JP7429344B2 - Collection method of halophilic microalgae - Google Patents
Collection method of halophilic microalgae Download PDFInfo
- Publication number
- JP7429344B2 JP7429344B2 JP2019204446A JP2019204446A JP7429344B2 JP 7429344 B2 JP7429344 B2 JP 7429344B2 JP 2019204446 A JP2019204446 A JP 2019204446A JP 2019204446 A JP2019204446 A JP 2019204446A JP 7429344 B2 JP7429344 B2 JP 7429344B2
- Authority
- JP
- Japan
- Prior art keywords
- microalgae
- halophilic
- container
- halophilic microalgae
- collecting
- 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.)
- Active
Links
- 238000000034 method Methods 0.000 title claims description 51
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 63
- 241000195634 Dunaliella Species 0.000 claims description 37
- 239000007788 liquid Substances 0.000 claims description 23
- 239000000126 substance Substances 0.000 claims description 23
- 239000013535 sea water Substances 0.000 claims description 21
- 230000000903 blocking effect Effects 0.000 claims description 12
- 238000004519 manufacturing process Methods 0.000 claims description 10
- 235000013343 vitamin Nutrition 0.000 claims description 8
- 229940088594 vitamin Drugs 0.000 claims description 8
- 229930003231 vitamin Natural products 0.000 claims description 8
- 239000011782 vitamin Substances 0.000 claims description 8
- 238000005119 centrifugation Methods 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 5
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 5
- 108090000623 proteins and genes Proteins 0.000 claims description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 5
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 5
- 102000015636 Oligopeptides Human genes 0.000 claims description 4
- 108010038807 Oligopeptides Proteins 0.000 claims description 4
- 229920001184 polypeptide Polymers 0.000 claims description 4
- 239000002243 precursor Substances 0.000 claims description 4
- 235000000346 sugar Nutrition 0.000 claims description 4
- 229930186217 Glycolipid Natural products 0.000 claims description 3
- 102000004895 Lipoproteins Human genes 0.000 claims description 3
- 108090001030 Lipoproteins Proteins 0.000 claims description 3
- 150000001720 carbohydrates Chemical class 0.000 claims description 3
- 235000014633 carbohydrates Nutrition 0.000 claims description 3
- 229930195733 hydrocarbon Natural products 0.000 claims description 3
- 150000002430 hydrocarbons Chemical class 0.000 claims description 3
- 150000003904 phospholipids Chemical class 0.000 claims description 3
- 239000002689 soil Substances 0.000 claims description 2
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 claims 1
- 239000004215 Carbon black (E152) Substances 0.000 claims 1
- 150000003839 salts Chemical class 0.000 description 23
- 238000011084 recovery Methods 0.000 description 22
- 238000004062 sedimentation Methods 0.000 description 21
- 210000004027 cell Anatomy 0.000 description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 15
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 14
- 238000012258 culturing Methods 0.000 description 11
- 239000003921 oil Substances 0.000 description 9
- 239000000446 fuel Substances 0.000 description 7
- 235000011187 glycerol Nutrition 0.000 description 7
- 238000000926 separation method Methods 0.000 description 7
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 6
- 238000011109 contamination Methods 0.000 description 6
- 238000001914 filtration Methods 0.000 description 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 6
- 239000005416 organic matter Substances 0.000 description 6
- 229910052760 oxygen Inorganic materials 0.000 description 6
- 239000001301 oxygen Substances 0.000 description 6
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 description 5
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 description 5
- 235000013734 beta-carotene Nutrition 0.000 description 5
- 239000011648 beta-carotene Substances 0.000 description 5
- 229960002747 betacarotene Drugs 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 238000005189 flocculation Methods 0.000 description 5
- 230000016615 flocculation Effects 0.000 description 5
- 239000000295 fuel oil Substances 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 241000195493 Cryptophyta Species 0.000 description 4
- 238000007796 conventional method Methods 0.000 description 4
- 238000001704 evaporation Methods 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 235000014593 oils and fats Nutrition 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 241000195633 Dunaliella salina Species 0.000 description 3
- 108010024636 Glutathione Proteins 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 239000001569 carbon dioxide Substances 0.000 description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 description 3
- 235000021466 carotenoid Nutrition 0.000 description 3
- 150000001747 carotenoids Chemical class 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000003925 fat Substances 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 150000008163 sugars Chemical class 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000209094 Oryza Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 230000003044 adaptive effect Effects 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000007667 floating Methods 0.000 description 2
- 239000008394 flocculating agent Substances 0.000 description 2
- 230000003311 flocculating effect Effects 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 230000003204 osmotic effect Effects 0.000 description 2
- 230000019935 photoinhibition Effects 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- -1 sodium ions Chemical class 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- 241000736718 Dunaliella bioculata Species 0.000 description 1
- 241001403474 Dunaliella primolecta Species 0.000 description 1
- 241000195632 Dunaliella tertiolecta Species 0.000 description 1
- 241001231664 Dunaliella viridis Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 238000010420 art technique Methods 0.000 description 1
- 238000011001 backwashing Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000005587 bubbling Effects 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000012824 chemical production Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000010908 decantation Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005868 electrolysis reaction Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000005188 flotation Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 238000012388 gravitational sedimentation Methods 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 239000003345 natural gas Substances 0.000 description 1
- 150000002823 nitrates Chemical class 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002787 reinforcement Effects 0.000 description 1
- 230000003014 reinforcing effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 150000004760 silicates Chemical class 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910001415 sodium ion Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000004078 waterproofing Methods 0.000 description 1
Classifications
-
- 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
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/59—Biological synthesis; Biological purification
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
本発明は、好塩性微細藻類の回収方法、及び、回収された好塩性微細藻類から有用有機物を得る方法に関する。 The present invention relates to a method for collecting halophilic microalgae and a method for obtaining useful organic substances from the collected halophilic microalgae.
太陽エネルギーや風力エネルギー等の再生可能エネルギーによってエネルギー需要の多くを賄うことが求められているが、再生可能エネルギーはエネルギー密度が低く、化石エネルギーからの早急な転換はハードルが高い。
そこで、微細藻類による燃料油生産によって、該ハードルを下げることが期待されている。
Renewable energy such as solar energy and wind energy is required to meet much of the energy demand, but renewable energy has a low energy density, making it difficult to quickly switch away from fossil energy.
Therefore, fuel oil production using microalgae is expected to lower this hurdle.
微細藻類による燃料油生産というアイデアは、既に提案されている（例えば、特許文献１、２）。しかし、現状を鑑みると、一部を除き実用化に関しては、多くの問題点が解決できていない。 The idea of producing fuel oil using microalgae has already been proposed (for example, Patent Documents 1 and 2). However, considering the current situation, many problems have not been solved in terms of practical application, with some exceptions.
また、培養・増殖した微細藻類から有用有機物を得ることも提案され、糖、ポリペプチド、ビタミン等、種々の有用物に関し、具体的な検討もなされている（例えば、特許文献３、４）。
特許文献３には、好塩性藍藻を、高塩濃度で定常期まで培養した後、低塩濃度で数時間～数日間培養して多糖類を生産させ、該多糖類を分離精製する方法が記載されている。
また、特許文献４には、水性藻類バイオマス懸濁液から、β－カロテンを溶媒抽出する方法が記載されている。
It has also been proposed to obtain useful organic substances from cultured and proliferated microalgae, and specific studies have been made regarding various useful substances such as sugars, polypeptides, and vitamins (for example, Patent Documents 3 and 4).
Patent Document 3 describes a method of culturing halophilic blue-green algae to a stationary phase at a high salt concentration, then culturing them at a low salt concentration for several hours to several days to produce polysaccharides, and separating and purifying the polysaccharides. Are listed.
Further, Patent Document 4 describes a method for solvent extraction of β-carotene from an aqueous algal biomass suspension.
一方、培養・増殖させた微細藻類を効率よく水中から回収する方法も検討されている（例えば、特許文献５～９）。 On the other hand, methods for efficiently recovering cultured and proliferated microalgae from water are also being studied (for example, Patent Documents 5 to 9).
特許文献５には、微細藻類が凝集しやすい表面ゼータ電位となるように、原水の塩類濃度を調整し、原水に無機凝集剤を添加して凝集反応を行わせる凝集工程と、該凝集工程で生成した凝集フロックを固液分離する加圧浮上分離工程とを有する、微細藻類の水からの分離回収方法が記載されている。
また、特許文献６には、原水に難溶性水酸化物を生成する金属塩を添加した後、該原水を難溶解性水酸化物が生成するｐＨに調整する水酸化物生成工程と、該難溶解性水酸化物によって微細藻類を凝集させる凝集工程と、生成した凝集フロックを固液分離する固液分離工程とを有する微細藻類の回収方法が記載されている。
Patent Document 5 describes a flocculation step in which the salt concentration of raw water is adjusted so that the surface zeta potential is such that microalgae can easily flocculate, and an inorganic flocculant is added to the raw water to cause a flocculation reaction, and in the flocculation step. A method for separating and recovering microalgae from water is described, which includes a pressure flotation separation step for solid-liquid separation of generated flocs.
Further, Patent Document 6 describes a hydroxide generation step of adding a metal salt that generates a hardly soluble hydroxide to raw water, and then adjusting the raw water to a pH at which the hardly soluble hydroxide is generated; A method for collecting microalgae is described that includes a flocculation step of flocculating microalgae with soluble hydroxide, and a solid-liquid separation step of solid-liquid separation of the generated flocs.
特許文献７には、微細藻類を透過させない分離膜を備えた多孔質フィルターを使用して、微細藻類を含む培養液から、濾過セル内に微細藻類を回収し、該微細藻類の成形体を作製し、逆洗を行って該成形体を排出させる微細藻類の回収・成形方法が記載されている。
また、特許文献８には、微細藻類の培養液の上方に気層が存在するようにしながら、該培養液で微細藻類を培養し、該培養液の液量を減らし、培養物を回収する、微細藻類の培養及び回収方法が記載されている。
Patent Document 7 discloses that microalgae are collected in a filtration cell from a culture solution containing microalgae using a porous filter equipped with a separation membrane that does not allow microalgae to pass through, and a molded body of the microalgae is produced. However, a method for collecting and molding microalgae is described in which the molded bodies are discharged by backwashing.
