WO2006090780A1 - 女性ホルモン物質分解性微生物およびその利用 - Google Patents
女性ホルモン物質分解性微生物およびその利用 Download PDFInfo
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- WO2006090780A1 WO2006090780A1 PCT/JP2006/303271 JP2006303271W WO2006090780A1 WO 2006090780 A1 WO2006090780 A1 WO 2006090780A1 JP 2006303271 W JP2006303271 W JP 2006303271W WO 2006090780 A1 WO2006090780 A1 WO 2006090780A1
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- WIPO (PCT)
- Prior art keywords
- female hormone
- microorganism
- hormone substance
- female
- microorganisms
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- 238000007710 freezing Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- FBPFZTCFMRRESA-GUCUJZIJSA-N galactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-GUCUJZIJSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 210000004392 genitalia Anatomy 0.000 description 1
- 125000000350 glycoloyl group Chemical group O=C([*])C([H])([H])O[H] 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 208000037824 growth disorder Diseases 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 238000005339 levitation Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000003992 organochlorine insecticide Substances 0.000 description 1
- 210000002394 ovarian follicle Anatomy 0.000 description 1
- 230000027758 ovulation cycle Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 229940049953 phenylacetate Drugs 0.000 description 1
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 230000000379 polymerizing effect Effects 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 230000014639 sexual reproduction Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 229940036555 thyroid hormone Drugs 0.000 description 1
- 239000005495 thyroid hormone Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 125000000026 trimethylsilyl group Chemical group [H]C([H])([H])[Si]([*])(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 238000003260 vortexing Methods 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
- 239000010938 white gold Substances 0.000 description 1
- 229910000832 white gold Inorganic materials 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- 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
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/305—Endocrine disruptive agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/365—Nocardia
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- 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
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/822—Microorganisms using bacteria or actinomycetales
Definitions
- the present invention relates to a female hormone substance-degrading microorganism and use thereof, and more specifically, a microorganism having the ability to degrade female hormone substances contained in domestic wastewater, wastewater from livestock breeding facilities, and the like.
- the present invention relates to a method and apparatus for biologically degrading female hormone substances using microorganisms.
- Endocrine disrupting chemicals that are thought to disrupt the endocrine system of living organisms have been a major issue since Colborn T., "Our Stolen Future” (1996)). It has become a social problem.
- the endocrine system in the living body is a system that normally maintains and regulates the development of life forms, the development of the genital organs, and the functions of various internal organs through the action of male hormones, female hormones, thyroid hormones, corticosteroids, etc. is there.
- Endocrine disrupting chemicals are a group of substances that cause damage to the endocrine system in these organisms, and are released in large quantities to nature through various human activities, causing many abnormalities in wildlife.
- 17 estradiol represented by the following formula (I) is secreted from ovarian follicles.
- This steroid hormone is the most bioactive substance among female hormone substances. Its secretion is controlled by the pituitary follicle-stimulating and luteinizing hormones.
- This 17-estradiol is also used as a drug for the treatment of amenorrhea, abnormal menstrual cycle, dysmenorrhea, uterine stunting and menopause.
- Estrone represented by the following formula ( ⁇ ⁇ ) is a kind of follicular hormone and is a metabolite of 17 j3_estradiol. It has strong estrogenic activity, appears in urine and is excreted from the body. Estrone is used as a female hormone agent to treat female sexual dysfunction, menopause, and prostate cancer.
- estriol represented by the following formula (III) is a female hormone that is a metabolite of 17-estradiol in vivo, and is produced via estrone and appears in urine. Moreover, this estriol has an estrus action.
- the female hormone substances as described above are also used in human medical drugs, and urine It is contained inside and discharged outside the body.
- these substances may be administered to livestock, they are also released into the environment from livestock breeding facilities.
- livestock breeding facilities because of these female hormone substances, wildlife and fish are polluted in rivers and lakes, for example, drainage treatment of female hormone substances in the wastewater before discharging the wastewater into the rivers, etc. It needs to be reduced at the facility.
- wastewater treatment methods at wastewater treatment facilities are divided into three types: physical treatment, chemical treatment, and biological treatment.
- the physical treatment specifically refers to a centrifugal separation method, a filtration method, a pressurized flotation separation method, an adsorption method, and the like
- a chemical treatment refers to the addition of a chemical drug or the like.
- Detoxification method for harmful substances electrodialysis method, ion exchange method, etc.
