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WO2013042340A1 - Procédé de récupération ou d'élimination de métal et procédé de production de lipides ou de pigment - Google Patents

Procédé de récupération ou d'élimination de métal et procédé de production de lipides ou de pigment Download PDF

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WO2013042340A1
WO2013042340A1 PCT/JP2012/005866 JP2012005866W WO2013042340A1 WO 2013042340 A1 WO2013042340 A1 WO 2013042340A1 JP 2012005866 W JP2012005866 W JP 2012005866W WO 2013042340 A1 WO2013042340 A1 WO 2013042340A1
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red algae
metal
cultured
concentration
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高郁 山本
歩 蓑田
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    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6463—Glycerides obtained from glyceride producing microorganisms, e.g. single cell oil
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
    • C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
    • 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
    • C12P3/00—Preparation of elements or inorganic compounds except carbon dioxide
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C—CHEMISTRY; METALLURGY
    • C22—METALLURGY; FERROUS OR NON-FERROUS ALLOYS; TREATMENT OF ALLOYS OR NON-FERROUS METALS
    • C22B—PRODUCTION AND REFINING OF METALS; PRETREATMENT OF RAW MATERIALS
    • C22B3/00—Extraction of metal compounds from ores or concentrates by wet processes
    • C22B3/18—Extraction of metal compounds from ores or concentrates by wet processes with the aid of microorganisms or enzymes, e.g. bacteria or algae
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P10/00—Technologies related to metal processing
    • Y02P10/20—Recycling

Definitions

  • the present invention relates to a method for recovering or removing a metal (metal ion) contained in a solution by culturing red algae in the solution, and a method for producing a lipid or a pigment.
  • a method using a living organism is known as a method that is less expensive than a chemical method. It has been.
  • a method using algae as a living organism in the removal and recovery of metal ions and the elution of metals has not been put into practical use.
  • algae can be used to remove and recover metal ions and elution of metals, and there is a possibility that useful substances such as biofuel can be efficiently produced using algae as a raw material. is there. That is, metal ions can be removed and recovered and metal can be eluted at low cost, and a biofuel feedstock can be produced.
  • Non-Patent Document 1 There is red algae as a group of algae, and Cyanidiaceae of this red algae is composed of three species of the genus Gardieria, Cyanidium, and Cyanidiosis. Details on the classification of Cyanidium are reported in Non-Patent Document 1 from a physiological and ecological viewpoint. In Non-Patent Document 1, it is reported that only red algae belonging to the genus Gardiella can grow using sugar and sugar alcohol.
  • Non-Patent Documents 2 to 4 and Patent Document 1 Various proposals have heretofore been made on methods for culturing cyanidium red algae and recovering metal ions contained in the solution, for example, Non-Patent Documents 2 to 4 and Patent Document 1.
  • Non-Patent Document 2 it is reported that a red algae belonging to the genus Gardieria collects metal corresponding to about 20% of the dry weight.
  • Non-Patent Document 3 it is reported that a red alga belonging to the genus Gardieria recovers approximately 98% of copper (6 ppm) contained in a medium under anaerobic conditions in 4 days.
  • Non-Patent Document 4 when a red algae belonging to the genus Gardieria is cultured for a week in a solution containing Ca, Mg, Fe, Cu, Al, Cr, Na and Ni as metal ions, the maximum is 32% Cu. It has been reported that Cr is recovered in the cell surface at a rate of 24% and Ni at a rate of 59%.
  • Patent Document 1 based on Non-Patent Document 4 reports alkali metal recovery and increased production of polysaccharides during metal addition by cyanidium red alga ATCC40080 strain isolated from nature.
  • Non-Patent Document 5 reports that hydrocarbons accumulated in red algae accumulate 19-25 alkene in the carbon chain.
  • TAG triacylglycerol
  • fatty acid methyl esters which are promising raw materials for biodiesel, in red algae.
  • alcohols long-chain alcohols
  • the present invention has been made in view of such circumstances, and a first object of the present invention is to provide a method for recovering metal from a solution containing metal ions by using cyanidium red algae with high efficiency. It is to provide.
  • a second object of the present invention is to provide a metal recovery method in which a metal contained as a solid in a solution is eluted (bioleaching) into a metal ion, and the metal ion is recovered.
  • the third object of the present invention is to effectively utilize the cultured red algae by obtaining a lipid useful as a raw material for biofuel or a pigment useful as a metal chelator from the cultured cyanidian red algae. is there.
  • the present inventors conducted various tests and made extensive studies. As a result, when cultivating cyanidium red algae in a solution, the concentration of red algae in the solution and Cl It has been found that by adjusting the concentration or adding acetic acid, the number of days required for metal recovery can be reduced and the recovery rate can be improved.
  • TAG triacylglycerol
  • fatty acid methyl esters and alcohols which are promising raw materials for biofuel
  • dye isolated from the cultured cyanidium red algae was useful as a metal chelator.
  • the present invention has been completed based on these findings.
  • the gist of the following (9) is a method for producing a pigment.
  • Cyanidium red algae are cultured in a solution whose cell concentration is adjusted within the range of 10 6 to 10 10 cells / ml, and metal ions contained in the solution are absorbed into the red algae and recovered. A method for recovering a metal.
  • Cyanidium red algae are cultured in a solution whose cell concentration is adjusted within the range of 10 6 to 10 10 cells / ml, and the metal ions contained in the solution are absorbed by the red algae and removed.
