[go: up one dir, main page]

WO2016181719A1 - Procédé permettant d'obtenir un produit purifié de caroténoïdes - Google Patents

Procédé permettant d'obtenir un produit purifié de caroténoïdes Download PDF

Info

Publication number
WO2016181719A1
WO2016181719A1 PCT/JP2016/060666 JP2016060666W WO2016181719A1 WO 2016181719 A1 WO2016181719 A1 WO 2016181719A1 JP 2016060666 W JP2016060666 W JP 2016060666W WO 2016181719 A1 WO2016181719 A1 WO 2016181719A1
Authority
WO
WIPO (PCT)
Prior art keywords
carotenoid
extract
obtaining
phycocyanin
purified product
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/JP2016/060666
Other languages
English (en)
Japanese (ja)
Inventor
健昌 陣内
康行 今井
久由 新井
平橋　智裕
伸生 小林
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.)
DIC Corp
Original Assignee
DIC Corp
Dainippon Ink and Chemicals Co Ltd
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 DIC Corp, Dainippon Ink and Chemicals Co Ltd filed Critical DIC Corp
Publication of WO2016181719A1 publication Critical patent/WO2016181719A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • A23L5/43—Addition of dyes or pigments, e.g. in combination with optical brighteners using naturally occurring organic dyes or pigments, their artificial duplicates or their derivatives
    • A23L5/44—Addition of dyes or pigments, e.g. in combination with optical brighteners using naturally occurring organic dyes or pigments, their artificial duplicates or their derivatives using carotenoids or xanthophylls
    • 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
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • 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
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67—Vitamins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C35/00—Compounds having at least one hydroxy or O-metal group bound to a carbon atom of a ring other than a six-membered aromatic ring
    • C07C35/21—Compounds having at least one hydroxy or O-metal group bound to a carbon atom of a ring other than a six-membered aromatic ring polycyclic, at least one hydroxy group bound to a non-condensed ring
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
    • 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