Further, Patent Document 8 discloses that microalgae are cultured in a culture solution while an air layer is present above the culture solution of microalgae, the volume of the culture solution is reduced, and the culture is recovered. A method for culturing and collecting microalgae is described.
また、特許文献９には、好塩性微細藻類に属する藻類の藻体を、タンパク質の変性による沈殿を利用して取得する好塩性微細藻類の回収方法が記載され、培養液中に、タンパク質を含む有機物溶液を添加して混合して静置し、沈殿物と上澄みとに分離し、該沈殿物に含まれる藻体を回収する好塩性微細藻類の回収方法が記載されている。 Further, Patent Document 9 describes a method for collecting halophilic microalgae, in which algal bodies of algae belonging to halophilic microalgae are obtained using precipitation due to protein denaturation. A method for collecting halophilic microalgae is described, in which an organic matter solution containing .
しかしながら、これらの技術は、化学的・物理学的に、培養液（微細藻類を有する原水）から、物体としての微細藻類を回収する方法であり、微細藻類が有する生物としての性質を利用して生物学的に濃縮する方法ではなかった。
また、後述するように、従来知られている濃縮・回収技術では、微細藻類を有する原水中から微細藻類を濃縮する回収率や効率（コストパフォーマンス）が悪かったり、濃縮・回収に要するエネルギーが多過ぎたりして、実用化までには、程遠いものであった（表１参照）。
However, these techniques are methods for chemically and physically recovering microalgae as objects from a culture solution (raw water containing microalgae), and utilize the properties of microalgae as living organisms. It was not a biologically concentrated method.
Furthermore, as will be discussed later, conventionally known concentration/recovery technologies have poor recovery rates and efficiency (cost performance) for concentrating microalgae from raw water containing microalgae, and require a large amount of energy for concentration/recovery. However, it was still far from being put into practical use (see Table 1).
そこで、石油や天然ガスに代わって、「微細藻類に含まれる燃料油を獲得すること」に関しては、コスト面、効率面等で十分ではなく、更なる改良が求められている。
また、「微細藻類から有用有機物を獲得すること」に関しても、消費必要エネルギー量の減少、微細藻類の回収率の向上等、コスト面（効率面）等から更なる改良が求められている。
Therefore, "obtaining fuel oil contained in microalgae" instead of oil or natural gas is not sufficient in terms of cost and efficiency, and further improvements are required.
Furthermore, with regard to "obtaining useful organic substances from microalgae," further improvements are required from a cost perspective (efficiency perspective), such as a reduction in the amount of energy required to be consumed and an improvement in the recovery rate of microalgae.
本発明は上記背景技術に鑑みてなされたものであり、その課題は、前記問題点を解決し、回収に必要なエネルギーが低く、回収率が高く、全体として効率の良い好塩性微細藻類の回収方法を提供することである。 The present invention has been made in view of the above-mentioned background art, and its object is to solve the above-mentioned problems and to produce halophilic microalgae that requires less energy for recovery, has a high recovery rate, and is highly efficient overall. The objective is to provide a collection method.
本発明者は、上記の課題を解決すべく鋭意検討を重ねた結果、池の水の中で好塩性微細藻類を培養し、該水を一旦回収容器に移し、その後、光を遮断することによって、該好塩性微細藻類が該容器の下に沈降することを見出して、本発明を完成するに至った。 As a result of intensive studies to solve the above-mentioned problems, the present inventor has developed a method of culturing halophilic microalgae in pond water, temporarily transferring the water to a collection container, and then blocking light. The inventors discovered that the halophilic microalgae settle under the container and completed the present invention.
すなわち、本発明は、以下の工程（１）（３）（４）（５）の全てをこの順に有することを特徴とする好塩性微細藻類の回収方法を提供するものである。
（１）池の水の中で好塩性微細藻類を培養する工程
（３）工程（１）で培養された好塩性微細藻類を、該池の水と共に回収して回収液を得る工程
（４）工程（３）で得られた回収液を容器に入れ、該容器内を遮光して、該好塩性微細藻類を該容器の底に沈降させる工程
（５）工程（４）で沈降させた好塩性微細藻類を回収する工程
That is, the present invention provides a method for collecting halophilic microalgae, which is characterized by having all of the following steps (1), (3), (4), and (5) in this order.
(1) A step of cultivating halophilic microalgae in pond water (3) A step of collecting the halophilic microalgae cultured in step (1) together with the pond water to obtain a recovered liquid ( 4) Put the collected liquid obtained in step (3) into a container, block the inside of the container from light, and let the halophilic microalgae settle to the bottom of the container (5) Step (4) Process of collecting halophilic microalgae
また、本発明は、上記容器の高さを上記池の深さより大きく設定し、該容器の水深を該池の水深より大きくして、好塩性微細藻類を該容器の底に沈降させる上記の好塩性微細藻類の回収方法を提供するものである。 The present invention also provides the above-mentioned method in which the height of the container is set larger than the depth of the pond, the water depth of the container is made larger than the water depth of the pond, and the halophilic microalgae settle to the bottom of the container. A method for collecting halophilic microalgae is provided.
また、本発明は、上記の「好塩性微細藻類の回収方法」を使用して回収した好塩性微細藻類から有用有機物を獲得することを特徴とする有用有機物の製造方法を提供するものである。 The present invention also provides a method for producing useful organic matter, which comprises obtaining useful organic matter from halophilic microalgae collected using the above-mentioned "method for collecting halophilic microalgae." be.
本発明によれば、前記問題点や解決し、好塩性微細藻類を培養・増殖させた原水中での濃縮や、培養・増殖された該好塩性微細藻類の回収に際し、該回収に必要なエネルギー量が低く、回収率が高く、消耗品を必要としないので変動費が低く抑えられ、全体として効率・コストパフォーマンスの良い回収方法を提供できる。 According to the present invention, the above-mentioned problems are solved, and when the halophilic microalgae are concentrated in the raw water in which they are cultured and propagated, and the halophilic microalgae are recovered, the halophilic microalgae are collected. The amount of energy required is low, the recovery rate is high, and variable costs are kept low because no consumables are required, making it possible to provide a recovery method with good overall efficiency and cost performance.
また、本発明によれば、濃縮・回収に際して、凝集剤、ｐＨ調整剤等の化学物質を必須とはせず、遠心分離、濾過（フィルター）、蒸発乾固、電圧印加等の物理的・機械的な操作や専用の装置も必須とはしないので、作業が簡便であり、種々の部材の交換が不要であるため変動費が抑制され、全体として極めて効率の良い好塩性微細藻類の回収方法を提供できる。 Furthermore, according to the present invention, during concentration and recovery, chemical substances such as flocculants and pH adjusters are not required, and physical and mechanical methods such as centrifugation, filtration (filter), evaporation to dryness, and voltage application are required. This is a method for collecting halophilic microalgae that is easy to operate as it does not require manual operations or dedicated equipment, and reduces variable costs as there is no need to replace various parts, and is extremely efficient overall. can be provided.
本発明は、「培養された微細藻類を含有する原水」から、化学的・物理学的に、物体としての微細藻類を濃縮するのではなく、回収しようとする微細藻類が有する生物としての性質を利用して生物学的に濃縮をする。従って、濃縮に要するエネルギーが低く、大規模な装置も部材の交換作業も必要とせず、また、添加した化学物質による雑菌混入等のコンタミネーションのおそれもない。 The present invention does not chemically and physically concentrate the microalgae as an object, but rather collects the biological properties of the microalgae to be recovered from "raw water containing cultured microalgae." It is used to biologically concentrate. Therefore, the energy required for concentration is low, there is no need for large-scale equipment or replacement work of parts, and there is no fear of contamination such as contamination by germs due to added chemical substances.
一般に、好塩性微細藻類を用いて、採算がとれるように、油脂（燃料）や有用有機物を得るためには、ある程度以上の生産規模が必要である。
本発明によれば、微細藻類を凝集させるための電力、遠心分離や濾過のための装置、浮上油回収装置等、生産規模が大きくなればなるほど大掛かりになる「エネルギーや装置」を必須とはせず、また、規模に比例して大量に必要になる凝集剤、ｐＨ調整剤等の化学物質の添加も必須としないので、コストパフォーマンスに優れ、公害を生じさせるおそれもない。また、初期投資も多くはかからないので、固定費も抑制される。
Generally, in order to obtain oils and fats (fuel) and useful organic substances using halophilic microalgae in a profitable manner, a production scale of at least a certain level is required.
According to the present invention, "energy and equipment" such as electric power for flocculating microalgae, equipment for centrifugal separation and filtration, and floating oil recovery equipment, which become larger as the scale of production increases, are not required. Moreover, since it is not necessary to add chemical substances such as flocculants and pH adjusters, which are required in large quantities in proportion to the scale, it is excellent in cost performance and there is no risk of causing pollution. In addition, since the initial investment is not large, fixed costs are also suppressed.
従って、それら長所があることによって、低コストで大規模化でき、その結果、本発明によって初めて、商業的に採算がとれるようになる可能性がある。 Therefore, due to these advantages, it is possible to increase the scale at low cost, and as a result, the present invention may become commercially profitable for the first time.
表１に、非特許文献１を参考にして（引用して）、従来技術に関し相互の比較を示す。
表１中、「ＴＳＳ」は、総浮遊物質（total suspended solids）を示す。
Table 1 shows a mutual comparison regarding the prior art, with reference to (quoting) Non-Patent Document 1.
In Table 1, "TSS" indicates total suspended solids.