  • a method for removing metal comprising:
  • Cyanidium red algae are cultured in a solution whose cell concentration is adjusted within the range of 10 6 to 10 10 cells / ml, and triacylglycerol, fatty acid methyl ester and alcohols are cultured from the cultured red algae.
  • a method for producing lipids, wherein one or more of them are obtained.
  • Cyanidium red algae are cultured in a solution whose cell concentration is adjusted within the range of 10 6 to 10 10 cells / ml, and 210 nm when UV-visible absorption spectrum is performed from the cultured red algae.
  • a method for producing a dye comprising obtaining a dye having absorption maximums at 249 nm, 393 nm, 495 nm, 528 nm, 565 nm, and 663 nm.
  • the metal recovery or removal method of the present invention can recover or remove metal with high efficiency by culturing cyanidium red algae at a high concentration.
  • the lipid production method of the present invention can obtain any one or more of TAG, fatty acid methyl ester, and alcohols that are useful as raw materials for biofuel from cyanidium red algae.
  • the dye production method of the present invention can provide a dye useful as a metal chelator.
  • FIG. 1 is a diagram showing the relationship between the number of days elapsed when red algae were cultured in a solution containing a solid metal and the ratio of lanthanoid in the supernatant or cell fraction, and FIG. FIG. 1 (b) shows the Dy ratio of the cell fraction of red algae, FIG. 1 (c) shows the Nd ratio of the supernatant, and FIG. 1 (d) shows the Nd ratio of the cell fraction.
  • FIG. 2 is a diagram showing the results of TLC analysis on the lipids of cultured red algae.
  • FIG. 3 is a diagram showing the results of gas chromatographic investigation of cultured red algae alcohols.
  • FIG. 4 is an HPLC chromatogram when the pigment is separated from red algae.
  • FIG. 1 is a diagram showing the relationship between the number of days elapsed when red algae were cultured in a solution containing a solid metal and the ratio of lanthanoid in the supernatant or cell fraction
  • FIG. FIG. 1 (b) shows the Dy ratio of
  • FIG. 5 is a diagram showing an ultraviolet-visible absorption spectrum of a pigment recovered from red algae.
  • FIG. 6 is a diagram showing an ultraviolet-visible absorption spectrum of a solution to which a pigment recovered from red algae is added.
  • FIG. 6A shows Cu 2+
  • FIG. 6B shows Dy 3+
  • FIG. is a La 3+
  • Figure 6 (d) is Cs +
  • FIG. 6 (e) is a Nd 3+
  • Fig 6 (f) are diagrams respectively illustrating a case of adding Au 3+.
  • cyanidian red algae are cultured in a solution in which the cell concentration is adjusted within the range of 10 6 to 10 10 cells / ml, and the metal ions contained in the solution are absorbed by red algae. And collecting it.
  • red algae When cultivating cyanidium red algae in a solution containing metal ions, the red algae absorb the metal ions. If the solution in which red algae is cultured is separated into a solution and red algae, the metal can be recovered together with the red algae.
  • the cell concentration of cyanidium red algae is adjusted within the range of 10 6 to 10 10 cells / ml. This is because if the cell concentration changes, the biological action caused by the cyanidian red algae cultured is changed, and as a result, the metal recovery efficiency varies. This cell concentration is a major factor affecting recovery efficiency in metal recovery by cyanidium red algae.
  • the cell concentration of cyanidian red algae is adjusted within the range of 10 6 to 10 10 cells / ml, that is, by culturing in a solution adjusted to a high cell concentration.
  • the number of days required for metal recovery can be reduced and the recovery rate can be improved.
  • the preferred range of the cell concentration varies depending on the type of metal ions to be recovered, although the reason is not clear. For example, when recovering gold ions or copper ions, metals can be recovered with high efficiency at any concentration as long as the cell concentration is in the range of 10 6 to 10 10 cells / ml. On the other hand, when recovering the ions of the lanthanoid element or recovering the iron ions, it is preferable to adjust the cell concentration range to be described later.
  • the metal that can be recovered is not particularly limited, and alkali metal elements, alkaline earth metal elements, transition metal elements (including rare earth elements), group 12 metal elements, group 13 metal elements, Group 14 metal elements can be recovered.
  • Cyanidian red algae can be cultured in fresh water or acidic aqueous solution, and can be cultured in acidic aqueous solution having a pH of 0.5 to 4.5. For this reason, for example, it is used for washing and cooling in a factory, and can be used for metal recovery from factory wastewater containing metal ions. Moreover, it is used for pickling etc. in a factory, and can be used for metal recovery from an acidic factory waste liquid containing metal ions. In addition, since cyanidian red algae have salt tolerance, they can be cultured even in an aqueous solution having a salt concentration of about 200 mM.
  • the metal recovery method of the present invention preferably uses a solution in which the Cl concentration is adjusted to less than 5 mM and / or acetic acid is added when cultivating red algae in the solution. This is because the metal can be recovered from the solution with higher efficiency.
  • the adjustment of the Cl concentration of the solution can be performed, for example, by reducing the Cl concentration by neutralization treatment.
  • the addition of acetic acid may be performed by adjusting the amount according to the cell concentration of red algae in order to obtain an effect of recovering the metal with higher efficiency. For example, if the cell concentration of red algae is 10 9 to 10 10 cells / ml, the acetic acid may be added to a concentration of 15 to 400 mM.