Definitions

  • the present invention relates to a method for obtaining a purified product of carotenoid, and particularly to a method having a saponification step for decomposing chlorophyll contained in an algal culture and a subsequent carotenoid precipitation step.
  • Blue-green algae are not only used as health foods but also as phycocyanin pigment sources because they contain a large amount of useful substances such as carotenoids, nucleic acid-related substances, amino acids, vitamins, minerals, and phycocyanin pigments.
  • Phycocyanin dyes usually have an absorption maximum wavelength at 618 nm and exhibit a water-soluble and vivid blue color. Therefore, they are used as water-soluble natural colorants for foods such as chewing gum and ice confectionery, or cosmetics such as eye shadows and lipsticks. It is used as a colorant for creams, eyeliners, shampoos, and emulsions.
  • Carotenoids have an antioxidant effect and are expected to have an antioxidant effect even after being taken into the body, so they are also added to various health foods and supplements.
  • zeaxanthin is widely used for the purpose of coloring cultured fish and improving the egg yolk quality of chicken eggs.
  • it is only two carotenoids that exist in the macular region of the human retina together with lutein, and it is AMD (aging-related). It is known to be related to a reduction in the risk of macular degeneration (Patent Document 1, Non-Patent Document 1).
  • Non-patent Document 2 zeaxanthin and lutein have been reported to have strong antitumor promoting properties. Recent reports have shown that it can play an important role in combating conditions that induce cardiovascular disease, atherosclerosis, skin cancer, ovarian cancer, and the like.
  • the phycocyanin dye can be produced, for example, from the following four steps (Patent Documents 2 and 3). 1) Water extraction process of cyanobacteria algae body components, 2) Centrifugation step, 3) Concentration process by ultrafiltration, 4) Drying process.
  • residual liquid the residue after separation of phycocyanin generated by this production process (hereinafter referred to as residual liquid) is still disposed of in spite of containing a large amount of nucleic acid-related substances, amino acids, carotenoid pigments, vitamins, minerals, etc. In many cases, this not only becomes a source of environmental pollution, but also leads to the waste of valuable biomass resources. Therefore, effective reuse of the residue is desired.
  • chlorophyll degradation products are also known to be harmful (Patent Document 4).
  • Chlorophyll contained in algae can cause photosensitivity, for example, cases of livestock damage due to feeding have been known for a long time, and excessive eating of chlorella tablets is also widely known to cause human injury. It became. This is directly attributed to pheophorbide produced by the decomposition of chlorophyll, and photosensitivity reaction reveals photosensitivity such as sunlight dermatitis, but this pheophorbide used white shark
  • LD 50 has a value of 45.5 mg / 100 g or more and MLD 50 has a value of 12 mg / 100 g or more (Non-patent Document 3).
  • chlorophyll tends to be acidic or pheophorbide a is easily generated by the coexistence of an organic solvent, there remains a problem in using carotenoids extracted from algae as a health food as they are.
  • Non-patent Document 4 In the purification of carotenoids derived from microalgae, especially zeaxanthin, separation from chlorophyll is difficult and the purification process is complicated. In order to overcome these points, fermentation production by the terrestrial bacterium Erwinia uredovora has been studied, but its yield is extremely low and cannot be practically used (Non-patent Document 4).
  • an object of the present invention is to provide an industrially useful production method for carotenoids that contributes to effective utilization of algal culture residue and does not contain harmful substances.
  • Another object of the present invention is to provide functional foods, feeds, cosmetics, food colors, and the like containing purified carotenoids.
  • a method for obtaining a purified product of carotenoid from a residue obtained by extracting the phycocyanin A saponification step using an alkaline substance for decomposing chlorophyll contained in the carotenoid extract (A) obtained after the steps of extracting and filtering the carotenoid from the residue, and a subsequent carotenoid precipitation step are provided.
  • a method for obtaining a purified carotenoid product characterized in that the conditions of the saponification step are as follows. (1) The alkaline substance is used in an amount corresponding to 1.9 to 7.6 mol of the alkaline substance relative to the solid mass (1 kg) of the carotenoid extract (A). (2) The temperature for saponification is 40 to 80 ° C.
  • the industrially useful manufacturing method which does not contain the harmful
  • the present invention includes the following items. 1.
  • a method for obtaining a purified product of carotenoid from a residue obtained by extracting phycocyanin after producing phycocyanin by culturing algae, A saponification step using an alkaline substance for decomposing chlorophyll contained in the carotenoid extract (A) obtained after the steps of extracting and filtering the carotenoid from the residue, and a subsequent carotenoid precipitation step are provided.
  • a method for obtaining a purified carotenoid product characterized in that the conditions of the saponification step are as follows.