従来技術は、何れも重大な短所がある。
それに対し、本発明は、従来技術と比較して、必要エネルギーが少なく；化学物質が混入せず；静置による単なる重力沈降に比べて遥かにプロセスが速く；電極を必要とする方法に比べて、必要エネルギーが低く、電極のメンテナンスや交換の必要もない。
All of the prior art techniques have significant shortcomings.
In contrast, the present invention requires less energy than conventional techniques; no chemicals are mixed; the process is much faster than simple gravitational sedimentation by standing still; and compared to methods requiring electrodes. , low energy requirements and no need for electrode maintenance or replacement.
本発明によれば、微細藻類に含まれる燃料油の獲得に関しては、コスト面、効率面等の点で十分である。
また、微細藻類から有用有機物の獲得に関しても、消費必要エネルギー量、微細藻類の回収率、コスト面、効率面等から十分である。
好塩性微細藻類から、採算がとれるように有用有機物や油脂（燃料）を得るためには、ある程度以上の生産規模が必要であるが、本発明によれば、大規模生産に好適に対応可能である（大規模生産にマッチングしている）。
According to the present invention, fuel oil contained in microalgae can be obtained in terms of cost and efficiency.
Furthermore, the acquisition of useful organic substances from microalgae is sufficient in terms of required energy consumption, recovery rate of microalgae, cost, efficiency, etc.
In order to profitably obtain useful organic substances and oils and fats (fuel) from halophilic microalgae, a production scale of at least a certain level is required, but according to the present invention, large-scale production can be suitably supported. (matching large-scale production).
以下、本発明について説明するが、本発明は、以下の具体的形態に限定されるものではなく、技術的思想の範囲内で任意に変形することができる。 The present invention will be described below, but the present invention is not limited to the following specific forms, and can be arbitrarily modified within the scope of the technical idea.
本発明は、以下の工程１、３、４及び５の全てをこの順に有することを特徴とする好塩性微細藻類の回収方法である。
（１）池の水の中で好塩性微細藻類を培養する工程
（３）工程（１）で培養された好塩性微細藻類を、該池の水と共に回収して回収液を得る工程
（４）工程（３）で得られた回収液を容器に入れ、該容器内を遮光して、該好塩性微細藻類を該容器の底に沈降させる工程
（５）工程（４）で沈降させた好塩性微細藻類を回収する工程
The present invention is a method for collecting halophilic microalgae, which is characterized by having all of the following steps 1, 3, 4, and 5 in this order.
(1) A step of cultivating halophilic microalgae in pond water (3) A step of collecting the halophilic microalgae cultured in step (1) together with the pond water to obtain a recovered liquid ( 4) Put the collected liquid obtained in step (3) into a container, block the inside of the container from light, and let the halophilic microalgae settle to the bottom of the container (5) Step (4) Process of collecting halophilic microalgae
＜工程１＞
＜＜池＞＞
工程１は、池の水の中で好塩性微細藻類を培養する工程である。
ここで、「池」とは、「狭い定義による池」には限定されず、自然界にあるもの若しくはそれを専用に加工・変更・手直し・区切り・補強等したもの、又は、人工的に作ったもの若しくは人工的に作ってあったものを専用に加工・変更・手直し・区切り・補強等したもの、等の何れでもよい。
<Step 1>
＜＜Pond＞＞
Step 1 is a step of culturing halophilic microalgae in pond water.
Here, the term "pond" is not limited to "ponds defined narrowly," but refers to things that exist in the natural world or those that have been specially processed, modified, reworked, divided, reinforced, etc., or those that are artificially created. It can be anything that has been specially processed, changed, reworked, divided, reinforced, etc. from a product or an artificially made product.
上記「自然界にあるもの若しくはそれを専用に加工等したもの」としては、例えば、海、川、湖沼、池等、又は、それらを専用に加工等したものが挙げられる。ここで、加工等（加工・変更・手直し・区切り・補強等）とは、具体的には、例えば、水を通さない仕切り等で小さく区切ったり、堰き止めたり、土手・縁等を補強したり、好塩性微細藻類を培養する際の好適な深さに埋め立てたり、内面を遮水加工したりすること等が挙げられる。 Examples of the above-mentioned "things found in the natural world or those that have been specially processed etc." include oceans, rivers, lakes, ponds, etc., and those that have been specially processed. Here, processing, etc. (processing, alteration, rework, division, reinforcement, etc.) specifically includes, for example, dividing into small sections with partitions that do not allow water to pass through, damming, reinforcing banks, edges, etc. , reclamation at a suitable depth for culturing halophilic microalgae, and water-proofing the inner surface.
上記「人工的に作ったもの若しくは人工的に作ってあったものを専用に加工・変更・手直し・区切り・補強等したもの」としては、具体的には、例えば、人工貯水池、プール、専用の培養容器、田圃・養殖池・人工池等；又は；それらを加工等（加工・変更・手直し・区切り・補強等）したもの等が挙げられる。ここでの「加工等」としては、具体的には、例えば、前記したもの等が挙げられる。以下、「加工等」と略記することがある。
日本では、休耕田が多いので、田圃等を加工等したものが、初期投資が抑制され、大きさ、深さ等が、本発明の「池」の条件にマッチングしている等の点から特に好ましい。
The above-mentioned "artificially made or artificially made objects that have been specially processed, modified, reworked, divided, reinforced, etc." include, for example, artificial reservoirs, pools, Examples include culture containers, rice fields, aquaculture ponds, artificial ponds, etc.; or; those that have been processed (processed, changed, modified, divided, reinforced, etc.). Here, "processing, etc." specifically includes, for example, those described above. Hereinafter, it may be abbreviated as "processing, etc."
In Japan, there are many fallow fields, so it is particularly preferable to use rice fields that have been processed, etc., because the initial investment is suppressed, and the size, depth, etc. match the conditions for the "pond" of the present invention. .
工程１における１つの「池」の面積は、該池が人工的に作ったものの場合には、１ｍ２以上１×１０６ｍ２以下であることが好ましく、１００ｍ２以上２．５×１０５ｍ２以下であることがより好ましい。
また、「池」が「自然界にあるもの若しくはそれを専用に加工等したもの」又は「人工的に作ってあったものを人工的に専用に加工等したもの」の場合には、１０ｍ２以上１００００ｍ２以下であることが好ましく、３０ｍ２以上３０００ｍ２以下であることがより好ましく、１００ｍ２以上１０００ｍ２以下であることが特に好ましい。
The area of one “pond” in Step 1 is preferably 1 m 2 or more and 1 × 10 6 m 2 or less, and 100 m 2 or more and 2.5 × 10 5 if the pond is artificially created. It is more preferable that it is less than m2 .
In addition, if the "pond" is "something that exists in the natural world or something that has been specially processed, etc." or "a pond that has been artificially created and has been artificially processed for exclusive use," the pond must be 10 m2 or more. It is preferably 10000 m 2 or less, more preferably 30 m 2 or more and 3000 m 2 or less, and particularly preferably 100 m 2 or more and 1000 m 2 or less.
１つの「池」の面積が上記下限以上であると、スケールメリットが得られる、作業性に優れる等の効果がある。
一方、１つの「池」の面積が上記上限以下であると、設備投資がかかり過ぎない、作業性が良い、無駄に広くない等の効果がある。
When the area of one "pond" is equal to or larger than the above-mentioned lower limit, there are effects such as obtaining economies of scale and excellent workability.
On the other hand, when the area of one "pond" is below the above upper limit, there are effects such as not requiring too much capital investment, good workability, and not being unnecessarily large.
「池」の水の体積又は／及び「池」の容積は、１ｍ３以上であることが好ましく、より好ましくは３ｍ３以上１００００ｍ３以下であり、更に好ましくは１０ｍ３以上３０００ｍ３以下であり、特に好ましくは３０ｍ３以上１０００ｍ３以下である。
１つの「池」の体積が上記下限以上であったり、上記上限以下であったりすると、前記した「池」の面積の場合と同様の理由で望ましい。
The volume of water in the "pond" and/or the volume of the "pond" is preferably 1 m 3 or more, more preferably 3 m 3 or more and 10,000 m 3 or less, even more preferably 10 m 3 or more and 3,000 m 3 or less, Particularly preferably, the area is 30 m 3 or more and 1000 m 3 or less.
It is desirable for the volume of one "pond" to be greater than or equal to the above lower limit or less than or equal to the above upper limit for the same reason as the area of the "pond" described above.
「池」の平均深さは、０．０５ｍ以上１．５ｍ以下が好ましく、０．１０ｍ以上１ｍ以下がより好ましい。
上記範囲であると、太陽光が好塩性微細藻類に当たり易い、干上がる恐れがない等の効果がある。
The average depth of the "pond" is preferably 0.05 m or more and 1.5 m or less, more preferably 0.10 m or more and 1 m or less.
Within the above range, the halophilic microalgae are easily exposed to sunlight and there is no risk of drying up.
＜＜水（培養のための原水）＞＞
培養は池の水の中で行われる。該水としては、好塩性微細藻類が増殖するようなものであれば、特に限定はないが、海水、海洋深層水、人工海水、又は、人工海洋深層水であることが好ましい。
海水は、平均的には塩分濃度が約３．５質量％であり、主に、ナトリウムイオン、マグネシウムイオン、カリウムイオン、カルシウムイオン等の陽イオン；塩素イオン、硫酸イオン等の陰イオンを含む。
<<Water (raw water for culture)>>
Cultivation takes place in pond water. The water is not particularly limited as long as it allows halophilic microalgae to proliferate, but seawater, deep ocean water, artificial seawater, or artificial deep ocean water is preferable.
Seawater has an average salt concentration of about 3.5% by mass, and mainly contains cations such as sodium ions, magnesium ions, potassium ions, and calcium ions; and anions such as chloride ions and sulfate ions.