  • the metal recovery method of the present invention can also recover a metal contained as a solid in a solution. If the solution contains solid metal, cyanidian red algae elute the metal contained as a solid in the solution by bioreeling to form metal ions, and further absorb the eluted metal ions by cyanidium red algae. Can be recovered. For this reason, the metal recovery method of the present invention can be used for a solution containing a solid metal, and for example, can be used for metal recovery from sludge containing a polishing liquid or a solid metal. In addition, cyanidian red algae grow in a dry state, so for example, by culturing cyanidian red algae on rocks (ores), the red algae absorb and collect the metal contained in the rocks as solids. can do.
  • Cyanidian red algae vary in the type of metal recovered with high efficiency depending on the culture conditions. For this reason, it is preferable that the metal recovery method of the present invention adjusts the culture conditions and selectively absorbs metal ions by red algae and recovers them.
  • the culture conditions the cell concentration, the light irradiation condition, the condition of the additive added to the solution, and the condition of aeration to the solution may be adjusted.
  • the light irradiation condition can be adjusted to a light condition for irradiating light to the solution or a dark condition for blocking light irradiation to the solution.
  • the conditions of the additive added to the solution can be adjusted to conditions for adding glucose, conditions for adding acetic acid, or conditions for not adding these additives.
  • Aeration conditions to the solution can be adjusted gas containing oxygen (e.g., air) aerobic conditions aeration, anaerobic conditions bubbling CO 2 gas, the anaerobic condition for venting the gas containing no CO 2 gas and oxygen.
  • CO 2 rich gas discharged from the factory can be used, thereby reducing CO 2 exhaust gas in the factory simultaneously with metal recovery.
  • a gas containing no CO 2 gas and oxygen is passed through the solution under anaerobic conditions, for example, a gas such as N 2 gas or Ar gas may be passed.
  • lanthanoid ions when lanthanoid ions are selectively recovered in a solution containing lanthanoid ions and other metal ions (except gold ions), the culture conditions are adjusted, and the cell concentration is 10 9 to 10 10 cells / ml,
  • anaerobic conditions CO 2 gas aeration or gas aeration not containing CO 2 gas and oxygen
  • dark conditions may be used in the solution to which acetic acid has been added.
  • the cell concentration of cyanidium red algae is 10 6 to 10 10. What is necessary is just to culture
  • the cell concentration of cyanidium red algae is 10 6 to 10 7 cells / ml, and glucose is added. What is necessary is just to culture
  • a solution in which the cell concentration of cyanidium red algae is 10 6 to 10 8 cells / ml it may be cultured in the anaerobic condition and the dark condition by gas vent containing no CO 2 gas and oxygen in the medium.
  • anaerobic conditions of gas aeration that does not contain CO 2 gas and oxygen if acetic acid is added regardless of light conditions or dark conditions, red algae grow using acetic acid as a carbon source.
  • the gas to be vented is irrespective of CO 2 gas and oxygen-free gas or CO 2 gas. The metal recovery efficiency is almost unchanged.
  • the anaerobic condition is a CO 2 gas aeration or a gas aeration that does not contain CO 2 gas and oxygen.
  • the anaerobic condition is a gas aeration that does not contain CO 2 gas and oxygen because it can be recovered without adding acetic acid.
  • gold ions are excluded from other metal ions, as shown in Examples described later, depending on cell concentration, light irradiation conditions, additive conditions added to the solution, and aeration conditions to the solution. This is because it is absorbed by red algae. Therefore, if you want to selectively recover another metal ion from a solution containing gold ions, specifically, if you want to selectively recover a lanthanoid from a solution containing gold ions and lanthanoid ions, the lanthanoid is recovered first. After selectively recovering gold by adjusting the conditions of light irradiation, the condition of additives added to the solution, and the condition of aeration to the solution under difficult conditions, another red algae is cultured under the above conditions to lanthanoid Can be recovered.
  • the metal recovery method of the present invention obtains at least one of TAG, fatty acid methyl ester, and alcohols from red algae used for metal recovery.
  • TAG fatty acid methyl ester
  • alcohol fatty acid methyl ester
  • the red algae used for metal recovery can be used effectively as a resource. The cost required for collection can be reduced.
  • the residue of algae after extraction of biofuel is currently a problem, but the residue after extraction of biofuel and useful substances can be used to recover easily recoverable metals such as gold, It is also possible to remove harmful metals from
  • the pigment contained in cyanidian red algae is useful for metal chelators.
  • glucose When glucose is added, it is released in a large amount to the outside of the cell, so that extraction work is not necessary and purification is easy. Therefore, the metal recovery method of the present invention obtains dyes having absorption maxima at 210 nm, 249 nm, 393 nm, 495 nm, 528 nm, 565 nm and 663 nm when UV-visible absorption spectra are performed from red algae used for metal recovery. Is preferred. It is possible to extract the pigment from the medium components of the culture solution and further use the cells for metal recovery. Thereby, red algae used for metal recovery can be effectively used as resources, and as a result, the cost required for metal recovery can be reduced.
  • cyanidium red algae in a solution having a total value of 200 mM of the concentration of each metal ion contained.
  • cyanidium is known to be resistant to aluminum concentrations of 200 mM, but has not been reported to be resistant to higher concentrations of metal ions. For this reason, when the total value of the concentration of each metal ion exceeds 200 mM, the concentration of the metal ion becomes excessively high, and there is a possibility that the red alga of cyanidium may be killed.