  • the alkaline substance is used in an amount corresponding to 1.9 to 7.6 mol of the alkaline substance relative to the solid mass (1 kg) of the carotenoid extract (A).
  • the temperature for saponification is 40 to 80 ° C. 2.
  • the alkaline substance is potassium hydroxide or sodium hydroxide; A method for obtaining a purified product of carotenoid as described in 1. 3. Furthermore, it has a step of extracting carotenoid with ethanol or a water-ethanol mixed solution before the saponification step. Or 2. A method for obtaining a purified product of carotenoid as described in 1. 4).
  • carotenoids are precipitated from the liquid obtained by the saponification step by adjusting the water / ethanol mass ratio in the range of 45/55 to 55/45.
  • ⁇ 3. A method for obtaining a carotenoid purified product according to any one of the above. 5.
  • Algae is a cyanobacterium containing spirulina or phycocyanin ⁇ 4.
  • the carotenoid is zeaxanthin, ⁇ -carotene, or myxoxanthophyll.
  • a food, feed, cosmetic, or food color containing carotenoid obtained by the method for obtaining a purified carotenoid product according to any one of the above.
  • the first step of the present invention is a step of obtaining an extract obtained by extracting phycocyanin in algae into an aqueous suspension.
  • Algae which can be used for the preparation of this extract include the genus Arthrospira, the genus Spirulina, the genus Aphanizomenon, the genus Fischerella, the genus Anabaena, the genus Nesmo (Genus Synechocystis), genus Synechococcus, genus Tolypothrix, genus Aphanothace, genus Mastigoclaus, genus Pleurocapsa To Arsulospira, which has been produced on a scale and has been confirmed to be safe Which is desirable.
  • Examples of the algae used in the present invention include raw algae and dried algae, but phycocyanin is easily extracted in the step of obtaining an extract obtained by extracting phycocyanin in algae into an aqueous suspension. Dry algae are preferred because the amount of phycocyanin that can be extracted is also stable.
  • Raw algae are harvested by a method such as centrifugation and filtration of algae cultured in water, and usually contain 70 to 90% by mass of water.
  • Algae are usually cultured in natural water or artificial light in water, but it is preferable to harvest cyanobacteria that are irradiated with light and undergoing photosynthesis. Especially for cyanobacteria grown in outdoor culture tanks under natural light, photosynthesis continues and the water temperature rises after 10 am, compared to cyanobacteria harvested at night or immediately after the start of light irradiation. More preferred are cyanobacteria harvested from sunset to sunset.
  • Examples of the dried algae include those obtained by freeze-drying or spray-drying raw algae cultured by the above method.
  • an extract is obtained in the first step, a calcium salt and phosphate are reacted in the extract in the second step to obtain calcium phosphate, and phycocyanin is added to the calcium phosphate.
  • a calcium salt and phosphate are reacted in the extract in the second step to obtain calcium phosphate
  • phycocyanin is added to the calcium phosphate.
  • the first step and the second step may be performed as follows. 1.
  • the first step is a step of preparing an aqueous suspension containing algae and a calcium salt, and obtaining an extract obtained by extracting phycocyanin in the algae into the aqueous suspension.
  • the first step is a step of preparing an aqueous suspension containing algae and phosphate, and obtaining an extract obtained by extracting phycocyanin in the algae into the aqueous suspension
  • the second step is the extract.
  • an extract obtained by extracting phycocyanin in algae into an aqueous suspension is obtained, and in the second step, phosphate and calcium salt are added to the extract to obtain calcium phosphate, and the calcium phosphate is added to the calcium phosphate.
  • the first step is a step of preparing an aqueous suspension containing algae, calcium salt and phosphate, and obtaining an extract obtained by extracting phycocyanin in the algae into the aqueous suspension. Then, the process proceeds to the second step as it is, and calcium phosphate is obtained with the extract, and adsorbate can be obtained by adsorbing phycocyanin impurities to the calcium phosphate. In addition, the 3. In the first step, calcium salt and / or phosphate may be added.
  • a water suspension containing algae and a calcium salt is prepared as a first step, and a step of obtaining an extract in which phycocyanin in the algae is extracted into the water suspension is obtained.
  • the extraction time of phycocyanin therein can be shortened, and an extract with less elution of phycocyanin impurities, particularly carotenoids, is preferable.
  • the following description of the first step and the second step is 1. The above method is assumed.
  • Examples of a method for obtaining an extract in the first step include a method of preparing an aqueous suspension containing algae and a calcium salt, and extracting the phycocyanin in the algae by maintaining the extract at 0 to 40 ° C. Is mentioned.
  • First method Add calcium salt to the algae-suspended aqueous solution.
  • Second method examples include a method of adding algae to an aqueous solution of calcium salt and suspending it, but the second method. Is preferred.
  • Examples of the calcium salt used in the preparation of the aqueous solution of the calcium salt used in the present invention include water-soluble calcium salts such as calcium chloride, calcium nitrate, and calcium nitrite, among which calcium chloride is preferable.