海洋深層水は、人工的な汚染がなく、低温であったため雑菌が少なく、太陽光が当たらなかったために生存植物性プランクトンの混入がない等の好塩性微細藻類を培養する上で好適な条件が整っている。更に、それらに加え、硝酸塩等での窒素（Ｎ）やリン酸塩等でのリン（Ｐ）やケイ酸塩等でのケイ素（Ｓｉ）等が豊富である。また、塩濃度が高いので、好塩性微細藻類の培養に障害となる菌等が生存し難い（少ない）。
そのため、海洋深層水は、本発明における好塩性微細藻類の培養のための水として特に好ましい。
Deep sea water has favorable conditions for culturing halophilic microalgae, such as no artificial pollution, low temperatures, so there are few bacteria, and lack of sunlight, so there is no contamination by living phytoplankton. are in place. Furthermore, in addition to these, it is rich in nitrogen (N) in nitrates, phosphorus (P) in phosphates, and silicon (Si) in silicates. In addition, since the salt concentration is high, it is difficult (few) for bacteria and the like that would interfere with the culture of halophilic microalgae to survive.
Therefore, deep ocean water is particularly preferred as water for culturing halophilic microalgae in the present invention.
海水及び／又は海洋深層水は、太陽熱等を利用して水を蒸発させて濃縮させることによって、好塩性微細藻類の培養に必要な窒素（Ｎ）源やリン（Ｐ）源の濃度を上げることができる。水の蒸発と共に（同時に）、海水や海洋深層水の食塩（塩化ナトリウム）濃度も（例えば約１５質量％まで）上がってしまうが、例えば、デュナリエラ（Dunaliella）属に属する微細藻類等の多くの（高度）好塩性微細藻類は、高い食塩（塩化ナトリウム）濃度（例えば約１５質量％）でも培養可能である。
その点からも、太陽熱を利用して、窒素（Ｎ）源やリン（Ｐ）源の濃度を上げることは好ましい。
また、高い食塩（塩化ナトリウム）濃度（例えば約１５質量％）になると、雑菌が繁殖し難くなり、コンタミネーション（汚染）が少なくなるので、好塩性微細藻類の増殖に好都合である。
By evaporating and concentrating seawater and/or deep ocean water using solar heat, etc., the concentration of nitrogen (N) and phosphorus (P) sources necessary for culturing halophilic microalgae is increased. be able to. As water evaporates (at the same time), the concentration of salt (sodium chloride) in seawater and deep ocean water also increases (for example, to about 15% by mass). Highly halophilic microalgae can be cultured even at high salt (sodium chloride) concentrations (for example, about 15% by mass).
From this point of view as well, it is preferable to use solar heat to increase the concentration of nitrogen (N) sources and phosphorus (P) sources.
Furthermore, a high salt (sodium chloride) concentration (for example, about 15% by mass) makes it difficult for bacteria to propagate and reduces contamination, which is favorable for the proliferation of halophilic microalgae.
海水や海洋深層水、及び、それらを、太陽熱等を利用して水を蒸発させて濃縮させた培養のための原水は、コスト的に安価であり、本発明にスケールメリットがある場合には、それを生かすためには特に好適である。 Seawater, deep ocean water, and raw water for cultivation that is made by evaporating water using solar heat or the like and concentrating it are inexpensive, and if the present invention has merits of scale, It is particularly suitable for taking advantage of this.
前記「人工海水や人工海洋深層水」は、最初から栄養分（必須元素等を含む）や塩（海水中の塩分を含む）を、全て人工的に加えて調製してもよいし、天然の海水や海洋深層水に、かかる栄養分や塩等を、人工的に追加配合して調製してもよい。 The above-mentioned "artificial seawater and artificial deep sea water" may be prepared by artificially adding nutrients (including essential elements, etc.) and salts (including salt in seawater) from the beginning, or may be prepared by adding all nutrients (including essential elements, etc.) and salt (including salt in seawater) artificially, or may be prepared by adding all nutrients (including essential elements, etc.) and salt (including salt in seawater) from the beginning, or may be prepared by adding natural seawater. It may also be prepared by artificially adding such nutrients, salts, etc. to water or deep ocean water.
＜＜好塩性微細藻類＞＞
本発明における好塩性微細藻類として、具体的には、例えば、デュナリエラ（Dunaliella）属に属する微細藻類等が挙げられる。１種又は２種以上の好塩性微細藻類を培養することができる。好塩性微細藻類としては、所謂「高度好塩性微細藻類」と言われているものも特に好ましい。
<<Halophilic microalgae>>
Specific examples of the halophilic microalgae in the present invention include microalgae belonging to the genus Dunaliella. One or more types of halophilic microalgae can be cultured. As the halophilic microalgae, so-called "highly halophilic microalgae" are also particularly preferred.
好塩性微細藻類を、特に高塩分濃度の海水中で培養することで、コンタミネーション（雑菌混入）のリクス低減を低コストによって実現することが可能となる。 By culturing halophilic microalgae, especially in seawater with high salinity, it is possible to reduce the risk of contamination (mixture of bacteria) at low cost.
培養の対象を「好塩性微細藻類」にする特徴としては、具体的には、例えば、以下が挙げられる。
好塩性微細藻類からは、炭水化物若しくは糖；オリゴペプチド、ポリペプチド若しくはタンパク質；ビタミン若しくはビタミン前駆体；又は；油脂、糖脂質、リン脂質、リポタンパク質若しくは炭化水素等の有用有機物を獲得できる；二酸化炭素以外の炭素源でも利用可能である；高度好塩性藻類（海洋性微細藻類）が多い；適応生育温度範囲が広い；強光阻害を受けない；等が挙げられる。また、好塩性微細藻類によって、大気中の二酸化炭素が資源化できる。
中でも、特に好ましい好塩性微細藻類として、上記の点等から、デュナリエラ（Dunaliella）属に属する微細藻類が挙げられる。
Specific examples of characteristics that make "halophilic microalgae" to be cultured include the following.
From halophilic microalgae, carbohydrates or sugars; oligopeptides, polypeptides, or proteins; vitamins or vitamin precursors; or; useful organic substances such as fats and oils, glycolipids, phospholipids, lipoproteins, or hydrocarbons; dioxide; Carbon sources other than carbon can also be used; there are many highly halophilic algae (marine microalgae); the adaptive growth temperature range is wide; and they are not subject to strong photoinhibition. In addition, halophilic microalgae can turn atmospheric carbon dioxide into a resource.
Among these, particularly preferable halophilic microalgae include microalgae belonging to the genus Dunaliella in view of the above points.
デュナリエラ（Dunaliella）属は、細胞壁をもたないので、油の抽出が容易である。
また、デュナリエラ（Dunaliella)属は、グリセリンを浸透圧調整のために細胞中に貯蔵しているため、該グリセリンを回収して有効利用することができる。なお、該グリセリンは溶媒や触媒等を含まない純粋なグリセリンであるため、化粧品や飲食品への使用から発酵基質としての使用にも利用価値が高い。
The Dunaliella genus does not have a cell wall, so oil can be extracted easily.
Furthermore, since the genus Dunaliella stores glycerin in its cells for adjusting osmotic pressure, the glycerin can be recovered and used effectively. In addition, since the glycerin is pure glycerin containing no solvent or catalyst, it has high utility value not only for use in cosmetics and food and drink products but also for use as a fermentation substrate.
また、デュナリエラ（Dunaliella）属は、バブリング（二酸化炭素のエアレーション）が不要であり、培養コストを抑えることができる。
また、デュナリエラ（Dunaliella）属は、増殖しながら脂質蓄積が可能な数少ない微細藻類である。
また、β－カロテンを蓄積することで、強光阻害を起こさずに培養可能である。
また、適応生育温度範囲が広いため、人為的な温度コントロールを行わずに培養可能なため、大幅に培養コストを低減することが可能となる。
In addition, the Dunaliella genus does not require bubbling (aeration of carbon dioxide), and culture costs can be reduced.
Additionally, the genus Dunaliella is one of the few microalgae that can accumulate lipids while proliferating.
Furthermore, by accumulating β-carotene, it is possible to culture without strong photoinhibition.
In addition, since the adaptive growth temperature range is wide, culture can be performed without artificial temperature control, making it possible to significantly reduce culture costs.
デュナリエラ（Dunaliella）属に属する微細藻類として、例えば、Dunaliella salina、Dunaliella viridis、Dunaliella bioculata、Dunaliella primolecta、Dunaliella tertiolecta、Dunaliella bardawil等が挙げられる。
生存可能温度範囲が４～６０℃と低温でも高温でも生存・生育することができる点、高浸透圧状況下で高温に耐えることができる点等から、デュナリエラ・サリナ（Dunaliella salina）を培養することが特に好ましい。本発明では、１種又は２種以上のデュナリエラ（Dunaliella）属に属する好塩性微細藻類を培養することができる。
Examples of microalgae belonging to the genus Dunaliella include Dunaliella salina, Dunaliella viridis, Dunaliella bioculata, Dunaliella primolecta, Dunaliella tertiolecta, Dunaliella bardawil, and the like.
Dunaliella salina is cultivated because it can survive and grow in both low and high temperatures, with a viable temperature range of 4 to 60 degrees Celsius, and can withstand high temperatures under high osmotic pressure conditions. is particularly preferred. In the present invention, one or more types of halophilic microalgae belonging to the genus Dunaliella can be cultured.
＜工程２＞
工程２は、本発明において、必須ではないが、上記工程（１）と下記工程（３）の間に行うことも好ましい。
工程２は、もし池の水が撹拌されていれば、該撹拌を停止して経時させ、上記好塩性微細藻類が、上記池の水の深さ方向の全体に亘って存在するか、又は、池の水の中の深さ方向の上側、中間若しくは下側の何れか１か所又は２か所に存在するかを確認する工程である。
<Step 2>
Although step 2 is not essential in the present invention, it is also preferable to perform it between the above step (1) and the following step (3).
In step 2, if the water in the pond is being stirred, the stirring is stopped and allowed to pass, and the halophilic microalgae are present throughout the depth of the water in the pond, or , is present in one or two locations at the top, middle, or bottom in the depth direction of the pond water.