  • cyanidian red algae that have absorbed metal and the metal can be performed, for example, by culturing red algae that have absorbed the metal in a medium by adjusting the culture conditions and releasing the metal from the red algae.
  • cyanidian red algae that have absorbed lanthanoids such as Dy, Nd, and La are aerobic in a solution having a cell concentration of about 10 8 cells / ml or less and with or without the addition of glucose.
  • absorbed lanthanoids are released. Since the release of metal occurs more efficiently under the aerobic condition and under the condition where red algae grows, it is possible to add organic substances such as glucose at a concentration at which red algae can grow, or to add a gas containing CO 2 at a high concentration. When a sufficient carbon source is supplied by aeration, the metal release rate increases. Thus, it is preferable to culture red algae to release metal, and to use red algae repeatedly for metal recovery, because the cost required for metal recovery can be reduced.
  • cyanidian red algae are cultured in a solution having a cell concentration adjusted within the range of 10 6 to 10 10 cells / ml, and the metal ions contained in the solution are absorbed into red algae. It is made to remove.
  • Such a method for removing a metal of the present invention can remove metal ions from a solution with high efficiency by causing red algae to absorb the metal ions contained in the solution, as in the above-described method for recovering a metal of the present invention.
  • the metal removal method of the present invention can employ the same embodiment as the metal recovery method of the present invention described above.
  • the lipid production method of the present invention involves culturing cyanidian red algae in a solution at a cell concentration of 10 6 to 10 10 cells / ml, and culturing triacylglycerol (TAG) from the cultured red algae. Any one or more of fatty acid methyl esters and alcohols are obtained. If the cell concentration is 10 6 to 10 10 cells / ml and the concentration is high, the energy used by red algae for growth decreases, and the amount of TAG, fatty acid methyl esters and alcohols accumulated in red algae increases. When the cell concentration is less than 10 6 cells / ml, the rate of energy used by red algae for growth increases, and the amount of energy converted and accumulated as TAG and alcohol decreases.
  • TAG triacylglycerol
  • the cell concentration is more preferably 10 8 to 10 10 cells / ml.
  • lipid production method of the present invention it is preferable to add sugar or acetic acid as a nutrient salt to the solution in order to further increase the amount of TAG, fatty acid methyl ester and alcohol accumulated in red algae.
  • sugar or acetic acid as a nutrient salt
  • other microorganisms such as fungi are mixed in the solution and proliferate, which may inhibit algae culture.
  • cyanidium red algae can be cultured in a solution containing metal ions. Therefore, in order to prevent the growth of other microorganisms such as mold, in a solution containing metals (metal ions) excluding iron. It is preferable to culture red algae.
  • the cyanidian red algae are cultured in the solution at a cell concentration of 10 6 to 10 10 cells / ml. From the red algae, a dye exhibiting absorption maximums at 210 nm, 249 nm, 393 nm, 495 nm, 528 nm, 565 nm and 663 nm when an ultraviolet-visible absorption spectrum is performed is obtained.
  • dyes that absorb at 210 nm, 249 nm, 393 nm, 495 nm, 528 nm, 565 nm, and 663 nm when performing an ultraviolet-visible absorption spectrum are useful as metal chelators.
  • the amount of dye obtained tends to increase as the number of cells increases, if the cell concentration is 10 6 to 10 10 cells / ml and the concentration is high, the amount of dye obtained can be increased. However, if the cell concentration exceeds 10 10 cells / ml, red algae become too dense in the solution, and red algae may die due to lack of light and nutrient salts.
  • the cell concentration is more preferably 10 8 to 10 10 cells / ml.
  • red algae of Cyanidia include red algae belonging to the genus Gardielia, genus cyanidium and genus cyanidiozone.
  • red algae of any of the above genus can be employed, and mutants and transformants thereof can also be employed.
  • Galdieria sulfuraria In the red algae belonging to the genus Gardieria, Galdieria sulfuraria, Galdieria sulfuraria-A, Galdiaria sulfuraria-B, Galdiaria sulfurdiaria M-8, Galeria radidia, Galdiaria prita.
  • cyanidium caldarium RK-1 and cyanidum caldarium Forma A can be used for cyanidium red algae.
  • Cyanidioschizon melolae can be used for red alga belonging to the genus Cyanidioscisone.
  • cyanidium red algae In cultivation of cyanidium red algae, more than 50 kinds of sugars, sugar alcohols, and organic acids (hydrocarbons having 2 to 4 carbon atoms) may be used.
  • nitrogen sources contained in sewage can be used as nutrients, and as with nitrogen sources contained in sewage, there is a possibility that sugar can be used in place of glucose and organic acid can be used in place of acetic acid. is there.
  • Cyanidian red algae can quickly change the state of cell color, the nature of metal recovery, the synthesis of TAG, fatty acid methyl esters and alcohols by changing the culture conditions. For this reason, the conditions until the cells are grown to the cell concentration defined in the present invention are the fastest growing conditions (for example, conditions under which an organic substance such as glucose is added and grown under aerobic conditions regardless of the light conditions), CO 2 It is possible to adopt a condition for fixing the glass with high efficiency (conditions for growing without adding organic substances under light conditions).