  • the concentration of the calcium salt in the aqueous suspension is preferably 0.1 to 10% by mass, more preferably 0.1 to 5% by mass, and still more preferably 0.5 to 3% by mass.
  • the aqueous solution of calcium salt is suspended in the range where the concentration of cyanobacteria is 0.1 to 20% by mass in terms of solid content. A range of 2 to 8% by mass is more preferable.
  • the suspension is preferably prepared in the range where the temperature of the aqueous suspension is 0 to 40 ° C, more preferably 0 to 35 ° C.
  • An aqueous suspension containing algae and calcium salt is prepared, and phycocyanin in the algae is extracted into the aqueous suspension to obtain an extract.
  • Phycocyanin is extracted from algae by standing still, but may be stirred if necessary.
  • the extraction time is preferably 1 to 48 hours, more preferably 1 to 20 hours.
  • phycocyanin in algae can be efficiently extracted into an aqueous suspension by adding a basic compound to the aqueous suspension or subjecting it to ultrasonic irradiation. Both the addition of the basic compound and the ultrasonic irradiation treatment may be performed, or only one of them may be performed. When both are performed, the basic compound may be added after ultrasonic irradiation, or the ultrasonic irradiation treatment may be performed after adding the basic compound, but the ultrasonic wave is added after adding the basic compound. It is preferable to perform irradiation treatment.
  • the irradiation method when performing the ultrasonic irradiation treatment is not limited as long as it can destroy algal cells and promote the transfer of phycocyanin into suspension, and examples include batch type and continuous type. However, a continuous type in which ultrasonic waves are continuously applied is preferable.
  • a continuous ultrasonic irradiation treatment apparatus for example, a multiple ultrasonic dispersion apparatus for production manufactured by Nippon Seiki Seisakusho Co., Ltd. may be used.
  • phosphate is added to the aqueous extract of phycocyanin obtained in the first step.
  • the phosphate may be added as a solid or in the form of an aqueous solution.
  • the phosphate include sodium phosphate such as sodium phosphate, sodium dihydrogen phosphate and disodium hydrogen phosphate; potassium phosphate such as potassium phosphate, potassium dihydrogen phosphate and dipotassium hydrogen phosphate;
  • magnesium phosphate water-soluble inorganic salts such as ammonium dihydrogen phosphate, among which sodium phosphate and potassium phosphate are preferable, sodium phosphate is particularly preferable, and disodium hydrogen phosphate is most preferable.
  • the phosphate is preferably added to the extract so that the phosphate concentration is 1 to 5% by mass in the extract, and is preferably added to the extract so that the concentration is 2 to 3% by mass. Is more preferable.
  • phosphate to the extract.
  • the phosphate reacts with the calcium salt in the water extract to cause precipitation of calcium phosphate and adsorbs and adsorbs chlorophyll and other contaminants contaminated with phycocyanin pigment on the calcium phosphate.
  • the purity of the phycocyanin pigment can be increased.
  • After adding the phosphate it may be allowed to stand or may be stirred if necessary.
  • the time required for the reaction (adsorption) of calcium ions and phosphate ions is preferably 2 to 10 hours, and more preferably 3 to 5 hours.
  • the pH when adsorbing phosphate and algae to obtain an adsorbate in the extract is preferably 4 to 8, more preferably 5 to 6, because the amount of phycocyanin obtained is increased.
  • the pH can be adjusted, for example, by adding a basic compound or an acidic compound to the extract.
  • the basic compound include alkali compounds such as sodium hydroxide, potassium hydroxide and lithium hydroxide; carbonates of alkali metals such as sodium carbonate, sodium hydrogen carbonate, potassium carbonate, potassium hydrogen carbonate and lithium carbonate; hydrogen carbonate Examples thereof include alkali metal hydrogen carbonates such as sodium, potassium hydrogen carbonate and lithium hydrogen carbonate; alkali metal acetates such as sodium acetate, potassium acetate and lithium acetate.
  • the acidic compound examples include citric acid, hydrochloric acid, lactic acid, acetic acid, and the like.
  • the pH of the extract can also be adjusted by adding a basic compound or acidic compound to the aqueous suspension in advance. If the pH of the aqueous suspension at this time is adjusted to 7 to 6, the pH of the extract is preferably 5 to 6.
  • algal residues and the adsorbate are removed from the extract after completion of the second step, but if the extract contains a chelating agent prior to the third step, the amount of recovered phycocyanin may be increased. Is particularly preferable. This is because phycocyanin includes phycocyanin C and allophycocyanin. Allophycocyanin is adsorbed on calcium phosphate, and allophycocyanin adsorbed on this calcium phosphate is removed together with calcium phosphate in the next third step, but before the third step. The inventor believes that when the chelating agent is contained in the extract, the chelating agent is adsorbed on the calcium phosphate, so that allophycocyanin is separated from the calcium phosphate and remains in the extract.
  • the chelating agent may be added before the third step.
  • the chelating agent may be added to the algae aqueous suspension in the first step, or may be added to the prepared extract. It may be added to the extract before obtaining the adsorbate in the process, or may be added after obtaining the adsorbate. In the present invention, it is preferably added at the time of suspension adjustment.