例えば、好塩性微細藻類が上層だけに多く存在すれば、上層だけを回収すればよいし、下層だけに多く存在すれば、上層だけを回収すればよいし、上層と下層に多く存在すれば、上層と下層だけを回収してもよいし、全部の層（全部の水）を回収してもよい。
そうすることによって、培養のための池にあった大量の水を減らして、すなわち、好塩性微細藻類の濃度を濃くした状態で容器に回収して回収液とすることができ、その後の回収コストを削減することができる。
For example, if halophilic microalgae are abundant only in the upper layer, you only need to collect the upper layer; if they are abundant only in the lower layer, you only need to collect the upper layer; , only the upper and lower layers may be collected, or all layers (all water) may be collected.
By doing so, it is possible to reduce the large amount of water that was in the culture pond and collect the halophilic microalgae in a container with a higher concentration as a recovery liquid, which can be used for subsequent recovery. Costs can be reduced.
ただ、下記する工程３と工程４の主たる目的（効果）は、遮光し易くすることや；深さの大きい容器に回収して、沈降させ易くする、又は、沈降物だけを回収し易くすることなので、その目的を達成するためには、池の水は全部回収することが好ましい。 However, the main purpose (effect) of Steps 3 and 4 below is to make it easier to block light; to collect it in a deep container and make it easier to settle, or to make it easier to collect only the sediment. Therefore, in order to achieve this purpose, it is preferable to recover all the water in the pond.
＜工程３＞
工程３は、上記工程１で培養された好塩性微細藻類を、該池の水と共に回収して回収液を得る工程である。
好塩性微細藻類を池の水と共に回収する方法としては、特に限定はないが、汲み出し、ポンプによる吸引、重力を利用しての流し出し、浮上油回収装置等の専用の装置による回収、デカンテーション、サイホンの原理（大気圧の利用）による抜き出し等が挙げられる。
工程２で、池から取り出す部分（深さ等）を確認して、池の水の一部又は実質的に全量を回収するかどうかを決めることもできる。
<Step 3>
Step 3 is a step in which the halophilic microalgae cultured in step 1 is collected together with the water in the pond to obtain a recovered liquid.
Methods for collecting halophilic microalgae along with pond water are not particularly limited, but include pumping, suction with a pump, draining using gravity, collection using a dedicated device such as a floating oil recovery device, and decanting. Examples include extraction using the siphon principle (using atmospheric pressure).
In step 2, the portion (depth, etc.) to be removed from the pond may also be determined to determine whether a portion or substantially all of the pond water is to be recovered.
全量回収すれば、回収液中の好塩性微細藻類の含有割合は、培養液中の（「池」の水中の）含有割合と同一になるし、濃度の高い場所だけを回収すれば、含有割合は高くなるが、工程３で得られる回収液中の好塩性微細藻類の含有割合は、１０３ｃｅｌｌ／ｍＬ以上１０１０ｃｅｌｌ／ｍＬ以下であることが好ましい。すなわち、このようになるまで培養することが好ましく、結果として、回収液中の好塩性微細藻類の濃度が、このような範囲になるように回収することが好ましい。 If the entire amount is collected, the content ratio of halophilic microalgae in the collected solution will be the same as the content ratio in the culture solution (in the "pond" water), and if only the areas with high concentrations are collected, the content of halophilic microalgae will be Although the proportion is high, the content of halophilic microalgae in the recovered liquid obtained in step 3 is preferably 10 3 cells/mL or more and 10 10 cells/mL or less. That is, it is preferable to culture until such a state is reached, and as a result, it is preferable to collect the halophilic microalgae so that the concentration of the halophilic microalgae in the collected liquid falls within this range.
＜工程４＞
工程４は、上記工程３で得られた回収液を容器に入れ、該容器内を遮光して、該好塩性微細藻類を該容器の底に沈降させる工程である。
工程４以降で用いる上記容器としては、特に限定はなく、「人工的に作ったもの若しくは人工的に作ってあったものを加工等したもの」、「自然界にあるもの若しくはそれを専用に加工等したもの」等が挙げられる。
<Step 4>
Step 4 is a step in which the collected liquid obtained in Step 3 is placed in a container, the inside of the container is shielded from light, and the halophilic microalgae are allowed to settle to the bottom of the container.
The containers used in Step 4 and beyond are not particularly limited, and may include "artificially made or processed artificially made" containers, "products found in the natural world or specially processed etc." Examples include "things that have been done".
工程４では、上記容器内を遮光して好塩性微細藻類を該容器の底に沈降させるので、該容器としては遮光し易いものが好ましい。
「容器内の遮光」は、容器を遮光材料で作るか、遮光材料で容器を覆うか、土中に掘られた穴若しくは空間を利用するか、トンネル若しくは洞窟を利用するか、遮光された屋内を利用することによってなされることが好ましい。
In step 4, the inside of the container is shielded from light and the halophilic microalgae are allowed to settle to the bottom of the container, so it is preferable that the container is easily shielded from light.
"Shading inside the container" means making the container with a light-blocking material, covering the container with a light-blocking material, using a hole or space dug in the soil, using a tunnel or cave, or using a light-blocking indoor space. Preferably, this is done by using.
大きなスケールで培養を行ったときには、上記容器も大きいものになるので、自然界のもの（又はそれを加工したもの）で遮光し易いものが好ましい。
一方、該容器に大きさを必要としないときは、容器を遮光材料で作るか、遮光材料で容器を覆うか、容器を遮光された屋内に収納すること等が好ましい。
When culturing is carried out on a large scale, the container is also large, so it is preferable to use a natural container (or one processed from it) that can easily block light.
On the other hand, when the size of the container is not required, it is preferable to make the container of a light-shielding material, cover the container with a light-shielding material, or store the container indoors where it is shielded from light.
上記容器の高さを上記池の深さより大きく設定し、該容器の水深を該池の水深より大きくして、好塩性微細藻類を該容器の底に沈降させることが、好塩性微細藻類が多く存在する液（深さ方向の場所）と殆ど存在しない液（深さ方向の場所）とに分け易い、特に濃度の高い底に近い場所から回収できる、遮光し易い、再撹拌され難い、光が届き難い等の点から好ましい。 Setting the height of the container to be larger than the depth of the pond, making the water depth of the container larger than the water depth of the pond, and causing the halophilic microalgae to settle to the bottom of the container, It is easy to separate the liquid into liquids where there is a lot of it (places in the depth direction) and liquids where there is little (places in the depth direction), it can be collected from places near the bottom where the concentration is particularly high, it is easy to shield from light, it is difficult to be stirred again, This is preferable because it is difficult for light to reach.
好ましい容器の深さ（高さ）は、池の深さ（高さ）の３倍以上３００倍以下が好ましく、７倍以上１２０倍以下がより好ましく、２０倍以上５０倍以下が特に好ましい。
具体的には、０．５ｍ以上２０ｍ以下が好ましく、１ｍ以上１０ｍ以下が特に好ましい。
なお、「池」の平均深さは、前記した通り、好塩性微細藻類の好適な培養を第一に考えて決められ、０．０５ｍ～１．５ｍが好ましく、０．１０ｍ～１ｍがより好ましい。従って、回収液の高さは、それに比べて、上記倍数の範囲で高いことが、底に近い場所の濃度を高くできてそこから回収できる、深さ方向の濃度差が深さ方向に拡大される、過度に平たくないので遮光し易い、その他上記した点等から好ましい。
The depth (height) of the container is preferably 3 to 300 times the depth (height) of the pond, more preferably 7 to 120 times, particularly preferably 20 to 50 times.
Specifically, the distance is preferably 0.5 m or more and 20 m or less, particularly preferably 1 m or more and 10 m or less.
In addition, as mentioned above, the average depth of the "pond" is determined by first considering the suitable culture of halophilic microalgae, and is preferably 0.05 m to 1.5 m, more preferably 0.10 m to 1 m. preferable. Therefore, if the height of the recovered liquid is higher than that in the above multiple range, the concentration near the bottom can be increased and the concentration can be recovered from there, and the concentration difference in the depth direction will be expanded in the depth direction. It is preferable because it is easy to block light because it is not excessively flat, and from the other points mentioned above.
工程４において、容器内を遮光して、好塩性微細藻類を容器に沈降させるための時間（以下、「静置時間」と略記することがある）は、３０分以上１か月以下が好ましく、４時間以上２０日以下がより好ましく、１日以上１５日以下が更に好ましく、５日以上１０日以下が特に好ましい。
静置時間が短過ぎると、沈降が十分でない、上層と下層で好塩性微細藻類の濃度に差が出ない、塩分濃度やｐＨによっては沈降の程度が水温に依存する等の場合がある。一方、静置時間が長過ぎると、単位時間当たりの好塩性微細藻類や有用有機物の生産効率が落ちる等の場合がある。
In step 4, the time for shading the inside of the container and allowing the halophilic microalgae to settle in the container (hereinafter sometimes abbreviated as "standing time") is preferably 30 minutes or more and 1 month or less. , more preferably 4 hours or more and 20 days or less, still more preferably 1 day or more and 15 days or less, particularly preferably 5 days or more and 10 days or less.
If the standing time is too short, sedimentation may not be sufficient, there may be no difference in the concentration of halophilic microalgae between the upper and lower layers, and depending on the salinity and pH, the degree of sedimentation may depend on water temperature. On the other hand, if the standing time is too long, the production efficiency of halophilic microalgae and useful organic matter per unit time may decrease.
すなわち、本発明は、上記工程４において、容器内を遮光してから、好塩性微細藻類を該容器の底に沈降させ、後記工程５において好塩性微細藻類を回収するまでの沈降時間が、３０分以上１か月以下である上記の好塩性微細藻類の回収方法でもある。 That is, in the present invention, the sedimentation time from shielding the inside of the container from light in step 4 to settling the halophilic microalgae to the bottom of the container and collecting the halophilic microalgae in step 5 described below is shortened. It is also the above-mentioned method for collecting halophilic microalgae, which takes 30 minutes or more and one month or less.