  • Cell concentration, Cl concentration and acetic acid addition test [Test method] The test was conducted by culturing red cyanobacteria in the solution and absorbing and removing the metal ions contained in the solution. In pre-culture for preparing red algae used in this test, glucose was added to the following solution and cultured under aerobic and dark conditions. In this test, the cyanidian red algae was Galdieria sulpharia, and the solution was a strongly acidic solution with ammonium sulfate ((NH 4 ) 2 SO 4 ) as the main component and pH 2.5.
  • the composition of the solution is as follows.
  • Solution composition (NH 4 ) 2 SO 4 : 2.62 g / l, KH 2 PO 4 : 0.54 g / l, MgSO 4 .7H 2 O: 0.5 g / l, CaCl 2 .2H 2 O: 0 .14 g / l, FeCl 3 .6H 2 O: 0.0008 g / l, Arnon's A6 metals: 1 ml / l
  • the solution having the above composition has a Cl concentration of 0.95 mM, and DyCl 3 ⁇ 6H 2 O, FeSO 4 ⁇ 7H 2 0 and metals as metals so that any of Dy, Fe and Cu concentrations is 100 ppm. Either CuSO 4 ⁇ 7H 2 O was added. Red algae were added to this solution at a cell concentration in the range of 10 6 to 10 10 cells / ml and cultured for 16 hours, and the culture temperature at that time was 40 to 42 ° C.
  • Test No. In 1-7 a solution having the above composition was used by adjusting the Cl concentration to be 5.0 mM.
  • Test No. In 1-1 to 1-5 and 1-7 to 1-9 acetic acid was added to a concentration of 400 mM.
  • the light irradiation condition was 40 ⁇ E
  • the aeration condition to the solution was an anaerobic condition in which N 2 gas was aerated at about 2 l / h.
  • test no. In 1-1 to 1-5 the cell concentration was adjusted within the range of 10 6 to 10 10 cells / ml, and among them, test nos. With the cell concentration of 10 9 to 10 10 cells / ml. The metal ratio of the cell fraction was good at 1-4 and 1-5. From these, it was confirmed that when recovering Dy belonging to the lanthanoid, the cell concentration is preferably 10 9 to 10 10 cells / ml.
  • Test No. 1-6 red algae were cultured in a solution without addition of acetic acid, and the conditions other than the addition of acetic acid were the same. Compared with 1-4, the metal ratio of the cell fraction decreased. From this, it was revealed that the efficiency of metal recovery can be improved by culturing red algae with a solution to which acetic acid has been added.
  • Test No. 1-7 the Cl concentration of the solution for cultivating red algae was 5.0 mM, and the conditions other than the Cl concentration of the solution were the same. Compared with 1-5, the metal ratio of the cell fraction decreased. From this, it became clear that the metal recovery efficiency can be improved by reducing the Cl concentration of the solution for cultivating red algae with the solution.
  • the culture temperature was 40 to 42 ° C.
  • the light irradiation was performed by irradiating the solution with a fluorescent lamp (about 40 ⁇ E) under the light condition and shielding the light irradiation under the dark condition.
  • the additive to the solution was any one of the conditions in which no additive was added, the conditions in which glucose was added to a concentration of 25 mM, and the conditions in which acetic acid was added to a concentration of 16 mM.
  • the aeration condition to the solution was either an aerobic condition in which air was vented at about 2 l / h or an anaerobic condition in which N 2 gas (purity 95.5%) was vented at about 2 l / h.
  • Table 2 shows light irradiation conditions, additive conditions, and aeration conditions for the solutions in each test.
  • the solution in which red algae were cultured was fractionated into a supernatant and a cell (red algae) fraction by centrifugation, and the concentration of each lanthanoid element was measured by ICP-MS for each of the supernatant and the cell fraction. Moreover, after isolate
  • Table 2 the ratio of each lanthanoid element shown in Table 2 is expressed as a percentage by dividing the concentration of each lanthanoid element measured in the supernatant or cell fraction by the concentration of each lanthanoid element in the solution before culture. It is.
  • test no. Test Nos. 2-1 to 2-4 Only in 2-4, the isolated red algae cells were stained red. For comparison, staining of red algae cells cultured in a solution containing no metal ions was confirmed with alizarin red S, and no staining was observed. In addition, Test No. After the red algae cultured in 2-4 were washed with ethylenediaminetetraacetic acid (EDTA), which is a metal chelating agent, staining was confirmed with alizarin red S, no staining was observed. From these, it was confirmed that by culturing red algae in the solution, the ions of the lanthanoid contained in the solution were absorbed by the cells of the red algae.
  • EDTA ethylenediaminetetraacetic acid
  • test no. In 2-4 the cells were cultured in a dark and anaerobic condition with a solution to which acetic acid had been added. , Nd and La ratios increased significantly. From this, test no. In 2-4, it was confirmed that almost 100% of Dy and Nd and about 80% of La were recovered. Therefore, when selectively recovering lanthanoid ions, it was confirmed that it was preferable to culture in a dark and anaerobic condition with a solution to which acetic acid was added.
  • Gold recovery test [test method] Test No. of this test In 3-1 to 3-6, gold (Au) was added to a solution having the composition shown in “1. Cell concentration, Cl concentration and acetic acid addition test” so that the concentration was 350 ppm. Then, Galdieria sulfuraria, a red alga belonging to the genus Gardieria, was added to this solution so that the cell concentration would be 10 8 cells / ml, and the cells were cultured for 24 hours. Test No. In 3-7 and 3-8, the gold concentration was 10 ppm, and the red algae was Cyanidischyzon melolae, a red algae belonging to the genus cyanidiozone.