  • the chelating agent examples include organic carboxylates such as sodium citrate, sodium oxalate, sodium tartrate, sodium gluconate; nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminopentaacetic acid (DTPA) Amino carbonates such as dihydroxyethyl glycine (DFG), triethanolamine (TEA), N- (2-hydroxyethyl) iminodiacetic acid (HEIDA), hydroxyethylenediaminetetraacetic acid (HEDTA) and the like; Examples thereof include ether carboxylates such as sodium carboxymethyltaltronate (CMT) and sodium carboxymethyloxysuccinate (CMOS). Of these, sodium citrate and sodium ethylenediaminetetraacetate are more preferable.
  • organic carboxylates such as sodium citrate, sodium oxalate, sodium tartrate, sodium gluconate
  • NTA nitrilotriacetic acid
  • EDTA
  • the amount of chelating agent added is preferably 5 to 100% by mass, more preferably 10 to 40% by mass based on the amount of calcium chloride used.
  • Algal residue can be obtained from the extract in the third step.
  • means for obtaining these include various methods, such as a filtration method using a filter medium such as filter paper and filter cloth, a decantation method performed by collecting the supernatant from the precipitate, and a centrifugation method. Of these, separation by centrifugation is preferable.
  • Centrifugation may be performed under conditions that can remove algae residues and adsorbates from the extract, but is preferably performed under a centrifugal acceleration of 1,000 to 30,000 G for 10 seconds to 2 hours, and a gravitational acceleration of 3 Centrifugation conditions of 1 to 30 minutes at 1,000 to 10,000 G are more preferred.
  • the obtained residue is preferably dried by a spray dryer method, a freeze-drying method or the like in order to prevent deterioration of the carotenoids contained, particularly zeaxanthin.
  • This extraction method can be performed by a known and commonly used method. For example, it is preferable to perform a solvent extraction method using ethanol or a water-ethanol mixed solution as an extraction solvent. Furthermore, when a water-ethanol mixed solution is used, it is preferably 99 wt% (mass%) to 80 wt% (mass%) in order to increase the extraction selectivity of zeaxanthin. If it is higher than the above range, ethanol and lipid will undergo transesterification to produce unpurified products such as fatty acid ethyl ester, and if it is lower, the zeaxanthin extraction efficiency will be reduced.
  • the temperature at the time of extraction can be in the range of 20 to 80 ° C., but in order to perform the extraction in a short time, it is preferable to carry out at 40 to 80 ° C. while stirring the solution uniformly. Is preferably extracted in a temperature range of 50 to 80 ° C.
  • This step can be performed by a known and commonly used method. For example, it can be filtered using a commercially available filter paper.
  • the solid component mass of the extract (A) is preferably 3.3 wt% to 33 wt% of the extract (A), more preferably 3.3 wt% to 13 wt%, and 5 wt% to 8.3 wt%. Particularly preferred.
  • the solid content mass is measured after drying the solvent at 105 ° C. after distilling off the solvent at a vacuum of 10 kPa or less while heating a part of the obtained extract at 40 ° C.
  • the saponification process of the obtained carotenoid extract (A) is performed.
  • the purpose of this step is to remove chlorophyll contained in the extract (A).
  • a treatment with an amount corresponding to 3.8 mol is particularly preferred.
  • the alkaline substance used in the present invention can be used without limitation as long as it is usually an alkaline substance capable of removing chlorophyll, but preferably potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, etc. Mention may be made of alkaline substances.
  • the mass of the alkaline substance to be used a numerical mass converted to a substantial amount of the alkaline substance is used.
  • the alkaline substance since commercially available potassium hydroxide has a content of about 85%, the mass of the numerical value converted from this is used.
  • the intended saponification can be performed in a short time at 30 ° C. or higher.
  • the solution is stirred so as to be uniform. However, it is preferably carried out at 40 to 80 ° C.
  • the time required for saponification is usually 30 minutes to 24 hours. When the time is shorter than this time, saponification of chlorophyll is insufficient, and when the time is longer, carotenoid is deteriorated.
  • a precipitation step for obtaining a purified carotenoid product is performed.
  • carotenoids are precipitated by adding water to the solution obtained after the saponification step and adjusting the water-ethanol concentration.
  • the water-ethanol concentration is preferably 70 wt% or less, and more preferably in the range of 45 wt% to 55 wt% when it is precipitated in a short time.
  • diatomaceous earth filtration, diatomaceous earth washing, and diatomaceous earth elution may be performed. That is, in this step, for example, a solution in which carotenoids are precipitated with 45 wt% ethanol water is filtered through diatomaceous earth, and then the filtrate is washed with 45 wt% ethanol water to elute impurities. Subsequently, the carotenoid eluate can be purified by eluting the filtrate with 99 wt% to 80 wt% ethanol water. This step has an effect that the purified carotenoid eluate is rich in zeaxanthin.
  • the carotenoid obtained from the present invention can be used as a functional product in foods, feeds, cosmetics, food colors, etc. and used for various applications.