工程４では、沈降させた部分の水中の好塩性微細藻類の濃度は、沈降させる前の回収液の平均濃度の、１０倍以上１００００倍以下にすることが好ましく、１００倍以上３０００倍以下にすることが特に好ましい。
濃度の絶対値で言うと、前記した通り、工程３で得られる回収液中の好塩性微細藻類の含有割合は、１０３ｃｅｌｌ／ｍＬ以上１０１０ｃｅｌｌ／ｍＬ以下であることが好ましいので、それぞれ、その濃度に、上記倍率を掛け合わせた濃度になるまで沈降させて、濃縮することが望ましい。
In step 4, the concentration of halophilic microalgae in the water in the sedimented portion is preferably 10 times or more and 10,000 times or less, and 100 times or more and 3,000 times or less, than the average concentration of the recovered liquid before sedimentation. It is particularly preferable to do so.
In terms of the absolute value of concentration, as mentioned above, the content ratio of halophilic microalgae in the collected liquid obtained in step 3 is preferably 10 3 cells/mL or more and 10 10 cells/mL or less, It is desirable to precipitate and concentrate until the respective concentration reaches a concentration obtained by multiplying the concentration by the above-mentioned magnification.
本発明は、好塩性微細藻類は、遮光することによって、すなわち、少なくとも太陽光を当てないことによって、容器の底に好適に沈降することを見出してなされたものである。
従って、工程４では、遮光が必須であるが、更に該遮光に加えて、ｐＨ調節、酸素（空気）の遮断、窒素（空気）の遮断等を行ってもよい。
The present invention was made based on the discovery that halophilic microalgae settle to the bottom of a container by shielding them from light, that is, by at least not exposing them to sunlight.
Therefore, in step 4, light shielding is essential, but in addition to the light shielding, pH adjustment, oxygen (air) blocking, nitrogen (air) blocking, etc. may be performed.
本発明は、上記工程４まで、好塩性微細藻類の回収に、遠心分離を用いない上記の好塩性微細藻類の回収方法でもある。遠心分離は、最も一般的な好塩性微細藻類の回収であるが、莫大なエネルギーを要する。本発明は、大量にある回収液に対して極めて不利な遠心分離法で好塩性微細藻類の回収を行わず、単に遮光するだけで好塩性微細藻類を濃縮できることを見出してなされたものである。 The present invention is also a method for collecting halophilic microalgae that does not use centrifugation in the recovery of halophilic microalgae up to step 4. Centrifugation is the most common method of recovering halophilic microalgae, but it requires a large amount of energy. The present invention was made based on the discovery that halophilic microalgae can be concentrated simply by blocking light, without collecting halophilic microalgae using centrifugation, which is extremely disadvantageous for large amounts of recovered liquid. be.
従って、本発明は、更に好ましくは、少なくとも上記工程４までは、好塩性微細藻類の回収に、実質的に、凝集剤を使用しないことであり、濾過をしないことであり、電解をかけることをしないことである。 Therefore, it is more preferable in the present invention that, at least up to step 4, the halophilic microalgae are collected without substantially using a flocculant, without filtration, and without applying electrolysis. It is important not to do this.
本発明をするに当たっては、静置中の種々の条件を振って検討したところ、驚くべきことに遮光が最も効果があった。かかる条件とは、遮光、酸素の遮断、窒素の遮断、温度変化、ｐＨ変化等である。 When carrying out the present invention, various conditions during standing were examined, and surprisingly, light shielding was the most effective. Such conditions include light shielding, oxygen shielding, nitrogen shielding, temperature change, pH change, and the like.
＜工程５＞
工程５は、上記工程４で沈降させた好塩性微細藻類を回収する工程である。
該回収の方法は、特に限定はされず、濾過；加熱、減圧、風乾等による水の蒸発乾固；遠心分離；凝集剤による凝集分離；容器下からの抜き取り；等が挙げられる。容器下から抜き取る場合は、工程４で使用する容器として、予め容器下方に抜取装置が具備されたものを使用することも好ましい。
<Step 5>
Step 5 is a step of recovering the halophilic microalgae sedimented in Step 4 above.
The method of recovery is not particularly limited, and examples include filtration; evaporation of water to dryness by heating, reduced pressure, air drying, etc.; centrifugation; flocculation separation using a flocculant; extraction from the bottom of the container; and the like. When extracting from the bottom of the container, it is also preferable to use a container that is previously equipped with a extraction device below the container as the container used in step 4.
また、沈降させた部分（好塩性微細藻類を高濃度で含む水）は、脱塩を行ってもよいし、真水を加えてもよいし、加熱をしてもよいし、好塩性微細藻類を殺してもよい。
また、コスト的に問題なければ、別容器を使用する等して、工程４と工程５を繰り返してもよい。繰り返す場合は、繰り返す度に、好塩性微細藻類の濃度等に応じて容器や上記の回収方法を変えてもよい。
In addition, the sedimented part (water containing a high concentration of halophilic microalgae) may be desalinated, fresh water may be added, heating may be performed, or the halophilic microalgae may be May kill algae.
Further, if there is no problem in terms of cost, steps 4 and 5 may be repeated by using a separate container. When repeating, the container and the above collection method may be changed each time depending on the concentration of halophilic microalgae.
表１等を含め前記した通り、濾過は消耗品であるフィルター（交換）の必要性、遠心分離はエネルギー大でコスト高、凝集剤使用は不純物の混入等から、容積の大きい工程４までは好ましくない。すなわち、本発明の工程１ないし４においては、必須ではないことは勿論のこと、むしろ好ましくない。
しかし、本発明によって工程４まで行って、容積が極めて少なくなった工程５の時点では、上記のような公知の回収方法を用いることもできる。処理体積が小さくなっているので、上記のような公知の回収方法の短所が重要ではなくなる。
As mentioned above, including Table 1, filtration requires a consumable filter (replacement), centrifugation requires a lot of energy and is expensive, and the use of a flocculant is preferable up to step 4, which involves large volumes, because of the contamination of impurities, etc. do not have. That is, in steps 1 to 4 of the present invention, it goes without saying that this is not essential, but is rather undesirable.
However, according to the present invention, at step 5, when the volume has become extremely small after reaching step 4, the above-mentioned known recovery method can also be used. Due to the smaller processing volume, the disadvantages of known recovery methods such as those mentioned above become less important.
工程５で回収された好塩性微細藻類は、そこから有用物を獲得したり、燃料として使用したりできる。 The halophilic microalgae collected in step 5 can be used to obtain useful substances or used as fuel.
＜回収した好塩性微細藻類の用途＞
本発明は、上記の好塩性微細藻類の回収方法を使用して回収した好塩性微細藻類から有用有機物を獲得することを特徴とする有用有機物の製造方法でもある。
該有用有機物として、具体的には、例えば、炭水化物若しくは糖；オリゴペプチド、ポリペプチド若しくはタンパク質；ビタミン若しくはビタミン前駆体；油脂、糖脂質、リン脂質若しくはリポタンパク質；又は；炭化水素；等が挙げられる。
<Uses of collected halophilic microalgae>
The present invention is also a method for producing useful organic matter, characterized in that useful organic matter is obtained from halophilic microalgae collected using the above-described method for collecting halophilic microalgae.
Specific examples of the useful organic substances include carbohydrates or sugars; oligopeptides, polypeptides, or proteins; vitamins or vitamin precursors; oils and fats, glycolipids, phospholipids, or lipoproteins; or; hydrocarbons; .
β－カロテン等のカロテノイド等を含むビタミン前駆体、グルタチオン等のオリゴペプチド、グリセリン等が、例えば、デュナリエラ（Dunaliella）属等の好塩性微細藻類から好適に獲得できる。 Vitamin precursors containing carotenoids such as β-carotene, oligopeptides such as glutathione, glycerin, etc. can be suitably obtained, for example, from halophilic microalgae such as the genus Dunaliella.
β－カロテンは、高い抗酸化作用を有し、例えば、医薬品、食品、食品添加物、サプリメント等に利用することができる。
グルタチオンは、高い抗酸化作用を有し、例えば、医薬品、食品、サプリメント、肥料、飼料等に利用することができる。
また、グリセリンは、例えば、化粧品、医薬品等の原料としても、燃料としても、好適に利用することができる。
油脂は、例えば、食料用油、飼料用油、燃料油等に利用することができる。
好塩性微細藻類には、良質なタンパク質が含まれているので、例えば、飼料、醗酵原料等として利用することができる。
β-carotene has a high antioxidant effect and can be used, for example, in medicines, foods, food additives, supplements, and the like.
Glutathione has a high antioxidant effect and can be used, for example, in medicines, foods, supplements, fertilizers, feeds, and the like.
Furthermore, glycerin can be suitably used, for example, as a raw material for cosmetics, medicines, etc., and as a fuel.
The fats and oils can be used, for example, as food oil, feed oil, fuel oil, and the like.
Since halophilic microalgae contain high-quality proteins, they can be used, for example, as feed, fermentation raw materials, and the like.
以下に、実施例及び比較例を挙げて本発明を更に具体的に説明するが、本発明は、その要旨を超えない限りこれらの実施例に限定されるものではない。 EXAMPLES The present invention will be described in more detail below with reference to Examples and Comparative Examples, but the present invention is not limited to these Examples unless the gist thereof is exceeded.
