 W / L, ie about 80% amplitude) for 10 min, with thermoregulation at 20 ° C and a pressure of around 2 bar.
• the extracts are then centrifuged for 10 min at 8000 rpm and then filtered by frontal filtration on Buchner (porosity 8 ⁇ ) in order to remove any particles in suspension.
• the extract obtained is of intense green color because of the presence of chlorophyll which has been extracted because of the intensity of the method.
• the phycocyanin is still present in the extract (1.47 g / 100 g MS) and the protein content is 46.19%.

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• 2017
  • 2017-06-15 FR FR1770630A patent/FR3067569B1/fr active Active
• 2018
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