  • Examples of methods for producing foods, feeds, cosmetics, edible pigments and the like as the functional products include publicly known methods.
  • zeaxanthin recovery rate calculation method By using the amount of zeaxanthin in each solution obtained, the zeaxanthin recovery rate in each step can be calculated. That is, the extraction process recovery rate can be calculated as Z2 / Z1 ⁇ 100%, the saponification process recovery rate can be calculated as Z3 / Z2 ⁇ 100%, and the purification process recovery rate can be calculated as Z4 / Z3 ⁇ 100%.
  • the suspension was stirred for 2 minutes and then allowed to stand at 20 ° C. for 5 hours.
  • This suspension was introduced into a continuous ultrasonic crusher (Nippon Seiki Seisakusho RUS-300TCVP type, frequency 19.5 kHz, ultrasonic irradiation unit volume 10 mL, output 300 W), and the suspension was 5 mL / second.
  • Ultrasonic treatment was performed at a speed to the ultrasonic irradiation unit to obtain an extract.
  • sodium dihydrogen phosphate was added to a concentration of 6.8 g / liter and disodium hydrogen phosphate to a concentration of 2.2 g / liter.
  • the mixture was introduced into a centrifuge and the gravitational acceleration was 10 Centrifugation was performed at 1,000 G for 15 minutes, and the phycocyanin residue and the phycocyanin solution were separated by filtration. Further, the phycocyanin residue was dried by a spray dryer method or the like to obtain a dried phycocyanin residue (blue residue).
  • Example 1 Zeaxanthin was extracted while stirring 300 g of the blue extraction residue in 900 g of a 95 wt% aqueous ethanol solution at 50 ° C. for 1 hour at 300 rpm. Solid-liquid separation was performed by filtration to obtain a filtrate and a filtration residue. The filtration residue was extracted again with a 95 wt% aqueous ethanol solution and added to the filtrate to obtain an extraction solution. The extract was concentrated under reduced pressure to 450 g using a rotary evaporator, and 30 g of a 25% KOH / EtOH solution (an equivalent amount of 3.8 mol of an alkaline substance was used relative to the solid mass (1 kg)) was added. Saponification was performed while stirring at 200 rpm at 50 ° C. for 1 hour to obtain a saponified extract having saponified chlorophyll water-solubilized.
  • the mixture was allowed to stand at 4 ° C. for 30 minutes or more to precipitate a red substance.
  • the saponified extract having the precipitate was passed through diatomaceous earth pre-coated to a thickness of 10 mm, and the precipitate and the liquid were separated by filtration and then washed with a 45 wt% aqueous ethanol solution to remove impurities.
  • the precipitate from which impurities were removed was dissolved in a 95 wt% aqueous ethanol solution to obtain a final purified product.
  • the extraction residue as a raw material and the extract, saponified extract, and final purified product obtained above were measured by UPLC, and each zeaxanthin recovery rate was calculated.
  • the chlorophyll content was calculated.
  • the dry mass was measured, and the zeaxanthin content in the solid content was calculated.
  • the measurement results were 80% zeaxanthin recovery, 13% zeaxanthin content, and 0.1% or less chlorophyll content.
  • Example 2 150 g of a 95 wt% aqueous ethanol solution was added to 50 g of the blue residue, and extracted with stirring at 78 ° C. for 1 hour. Solid-liquid separation was performed by filtration to obtain a filtrate and a filtration residue. The filtration residue was extracted again with a 95 wt% aqueous ethanol solution and added to the filtrate to obtain an extraction solution. A 95 wt% aqueous ethanol solution was added to the extract to make 500 g. 50 g was collected, and 0.25 g of 25% KOH / EtOH was added (the amount corresponding to 1.9 mol of the alkaline substance relative to the solid mass (1 kg) was used), and saponified while stirring at 78 ° C. for 1 hour. The saponified extract had a chlorophyll content of 0.1% or less and a zeaxanthin recovery rate of 85%.
  • Example 3 150 g of a 95 wt% aqueous ethanol solution was added to 50 g of the blue residue, and extracted with stirring at 78 ° C. for 1 hour. Solid-liquid separation was performed by filtration to obtain a filtrate and a filtration residue. The filtration residue was extracted again with a 95 wt% aqueous ethanol solution and added to the filtrate to obtain an extraction solution. A 95 wt% aqueous ethanol solution was added to the extract to make 500 g. 50 g was collected, and 1.0 g of 25% KOH / EtOH was added (alkaline substance equivalent to 7.6 mol was used with respect to the solid mass (1 kg)), and saponified while stirring at 78 ° C. for 1 hour. The saponified extract had a chlorophyll content of 0.01% and a zeaxanthin recovery rate of 77%.
  • the filtration residue was extracted again with a 95 wt% aqueous ethanol solution and added to the filtrate to obtain an extraction saponification solution.
  • the saponified extract had a chlorophyll content of 0.83% and a zeaxanthin recovery rate of 84%.
  • the purified product such as carotenoid obtained in the present invention can be used as, for example, functional foods, feeds, cosmetics, food colors, etc. contained in foods, feeds, cosmetics, food colors and the like.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Polymers & Plastics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Birds (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Botany (AREA)
  • Mycology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Cosmetics (AREA)

Abstract

Le problème abordé par la présente invention est de pourvoir à un procédé industriellement utile pour contribuer à l'utilisation efficace du liquide résiduel d'une culture algale et à la production de caroténoïdes exempts de substances dangereuses. La solution selon l'invention porte sur un procédé permettant d'obtenir un produit purifié de caroténoïdes à partir d'un résidu dont la phycocyanine a été extraite après production de ladite phycocyanine par culture algale, le procédé permettant d'obtenir un produit purifié de caroténoïdes étant caractérisé en ce qu'il comporte une étape de saponification consistant à utiliser une substance alcaline pour décomposer la chlorophylle contenue dans un extrait de caroténoïdes (A) obtenu après une étape d'extraction de caroténoïdes à partir du résidu et de filtration, et une étape de précipitation des caroténoïdes mise en œuvre ultérieurement, les conditions de l'étape de saponification étant les suivantes : (1) de 1,9 à 7,6 équivalents molaires d'une substance alcaline sont utilisés par rapport à la masse solide (1 kg) de l'extrait de caroténoïdes (A) ; et (2) la température à laquelle la saponification est mise en œuvre est de 40 à 80°C.
PCT/JP2016/060666 2015-05-12 2016-03-31 Procédé permettant d'obtenir un produit purifié de caroténoïdes Ceased WO2016181719A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2015097417 2015-05-12
JP2015-097417 2015-05-12

Publications (1)

Publication Number Publication Date
WO2016181719A1 true WO2016181719A1 (fr) 2016-11-17

Family

ID=57248135

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2016/060666 Ceased WO2016181719A1 (fr) 2015-05-12 2016-03-31 Procédé permettant d'obtenir un produit purifié de caroténoïdes

Country Status (1)

Country Link
WO (1) WO2016181719A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017199830A1 (fr) * 2016-05-18 2017-11-23 Dic株式会社 Procédé d'obtention du produit purifié de caroténoïdes
CN108623675A (zh) * 2017-03-21 2018-10-09 江苏海睿生物科技有限公司 鲜螺旋藻中藻蓝蛋白、叶绿素和螺旋藻多糖的提取工艺
CN112931870A (zh) * 2021-03-25 2021-06-11 云南爱尔康生物技术有限公司 一种藻类来源的类胡萝卜素抗氧化食品添加剂

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4439629A (en) * 1980-11-20 1984-03-27 Hoffmann-La Roche Inc. Extraction process for beta-carotene
JPH01123865A (ja) * 1987-11-10 1989-05-16 Dainippon Ink & Chem Inc 藍藻からのフィコシアニン含有青色顔料の製造方法及びその顔料を含む化粧品
JPH0616519A (ja) * 1992-07-02 1994-01-25 Dainippon Ink & Chem Inc 農園芸用肥料
JP2000175696A (ja) * 1998-12-14 2000-06-27 Yoshio Tanaka ドナリエラ藻体の抽出方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4439629A (en) * 1980-11-20 1984-03-27 Hoffmann-La Roche Inc. Extraction process for beta-carotene
JPH01123865A (ja) * 1987-11-10 1989-05-16 Dainippon Ink & Chem Inc 藍藻からのフィコシアニン含有青色顔料の製造方法及びその顔料を含む化粧品
JPH0616519A (ja) * 1992-07-02 1994-01-25 Dainippon Ink & Chem Inc 農園芸用肥料
JP2000175696A (ja) * 1998-12-14 2000-06-27 Yoshio Tanaka ドナリエラ藻体の抽出方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
HIROYUKI KAMOGAWA: "Carotenoid Contents in Chlorella pyrenoidosa and Concentration by Saponification", BULLETIN OF FISHERIES SCIENCES, vol. 62, 3, 2012, pages 83 - 88 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017199830A1 (fr) * 2016-05-18 2017-11-23 Dic株式会社 Procédé d'obtention du produit purifié de caroténoïdes
CN108623675A (zh) * 2017-03-21 2018-10-09 江苏海睿生物科技有限公司 鲜螺旋藻中藻蓝蛋白、叶绿素和螺旋藻多糖的提取工艺
CN112931870A (zh) * 2021-03-25 2021-06-11 云南爱尔康生物技术有限公司 一种藻类来源的类胡萝卜素抗氧化食品添加剂

Similar Documents

Publication Publication Date Title
JP4969370B2 (ja) カロテノイドの製造方法
JP5795572B2 (ja) 種々の植物からカロテノイド結晶を分離する方法
AU2005331246B2 (en) Isolation and purification of carotenoids from Marigold flowers
JP6245170B2 (ja) フコキサンチン含有組成物の製造方法
US9682932B2 (en) Process for production of high purity beta-carotene and lycopene crystals from fungal biomass
WO2012047120A1 (fr) Production microbienne hétérotrophique de pigments xanthophylle
WO2016181719A1 (fr) Procédé permettant d'obtenir un produit purifié de caroténoïdes
US8481769B2 (en) Isolation and purification of cartenoids from marigold flowers
CN109232345A (zh) 一种从含叶黄素二酯的植物油树脂中提取分离叶黄素晶体的方法
EP2571942B1 (fr) Procédé d'isolement et de purification de caroténoïdes
WO2021172372A1 (fr) Composition de cis-xanthophylle et son utilisation
WO2017199830A1 (fr) Procédé d'obtention du produit purifié de caroténoïdes
JP2004208504A (ja) アスタキサンチン含有酵母抽出物の製造方法
JP5116982B2 (ja) カロテノイドの製造方法
JP5116994B2 (ja) カロテノイドの製造方法
JP2015204764A (ja) 無色カロテノイドの製造方法
JP2009201503A (ja) アスタキサンチン含有抽出物の香味改善方法
JP2012070717A (ja) アスタキサンチン結晶の製造方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16792444

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16792444

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: JP