  • the culture temperature was 40 to 42 ° C.
  • the light irradiation was performed by irradiating the solution with a fluorescent lamp (about 40 ⁇ E) under the light condition and shielding the light irradiation under the dark condition.
  • the additive to the solution was any one of the conditions in which no additive was added, the conditions in which glucose was added to a concentration of 25 mM, and the conditions in which acetic acid was added to a concentration of 16 mM.
  • the aeration conditions for the solution were aerobic conditions for venting the atmosphere at about 2 l / h, anaerobic conditions for venting CO 2 gas (purity 98%) at about 2 l / h, and N 2 gas (purity 99.99%).
  • Table 3 shows the gold concentration of the solution before culturing, the type of red algae, the light irradiation conditions, the additives and the aeration conditions for the solutions in each test.
  • the color of the solution after the culture was visually observed to confirm a change in the color of the solution.
  • the solution in which red algae is cultured is separated into a supernatant and a cell (red algae) fraction by centrifugation, and the red algae are collected and observed with a microscope, and gold fine particles are deposited on the surface layer of the cells. The presence or absence of discoloration to purple due to was confirmed.
  • the gold concentration of the supernatant was measured by ICP-MS. The recovery rate was calculated by dividing the difference between the gold concentration of the pre-culture solution and the gold concentration of the supernatant by the gold concentration of the pre-culture solution. Table 3 also shows the color of the solution after culture, the presence or absence of discoloration of the cells, and the calculated recovery rate.
  • Bioleaching test of lanthanoid [Test method] In this test, 50 mL of a solution having the composition shown in “1. Cell concentration, Cl concentration and acetic acid addition test” was added to granular and water-insoluble Dy and Nd dioxide (Dy: 14.7% by mass and Nd: 85). . Containing 3% by mass), and to this solution was added Galdieria sulfuraria, a red alga belonging to the genus Gardieria, so that the cell concentration would be 10 9 to 10 10 cells / ml, and the cells were cultured for 10 days. .
  • the culture temperature was 40 to 42 ° C.
  • the light irradiation was performed by irradiating the solution with a fluorescent lamp (about 40 ⁇ E) under the light condition and shielding the light irradiation under the dark condition.
  • the additive to the solution was one of the conditions in which no additive was added, the condition in which glucose was added to a concentration of 25 mM, and the condition in which acetic acid was added to a concentration of 150 mM.
  • the aeration condition to the solution was either an aerobic condition in which air was vented at about 2 l / h or an anaerobic condition in which N 2 gas (purity 95.5%) was vented at about 2 l / h.
  • Table 4 shows the light irradiation conditions, the additives and the aeration conditions to the solution in each test.
  • the mass of Dy and Nd was measured by ICP-MS for the supernatant and the cell fraction, respectively.
  • the measured mass of Dy or Nd was divided by the mass of Dy or Nd contained in the dioxide added to the solution and expressed as a percentage, which was defined as the Dy or Nd ratio.
  • FIG. 1 is a diagram showing the relationship between the number of days elapsed when red algae were cultured in a solution containing a solid metal and the ratio of lanthanoid in the supernatant or cell fraction
  • FIG. FIG. 1 (b) shows the Dy ratio of the cell fraction of red algae
  • FIG. 1 (c) shows the Nd ratio of the supernatant
  • FIG. 1 (d) shows the Nd ratio of the cell fraction.
  • Test No. 4-1 culturing was carried out in a solution containing no additive under light conditions and aerobic conditions, and the Dy and Nd ratios of the supernatant increased with the passage of days.
  • Test No. 4-2 the solution was added with glucose and cultured under light and aerobic conditions, and the Dy and Nd ratios of the supernatant increased with the passage of days.
  • the Dy and Nd ratio of the cell fraction is No. In any test of 4-1 to 4-4, it increased with the passage of days. In particular, no. In 4-4, the cells were cultured in a dark and anaerobic condition with a solution to which acetic acid had been added, and the Dy and Nd ratios of red algae cells increased markedly with the passage of days.
  • Bioleaching test for neodymium magnet waste [test method] In this test, 50 mg of a granular and water-insoluble neodymium magnet waste material is added to 50 ml of the solution having the composition shown in “1. Cell concentration, Cl concentration and acetic acid addition test”, and this solution is a red alga belonging to the genus Gardieria. Galdieria sulpharia was added to a cell concentration of 10 9 to 10 10 cells / ml, and cultured for 4 days.
  • the main composition of neodymium magnet waste is as follows. Main composition of neodymium magnet waste: Nd: 19.91% by mass, Dy: 4.41% by mass and Fe: 53.87% by mass
  • the culture temperature was 40 to 42 ° C.
  • the light irradiation was performed by irradiating the solution with a fluorescent lamp (about 40 ⁇ E) under the light condition and shielding the light irradiation under the dark condition.
  • the additive to the solution was one of the conditions in which no additive was added, the condition in which glucose was added to a concentration of 25 mM, and the condition in which acetic acid was added to a concentration of 150 mM.
  • the aeration condition to the solution was either an aerobic condition in which air was vented at about 2 l / h or an anaerobic condition in which N 2 gas (purity 99.99%) was vented at about 2 l / h.
  • Table 5 shows the light irradiation condition, the additive and the aeration condition to the solution in each test.