実施例１
図１（左）に示すように、高さ９ｃｍ、底面積１０ｃｍ２、容積７０ｃｍ３の容器に、海水を入れ、デュナリエラ（Dunaliella）属に属する好塩性微細藻類を、１．３×１０６ cells／ｍＬとなるように入れた。
なお、通常、広大な池の中で培養されたデュナリエラは、該池の海水をデュナリエラごと全て回収したとして（又は均一になるように撹拌してから回収したとして）、平均で、約１．３×１０６ cells／ｍＬ程度となるので、それを勘案して、上記初期濃度に設定した。
Example 1
As shown in Figure 1 (left), a container with a height of 9 cm, a bottom area of 10 cm 2 , and a volume of 70 cm 3 is filled with seawater, and 1.3 × 10 6 of halophilic microalgae belonging to the genus Dunaliella are added to the container. cells/mL.
Generally, Dunaliella cultivated in a vast pond has an average yield of about 1.3 ml, assuming that all of the seawater in the pond is collected together with Dunaliella (or that it is collected after stirring to make it uniform). Since the concentration was approximately 10 6 cells/mL, the above initial concentration was set in consideration of this.
その後、容器の周囲を遮光して、７日間静置した。７日後の写真を図１（右）に示す。
次いで、容器の真ん中から、ピペットで、測定サンプルを抜き出して、濃度を測定したところ、２．１×１０４ cells／ｍＬであった。容器の真ん中のデュナリエラの濃度は、約２桁下がったことになり、十分なデュナリエラの沈降が見られた。
Thereafter, the surroundings of the container were shielded from light and allowed to stand for 7 days. A photograph after 7 days is shown in Figure 1 (right).
Next, the measurement sample was extracted from the center of the container with a pipette, and the concentration was measured, and it was found to be 2.1×10 4 cells/mL. The concentration of Dunaliella in the middle of the container decreased by about two orders of magnitude, indicating that sufficient sedimentation of Dunaliella was observed.
その後、容器の下から体積で（すなわち高さで）、１／３０だけを残して、デカンテーションで上澄みを除去した。７日間、全く外からエネルギーを加えることなく、全体積を１／３０にできた。
すなわち、海水中のデュナリエラの濃度を３０倍にすることができたので、その後のデュナリエラのみの回収が極めて容易になった。
The supernatant was then removed by decantation, leaving only 1/30 of the volume (ie, height) from the bottom of the container. For 7 days, we were able to reduce the total volume to 1/30 without applying any external energy.
That is, since the concentration of Dunaliella in seawater could be increased 30 times, subsequent recovery of only Dunaliella became extremely easy.
デュナリエラを含め、殆ど全ての属の微細藻類が光合成を行うが、光合成には二酸化炭素と水と光が必要である。
デュナリエラは、遮光することで、光合成ができなくなり動かなった。そのことによって、沈降したと考えられた。
デュナリエラのような好塩性微細藻類の場合、暗くすることは、沈降、その後の微細藻類のみの回収等にとって極めて効果的であることが分かった。
Almost all genera of microalgae, including Dunaliella, perform photosynthesis, which requires carbon dioxide, water, and light.
By blocking light, Dunaliella became unable to photosynthesize and became motionless. This is thought to have caused the sedimentation.
In the case of halophilic microalgae such as Dunaliella, it was found that darkness is extremely effective for sedimentation and subsequent recovery of only the microalgae.
比較例１
実施例１と同様に、１．３×１０６ cells／ｍＬの濃度の均一分散のデュナリエラ含有海水を用い、遮光はせず、酸素（Ｏ２）を遮断して（空気を遮断して）、７２日間（約２か月）静置した。
図２にその結果を示す。なお、図２の縦軸の「１．００Ｅ＋０５」等は、「１．００×１０５」等を示す。
Comparative example 1
As in Example 1, uniformly dispersed Dunaliella-containing seawater with a concentration of 1.3 x 10 6 cells/mL was used, and oxygen (O 2 ) was blocked (air was blocked) without shielding light. It was left standing for 72 days (about 2 months).
Figure 2 shows the results. Note that "1.00E+05" and the like on the vertical axis in FIG. 2 indicate "1.00×10 5 " and the like.
容器の真ん中でサンプリングし、その濃度が、６．７×１０４ cells／ｍＬに下がったが、それには、７２日間（約２か月）も要した（図２参照）。 When sampled in the middle of the container, the concentration decreased to 6.7×10 4 cells/mL, but this took 72 days (about 2 months) (see FIG. 2).
実施例２
比較例１において、酸素（Ｏ２）を遮断して（空気を遮断して）、更に、遮光もして、静置した。図２に、その結果を比較例１と合わせて示す。
７日間、遮光して静置することで、２．０×１０４ cells／ｍＬにまで下がったことが確認できた（図２参照）。
また、沈降には、酸素（Ｏ２）の遮断より、遮光の方が有効であることが分かった。
Example 2
In Comparative Example 1, the sample was allowed to stand still while blocking oxygen (O 2 ) (blocking air) and further blocking light. FIG. 2 shows the results together with Comparative Example 1.
By allowing the cells to stand still in the dark for 7 days, it was confirmed that the concentration had dropped to 2.0×10 4 cells/mL (see FIG. 2).
It was also found that blocking light is more effective for sedimentation than blocking oxygen (O 2 ).
実施例３
海水の塩分濃度を、すなわち、培養液の塩分濃度及び回収液の塩分濃度を、共に、１２．５質量％、及び、１７．５質量％と振って、沈降の程度を観察した。
上記２種類の塩分濃度で、１週間、デュナリエラを培養した。その後、遮光して、沈降実験（４時間と１８時間後）を行った。結果を図３に示す。
Example 3
The degree of sedimentation was observed while changing the salt concentration of the seawater, that is, the salt concentration of the culture solution and the salt concentration of the recovered solution, to 12.5% by mass and 17.5% by mass.
Dunaliella was cultured for one week at the above two types of salt concentrations. Thereafter, a sedimentation experiment (4 hours and 18 hours later) was conducted while shielding from light. The results are shown in Figure 3.
遮光したので、遮光したことによって、塩分濃度によらずデュナリエラが沈降した（図３）。
ただし、１２．５質量％の塩分濃度の場合、全ての温度（４℃～５０℃）で同じように沈降したが（図３（上の棒グラフ））、塩分濃度が１７．５質量％の場合は、「２０℃～３０℃、及び、４０℃以上」では同じように沈降したが、それ以外の温度では沈降速度が遅くなった（図３（下の棒グラフ））。
なお、図３、４における「沈殿率（Deposition percentage）」は、最初と最後の濃度を常法に従って測定して、１００×[final cell count]／[initial cell count] を計算して求めた。
Since light was blocked, Dunaliella sedimented regardless of the salinity concentration (Figure 3).
However, when the salinity concentration was 12.5% by mass, sedimentation occurred in the same way at all temperatures (4°C to 50°C) (Figure 3 (upper bar graph)), but when the salinity concentration was 17.5% by mass sedimented in the same way at ``20°C to 30°C and 40°C or higher,'' but the sedimentation rate became slower at other temperatures (Figure 3 (bottom bar graph)).
The "deposition percentage" in FIGS. 3 and 4 was determined by measuring the initial and final concentrations according to a conventional method and calculating 100×[final cell count]/[initial cell count].
実施例４
デュナリエラから獲得できる有用有機物であるカロテノイドは、窒素（Ｎ２）を遮断してデュナリエラを培養すると、デュナリエラ内に高い濃度で貯まることが知られている。
そこで、窒素（Ｎ２）を遮断して、実施例３と同様にして、デュナリエラを培養し、遮光して静置させ、沈降の仕方を観察した。結果を図４に示す。
Example 4
Carotenoids, which are useful organic substances that can be obtained from Dunaliella, are known to accumulate at a high concentration in Dunaliella when Dunaliella is cultured with nitrogen (N 2 ) cut off.
Therefore, Dunaliella was cultured in the same manner as in Example 3 with nitrogen (N 2 ) cut off, allowed to stand still in the dark, and the manner of sedimentation was observed. The results are shown in Figure 4.
窒素（Ｎ２）を遮断していない、実施例３とほぼ同様の結果が得られた（図４）。
すなわち、遮光したことによって、塩分濃度によらず、デュナリエラが沈降した（図４）。
ただし、１２．５質量％の塩分濃度の場合、全ての温度（４℃～５０℃）で同じように沈降したが（図４（上の棒グラフ）、塩分濃度が１７．５質量％の場合は、「２０℃～３０℃、及び、４０℃以上」では同じように沈降したが、それ以外の温度では、沈降速度が遅くなった（図４（下の棒グラフ）。
Almost the same results as in Example 3, in which nitrogen (N 2 ) was not blocked, were obtained (FIG. 4).
That is, by blocking light, Dunaliella sedimented regardless of the salinity concentration (Figure 4).
However, when the salinity concentration was 12.5% by mass, sedimentation occurred in the same way at all temperatures (4°C to 50°C) (Fig. 4 (upper bar graph)), but when the salinity concentration was 17.5% by mass, , "20°C to 30°C and 40°C or higher", the sedimentation rate was the same, but at other temperatures, the sedimentation rate became slower (Figure 4 (bottom bar graph)).
実施例５
縦２０ｍ、横２０ｍ、深さ０．２０ｍ（２０ｃｍ）の屋外の田圃を利用した池に、海洋深層水（塩分濃度３．５質量％）を入れ、約１５℃～約２５℃に保ちつつ、デュナリエラ（Dunaliella）属に属する好塩性微細藻類を２週間培養する。
一般に、上層と下層の濃度が高くなり、中層の濃度が低くなるが、撹拌後に全体の濃度を測定すると、１．０×１０６ cells／ｍＬとなる。又は、ほぼ該濃度となるように、培養日数等を調整して培養する。
Example 5
Deep ocean water (salt concentration 3.5% by mass) is poured into an outdoor pond measuring 20m long, 20m wide, and 0.20m (20cm) deep, and kept at a temperature of about 15°C to about 25°C. Halophilic microalgae belonging to the genus Dunaliella are cultured for two weeks.
Generally, the concentrations of the upper and lower layers are high and the concentrations of the middle layer are low, but when the overall concentration is measured after stirring, it is 1.0×10 6 cells/mL. Alternatively, culture is performed by adjusting the number of culture days etc. so that the concentration is approximately the same.