- biological treatment uses microorganisms to decompose and remove organic substances in wastewater, and can also be used to treat substances that are difficult to treat with physical and chemical treatments. It is valid.
- biological treatment is a method used in many wastewater treatment facilities in recent years, and is generally divided into the following three stages. That is, pretreatment, biological oxidation treatment, and sludge treatment.
- Pre-treatment includes screens, sand basins, sedimentation basins, levitation tanks, etc. These devices remove solid particles with a large particle size and inorganic suspended solids contained in the wastewater. In addition, it helps to reduce the burden of organic substances on bio-oxidation facilities.
- microorganisms are used.
- Methods for using microorganisms at that time include a method of growing microorganisms by attaching them to the surface of a solid support, and a group of microorganisms in a liquid.
- the former is usually carried out by a fixed bed type apparatus such as a trickling filter method, and the latter is carried out by a fluidized bed type apparatus such as an activated sludge method.
- a binding immobilization method is employed as a method for immobilizing microorganisms.
- the bond immobilization method is a method in which microorganisms are attached to an insoluble carrier by shared bond, ionic bond, hydrogen bond, physical adsorption, etc.
- the comprehensive immobilization method is a method for polymerizing or associating low molecular weight compounds.
- the carrier include cellulose carrier, cellulose carrier, and granular activated carbon.
- Polyvinylinol alcohol (PVA), carrageenan, etc. are used as the latter material.
- Rhodococcus bacteria and Sphingomonas bacteria that degrade 17- ⁇ estradiol and the like.
- these female hormone-degrading bacteria show a favorable resolution in a situation where relatively low concentrations of female hormones continue to flow, primary high-concentration female hormones are In case of inflow, it was not enough.
- Patent Document 1 Japanese Patent Laid-Open No. 2001-333767
- Patent Document 2 JP-A-11 341978
- Patent Document 3 Japanese Patent Application Laid-Open No. 2003-52356
- Patent Document 4 Japanese Patent Laid-Open No. 2004-65008
- the present invention has been made in view of the above-described situation, and provides microorganisms having the resolution of female hormone substances, domestic wastewater containing female hormone substances using the microorganisms, drainage from livestock farms, etc. It is a simple and efficient way to decompose female hormone substances from It is an object of the present invention to provide an apparatus.
- the present inventor has found a microorganism having the ability to effectively degrade female hormone substances by using a screening method described later. Further, the present inventors have found that the female hormone substance present in the waste water can be easily and efficiently decomposed by using these, and the present invention has been completed.
- the present invention relates to a genus Pseudaminobacter
- the present invention is a method for decomposing a female hormone substance, characterized in that any of the microorganisms described above is used to biologically decompose the female hormone substance.
- the present invention provides an apparatus for decomposing female hormone substances, comprising means for supporting the above-mentioned female hormone substance-degrading microorganisms and means for bringing the microorganisms into contact with waste water or soil. is there.
- the present invention is the use of the female hormone substance-decomposing microorganism described above for the degradation of female hormones.
- the female hormone substance-degrading microorganism of the present invention has a resolution of a female hormone substance such as 17 ⁇ estradiol, estrone, or estriol.
- the female hormone-degrading microorganism of the present invention (hereinafter simply referred to as “the microorganism of the present invention”) is an in vivo female such as 17 j3 _estradiol, estrone or estriol represented by the above formulas ( ⁇ ) to ( ⁇ ).
- Female hormonal substances such as hormonal substances or their metabolites (hereinafter referred to as this These are collectively called “female hormonal substances”). These female hormonal substances are difficult to remove chemically because they cannot be removed by ordinary biological oxidation treatment and are stable in chemical structure.
- microorganism of the present invention having the above properties includes, for example, pseudoaminobacter
- Examples include those belonging to the genus (Pseudaminobacter), Gordonia, Nocardia, Zoogloea, Pandoraea, Cryptococcus or Trichosporon. . More specifically, one or more microorganisms belonging to the following group A can be mentioned.
- microorganism of the present invention can be obtained by performing multi-stage screening using a female hormone substance from various microorganisms present in activated sludge such as sewage treatment plants and return sludge.
- the female hormone substance is used for the microorganisms contained in the activated sludge.
- the female hormone substance is separated from those that are not.
- This agglomerated culture is carried out by using sludges such as activated sludge and return sludge, and each MDG medium (Modified DOMINIC & GRAHAM 'S medium) containing, for example, female hormone substances in the composition shown in Table 1 below. It is preferable to use a culture medium that contains the trace elements shown in Table 2). Cultivation is preferably performed, for example, at about 28 ° C for about 5 generations per week for 1 week with vigorous shaking in a test tube or the like.
- Microorganisms contained in the enrichment culture solution selected as described above may be prepared by using a common medium used for microorganism separation, for example, a commercially available medium such as ISP medium, RA medium, or YM medium.
- a common medium used for microorganism separation for example, a commercially available medium such as ISP medium, RA medium, or YM medium.
- Each of the isolated microorganisms is further cultured in a medium containing a female hormone substance, and the residual amount of the female hormone substance in the cultured medium is measured by, for example, a thin layer chromatograph or a gas chromatograph. By doing so, it is possible to screen microorganisms with high resolution of female hormone substances. By performing this screening multiple times, female hormones Microorganisms with high quality resolution can be obtained.
- the microorganism of the present invention obtained by force is obtained when the microorganisms 10 7 to 10 8 / L are allowed to act on sludge, sewage, medium, etc. containing a female hormone substance at a concentration of 100 mg / L. After being cultured for 5 hours, 65% or more, preferably 80% or more, of the female hormone substance contained in the medium is degraded. In particular, the microorganism of the present invention degrades 17 / 3-estradiol among female hormone substances by 90% or more, preferably 99% or more in 8 hours.
- the temperature at which the microorganism of the present invention acts is 25-30 ° C, preferably 27-28 ° C.
- Physiological properties It is positive for oxidase and catalase.
- the growth temperature is 20-40 ° C, and it does not grow even at 7 ° C at 10 ° C and 45 ° C.
- D-Glucose, D-Mannitol, Dulcitol, Melibiose are used aerobically. It also has an anabolic effect on D-glucose, D_maltose, D-ribose, D-xylose and the like.
- the cell wall peptide darican is A1 type and has many glycolyl types.
- MK-9 H
- Cellular fatty acids are linear-monounsaturated and contain 10Me_18: 0. Odd number The acid content varies from species to species. As phospholipids, PE is detected and DPG, PG, PI, PIMs and some glycolipids are also found.
- the optimal temperature for growth is 28-37 ° C.
- Absolute aerobic, non-motile grows in mycelium and shows weak acidity. Spores do not form, and the elongated mycelium breaks with the culture time to become short-form fungi or pseudococcal fungi.
- Cell wall peptidoglycan is A1 ⁇ -type and has many glycolyl-types, arabinose and galactose
- Cellular fatty acids are linear monounsaturated and include 16: 0, 18: 0, 18: 1 and 10Me_18: 0.
- PE is detected, DPG, PG, PI,
- Growth temperature is 20-37 ° C.
- Nocardia spi-sheath containing Nocardia 'asteroides' (Nocardia sp) described in the literature (Japan Society for Actinomycetes, “Classification and Identification of Actinomycetes”, Japan Society for the Study: 183-184 (2001)) .) was the same. [0044] Further, among the above-mentioned microorganisms, the microbiological properties of Zooglare 'Spisheath' TMJ-II strain were examined.
- Absolute aerobic and non-motile It often grows in mycelium and forms white to light yellow colonies. Included in Gram-negative bacteria / 3—Proteobacteria ⁇ -subdivision. It exists by forming flocs in sludge.
- Growth temperature is 20-37 ° C.
- the optimum temperature for growth is 42 ° C.
- Pandoraea norimbergensis comb Soundea norimbergensis comb.Same as (Pandraea sp.), Including Pandorea pnomenusa, reported in Pandoraea norimbergensis comb.Nov. there were.
- basidiode As a sexual generation, a basidiode is produced and basidiospores are exogenous. Often forms white colonies. Colonies develop quickly and give off a soft strong or dull luster. Budding-type conidia are unicellular and are spherical, oval, and multipolar. Physiological properties:
- Growth temperature is 20-37 ° C.
- Budding conidia are unicellular and vary in shape. Segmental conidia are unicellular and elongate. Physiological properties:
- Sucrose, Lactose, sorbitol (Sorbitol), and glucose (Glucose) are positive, and ropinulinate (Laevulinate) is negative.
- Growth temperature is 20-37 ° C.
- Trichosporon sp. 1 to 3 including Trichosporon sp.
- microorganism of the present invention described above can be used for decomposing female hormone substances in wastewater or soil, alone or in combination of a plurality of types.
- the microorganism of the present invention in order to decompose the female hormone substance in the wastewater using the microorganism of the present invention, it is preferable to use the microorganism of the present invention between the first sedimentation basin and the final sedimentation basin in wastewater treatment. It is particularly preferable to use it in the secondary treatment step during the oxidation treatment.
- a method of adhering and growing microorganisms on the surface of the solid support is preferable to a method of suspending microorganisms in a liquid. This is because by attaching microorganisms to the surface of the solid support, the microorganisms can be prevented from flowing out, and a concentrated bacterial solution can be obtained.
- porous cellulose is preferable when used in the secondary treatment process, which includes VA VA (polybulal alcohol) and porous cellulose.
- VA VA polybulal alcohol
- a bond immobilization method using polypropylene, ceramic, or the like a comprehensive immobilization method such as PVA-freezing method, and the like can be mentioned. Of these, the bond immobilization method is preferable.
- the present invention microorganisms in wastewater treatment, mass culture by introducing from 10 8 microbial broth obtained by proliferation directly wastewater treatment layer 10 9 cells / mL or processing layer microorganisms immobilized carrier Can be used by
- the immobilization carrier is prepared by directly feeding a certain amount of the carrier into a device cultured in a large amount with a jar mentor, etc., operating the culture device as it is for 2 to 3 days, and stirring to fix the microorganism to the carrier. .
- this immobilization carrier is added to the treatment tank, it should be lifted up and stirred in the treatment layer.
- the degradation of female hormone substances in soil using the microorganism of the present invention is carried out by centrifuging a microorganism grown from 10 8 to 10 9 cells / mL in a large-scale culture with a jar armor. If necessary, it can be carried out by suspending it in physiological saline or the like and spraying it directly onto the contaminated soil.
- the microorganism of the present invention may be adjusted and sprayed on the surface of the soil to be treated and the excavated portion so as to be about 10 9 to 10 1Q per lm 3 .
- an apparatus comprising means for supporting the microorganism of the present invention and means for bringing the microorganism into contact with waste water or soil.
- a bioreactor apparatus such as an air lift type column packed type or a lag flow type can be used.
- Such a device can be used even during the secondary treatment process of wastewater treatment, and can efficiently decompose female hormone substances in wastewater and soil.
- this enrichment culture was subjected to shaking culture at 28 ° C for 1 week. After completion of the culture, 1 mL was collected from the culture solution, added to a test tube together with 10 mL of a new MDG medium, and cultured with shaking for 1 week. This was repeated 5 times and 5 passages were performed.
- ISP medium Difco277010
- RA medium Difco218263
- YM medium Difco271120: both manufactured by Difco.
- the solution was dropped into the petri dish and cultured. This isolated a single colony in each slant.
- the enrichment culture solution of (1) above was developed and analyzed directly by thin layer chromatography (TLC), and the substrate estradiol (hereinafter referred to as "E2”) ) was selected (primary screening).
- TLC thin layer chromatography
- E2 substrate estradiol
- Tables 5 and 6 show the degradation activity of E2 and the number of isolates when each enriched culture broth was developed by TLC.
- the amount of 17 / 3_estradiol remaining in the culture medium after the culture is low. Selected in the next step.
- each of 111 microorganisms was streaked on a YM plate medium. Then, the white gold ears were inoculated into each culture solution added with 17-estradiol to be 100 mg / L. One day later, the entire amount of the grown culture was subjected to solid phase extraction with a C18 solid phase column (manufactured by Waters) and spotted on a thin layer chromatograph. Then, we selected 23 strains in which 17 estradiol was completely digested.
- each of the 23 strains of microorganisms was streaked on a YM plate medium and cultured in a test tube in the same manner as in the primary screening.
- the culture was terminated at 0, 3, 5, 8, and 24 hours from the start of the culture, and each was immediately subjected to solid phase extraction.
- the entire amount of each culture solution was retained on a C18 solid phase column (manufactured by Waters), and then eluted with methanol. An appropriate amount of the eluate was collected, and a surrogate substance (17 / 3-estradiol _d) was added.
- a TMS (trimethylsilyl) derivative was derived with BSTFA (N, O-bistrimethylsilyltrifluoroacetamide), dried again, dissolved in n-hexane, The test solution was analyzed with a gas chromatography mass spectrometer (GC—MS). The concentration of female hormone substances after each time of cultivation from the start of cultivation was measured, and the amount of degradation was measured.
- the analytical conditions for GC-MS are shown in Table 4.
- DNA was extracted from each of the above 9 strains of purely cultured microorganisms using the Shihi benzinole method. That is, after culturing at 30 ° C for 3 days using YM slope agar medium, collected microorganisms and 250 ⁇ L of DNA extraction buffer (100 mmol / L Tris_HCl, 40 mmol / L EDTA, pH 9.0), 200 ⁇ L of benzino chloride and 50 ⁇ L of 10% SDS were prepared and shaken vigorously at 50 ° C for 30 minutes.
- DNA extraction buffer 100 mmol / L Tris_HCl, 40 mmol / L EDTA, pH 9.0
- the following 8F and 15R were used for 16S, and the following Y1F and Y1770R were used for 18S.
- the total amount is 50 / i L, 10mmol / L Tris—HC1 (pH8.3), 50mmol / L KC1, 1.5mmol / L MgCl, 200 / i L dNTP mixture, 1U Taq
- DNA polymerase (manufactured by Takara) 50 ng of vertical DNA and amplification primer 0.4 ⁇ mol / L
- the PCR reaction was carried out using DNA thermal cycler PTC200 (MJ Research) for 25 cycles of 94 ° C for 20 seconds, 55 ° C for 15 seconds, and 72 ° C for 180 seconds. Thereafter, the amplification product was purified using a micro-PCR (Millipore).
- Y1770R (5 '-CTACGGAAACCTTGTTACGAC-3')
- the 18S rDNA base sequence can be decoded using the ABI PRISM TM Dye Terminator Kit (Perkin Elmer) using the ABI PRISM 373A
- the sequencer (ABI) was used. Table 9 shows the primers used for 16S and 18S decoding. AutoAssembler TM (Perkin Elmer) was used for sequence alignment.
- strains that were considered to be yeast from the fungal form were determined using the 18S rDNA sequence, and the other strains were determined using the 16S rDNA sequence.
- a homology search of the determined sequence with the IJ sequence on the database revealed that 4 of the 23 isolates were identified as Gram-positive High G + C gnoleps, 1 strain S a-Proteobacteria, 2 strains S ⁇ — Proteobacteria and two strains were found to be yeast (Table 10). So each Dal
- TAI-I5 strain Obtain commercially available microbial immobilization carriers, and identify TAI-I5 strain on each carrier. The degree of fixing was examined.
- As the material for the immobilization carrier polypropylene, ceramic, alumina silica, and polybulol alcohol-immobilized activated carbon were used.
- the TAI-15 strain was grown to 10 7 cells / mL by mass culture using a jar mentor, and then about 150 g of the above carrier was added per liter, and each carrier was shaken for 24 hours. The stock was fixed. The carrier was taken out 24 hours after the addition, and after lightly wiping off the moisture on the filter paper, the weight of the carrier was measured.
- the carrier was put into physiological saline, sufficiently stirred by vortexing and then treated with ultrasound for about 5 minutes to elute the TAI-15 strain from the carrier.
- This eluate is diluted appropriately with physiological saline, applied to an agar medium (petri dish) prepared with YG medium (Table 11), cultured at 28 ° C, and the number of viable bacteria per carrier weight is measured. did. The results are shown in Table 12.
- the 17b-estradiol degradation activity was examined using activated sludge from a treatment plant actually operating in Ibaraki Prefecture.
- Polypropylene carrier was sterilized at 121 ° C for 10 minutes and cultured at 10 7 cells / mL. The stock was introduced at a rate of 150 g per liter. After 24 hours of shaking culture, the TAI-I5 strain was fixed on a polypropylene carrier. The microorganism-immobilized carrier was adjusted to 50 g per liter of sludge and charged into a sludge tank.
- 17 j3_estradiol was prepared at a concentration of O. lmg / L and added to the sludge tank containing this polypropylene carrier, and the 17-estradiol concentration after 8 hours was measured. The results are shown in Table 13. It was revealed that the measured concentration after 8 hours decomposed almost 100% of the added amount.
- the female hormone substance-degrading microorganism of the present invention can effectively dissect female hormones in a short time.
- FIG. 1 is a drawing showing the time course of the residual amount of 17 ⁇ estradiol.
- FIG. 2 is a drawing showing the time course of the residual amount of estrone.
- FIG. 3 is a drawing showing the change over time of the residual amount of estriol.
- Fig. 4 shows the strain using the neighbor binding method based on the 16S rDNA base sequence (about 1450 bases) of the isolate belonging to the Gram-positive High G + C group that is closely related to the isolate.
- Figures are Bootstrap values from 100 iterations. The lower left bar indicates 5% base substitution distance).
- Fig. 5 shows the system using the neighbor binding method based on the 16S rDNA base sequence (about 1450 bases) of the isolate belonging to Proteobacteria a-subdivision that is closely related to the isolate. It is a drawing showing the governance (the number is the Bootstrap value after 100 iterations. The lower left bar is 5)
- FIG. 6 is a drawing showing a tree using a neighbor-joining method based on the 16S rDNA base sequence (about 1450 bases) of the isolate belonging to Proteobacteria ⁇ subdivision, which is closely related to the isolate. (The number is the Bootstrap value from 100 iterations. The lower left bar is 5)
- FIG. 7 is a drawing showing a phylogenetic tree using the neighbor joining method based on the 18S r DNA base sequence (about 1750 bases) of the isolate and the yeast closely related to the isolate (the numbers are Bootstrap value after 100 iterations (lower left bar indicates 3% base substitution distance).
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JP2007504770A JP5027649B2 (ja) | 2005-02-23 | 2006-02-23 | 女性ホルモン物質分解性微生物およびその利用 |
EP06714411A EP1857541B1 (en) | 2005-02-23 | 2006-02-23 | Estrogenic substance degradable microorganism and use thereof |
ES06714411T ES2381566T3 (es) | 2005-02-23 | 2006-02-23 | Microorganismo que puede degradar una sustancia estrogénica y uso del mismo |
GB0716400A GB2437692B (en) | 2005-02-23 | 2006-02-23 | Estrogenic substance degrading microorganism and use thereof |
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CN100551843C (zh) * | 2006-10-25 | 2009-10-21 | 中国科学院大连化学物理研究所 | 一种催化湿式氧化降解雌激素污染物的方法 |
CN102559542A (zh) * | 2011-12-14 | 2012-07-11 | 首创爱华(天津)市政环境工程有限公司 | 低温条件下去除污水中氨氮的菌Pandoraea sp.及分离培养方法 |
CN112410245A (zh) * | 2020-10-13 | 2021-02-26 | 哈尔滨雁成生物科技有限公司 | 一种低温堆肥微生物复合菌剂及其制备方法和应用 |
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CN116218734B (zh) * | 2023-03-13 | 2023-10-13 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | 一株左炔诺孕酮降解菌及其应用 |
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CN100551843C (zh) * | 2006-10-25 | 2009-10-21 | 中国科学院大连化学物理研究所 | 一种催化湿式氧化降解雌激素污染物的方法 |
CN102559542A (zh) * | 2011-12-14 | 2012-07-11 | 首创爱华(天津)市政环境工程有限公司 | 低温条件下去除污水中氨氮的菌Pandoraea sp.及分离培养方法 |
CN102559542B (zh) * | 2011-12-14 | 2013-06-05 | 首创爱华(天津)市政环境工程有限公司 | 低温条件下去除污水中氨氮的菌Pandoraea sp.及分离培养方法 |
CN112410245A (zh) * | 2020-10-13 | 2021-02-26 | 哈尔滨雁成生物科技有限公司 | 一种低温堆肥微生物复合菌剂及其制备方法和应用 |
CN112410245B (zh) * | 2020-10-13 | 2024-03-08 | 哈尔滨雁成生物科技有限公司 | 一种低温堆肥微生物复合菌剂及其制备方法和应用 |
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EP1857541A4 (en) | 2009-06-03 |
PT1857541E (pt) | 2012-05-21 |
EP1857541A1 (en) | 2007-11-21 |
EP1857541B1 (en) | 2012-04-11 |
ES2381566T3 (es) | 2012-05-29 |
JPWO2006090780A1 (ja) | 2008-07-24 |
GB2437692B (en) | 2009-09-30 |
GB2437692A (en) | 2007-10-31 |
GB0716400D0 (en) | 2007-10-03 |
US8163534B2 (en) | 2012-04-24 |
JP5027649B2 (ja) | 2012-09-19 |
US20090029442A1 (en) | 2009-01-29 |
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