  • test no In any of the tests of 5-1 to 5-4, the ratio of Dy, Nd, and Fe is increased, and cyanidium red algae can be recovered by eluting Dy, Nd, and Fe from neodymium magnet waste. It could be confirmed.
  • test no. In 5-3 culture was performed in a dark and aerobic condition with a solution to which glucose was added, and the Fe ratio of the cell fraction was significantly increased.
  • Test No. When the solution in which the red algae were cultured before the separation of the red algae in 5-3 was confirmed, the solution that had been green before the culture was changed to brown.
  • Test No. When a part of red algae was collected from the cell fraction of 5-3 and confirmed with a microscope, a brownish brown mass was confirmed on the cell surface layer. Even if the red algae used in other tests were observed with a microscope, a brownish brown lump was not confirmed. Therefore, this lump was presumed to be iron, and the cyanidium red algae shown in Non-Patent Document 6 was used. It is speculated that this is different from the formation of a structure that accumulates metal in the cells.
  • test no. In No. 5-4 the Dy and Nd ratios of red algae cells were remarkably high when cultured in a dark and anaerobic condition with a solution supplemented with acetic acid. From these, when cultivated in a dark and anaerobic condition with a solution containing acetic acid, cyanidium red algae can elute lanthanoids from neodymium magnet waste by bioleaching and absorb the eluted lanthanoids with high efficiency. Became clear.
  • TAG detection test In this test, the cyanidium red algae cultured in Example 2 of the present invention in “1. Cell concentration, Cl concentration and acetic acid addition test” were first collected and reddish with Nile red reagent (Nile red reagent). Algal cells were stained. As a result, a plurality of red algae cells were spotted in a fluorescent yellow color, that is, lipid droplets containing TAG were observed. Next, the composition of red algae lipids was examined by TLC analysis (thin layer chromatography analysis).
  • FIG. 2 is a diagram showing the results of TLC analysis on the lipids of cultured red algae.
  • WAX means alkene
  • TAG means triacylglycerol
  • FA means free fatty acid
  • DG means diacylglycerol. From FIG. 2, it was confirmed that spots corresponding to TAG were present in red algae cells, that is, it was confirmed that TAG was accumulated in red algae cells. As a result of TLC analysis, the proportion of TAG in the lipid accumulated in red algae was about 30% by mass.
  • TAG was extracted from the silica gel of the spot portion corresponding to the TAG of the silica gel (TLC) plate, and analyzed by gas chromatography. The results are shown in Table 6. For comparison, general compositions of plant-derived rapeseed oil and palm oil are also shown in Table 6.
  • TAG accumulated in red algae accounted for about 40 mass% in total of saturated fatty acids having no unsaturated bonds (the number of unsaturated bonds is 0).
  • palm oil containing a lot of saturated fatty acids having no unsaturated bonds is more likely to produce NOx during combustion in biofuel obtained when used as a raw material than rapeseed oil containing many saturated fatty acids having unsaturated bonds. It is known that the amount is small.
  • TAG accumulated in red algae contains a lot of saturated fatty acids having no unsaturated bonds as in palm oil
  • TAG obtained by the metal recovery method or lipid production method of the present invention is: It was confirmed that it is suitable for biofuel.
  • Detection test of alcohols first, cyanidium red algae cultured in Example 2 of the present invention in “1. Cell concentration, Cl concentration and acetic acid addition test” were collected and analyzed by gas chromatography. The composition was investigated for the species.
  • FIG. 3 is a diagram showing the results of gas chromatographic investigation of cultured red algae alcohols. From FIG. 3, it was confirmed that phytol was detected as a long-chain alcohol in cells of red algae, and fatty acid methyl esters were detected along with fatty acids, that is, primary alcohol was accumulated in cells of red algae.
  • fatty acid methyl esters are synthesized from waste oil or plant-derived TAG and used as biodiesel, but a large amount of glycerol, which is a by-product of the synthesis, is problematic. Since accumulation of fatty acid methyl esters is observed in cells this time, synthesis of fatty acid methyl esters in cells of red algae leads to the solution of the above problems.
  • Pigment Isolation and Metal Removal Test In this test, first, the cyanidium red algae cultured in Example 2 of the present invention in “1. Cell concentration, Cl concentration and acetic acid addition test” were collected, It was washed with a chelating agent, ethylenediaminetetraacetic acid (EDTA). The washed red algae were mixed with the ethyl acetate solution and then separated to recover the pigment in the ethyl acetate fraction. The ethyl acetate fraction from which the dye was recovered was concentrated to dryness, and then the methanol-soluble fraction was separated by HPLC. The HPLC separation conditions were ODS columns and acetonitrile (10-60% gradient, flow rate 1 ml / min).
  • FIG. 4 is an HPLC chromatogram when the pigment is separated from red algae.
  • the pigment recovered from red algae was isolated as a 23 minute fraction.
  • the isolated dye was observed under natural light, it showed a reddish purple color and fluorescent under UV light.
  • the ultraviolet-visible absorption spectrum of the collected red algae pigment was examined.
  • the pigment separated from red algae was added to 90% by mass of methanol, and this solution was examined by DU800 manufactured by Beckman Coulter.
  • FIG. 5 is a diagram showing an ultraviolet-visible absorption spectrum of a pigment recovered from red algae. It is confirmed from FIG. 5 that the pigments collected from red algae show absorption maxima at 210 nm, 249 nm, 393 nm, 495 nm, 528 nm, 565 nm and 663 nm.
  • Flavonoids are known to undergo spectral changes by chelating metals.
  • the test conditions were 100 ppm Fe 2+ (FeSO 4 .7H 2 0), Cu 2+ (CuSO 4 ), Nd 3+ (NdCl 3 ), Dy 3+ (DyCl 3 ), La 3+ (LaCl 3 ) in 90% by mass methanol.
  • Au 3+ (AuCl 3 ) and Cs + (CsCl) were added, and 1 mM EDTA was added.
  • the UV-visible absorption spectrum of the solution to which the pigment recovered from red algae was added was examined using DU800 manufactured by Beckman Coulter.
  • FIG. 6 is a diagram showing an ultraviolet-visible absorption spectrum of a solution to which a pigment recovered from red algae is added.
  • FIG. 6A shows Cu 2+
  • FIG. 6B shows Dy 3+
  • FIG. is a La 3+
  • Figure 6 (d) is Cs +
  • FIG. 6 (e) is a Nd 3+
  • Fig 6 (f) are diagrams respectively illustrating a case of adding Au 3+. From FIG. 6, the metal bound to the pigment collected from red algae slightly shifted the height and position of the peaks at 393 nm and 565 nm, and the peaks at 495 nm and 663 nm disappeared.
  • the metal recovery or removal method of the present invention can recover or remove metal with high efficiency by culturing cyanidium red algae at a high concentration. Therefore, if the method for recovering or removing metal according to the present invention is applied to recovering or removing metal ions from industrial waste water or waste liquid containing metal ions, the cost required for recovery or removal can be reduced.
  • the lipid production method of the present invention can obtain any one or more of TAG, fatty acid methyl ester and alcohols useful as a raw material for biofuel from cyanidium red algae.
  • the dye production method of the present invention can provide a dye useful as a metal chelator.

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Abstract

La présente invention concerne un procédé de récupération de métal grâce auquel il est possible de récupérer avec une grande efficacité des métaux (ions métalliques) contenus dans une solution par mise en culture d'algues rouges faisant partie de l'ordre des Cyanidiales dans une solution, dans laquelle leur concentration cellulaire a été amenée dans une plage allant de 106 à 1010 cellules/ml, et à récupérer les ions métalliques contenus dans la solution qui ont été absorbés par les algues rouges. Dans ce cas, la solution utilisée de préférence pour la culture d'algues rouges dans une solution est une solution ayant été amenée à une concentration de Cl inférieure à 5 mM et/ou à laquelle de l'acide acétique a été ajouté. De plus, une partie ou l'ensemble des ions métalliques contenus dans la solution peuvent être des ions métalliques ayant été élués des métaux contenus en tant que solide dans la solution. Cela signifie qu'il est possible d'utiliser la biolixiviation afin d'amener les métaux contenus en tant que solide dans la solution à être élués en tant qu'ions métalliques puis à récupérer les ions métalliques élués.
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WO2018155687A1 (fr) * 2017-02-27 2018-08-30 株式会社ガルデリア Agent de récupération de métal, agent de récupération de composé métallique et procédé de récupération de métal ou de composé métallique
WO2021077027A1 (fr) * 2019-10-17 2021-04-22 The Regents Of The University Of California Acides gras et polymères d'origine biologique
WO2022049790A1 (fr) * 2020-09-03 2022-03-10 株式会社ガルデリア Procédé de récupération d'or et kit de récupération d'or
WO2024053487A1 (fr) * 2022-09-06 2024-03-14 株式会社Ihi Agent de récupération de métal, élément de récupération de métal, agent liquide de récupération de métal et procédé de récupération de métal

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JP6359314B2 (ja) * 2014-03-31 2018-07-18 学校法人立教学院 紅藻シアニジウム目のための脂質生産用培地組成物および脂質生産方法
FR3041653B1 (fr) * 2015-09-25 2017-12-29 Fermentalg Procede de culture d'algues, particulierement d'algues rouges unicellulaires (arus)
JP7130278B1 (ja) * 2021-04-27 2022-09-05 株式会社ガルデリア 金属の回収方法
US20240384368A1 (en) * 2021-08-31 2024-11-21 Ihi Corporation Metal recovery material, and method for recovering metal from solution including metal ion or metal complex ion
AU2022337471B2 (en) * 2021-08-31 2024-05-09 Ihi Corporation Method for recovering metal from metal element-containing substance

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WO2017111092A1 (fr) * 2015-12-22 2017-06-29 株式会社ガルデリア Agent pour la récupération sélective de métal, procédé de récupération de métal et procédé d'élution de métal
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WO2018155687A1 (fr) * 2017-02-27 2018-08-30 株式会社ガルデリア Agent de récupération de métal, agent de récupération de composé métallique et procédé de récupération de métal ou de composé métallique
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WO2021077027A1 (fr) * 2019-10-17 2021-04-22 The Regents Of The University Of California Acides gras et polymères d'origine biologique
WO2022049790A1 (fr) * 2020-09-03 2022-03-10 株式会社ガルデリア Procédé de récupération d'or et kit de récupération d'or
JP2022045929A (ja) * 2020-09-03 2022-03-23 株式会社ガルデリア 金の回収方法
WO2024053487A1 (fr) * 2022-09-06 2024-03-14 株式会社Ihi Agent de récupération de métal, élément de récupération de métal, agent liquide de récupération de métal et procédé de récupération de métal

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