１．０×１０６ cells／ｍＬでデュナリエラを含む海洋深層水６０ｍ３を、縦４ｍ、横４ｍ、深さ４ｍの容器に移し替える。深さは、培養池の２７倍（４ｍ／０．２０ｍ≒２０）になったことになる。 60 m 3 of deep ocean water containing Dunaliella at 1.0×10 6 cells/mL is transferred to a container measuring 4 m long, 4 m wide, and 4 m deep. The depth was 27 times that of the culture pond (4m/0.20m≒20).
３日間、酸素（Ｏ２）（空気）あり、窒素（Ｎ２）ありの通常の大気の状態で静置した。静置の間、該容器は完全に遮光する。
静置して、３日後、デュナリエラが沈降したので、上澄みを除去し、沈降したデュナリエラを回収する。
It was left standing for 3 days in a normal atmospheric condition with oxygen (O 2 ) (air) and nitrogen (N 2 ). During standing, the container is completely protected from light.
After 3 days of standing, the Dunaliella sedimented, so the supernatant was removed and the sedimented Dunaliella was collected.
回収したデュナリエラからは、β－カロテン等のカロテノイド、グルタチオン等の（オリゴ）ペプチド、グリセリン等の有用有機物が得られる。
また、油脂も獲得でき、燃料としても使用できる。
この方法では、得られる有用有機物や油脂に関し、従来法に比べて、回収率、必要エネルギー、それらの比（すなわち、コストパフォーマンス（効率））が、表１に示した従来法の何れに比べても優れている。
Useful organic substances such as carotenoids such as β-carotene, (oligo)peptides such as glutathione, and glycerin can be obtained from the recovered Dunaliella.
Oils and fats can also be obtained and used as fuel.
With this method, the recovery rate, required energy, and their ratio (i.e., cost performance (efficiency)) of the useful organic substances and fats and oils obtained are lower than those of the conventional methods shown in Table 1. is also excellent.
実施例６
「従来法をまとめた前記表１」と同様の尺度（測定方法、基準）で、本発明の「好塩性微細藻類の回収方法」を比較した。結果を以下の表２に示す。
Example 6
The "Method for collecting halophilic microalgae" of the present invention was compared using the same scale (measurement method, standard) as in "Table 1 summarizing conventional methods". The results are shown in Table 2 below.
本発明の好塩性微細藻類の回収方法は、回収に必要なエネルギーが低く、回収率が高く、全体として低コストで、効率が良いので、本発明で回収された好塩性微細藻類から獲得された有用有機物や燃料等は、化学品製造分野、化学品使用分野、エネルギー産生分野等に広く利用されるものである。
The method for collecting halophilic microalgae of the present invention requires low energy for collection, has a high recovery rate, is low in cost overall, and is efficient, so it is possible to obtain The useful organic substances and fuels produced are widely used in the fields of chemical production, chemical use, energy production, etc.
Claims (10)
（１）池の水の中で該好塩性微細藻類を培養する工程
（３）上記工程１で培養された好塩性微細藻類を、該池の水と共に回収して回収液を得る工程
（４）上記工程３で得られた回収液を容器に入れ、該容器の高さを該池の深さより大きく設定し、該容器の水深を該池の水深より大きくして、該容器内を遮光して、該好塩性微細藻類を該容器の底に沈降させる工程
（５）上記工程４で沈降させた好塩性微細藻類を回収する工程 A method for collecting halophilic microalgae, comprising all of the following steps 1, 3, 4, and 5 in this order, wherein the halophilic microalgae belong to the genus Dunaliella .
(1) A step of cultivating the halophilic microalgae in pond water (3) A step of collecting the halophilic microalgae cultured in step 1 above together with the pond water to obtain a recovered liquid ( 4) Put the collected liquid obtained in step 3 above into a container, set the height of the container to be greater than the depth of the pond, make the water depth of the container greater than the depth of the pond, and shield the inside of the container from light. and settling the halophilic microalgae to the bottom of the container (5) collecting the halophilic microalgae sedimented in step 4 above.
The production of a useful organic substance according to claim 9 , wherein the useful organic substance is a carbohydrate or sugar; an oligopeptide, a polypeptide, or a protein; a vitamin or a vitamin precursor; an oil or fat, a glycolipid, a phospholipid, or a lipoprotein; or a hydrocarbon. Method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2019204446A JP7429344B2 (en) | 2019-11-12 | 2019-11-12 | Collection method of halophilic microalgae |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2019204446A JP7429344B2 (en) | 2019-11-12 | 2019-11-12 | Collection method of halophilic microalgae |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2021073931A JP2021073931A (en) | 2021-05-20 |
| JP7429344B2 true JP7429344B2 (en) | 2024-02-08 |
Family
ID=75898627
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019204446A Active JP7429344B2 (en) | 2019-11-12 | 2019-11-12 | Collection method of halophilic microalgae |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP7429344B2 (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003325165A (en) | 2002-05-09 | 2003-11-18 | Microalgae Corporation | Method for culturing and producing dunaliella and agent which used for thalassotherapy containing dunaliella obtained by the culturing and producing method as main ingredient |
| JP2004521647A (en) | 1999-11-11 | 2004-07-22 | プロアルゲン バイオテック リミテッド | Novel medium for the production of β-carotene and other carotenoids from Donariella salina (ARL5) and strains of Donaliella salina for the production of carotene using the novel medium |
| CN104232489A (en) | 2013-06-09 | 2014-12-24 | 中国石油化工股份有限公司 | Microalgae culturing and harvesting method |
| KR20150014801A (en) | 2013-07-30 | 2015-02-09 | 공석태 | ccc |
| JP2015053863A (en) | 2013-09-10 | 2015-03-23 | 株式会社日立製作所 | Separation system of photosynthetic microorganisms, and separation method of photosynthetic microorganisms |
| CN104593262A (en) | 2015-01-06 | 2015-05-06 | 临沂大学 | Series cultivation and rapid collection method for marine microalgae |
-
2019
- 2019-11-12 JP JP2019204446A patent/JP7429344B2/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004521647A (en) | 1999-11-11 | 2004-07-22 | プロアルゲン バイオテック リミテッド | Novel medium for the production of β-carotene and other carotenoids from Donariella salina (ARL5) and strains of Donaliella salina for the production of carotene using the novel medium |
| JP2003325165A (en) | 2002-05-09 | 2003-11-18 | Microalgae Corporation | Method for culturing and producing dunaliella and agent which used for thalassotherapy containing dunaliella obtained by the culturing and producing method as main ingredient |
| CN104232489A (en) | 2013-06-09 | 2014-12-24 | 中国石油化工股份有限公司 | Microalgae culturing and harvesting method |
| KR20150014801A (en) | 2013-07-30 | 2015-02-09 | 공석태 | ccc |
| JP2015053863A (en) | 2013-09-10 | 2015-03-23 | 株式会社日立製作所 | Separation system of photosynthetic microorganisms, and separation method of photosynthetic microorganisms |
| CN104593262A (en) | 2015-01-06 | 2015-05-06 | 临沂大学 | Series cultivation and rapid collection method for marine microalgae |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2021073931A (en) | 2021-05-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Udayan et al. | Mass cultivation and harvesting of microalgal biomass: Current trends and future perspectives | |
| US4115949A (en) | Production of glycerol from algae | |
| Besson et al. | High-pH-induced flocculation–flotation of the hypersaline microalga Dunaliella salina | |
| Arbib et al. | Long term outdoor operation of a tubular airlift pilot photobioreactor and a high rate algal pond as tertiary treatment of urban wastewater | |
| Milledge et al. | A review of the harvesting of micro-algae for biofuel production | |
| Uduman et al. | Dewatering of microalgal cultures: a major bottleneck to algae-based fuels | |
| Chen et al. | Cultivation, photobioreactor design and harvesting of microalgae for biodiesel production: a critical review | |
| Saeid et al. | Toward production of microalgae in photobioreactors under temperate climate | |
| Zhu et al. | Seawater desalination concentrate for cultivation of Dunaliella salina with floating photobioreactor to produce β-carotene | |
| JP5815931B2 (en) | Useful substance production system by culturing algae | |
| Japar et al. | Application of different techniques to harvest microalgae | |
| Das et al. | The effect of culture salinity on the harvesting of microalgae biomass using pilot-scale tangential-flow-filter membrane | |
| Das et al. | Long-term semi-continuous cultivation of a halo-tolerant Tetraselmis sp. using recycled growth media | |
| Shirazi et al. | Simultaneous biomass production and water desalination concentrate treatment by using microalgae | |
| Mohan et al. | A sustainable process train for a marine microalga-mediated biomass production and CO2 capture: A pilot-scale cultivation of Nannochloropsis salina in open raceway ponds and harvesting through electropreciflocculation | |
| WO2010046777A2 (en) | Method for the efficient and continuous growth and harvesting of nutrient-rich phytoplankton and methods of using the same | |
| Chatsungnoen et al. | Continuous flocculation-sedimentation for harvesting Nannochloropsis salina biomass | |
| Mixson et al. | pH-induced flocculation, indirect electrocoagulation, and hollow fiber filtration techniques for harvesting the saltwater microalga Dunaliella | |
| Sreekumar et al. | Lipid enhancement in microalgae by temporal phase separation: Use of indigenous sources of nutrients | |
| Gautam et al. | Application of halophilic algae for water desalination | |
| CN103103133A (en) | Method for harvesting dunaliella salina | |
| JP7429344B2 (en) | Collection method of halophilic microalgae | |
| JP5569874B2 (en) | Microalgae culture method | |
| Satpati et al. | Recent development in microalgal cultivation and biomass harvesting methods | |
| Joy et al. | Microalgal Biomass: Introduction and Production Methods |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20221107 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20230926 |
|
| A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20230929 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20231114 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20231205 |
|
| A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20231214 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20231214 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20231214 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 7429344 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |