TW200925272A - Microbial reagents for scavenging dioxin pollutants present in a contaminated medium and methods of using the same - Google Patents

Abstract

Disclosed herein is a microbial reagent for scavenging dioxins, dioxin-like compounds and/or polycyclic aromatic hydrocarbons present in a contaminated medium, the reagent comprising Pseudomonas mendocina NSYSU deposited in the Biosource Collection and Research Center (BCRC) of the Food Industry Research and Development Institute (FIRDI) under accession number BCRC 910356. The microbial reagent has the abilities to degrade dioxins, dioxin-like compounds and polycyclic aromatic hydrocarbons. Also disclosed herein are methods and devices that utilize the microbial reagent to scavenge dioxin, dioxin-like compounds and/or polycyclic aromatic hydrocarbons present in a contaminated medium.

Description

200925272 IX. Description of the Invention: [Technical Field 3 of the Invention] Field of the Invention 5 ❹ 10 15 ❹ The present invention relates to a dioxins for scavenging in a contaminated medium, A microbial reagent of dioxin-like compounds and/or polycyclic aromatic hydrocarbons, which contains an accession number BCRC 910356 deposited in the food industry Psewi/omowa·? mewc/oc/wa NSYSU of the Institute for Biological Resource Conservation and Research (BCRC of FIRDI). The microbial agent has the ability to degrading the following environmental pollutants: dioxin, dioxin, and polycyclic aromatic hydrocarbons. The invention also relates to methods and apparatus for using such microbial agents to remove such environmental contaminants present in a contaminated medium. L Prior Art 3 Background of the Invention Due to the excessive development of civilization, many tangible or intangible environmental pollutants, including dioxin and dioxins, polycyclic aromatic hydrocarbons (PAHs), and chlorinated phenols, have repeatedly Invade human society living in a civilized world. Among these environmental pollutants, dioxins are considered to be the most toxic chemicals in humans and animals today. In the past two decades of research, it has been found that Dioxin not only affects the body's fertility, causes developmental disorders, destroys the immune system, and interferes with the endocrine and even carcinogenicity (DB McGregor eia/· (1998), ifea Xian Penspeci., Apr; 106 Suppl 2: 755-760). However, dioxin is a substance that is not easily decomposed, and it will continue to accumulate and exist in nature and living organisms, forming a serious potential hazard to the entire environment. In particular, because Dioxin is very stable under normal conditions, even in hot, acid or alkaline environments, when it is released into the environment and enters the food chain, it will become more concentrated as the food chain climbs. The higher. The chemical structure of Dai® Ossin is similar to that of human hormones (E丄. 10 Gregoraszczuk (2002), Cad. Saude. Publica” 18(2): 453-462), so when it enters via the food chain After the human body, on the one hand, it will form a "false hormone" to produce a hormone-like effect, on the other hand, it will affect the hormone content in the body, both of which will interfere with the original endocrine mechanism of the human body, and It seriously affects the body's metabolism and interferes with the balance of Heilongjiang, which leads to physical damage. Qin In addition, Dioxin has also been shown to be a powerful tumor-promoter that causes carcinogenicity in animals, including increased incidence of liver cancer, lung tumors, and skin tumors in rats. The US Environmental Protection Agency (US_EPA) and the World Health Organization (WHO) classify Dioxin as a possible human carcinogen, while the International Agency for Research on Cancer (IARC) has the most toxic 2,3 in 1997. 7,8-tetraqidibenzo-p-dioxin (2,3,7,8-tetrachlorodibenzo-/?-dioxin, 2,3,7,8-TCDD) is classified as "a human-derived carcinogen, (DB McGregor is α/· (1998), same as above.) 200925272 Dioxin is a collective term for a group of compounds formed by the joining of one or two oxygen atoms to two benzene rings, and such compounds are functionalized aromatics. A class of halogenated aromatic hydrocarbons (HAHs). Dioxin can be divided into two series of coplanar tricyclic compounds, respectively 5 ❹ 10 15 ❹. (1) polychlorinated dibenzo-/7-dioxins , PCDDs), such compounds have a chemical formula of C12H8_n02Cln, where n = l~8; and (2) polychlorinated dibenzofurans (PCDFs), such compounds have a chemical formula of C12H8_nOCln 'where n= 1~8 When a different number of gas atoms are attached to the eight substitution sites of the benzene ring, PCDDs have 75 isomers' and PCDFs have 135 isomers. In these 210 dioxins isomers, 2,3,7,8-TCDD is the most toxic, so it is also called "century poison." PCDDs and PCDFs are aromatic compounds with almost planar structure. These two compounds have similar physical properties and chemical properties, and also have a very stable organic molecular structure' and have high melting point, high boiling point, and lipophilicity (1) And the property of being difficult to decompose, so there are such compounds in various environmental media (such as air, soil, water and food) (W. Parzefall (2002), Foot/ CAe/w. 7bjaCO /·, 40(8): 1185-9). In addition, dioxin has a property of being slightly soluble in most organic solvents and hardly soluble in water, at 25. (The solubility in water is approximately between 7. 4 χ 10.8 and 4.2 x 〇 -4 mg/L, and the solubility decreases as the number of gas atoms on the benzene ring increases. Except that it is exposed to isoxin In the hospital and ultraviolet light, its chemical properties will be changed. 'Dioxin is highly qualitative for heat, acid and alkali. 200925272 is very high, so once it is produced in nature, it is difficult to be decomposed and will continue to accumulate in nature. 5 ❹ 10 15 ❹ 20 dioxin-like compounds are a group of compounds with similar chemical and physicochemical properties to 2,3,7,8-TCDD, including: Polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polybrominated dibenzo-/>-dioxins (PBDDs), and polybrominated dibenzopyrenes (polychlorinated biphenyls (PCBs)) Polybrominated dibenzofurans, PBDFs, etc. Dioxin compounds have good thermal stability, high lipophilicity and non-decomposable properties, so these compounds will continue to accumulate and exist in However, in the environment, it is a serious hazard to organisms. Polycyclic biphenyls (PCBs) are a general term for a group of compounds formed by linking two benzene rings by a covalent bond. * ε12Η10-η(:1η, where n=l~10. PCBs can be further divided into two types, planar and non-planar. Depending on the number of chlorine atoms connected to the benzene ring Depending on the location, PCBs have a total of 209 isomers. PCBs have been found to be potentially carcinogenic, so the United Nations Environment PiOgramme (UNEP) has classified it with Dioxin as a priority that needs to be prioritized. Persistent Organic Pollutants (POPs). Polybrominated diphenyl ethers (PBDEs) are a group of compounds in which only one oxygen atom is bonded between two benzene rings. ^HunBrnO, where n=l~1〇. According to the number and position of the bromine atom 8 connected on the benzene ring, there are 209 kinds of isomers in PBDEs. Studies have shown that PBDEs will Disturbance of thyroid hormone secretion (thyroid hormone), and organism and cause neurological toxicity and carcinogenicity (T.A. McDonald (2002),

Chemosphere, 46..Ί45-755). 5 ❹ 10 15 ❹ PBDDs and PBDFs are collectively referred to as brominated dioxins because of their similar chemical structure to PCDDs/PCDFs. Depending on the number and position of the bromine atoms attached to the phenyl ring, PBDDs have 75 isomers, while PBDFs have 135 isomers. PBDDs/PBDFs have higher molecular weights than PCDDs/PCDFs and have high melting point, low vapor pressure and low water solubility. The physicochemical properties, persistence and toxicity of PBDDs/PBDFs are similar to those of PCDDs/PCDFs, and therefore cause serious harm to the natural environment and humans. Polycyclic aromatic hydrocarbons (PAHs) are a general term for a group of compounds formed by the joining of two or more benzene rings, and can be further classified into low molecular weight (LMW) PAHs depending on the number of benzene rings. (having 2 to 3 benzene rings) and high molecular weight (HMW) PAHs (having 4 or more benzene rings). pAHs have a souther n-octanol/water partition eoeffieient' Kow (in particular, the value increases with the number of benzene rings) '疋 is a hydrophobic organic compound (hydrophobic organic compounds) ) 'Therefore, it can only be dissolved in non-polar or low-polar organic solvents. In addition, 'PAHs have high melting point and high boiling point, and the melting point and the point of the Buddha increase with the number of stupid rings, so it will continue Cumulative 20 200925272 and exists in nature and in organisms. To date, more than 30 PAHs have been shown to be highly carcinogenic, making them the largest group of known carcinogenic compounds. 5 Ο 10 15 ❹ 20 Chlorinated phenols are generally highly toxic and one of the most widely distributed environmental pollutants, while pentachlorophenol (PCP) is one of the most stable and harmful of phenols. . Pentaphenol (PCP) is an aromatic compound containing a hydroxyl group and five gas atoms attached to an aromatic ring with a molecular weight of 266.35 and a melting point of 190°. C, the solubility in water at 26.7 ° C is 14 mg / L. Pentaphenol is difficult to decompose and has a carcinogenic risk in nature. Its minimum lethal dose (LDl.) for humans is 29 mg/kg. Therefore, the production and use of five gas hopes are strictly controlled in countries all over the world. Since environmental pollutants have seriously threatened human health and caused damage to the ecological environment, how to effectively treat environmental pollutants has become the focus of attention and research in the world. Currently known methods for treating environmental pollutants include solidification, removal method, incineration, activated carbon adsorption, and catalytic reduction. , photolysis, and bioremediation. Bioremediation is the use of microbial biodegradative activities to remove environmental pollutants and recalcitrant xenobiotics, and in all bioremediation techniques, biological additions 10 200925272 (bioaugmentation), which adds bacteria to a contaminated environment to enhance the degradation of contaminants, is particularly important because indigenous microorganisms cannot degrade toxins that are difficult to decompose 5 Ο 10 15 ❹ 20 In the case of 'it may be the only way to achieve the purpose of biological reproduction. The biological rejuvenation method has the advantages of low cost, no secondary toxic by-products in the process of degrading environmental pollutants, and secondary pollution (in Wm). Therefore, bioremediation has been widely applied to the remediation of contaminated sites, and the isolation and screening of microorganisms suitable for use in bioremediation is an extremely important research and development topic. In Habe et al. (2QQI), Appl. Microbiol. Biotechnol" 56:788-795, H. Habe et al. developed a carbazole-utilizing bacterium [fake] A strain of CAV] is used to bioremediate dioxin-contaminated soil. Their experimental results show that Pseudomonas strain CA10 has a degrading of dioxin with 4 to 7 gas atoms ( Including the potential of the most toxic 2,3,7,8-TCDD). Later, in Ma. A. (2003), 乂Mo/· 326:21-33, K. Maeda et al. confirmed that the strain was a food. Pseudomonas resin strain CA10. In HB Hong ei α/· (2004), Bioi/egrai/aizo”, 15: 303-313, HB Hong et al. by selective enrichment techniques A dioxin-degrading strain of Pseudomonas vaginalis (Sewi/omcmas verowii) PH-03 was isolated from the contaminated soil. The strain is grown on dibenzo-p_戴奥11 200925272 5 ❹ 10 15 20 dibenzo-/?-dioxin (DD) and dibenzofuran (DF) as the sole source of carbon, which can also be metabolized. 1-chlorodibenzo-/?-dioxin, 2-oxodibenzo-; 2-chlorodibenzo-/?-dioxin and other dioxin metabolites [eg water Salicylic acid and catechol. In M. Bunge ei α/. (2003), iVa/wre, 421:357-360, M.

The results of Bunge et al. showed that a chlorobenzene-dehalorespiring bacterium (De/iiz/ococcoWe·? #) strain CBDB1 was able to bind certain dioxins. [eg '1,2,3-triseo-dichlorodibenzo-/?-dioxin, 1,2,3-TrCDD), l,2,4-TrCDD, l,2, 3,4-TeCDD and 1,2,3,7,8-pentachlorodibenzo-#-dioxin (l,2,3,7,8-pentachlorodibenzo-j9-dioxin, l,2,3,7,8 -PeCDD), etc.] Reductively degassing, thus the strain is potentially available for bioremediation of contaminated sites containing artificial PCDD (anthropogenic PCDD).

JP2002300873 discloses a strain of Pseudomonas mendocereus FEM P187187 which is isolated from an oil field and capable of degrading organochlorine aromatic compounds, and the use of the strain for the degradation treatment of organochlorine aromatic compounds The method, wherein the organochlorine aromatic compound may be dioxin (eg, PCDDs, PCDFs, and coplanar polychlorinated biphenyls, Co-PCBs, etc.). 12 200925272 A master's thesis by Cai Qitang of the National Institute of Biological Sciences of Sun Yat-Sen University [name: "Characterization of bacteria degrading pentachlorophenol"] reveals a soil contaminated with pentaphenol The strain was isolated and screened and can be used as a 5 e 10 15 ❹ 20 carbon source. This strain was identified as Pseudomonas mendocina NSYSU ° at National Sun Yat-Sen University. A master's thesis by Su Zonglin from the Department of Marine Environment and Engineering [Name: "Biodegradation of PAHs in Diesel Fuel by i in Salty Environment"] reveals a strain from It is isolated from a mixed flora that has been domesticated for a long time in a diesel-containing aerobic biofilter bed and is capable of degrading diesel/isolated strains. The master's thesis also explored the effect of the strain on the degradation of polycyclic aromatic hydrocarbons (PAHs), and showed that the strain had the best degradation effect on naphthalene, and the degradation effect on onion (anthracene) was poor. There is no degradation effect for fluoranthene. Therefore, the strain was inferred to have no ability to degrade PAHs having 4 or more aromatic rings. Although the above-mentioned literature reports and patent pre-existing cases exist, there is still a need in the art to screen for microorganisms that can degrade environmental pollutants (especially dioxin) for environmental protection. After research, the applicant unexpectedly discovered a bacterial isolate isolated from the soil contaminated by the party Dioxin of the Sinopec Anshun Plant (Tainan City, Taiwan) (it was later identified as a Pseudomonas Mendoc) NSYSU) In addition to 13 200925272, it has the ability to degrade five gas phenols, and also has the ability to degrade environmental pollutants such as dioxin, dioxin and polycyclic aromatic hydrocarbons (PAHs). Therefore, the plant has great potential in the application of environmental pollution. 5 Ο 10 15 20 SUMMARY OF THE INVENTION Summary of the Invention Accordingly, in a first aspect, the present invention provides a method for removing dioxin, dioxin, and/or polycyclic aromatic hydrocarbons present in a fork-contaminated medium. The microbial reagent, which contains the storage number BCRC 9 deleted 6 is deposited in the food 1# Development Research Resources Resource Conservation and Research Center (BCRC 〇f FIRDI) Pseudomonas syriae NSYSU. In a second aspect, the present invention provides a method for removing a dioxin, a dioxin-like compound and/or a polycyclic aromatic stone antihydrogen compound present in a contaminated media towel, comprising: using a a microbial agent to treat the contaminated medium such that dioxin, dioxin, and/or polycyclic aromatic hydrocarbons present in the contaminated medium are subjected to Mendoza pseudomons in the microbial reagent_NSYSU Degraded and disappeared. The above and other objects, features and advantages of the present invention will become apparent from DETAILED DESCRIPTION OF THE INVENTION For the purposes of this specification, it will be clearly understood that the words "comprising" means "including but not limited to" and the words "including 14 200925272 (comprises)" have a correspondence The meaning of what is to be understood is that if any of the previous publications is quoted here, the reference does not constitute an acknowledgement that in Taiwan or any other country, the publication forms a common general in the art. Part of the knowledge. 5 Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by those skilled in the art to which the invention pertains. In order to effectively deal with environmental pollutants (envir〇nmental) Pollutants), to avoid serious damage to the environment and ecology, have invested a lot of manpower and financial resources to find a solution. Among the current environmental pollution treatment methods, the biological remediation method has low cost and is in a degrading environment. No toxic by-products are produced during the process of pollutants, causing secondary pollution, and In situ (~ their 7 ") the advantage operation, it is widely used in remediation of contaminated sites (contaminated site) is. In order to screen out microorganisms suitable for supply to the bioremediation method, 15 people were asked to study a bacterial isolate having the ability to degrade pentaphenol from the soil contaminated by Dioxin from Sinopec Anshun Plant. The bacterial isolate was identified as Pseudomonas mendocina NSYSU by preliminary test and characterization of 16S rDNA sequence analysis, and was deposited in Taiwan on July 6, 2007. The Bioresource Conservation and Research Center (BCRC of FIRDI) of the Industrial Development Institute, with the registration number BCRC 910356. Applicants have found that Pseudomonas Mendocs NSYSU has the ability to degrade pentoxide, dioxins, and polycyclic aromatic hydrocarbons (PAHs) in addition to its ability to degrade pentoxide. In addition to this 15 200925272, the bacterial isolate also has good mercury resistance and is resistant to damage from mercury ions present in contaminated environments. Accordingly, the present invention provides a microbial agent for removing dioxin, dioxin-like compounds and/or polycyclic aromatic hydrocarbon hydrides (PAHs) present in a contaminated medium, comprising a storage number BCRC 910356 It is deposited in the BCRC of FIRDI's Bioresource Conservation and Research Center (BCRC of FIRDI) by Pseudomonas sinensis (9) and NSYSU. According to the present invention, the contaminated medium is a solid, Environmental medium in liquid or gaseous state 10, and includes 'but is not limited to: soil, sludge, sediment, groundwater, waste water, and exhaust. The contaminated medium is selected from the group consisting of agricultural land (eg, field, orchard land, grazing grassland, etc.), drinking water source (eg, well water), fish culture pond, 15 factory wastewater, Domestic sewage and sludge from sewage treatment plants. As used herein, the term "degradation" means the metabolic decomposition of a compound into a less complex molecule. As used herein, the term "Dioxin" means a group comprising polychlorinated dibenzo-/>-dioxins (PCDDs) 20 and polychlorinated dibenzofurans (PCDFs). The microorganism reagent according to the present invention can degrade a dioxin having 4 to 8 gas atoms. In a preferred embodiment of the present invention, the dioxin is selected from the group consisting of: 2, 3 , 7,8-four-gas dibenzo-dioxin, 16 200925272 ' ' ' '8- five gas dibenzo-dioxin, 2,3,4,7,8_ five gas two stupid and _^_Dioxin, 1,2 ,3,4,7,8-hexachlorodibenzo-dioxine, 12,3,6,7,8-six gas two and A planting Osin, 2,3,4,6,7,8_6 Chlorobibenzo-_ household_Daixin, '8,9-six-gas dibenzo- _ dioxin, 1,2,3,4,6,7,8-seven gas dibenzopyrene dioxin, l2 , 3,4,7,8,9·seven gas dibenzo-/?-dioxine, octachlorodiphenyl, oxin, 2,3,7,8-tetrachlorodibenzofuran, ι, 2,3,7,8-five gas, stupid, 1,2,3,4,7,8-hexachlorodibenzofuran, 1,2,3,6,7,8-hexa-diphenyl 0 husband, 1

10 15

20,2,3,7,8,9-hexa-dibenzofuran, 1,2,3,4,6,7,8-seven-gas dibenzophenanol, ', eight gas dibenzofuran, And their combination. In a more preferred embodiment of the invention, the dioxin is octa-dibenzo-π-dioxine (D) and/or octa-dibenzofuran (〇CDF). The dioxin-like hydrazine which can be degraded according to the present invention is selected from the group consisting of brominated dioxin, polybrominated diphenyl ether (Es), polychlorinated biphenyl (PCBS), and their combination. As used herein, the term "brominated dioxin" means a group of polybrominated dibenzo-/?-dioxins, and polybrominated dibenzofurans (PBDFs). The microbial agent according to the present invention can degrade a deodorized dioxin having 4 to 8 bromine atoms. In the preferred embodiment of the present invention, the desert dioxin is selected from the group consisting of: 2,3,7,8 four desert dibenzo- corpse _ wear f Xin 1 '2,3,7,8-pentabromo 2 stupid _ corpse _ dioxin, 1,2,3 4/6,7,8 _ hexabromo 2 stupid and / gt;- dioxin, ^^ hexaphenodibenzox dioxin, ^^ ' 2,3,4,7,8-J. 17 200925272 bromodibenzofuran, and combinations thereof. As used herein, the term "polybrominated diphenyl ether," means a group of compounds having from 1 to 10 discrete atoms attached to the diphenyl mystery. 5 10

15 Q 20 The microbial reagent according to the present invention can degrade a polybrominated diphenyl ether containing 2 to 1 Torr of a bromine atom. In a preferred embodiment of the present invention, the polybrominated diphenyl is selected from the group consisting of 2,4-dibromobiphenyl, 4,4, _, and odorous diphenyl ether. 2,2',4-triple diphenyl test, 2,4,4'-tribromodiphenyl ether, 2,2',4,5'-tetrabromodiphenyl ether, 2,3',4', 6-tetrabromodiphenyl ether, 2,2,4,4,-tetrabromodiphenyl ether, 2,3',4,4'-tetrabromodiphenyl ether, 3,3,,4,4,- Tetrabromodiphenyl ether, 2,2',4,4',6-pentabromodiphenyl ether, 2,3',4,4,6-pentabromodiphenyl ether, 2,2',4,4 ',5-pentabromodiphenyl ether, 2,2',3,4,4'-pentabromodiphenyl ether, 3,3',4,4',5-pentabromodiphenyl ether, 2,2' ,4,4',5,6'-hexabromodiphenyl ether, 2,2',4,4,5,5'-hexabromodiphenyl ether, 2,2',3,4,4\6 HexaBDE, 2,2,,3,4,4,6'-hexabromodiphenyl ether, 2,2',3,4,4',5'-hexabromodiphenyl ether, 2 ,3,3,,4,4,,5-hexabromodiphenyl ether, 2,2',3,4,4',6,6'-heptabromodiphenyl ether, 2,2',3,4 , 4',5,,6-heptabromodiphenyl ether, 2,3,3',4,4',5',6-heptabromodiphenyl ether, 2,2,,3,3',4, 4,6,6,-octaBDE, 2,2',3,4,4',5,5',6- OctaBDE, 2,2',3,3',4,4',5,6,-octaBDE, 2,2',3,3',4,5,5,6 6,6-nonabrominated diphenyl ether, 2,2',3,3,4,4',5,6,6'-nonabrominated diphenyl ether, decabromodiphenyl ether, and combinations thereof. In a more preferred embodiment of the present invention, the polybrominated diphenyl ether is selected from the group consisting of 2,2',4,4,6-pentabromodiphenyl ether, 2,3, , 4,4',6-pentabromodiphenyl ether, 2,2·,4,4',5-pentabromodiphenyl ether, 2,2,3,4,4'·pentabromodiphenyl ether, 3,3,4,4,5-five desert diphenyl mystery, 2,2',4,4',5,6'-hexabromodiphenyl ether, 2,2·,4,4·,5,5 · HexaBDE, 2,2,3,4,4,6-hexabromodiphenyl ether, 18 200925272 5 ❹ 10 2,2',3,4,4',6'-hexabromo Diphenyl ether, 2,2',3 4 ,5, hexabromodiphenyl ether, 2,3,3,4,4,5-hexabromodiphenyl ether, and their grades as used herein , the term "multi-gas flocculation," means a group of compounds having from 1 to 10 gas atoms attached to biphenyl. The microbial reagent according to the present invention can be used to decontaminate a group containing 4 to 7 germanium atoms. Chlorinated biphenyl. In the present invention / preferred embodiment _ ', is selected from the group consisting of: 3, 3', 4, 4 tetrahydrobiphenyl, 2, 3, 3', 4, 4 '-Pentachlor chloride stupid,...' ζ, 3,4,4',5-five-gas biphenyl, Μ'4'4Ή«υ''3'4'4^^ Stupid, 3,3|, 4, 4,, 5_ five-phase biphenyl, 2,3,3',4,4',5·six gas stupid, 2,3 v ^ '3,4,氕5 '-Six-gas biphenyl, 2,3',4,4',5,5'-hexa-biphenyl, 3,3',4 4,5,5-hexabenzene, 2,3,3, , 4, 4, 5, 5, - heptaphene biphenyl, and their oxime, and in a more preferred embodiment of the invention, the amidine & ^ polyglycol is selected from the following Group: 2,3,4,4,,5-five gas, stupid, polychlorinated biphenyl, tetrahydrobiphenyl, 3,4,4,,5-

15G,4,4'5-penta-biphenyl, 2',3,4,4',5-pentachlorobiphenyl, 2,3,3,,4 4| $ 2,3,3',4, 4',5'-hexa-biphenyl, 2,3',4,4' 20 six-biphenyl, 5'·hexa-biphenyl, 2,3,3,,4,4,,5,5 , - seven gas biphenyl, and their cockroaches and people. As used herein, the term "polymetallic aromatic. hydrocarbon" means that a group of persons formed by joining two or more benzene rings can be degraded according to the microorganism of the present invention; there are 2 to 6 a polycyclic aromatic hydrocarbon of a benzene ring. In a preferred embodiment of the present invention, the polycyclic aromatic carbonitride is selected from the group consisting of naphthalene, acenaphthylene, acenaphthene, fluorene. ), phenanthrene, anthracene, propadiene 19 200925272 (fluoranthene), pyrene (pyrene), benzo (a) onion [benzo(a) anthracene], brook (chrysene), benzo ( b) benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(8)芘[benz〇(a)pyrene],茚5 l,2,3-cd)芘[indeno(l,2,3-cd)pyrene], dibenzo(a,h) onion [dibenzo(a,h)anthracene], benzo(g,h,i ) Flower '[benz〇(g,h,i)perylene] 'and combinations thereof. In a more preferred embodiment of the invention, the polycyclic aromatic hydrocarbon is naphthalene. The microbial agent according to the present invention may further comprise at least one microorganism which can remove environmental pollutants. The microorganism capable of removing environmental pollutants includes, but is not limited to, microorganisms of the genus Pseudomonas, microorganisms of the genus Dehalobacteria (I^/m/ococcoWes), micro-branches of the genus Candida, red Microtubules of the genus (j^j0ijococcus), microorganisms of the genus Aeromonas (/erowowiw), 15 microorganisms of the genus Rhizobium, microorganisms of the genus Leptomon (ASp/iiwgomowas), and Arthrobacter A microorganism belonging to the heart okckr), a microorganism of the genus Froatella (such as (9) η·α), a microorganism of the genus Flavobacterium Cumavocwww, and a microtubule of Bacillus. In a preferred embodiment of the present invention, the microorganism capable of removing environmental pollutants is selected from the group consisting of Pseudomonas resin (hew and / «〇 « still rw / wovorawi) strain CA10, Pseudomonas veronz·/ PH-03, Decamago species CDe/^/ococcozWa pregnancy.) strain CBDB1, Pseudomonas mendocina FERM P- 18187, Candida 20 200925272, and combinations thereof. 5 Ο 10 15 ❹ 20 The microbial agent according to the present invention may optionally contain nutrients which are beneficial for the growth of microorganisms, such as glycerol, riboflavin, casein, polypeptone. ), meat extract, soybean cake, yeast extract, cellulose, glucose, corn extract, whey powder, starch, vitamins [ Such as thiamine, biotin, nicotinic acid amide or pantothenic acid (〇 & 1 at 111110 & 11 (^1^6)], or containing enzymes such as starch An amylase, a protease or a lipase. The microbial agent according to the present invention can be manufactured into a form suitable for use by techniques well known to those skilled in the art, including, but not limited to: culturing a culture solution, a suspension, a granule, a powder, a tablet, a pill, a capsule, a slurry, and the like. In addition, the microbial reagent can also be used. Fixed (imm Obilized) is used on an insoluble support. The microbial agent according to the present invention may further comprise a biocompatible carrier. In a preferred embodiment of the invention The Pseudomonas Mendocs NSYSU in the microbial agent is entrapped in the biocompatible carrier. The biocompatible carrier comprises, but is not limited to: silica gel, starch , agar, chitin, chitosan, p〇iyVinyl alcohol, alginic acid, polyacrylic acid (p〇iyacryiainide), carrageenan 21 200925272 (carrageenan), agarose, gelatin, cellulose, cellulose acetate, dextran, and collagen. 5 Ο 10 15 ❹ 20 Another in the present invention In a preferred embodiment, Pseudomonas mendocensis NSYSU in the microbial agent is supported on the biocompatible carrier. The biocompatible carrier includes, but is not limited to, glass, ceramic, metal oxide, activated carbon, kaolinite, bentonite, zeolite (zeolite) ), aluminum, anthracite, glutaraldehyde, polyacrylic acid, p〇iyurethane, polyvinyl chloride, ion exchange resin (i Exchangen exchange resin), epoxy resin (ep0Xy resin), photoplastic resin (ph〇t〇seUing resin), polyester (polyester) and polystyrene (p〇iystyrene).

The microbial agent according to the present invention can also be produced by a technique known to those skilled in the art for removing dioxin, dioxin, and/or polycyclic aromatic hydrocarbons present in a contaminated medium. Bioreactor or device. For the manufacture of a bioreactor, reference is made to, for example, US 5,279, 963, US 5, 258, 303, US 5, 520, 520, US 5, 494, 574, US 60 030 533, US 2003/0008 381 Al, US 2006/0270024 Al, EP 0 609 399 Bl, EP 0 867 238, and K. Ishii and T. Fumichi, (2007), (7)·α/5,148(3): 693-700. The present invention also provides a method for removing dioxin, dioxin-like compounds and/or polycyclic aromatic hydrocarbons present in a contaminated medium, comprising: a microbial agent as described above for treating the subject 22 200925272 Contaminated, dioxin, dioxin-like, and/or polycyclic aromatic hydrocarbons present in the _ media towel are degraded by the Η多萨假单(10) in the microbial reagent and disappear. 5 ❹ 10 15 ❹ 20 In the method according to the invention, the microbial agent can be used in combination with at least a microorganism selected from the group consisting of environmentally identifiable contaminants. Microorganisms of the genus Staphylococcus, micro-records of the genus Candida, the property of the genus Astragalus, microorganisms of the genus Gastrogen, microorganisms of the genus Rhizobium, microorganisms of the genus Lysosporium, microorganisms of the genus Arthrobacter, Microorganisms of the genus E. genus, microorganisms of the genus Yellow genus, and microorganisms of the spore rod. Preferably, the microorganism capable of removing environmental pollutants is selected from the group consisting of: Pseudomonas sphaeroides strain CA10, Pseudomonas vaginalis deletion 3, Desococcus species strain CBDB1, gate Pseudomonas donovar FERM pi8187, and vhwawaiA", and combinations thereof. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be further described with respect to the following embodiments, but it should be understood that the embodiments are merely illustrative and not to be construed as the invention The implementation of the restrictions. Example 1. Separation of the bacteria box with the ability to degrade pentachlorophenol. Screening experimental materials: 1. The actual soil sample used in the experiment was taken from the Sinopec Anshun Plant (Tainan City 'Taiwan) by Dioxin pollution. Soil. 2. A defined liquid medium for culturing the isolated microorganisms in the experiment 23 200925272 (defined broth medium) having the formulation shown in Table 1 below. Table 1. Formulations for limiting liquid medium. Ingredients: NH4C1 Na2HP04 · 12H2〇KH2P〇4 MgS04 · 7H20 CaCl2 trace metal solution* pentachlorophenol deionized water 0.5 g ~ 3.2 g 0.8 g 0.2 g 0.01 g 1 mL The amount is adjusted to 1 L depending on the experiment. The pH is adjusted to 7.20~7.40 using 2 N H2S04. * · Like the person to think · ν? from the approval of the β β ε T? _cr \ . π χ λ r \ ^ λ ^ : The preparation of trace metal solution is 5 gFeS04 · 7H20, 4 gZnS04 · 7H2 〇, 0.2 g MnCl2. 4H2O, 0.5 g C〇C12.6H20, 0.1 g NiC1.6H2O, 0.15 g H3B〇3 and 2.5 g EDTA were combined with 1 L of deionized water. Ο 5 3. The defined agar medium used to culture the isolated microorganisms in the experiment was prepared by adding the nomine agar (Difco) (15 g/L) to the defined liquid medium. Was made. The fine η separation spider with the ability to degrade jade gas is selected from the following: 10 Ο 15 1. Add 1 gram of the actual soil sample to a 100 mL qualified liquid medium (containing l〇mg/L five gas phenol) The culture flask was then cultured in a constant temperature shaking incubator (30 ° C, 160 rpm) for 14 days. 2. Remove 1 mL of the culture obtained in step 1 and add it to a flask containing 99 mL of defined liquid medium (containing 20 mg/L of five gas), followed by a constant temperature shaking incubator (30). The culture was carried out in °C, 160 rpm for 14 days. 3. 1 mL of the culture obtained in Step 2 was taken and the operation of Step 2 was repeated. However, the culture was carried out by using a defined liquid medium containing 30 mg/L of pentaphenol. 24 20 200925272 4. Remove 1 mL of the culture obtained in Step 3 and repeat the operation of Step 2, but here, use a defined liquid medium containing 40 mg/L of pentaphenol to culture. 5 ❹ 10 15 ❹ 20 5. The culture obtained in step 4 was applied to a defined agar medium (containing 40 mg/L of pentaphenol) by a four-zone scribing method (f〇ur-quadrant streak method). The culture was carried out in an incubator (3 Torr.) for 1 day to obtain a bacterial isolate having the ability to degrade five gas. Example 2. Characterization of a fine sputum isolate having pentachlorophenol degradation ability The following preliminary test and 16SrDNA sequence analysis were carried out to confirm the species of the bacterial isolate obtained in the above Example 1. A. Preliminary test: This test is a reference J. Cappuccino and N. Sherman (2007)

Microbiology: A laboratory manual, 8th ed., Benjamin/ Cummings Science Publishing, California, including the following test items: Gram staining, morphological observations, motility, temperature growth Test (4 C, 42 C), phenylethyl alcohol agar (PEA) growth test, McConnex agar growth test, Oxidation-Fermentation test, glucose fermentation test, Lactose fermentation test, sucrose fermentation test, starch hydrolysis test, citrate utilization test, phenylamine test, H2S production test, 1^03_ reduction test, indole production test, methyl Red-Voges Proskauer (MR-VP) test, urease activity test, catalase activity test, oxidase activity test 25 200925272 test and gelatin Hydrolysis test, etc. Results: The fineness of the five-gas phenol degradation ability transmitted by the αβ丄 in the above Example 1

Preliminary test results of the bacterial isolates are shown in Table 2 below. As can be seen from Table 2, the isolate is a Gram-negative bacillus, has mobility (movabie), grows well under hunger, can grow on stupid ethyl alcohol loam and MacKachai reconstituted fat, can take care of (four) sugar and turn The citrate, catalase and oxidase activity tests were all positive. According to the results obtained, the person who was initially lying on the bacterial isolate was belonging to the genus Pseudomonas mpse "d_nas". Table 2. The fine test items obtained in Example 1 η-- ΤϊΧϋ; '-- — Type observation naked movement / growth at 4 ° C _ growth at 42 ° C + phenethyl alcohol agar growth test + MacKangkai agar growth test + oxidation - fermentation test 〇 / _ glucose fermentation test + (production Acid) lactose fermentation test _ sucrose fermentation test - starch hydrolysis test -- code 1 «, day box fruit citrate utilization test + aniline test - H2s production test - N 〇 3 reduction test + 吲哚 production test monomethyl Red test-Berbo's test-urea activity test _ catalase activity test + oxidase activity test + gelatin hydrolysis test one note: + 'positive reaction; one, negative reaction; hydrazine, oxidation. B, 16S rDNA sequence analysis : For the bacterial isolate obtained in the above Example 1, the genomic DNA was extracted by the following procedure: 1.5 mL of the bacterial solution [cell density greater than 0.3 (OD_)] was placed in a microcentrifuge tube (microtube) Centrifuge at 12,000 rpm for 1 minute. After removing the supernatant, add 100 μίΤΕ buffer to disperse the cells' followed by 100 26 15 200925272 pL lysinzyme (50 mg/ (mL) and mix well, then apply at 37 ° (: water bath for 30 minutes. Then add '350 pLTE buffer, 30 μί 10% SDS and 5 pL proteinase K (20 mg/mL) and give The mixture was uniformly mixed. The resulting mixture was placed in a water bath at 56 ° C for 15 hours, then 100 μί 5 M NaCl solution was added and mixed well. The resulting mixture was added to 650 pL of cold phenol and given The mixture was shaken vigorously, and then centrifuged at 14,000 rpm for 5 minutes to cause partitioning of the aqueous layer and the organic layer. Thereafter, the supernatant was transferred to a new microcentrifuge tube and the distribution was repeated. Separation 10 was treated several times until the white matter between the aqueous layer and the organic layer in the microcentrifuge tube disappeared. Then, 600 pL of ice-cold chloroform/isoamylalcohol (IAA) was used (v/v= 24:l) Join The microcentrifuge tube containing the finally collected supernatant was mixed and homogenized, followed by centrifugation at 14, rpm for 5 minutes. Thereafter, 500 pL of the supernatant was transferred to 15 to a new microcentrifuge. In the tube, slowly add 1,000 pL of 95% ice ethanol and mix gently, then centrifuge at 14,000 rpm for 2 minutes. After removing the supernatant, the pellet was washed with 1 〇〇 pL of 70% ice ethanol, and then the pellet was air-dried at room temperature. Finally, 30 to 50 pL of TE buffer and 2 pL of RNAase (50 mg/mL) were added to disperse the pellet, and placed in a 37 ° C water bath for 30 minutes, thereby obtaining the bacterial isolate. Genomic DNA. Using the obtained genomic DNA as a template and using a set of 16S rRNA genes directed against bacteria to design a universal primer pair F1 and R1 with the nucleotide sequence shown below. 27 200925272 Polymerase chain reaction (PCR) of the operating conditions shown in Table 3 below: Forward primer F1 5'-attgaacgctggcggcaggc-3' (SEQ ID NO: 1) Reverse primer R1 5 5'-cccagtcatgaatcactccg-3' (sequence Identification number: 2) Table 3. Reaction conditions of PCR Content volume (genomic DNA of μ〇pentaphenol-degrading strain (5 ng/pL) 10 Forward primer FI (25 μΜ) 1 Reverse primer R1 (25 μΜ) 1 nucleotide (dNTPs) (50mM) 2 reaction buffer reaction buffer) (lOX) 10 DMA polymerase DNApolymerase) (2.5 υ / μί) 0.5 secondary deionized water (dd H20) 75.5 Operating conditions: at 94 ° C The denaturation was carried out for 30 seconds, the primer annealing was carried out at 52 ° C for 30 seconds, and the elongation reaction was carried out at 72 ° C for 90 seconds for a total of 30 cycles (cycles). ). The amplified product (ampiifjed product) obtained after the completion of the PCR was confirmed by agarose gel electrophoresis, and then purified and recovered by QIAquick PCR Purification kit (Qiagen), and the consortium was entrusted to the consortium. The Corporate Biotechnology Development Center (Taipei, Taipei) conducted the sequencing of the recovered PCR amplification products, and the resulting sequencing results were analyzed by the Gene Blast software provided on the NCBI website. Results: The results of 16S rDNA sequence analysis of the bacterial isolate 15 having the pentagas-degrading ability obtained in Example 1 are shown in Fig. 1. The 16S rDNA sequence (SEQ ID NO: 3) of the bacterial isolate was found to be 16S with Pseudomonas mendocs 1^8¥31] after comparison with the gene-shell library in the NCBl website. There is up to 99% identity between rDNA sequences. 28 200925272 5 G 10 15 ❹ 20 Combining the above characterization results, the bacterial isolate having the pentachlorophenol-degrading ability obtained in the above Example 1 was identified as "Pseudomonas Mendocs NSYSU"' and It was deposited at the Bioresource Conservation and Research Center (BCRCofFIRDI) of the Food Industry Development Research Institute, No. 331, Food Road, Hsinchu City, Taiwan on July 6, 2007. The registration number is BCRC 910356. Example 3. Hg2+ tolerance test of NSYSU, the bacterial isolate obtained in the above Example 1 was carried out in a nutrient broth (Difco 003-01) (ie Mendoza) Sub-culture of Pseudomonas NSYSU - After that, the aliquots of the bacterial culture (10 mL) were mixed separately to 1 〇mL containing different concentrations [〇, 1〇, 20 , 30, 40, 50, 60, 70, 100, 150, 200, 250, 300, 350 and 400 ppm (w/w) of HgCl2 nutrient broth medium in a constant temperature shaking incubator (30 ° C The culture was carried out in 120 rpm for 2 days. After that, it was taken with the naked eye and the microscope. The growth of the bacterial isolate was observed. It was observed by the naked eye that the nutrient broth containing 〇~60 ppm (w/w) HgCl2 was cloudy or suspended in the medium. However, the presence of Pseudomonas mendocs NSYSU was observed under the microscope. However, the nutrient broth containing 70卩111 (\^/>¥) above 1匚匚12 remained The state of clarification, and the presence of bacterial cells could not be observed under the microscope. The results of this experiment show that the bacterial isolate obtained in the above Example 1 can resist the mercury ions present in the contaminated environment 'because the bacterial isolate For mercury ions, there is a high tolerance of not more than 70 29 200925272 ppm (w/w). Example 4: Evaluation of the ability to degrade Dyson from Pseudomonas mendocs NSYSU in order to confirm the above example Whether the bacterial isolate obtained in 1 has the ability to degrade dioxin, the following experiment was carried out. 5 Experimental materials: 1. Preparation of dioxin standard: n-n〇nane was used as a solvent in a suitable container A composition of 17 dioxin/furan compounds [provided by the University of Science and Technology's Ultra Micro Center] was prepared according to the ingredients listed in Table 4 below. ' Table 4. Composition of dioxin standards _ Compound name concentration (pg/pL) 2,3,7,8-TeCDF 5 _ 1,2,3,7,8-PeCDF 15 _ 2,3,4,7,8-PeCDF 15 1,2,3 ,4,7,8-HxCDF 20 _ 1,2,3,6,7,8-HxCDF 20 _ 2,3,4,6,7,8-HxCDF 20 _1,2,3,7,8,9 -HxCDF 20 — 1,2,3,4,6,7,8-HpCDF 20 l,2,3,4,7,8,9-HpCDF Wide 20 _ OCDF 50 __2,3,7,8-TeCDD 5 1,2,3,7,8-PeCDD 20 _ l,2,3,4,7,8-HxCDD 15 _ 1,2,3,6,7,8-HxCDD 15 1,2,3,7, 8,9-HxCDD 20 __1,2,3,4,6,7,8-HpCDD 20 _ OCDD 50 2·Preparation of Pseudomonas species of Pseudomonas species: 3 g of nutrient broth medium powder Dissolved in 1 mL of water 30 200925272 ' and dispensed into a covered glass tube (6 mL per tube), then autoclaved at 121 ° C (vapor pressure approximately 1.5 lb / in 2) ) After 15 minutes, it will be used. Pseudomonas mendocs NSYSU was inoculated into the above sterilized nutrient meat 5 soup medium and cultured in a strange temperature shaking incubator (30 ° C, 120 rpm) for 3 days. The actual method:

10 15 Add 50 μM Dioxin standards to 6 1 mL mL covered glass tubes, followed by 1 mL of Pseudomonas Mendocs NSYSU strain culture medium. The 6 tubes were placed in a constant temperature shaking incubator (30 ° C, 120 rpm) for culture, and 1 tube was taken at the 1st, 1st, 20th, 30th, 40th and 50th days to achieve high resolution. High resolution gas chromatography (HRGC, HP6970) / high resolution mass spectrometry (HRMS, Micromass)

Autospec Ultimate) for concentration analysis of dioxin/furan compounds. The experiment was repeated twice. Results·· It was found from two experimental results that Pseudomonas mendocs NSYSU can degrade 17 kinds of dioxin/snack compounds as listed in Table 4, and the degradation effect of 〇cdf 20 and 0CDD is most obvious. Therefore, the applicant further clarifies the experimental results of 〇cdf and OCDD. Referring to Figure 2 and Table 5, the concentration of OCDF and OCDD will gradually decrease with the increase of culture time, especially 'when the culture reaches the 40th day, the concentration of OCDF and OCDD is reduced to about 50 to 60% of the original concentration. . This result shows: Pseudomonas mendocs 31 200925272 NSYSU has a good ability to degrade 〇cdf and OCDD, and the degradation effect will become more and more obvious with the increase of the writing time. Table 5. Effect of Pseudomonas Mendocs NSYSU on the OCDF and OCDD in Degradation of Dioxin Standards OCDF (ρβ/μί) OCDF (%) OCDD (ρβ/μί) OCDD (%) Day 51.4583079 100* 66.99054405 100* Day 10 39.6764098 77.104 44.61958880 66.605 Day 1 of the first experiment 37.0784850 72.056 41.67901485 62.216 Day 30 33.8968179 65.873 36.92067188 55.113 Day 40 26.6726693 51.834 34.91567785 52.12 — ^ Day 50 23.2440963 45.19 23.58496628 35.206 No. 0 Day 60.5318687 100* 61.0062094 100* Day 10 48.698722 80.451 56.4773418 92.577 2nd experiment 20th day 40.0898431 66.229 55.9026433 91.635 Day 30 40.0434161 66.152 49.31205965 80.831 Day 40 30.7501811 50.8 37.5072404 61.481 _ 50th day 26.9864034 44.582 36.190191 59.322 In Dijon The measured OCDF and OCDD concentrations were defined as 100 〇/〇, respectively. Example s. Evaluation of the ability of Pseudomonas syriae NSYSU to remove dioxin in soil samples. To confirm whether the bacterial isolate obtained in the above Example 具有 has the ability to remove dioxin present in contaminated soil. The following actual test is carried out. Experimental materials: 1. Preparation of self-made soil samples: Bai Xian, obtained from the basketball field of the Institute of Engineering and Management of Zhengxiu University of Science and Technology, the soil that has not been dyed by Dioxin is sifted through the mesh (4) mesh to make the particles of the soil 15 It is about 0.149mm. Next, 100 grams of the screened soil was placed in a sample bottle with a foam on the bottom, and the sample was placed in a sample and prepared for t. Thereafter, a cable made of polystyrene (polyvinyl (di) 32, 200925272 PVC) was burned in a small incinerator for 30 minutes, and then all the exhaust gas generated after burning the cable was introduced into the sampling device. 'The exhaust gas was brought into contact with the soil in the sample bottle for 80 minutes, whereby a homemade soil sample was obtained. 5 〇 10 15 20 2. The Pseudomonas mendocs NSYSU strain culture solution used in this experiment was prepared by the method described in Example 4 above. Experimental methods: A. Removal of dioxin from soil samples: 36 grams of the actual soil sample as described in the "Experimental Materials" of Example 1 above was dispensed equally into 18 10 mL covered glass tubes. So that each tube contains 2 grams of the actual soil sample. After that, 9 tubes were randomly selected and 6 mL of Pseudomonas mendocs NSYSU strain culture solution was added as the experimental group, and the remaining 9 tubes were separately added with 6 mL of sterilized nutrient broth medium. (nutrjent broth, Difco 003-01) used as a control group. Next, the 18 tubes were placed in a constant temperature shaking incubator (30 ° C, 120 rpm) for culture, and taken out on the first, first, 17, 24, 31, 38, 45, 52, and 59 days, respectively. The experimental group test tube and one control tube were used for the following Dioxin concentration analysis. In addition, the homemade soil samples were also used for the same experiment. B. Analysis of Dioxin concentration: 1. Extraction step: Take 2 g of soil sample and put it into a cleaned Thimble Filter (Toy〇R〇shiKaisha, Ltd.) Inside, then round the paper; put the paper into the Soxhlet extractor return tube (s〇xhlet), then add 33 200925272 2 g of anhydrous sodium sulfate (anhydrous sodium sulfate), and take 150 mL of toluene (toluene) ) In a 250 mL flat-bottomed flask, the sample 5 Ο 10 15 Ο 20 was subjected to an extraction treatment with refluxing for 24 hours. Thereafter, the toluene extract was concentrated to about 1 mL by depressurization under reduced pressure for subsequent processing. 2. Acid-wash step: Add the toluene extract from step 1 to a 24 mL vial with a Teflon-lined bottle cap, followed by 7 mL of n-hexane (n- Hexane) and shake the vial for about 5 seconds, then add 4 mL of concentrated sulfuric acid (98%) to pickle and shake the vial vigorously for about 20 seconds. Thereafter, centrifugation (2000 rpm, 2 minutes) was carried out to produce a layered layer and a sulfuric acid layer was collected, and the layer which had been fired was then pickled with sulfuric acid until the sulfate layer became white (no more than 3 picklings). The n-hexane layer was collected in a 50 mL sample vial. Each sulfuric acid layer was extracted twice with 7 mL of hexane. All n-hexane layers were collected in the 50 m L vial and concentrated under reduced pressure to approximately 1 mL for subsequent processing. 3. Purification step using a multi-layered silica gel column · Filling a multi-layered rubber tube column (0.5 cm in diameter and 20 cm long) with a glass wool (glass) Wool), followed by 0.5 g of silica gel, 0.5 g of silver nitrate gel (AgN03-silica gel), 0.5 g of tannin extract, and 5 g of sodium hydroxide gel (NaOH-silica gel) , 0.5 g of silicone, 5 g of sulfuric acid-silica gel, 0.5 g of silicone and 0.5 g of anhydrous sodium sulfate, and pressed with glass rod 34 200925272 during packing real. Fill the column with 30 mL of pre-wash and discard the wash solution. 5 ❹ 10 15 ❹ 20 Transfer 1 mL of the acid-washed product obtained in Step 2 to the pre-washed multi-layer rubber hose column. After all the products to be pickled enter the column, clean it with 丨mL of hexane. The sample of the pickled product was run a total of 3 times, and the positively washed liquid was transferred to the column. Next, elute the column twice with 5 mL of n-hexane, then elute the column with 120 mL of n-hexane, and collect the eluate into a 300 mL Erlenmeyer flask. . The collected eluate was blown to near dryness with a nitrogen purge. The purified product thus obtained contains dioxin, non-planar polychlorinated biphenyl compounds and planar polychlorinated biphenyl compounds ° if desired to analyze the product contained in the purified product The total concentration of the above three types of compounds can be dissolved in DMSO and processed using a high-resolution gas chromatograph (HRGC, HP6970) / high resolution mass spectrometer (HRMS, Micromass Autospec Ultimate). Detection of the total concentration of the compound. If the individual concentrations of dioxin, non-planar poly-biphenyl compounds and planar poly-biphenyl compounds contained in the purified product are to be analyzed, the purified product must be further subjected to the following acidic alumina column and activated carbon tube. Column purification step, which separates dioxin, non-planar poly-biphenyl compounds and planar poly-biphenyl compounds, and then uses high-resolution gas chromatograph (HRGC, HP6970) / high-resolution mass spectrometry Instrument (HRMS, 35 200925272

Micromass Autospec Ultimate) for concentration analysis. 4. Purification step using an acidic alumina column: 5 Ο 10 15 ❹ 20 Fill the glass wool with a pointed bottom at the acid aluminum oxide column (0.5 cm in diameter and 20 cm in length). 1 g of tannin, 6 g of acid alumina, 丨g gum and 丨g anhydrous sodium sulfate were sequentially filled. The packed column was pre-washed with 20 mL of n-hexane and the wash was discarded. The eluate eluted from the multi-layer rubber column in step 3 was purged to 1 mL after being purged with nitrogen, and then transferred to a pre-washed acid alumina column, and then eluted with 5 mL of n-hexane. Two times, the column was eluted with 9 hexanes of n-hexane and the eluate was collected into a 15 〇 mL conical flask. The eluate thus obtained contains a non-planar multi-gas biphenyl compound. To analyze the concentration of the non-planar poly-biphenyl compound contained in the eluate, the eluate can be purged to near-dry with nitrogen, then dissolved in DMSO' and using high-resolution gas chromatography. The instrument (HRGC, HP6970) / high resolution mass spectrometer (HRMS, Micromass Autospec Ultimate) was used to perform concentration analysis of the compound. After completion of the self-cleaning, the acidic alumina column is further eluted with 20 mL of methylene chloride/n-hexane (20/80, v/v). The eluate was collected in a 50 mL vial and slowly purged to near dryness with nitrogen. The resulting eluate contained a mixture of dioxin and a planar multi-gas compound. The eluate was dissolved in 1 mL of n-hexane and placed in a vial for the following active residue column purification step. 36 200925272 5. Purification step using activated carbon pipe column: Fill the glass wool with a pointed bottom at the activated carbon/celite column (diameter 5·5 cm, length 20 cm). Then, 〇·5 g Shixi gum, 0.5 g activated carbon/Shiyakutu soil G8/82, v/v) and 0.5 g of Shiqi gum were sequentially filled, and 5 was compacted with a glass rod at the time of filling. Thereafter, the packed column is sequentially 5 mL each of methanol, toluene, digastone / decyl alcohol / benzene (75/20/5 'v/v/v) , cyclohexane/methylene chloride (50/50 'v/v) and n-hexane were pre-washed and the lotion was discarded. 10 Transfer the 1 mL of n-hexane solution obtained from the above-mentioned step 4 after washing with dichlorohydrazine/n-hexane to the activated carbon/diatomite column, and wait until the n-hexane solution has completely entered the column. The vial was washed 3 times with 1 mL of n-hexane and the n-hexane wash was transferred to the column. Then, elute the live 15-carbon/Shixiazao soil column with 2 mL of cyclohexane/dichlorodecane (50/50, v/v) for 4 times, and then 1 mL of dichloromethane. The methanol/toluene (75/20/5, v/v/v) elution column was used twice in total, and all the eluate was combined in a 50 mL sample vial. The resulting eluate contained a flat type. Multi-gas biphenyl compounds. To analyze the concentration of the planar polychlorinated biphenyl compound, the wash 20 can be purged to near dryness with nitrogen, then dissolved in DMSO, and using a high-resolution gas chromatograph (HRGC, HP6970)/ High resolution mass spectrometer (HRMS,

Micromass Autospec Ultimate) for concentration analysis of compounds. After the above elution is completed, the activated carbon/diatomite column is further eluted with 35 mL of toluene, and the washed matter is collected in a 150 mL conical flask 37 200925272, and the washing obtained in this step is obtained. The output contains Dioxin. Right to analyze Dioxin separately, the collected toluene wash can be purged to near dryness with nitrogen, then dissolved in DMSO, and using a high-resolution gas chromatograph (HRGC, HP6970) / high-resolution mass spectrometer ( HRMS, • 5 MlCn) massAut〇spec Ultimate) for concentration analysis of dioxin. Results: Tables 6 and 7 below show the results of Deoxin in the soil samples of Pseudomonas menedus NSYSU and the self-made soil samples, respectively. The dioxins tested were OCDF and OCDD. 1〇 It can be seen from Table 6 that there is a soil sample of the addition of Pseudomonas mendocs NSYSU. The concentration of sputum, CDF and OCDD will gradually decrease with the increase of writing time. In particular, when cultured to the 31st day, the concentration of 〇CDF and 〇CDD is reduced to about 47% of the original concentration. 5〇%, and at day 59, the concentrations of OCDF and OCDD were reduced to about 12% of the original concentration, 5 and 13%, respectively (see Figure 3). In contrast, in the actual soil samples without the addition of Pseudomonas Mendocs 〇 NSYSU, the concentrations of 〇CDF and 〇CDD did not change significantly with " time. In addition, as can be seen from Table 7, in the self-made soil sample + with the addition of Pseudomonas mendocs NSYSU, the concentration of 〇CDF and 〇CDD gradually decreased with the increase of the writing time, especially when cultured to the 31st At daytime, the 〇CDD's volatility was reduced to approximately 46% of the original concentration, while at day 45, the 〇CDF concentration was reduced to approximately 44% of the original concentration. If culture is continued until day 59, the concentrations of OCDF and OCDD are reduced to approximately 22% and 15%, respectively, of the original agronomy (see Figure 4). In contrast, in self-made soil samples without and with the addition of Pseudomonas mendocs 38 200925272 NSYSU isolates, the concentrations of OCDF and OCDD did not change significantly over time. From the above results, it can be seen that Pseudomonas mendocs NSYSU has the ability to remove OCDF and OCDD in the soil contaminated by dioxin, and the scavenging effect of the bacterium 5 isolate becomes more and more obvious with the increase of the writing time.

39 200925272 Table 6. Effect of Pseudomonas Mendocs NSYSU on the time of OCDF and OCDD removal in plant soil samples. OCDF OCDF OCDD OCDD (pg/g) (%) (pg/g) (% Day 0 382912.172 100.000* 594077.817 100.000* Day 10 382292.942 99.838 533244.153 89.760 Day 17 373762.042 97.610 491770.787 82.779 Day 24 210979.667 55.099 523657.619 88.146 Experimental Group Day 31 181924.845 47.511 301537.269 50.757 Day 38 175315.695 45.785 282277.313 47.515 Day 45 152463.249 39.817 210259.118 35.393 Day 52 128604.841 33.586 154345.444 25.981 Day 59 48174.406 12.581 78360.575 13.190 Day 0 382912.172 100.000* 594077.817 100.000* Day 10 384922.532 100.52502 593866.56 99.96443951 Day 17 386523.552 100.94314 594026.8 99.9914124 Control Day 24 384920.154 100.5244 594080.85 100.0005105 Day 31 382802.155 99.971268 594075.87 99.99967227 Day 38 381912.226 99.738858 594104.76 100.0045353 Day 45 382546.521 99.904508 593067.57 99.82994703 Day 52 382491 .526 99.890146 593977.996 99.98319732 Day 59 382886.156 99.993206 594567.852 100.0824867: The OCDF and OCDD concentrations measured on Days of Day are defined as 100%, respectively.

Table 7. Effect of Pseudomonas Mendocs NSYSU on the time of removal of OCDF and OCDD in homemade soil samples with OCDF OCDF OCDD OCDD (pg/g) (%) (pg/g) (%) Day 267.121 100.000* 263.988 100.000* Day 10 205.115 76.787 202.954 76.880 Day 17 191.939 71.855 199.889 75.719 Experimental group day 24 192.770 72.166 183.470 69.499 Day 31 194.943 72.979 121.489 46.021 Day 38 180.600 67.610 113.409 42.960 Day 45 118.047 44.192 47,069 17.830 Day 52 91.860 34.389 64.824 24.556 Day 59 61.146 22.891 41.923 15.881 Day 0 267.121 100.000* 263.988 100.000* Day 10 268.203 100.40506 264.866 100.3325909 Day 17 265.621 99.438457 263.886 99.96136188 Day 24 266.665 99.829291 264.125 100.0518963 Control Day 31 265.957 99.564242 264.988 100.3788051 Day 38 268.211 100.40805 262.999 99.62536176 Day 45 267.556 100.16285 264.011 100.0087125 Day 52 266.997 99.953579 264.114 100.0477294 Day 59 266.665 99.829291 264.556 100.2151613: measured on the third day The OCDF and OCDD concentrations were defined as 100%, respectively. 40 200925272 Example 6 Evaluation of the ability of Mendoza Pseudomonas® NSYSU to degrade brominated dioxin In order to confirm whether the bacterial isolate obtained in the above Example 1 has the ability to degrade brominated dioxin, the following experiment was carried out. 5 Experimental Materials·· 1. > Preparation of Stinky Dioxin Standard: In a suitable container, use n-decane as solvent and formulate four kinds of polybrominated diphenyls according to the ingredients listed in Table 8 below. And dioxin (PBDDs) and three polybrominated dibenzofurans (pBDFs) [they are supplied by the University of Science and Technology 10 University Micro-Micro Center] brominated Dioxin standards. Table 8. Composition of Brominated Dioxin Standards Compound Name Concentration (Pg/gL) 2,3,7,8-TeBDF 15 ^2,3,7,8-PeBDF 60 2>3,4,7,8-PeBDF 60 2,3,7,8-TeBDD 15 1,2,3,7,8-PeBDD 60 1,2,3,4/6,7,8-HxBDD 100 “2,3,7,8,9- HxBDD 50 2. The Pseudomonas mendocs NSYSU strain culture solution used in this experiment was prepared by the method described in the above Example 4. Experimental method: 20 卟 brominated dioxin standard was added to 5 In a 10 mL covered glass test tube, add 5 mL of Pseudomonas mendocs NSYSU strain culture solution to each tube. Place the 5 tubes in a constant temperature shaking incubator (30 ° C, Cultured in 120 i*pm), one tube was taken on Days 3, 3, 6, 9 and 12, respectively, with high-resolution gas chromatograph (HRGC, 41 200925272 HP6970) / High-resolution mass spectrometer (HRMS, Micromass Autospec Ultimate) was used to analyze the concentration of PBDDs and PBDFs. The experiment was repeated twice.Results: 5 From Table 9, it can be seen that the concentration of PBDDs and PBDFs contained in the brominated dioxin standard will vary with the incubation time. The growth is gradually decreasing, especially The concentration of PBDDs and PBDFs decreased to about 13 to 38% of the original concentration when cultured until day 12. This result showed that Pseudomonas mendocs 〇NSYSU has good ability to degrade PBDDs and PBdfs. And the effect of degradation 10 will become more and more obvious as the time of writing increases. 〇42 200925272 ο ο

>9. π 瞵脔 瞵脔 荠 & M > JSYSU 厣 吝 吝 a public 14 雄琴承澉 PBDDs listening PBDFSS5i: > 43 200925272 Example 7. Pseudomonas Mendoca NSYSU isolate degrading polybrominated diphenyl Evaluation of Ether (PBDEs) Capability Test Materials: 1. Preparation of PBDE standards: 配制 Prepare a peptizer in a suitable container using n-decane as a solvent and according to the ingredients listed in Table 10 below. Twenty-two polybrominated diphenyl ether compounds [they are supplied by the University of Science and Technology's Ultra Micro Center] PBDE standards. Table 10. Composition of polybrominated diphenyl ether standards. Compound name concentration (pg/HL) 2,4-DiBDE 100 4,4,-DiBDE 150 2,2',4-TrBDE 150 2,4,4'-TrBDE 150 2,2',4,5'-TeBDE 125 2,3',4',6-TeBDE 150 2,2',4,4'-TeBDE 125 2,3',4,4'-TeBDE 125 3 ,3',4,4'-TeBDE 150 2,2',4,4',6-PeBDE 150 2,3',4,4',6-PeBDE 150 2,2',4,4',5 -PeBDE 150 2,2',3,4,4'-PeBDE 150 3,3',4,4',5-PeBDE 175 2,2',4,4',5,6'-HxBDE 325 2, 2',4,4',5,5'-HxBDE 325 2,2',3,4,4',6-HxBDE 325 2,2',3,4,4',6'-HxBDE 400 2, 2',3,4,4',5'-HxBDE 325 2,3,3',4,4',5-HxBDE 350 2,2',3,4,4',6,6'-HpBDE 325 2,2',3,4,4',5',6-HpBDE 325 2,3,3',4,4',5',6-HpBDE 350 450 2,2',3,4,4' ,5,5',6-OcBDE 350 2,2',3,3',4,4',5,6'-OcBDE 325 2,2',3,3',4,5,5',6 ,6'-NoBDE 950 2,2,,3,3',4,4',5,6,6'-NoBDE 850 DeBDE 900 ❿ 44 200925272 2. Pseudomonas Mendoca NSYSU used in this experiment The culture solution was prepared by referring to the method described in Example 4 above. Experimental method: 20 μί PBDE standards were separately added to 5 mLi 5 covered glass tubes, followed by 5 mL of Pseudomonas mendocs NSYSU strain culture solution. The 5 tubes were placed in a constant temperature shaking incubator (3 (TC, 120 rpm) for culture, and 1 tube was taken at the 3rd, 3rd, 6th, 9th and 12th day to form a high-resolution gas phase layer. The analyzer (HRGC, Ο w HP6970) / high-resolution mass spectrometer (HRMS, Micromass Autospec 10 Ultimate) was used to carry out the concentration analysis of the PBDE compound. The experiment was repeated twice. - As can be seen from Table 11, the PBDEs were The concentration will gradually decrease as the culture time increases. In particular, when the culture reaches the 12th day, the concentration of PBDEs drops by 15 to about 12 to 45% of the original concentration, and the degradation effect of PeBDEs and HxBDEs is the most. Preferably, their concentrations can be reduced to about 12 to 23% of the original concentration. This result shows that Pseudomonas mendocs NSYSU has good ability to degrade PBDEs, and the degradation effect will increase with the time of use. And more and more obvious. 20 45 200925272 ο

> lrzs you pay ZSISU It^^ SB^s cool 莩; public PBDEs customer 涔 > 46 200925272 2,-,3,4,-,--HXWDE (pg/μ- -2-3> -,--HXBDE(%) 2.2- 3,4,4-6--κΧΒΟΕ (Pg-L)2.2- ·4,4-,6--ΗΧωϋΕ _s_ 2.2- ,3,4,4-,6- Hxbde (Pg-L)2.2-,-4,4-,6-HXBDE _(%) 2,-,4,-,5,5'KXBDE (Pg-L) 2,2-,4,4-5 ,--HXBDE _s_ 2.2- >4-5,--HXBDE (SL)2.2- >4----HXBDE _(%) 3.3- , -4-, 'peBDE(pgir)3.3- >4 -5-peBDE(%) ·-,3,---peBDE(pg/μ-2,2-,3,4,4--peBDE(%) 2.2- -4-5-peBDE (Pg-L) 2.2- >4-,5-peBDE(%) 2,3->4-,6-peBDE (Pg-L)·-,4,4-,6-PeBDE (%) 2.2- 4,4- 6-peBDE (Pg-L) 2.2- , 4,4-, 'peroDE(%) 335.5122 loo* 423.623 loo* 33-337 loo* 323.7856 loo* 32^9675 100* 175.5492 loo* 159.4585 loo* 152.2916loo* 153.5181 Loo* 15^.7361°-00* 335.5122 100 421.3596 99.46572 334.337 100 2^5-571 2.40526 253.H2 78.50393

Is.6938 92.677s 155.2691 97.37269 1S.7974 99.01887 141.3799 92b9329 113.5-0° 72.93828 oo 329.8263 98.30532 231.6497 69.043s 69.520710 20.72079 335.5122 100* 3S.8899 90.57492 253.3196 75.5S36 132.413 39.4659433 Private 5.° private ^6 13-2561 423.623 100

291.S300 68.701U 88.65336 2°92742 423.623 loo* 385.213°0 90.93317 327.2983 77.2617 174.§82 41.097431 60.96761 14.39195 333.3773 99.712s 323.*7856 100 267.2==86 82.76022 175.5492 100 15" Private 585 100 152.2916 100 153.5181 100 118 ·3^7^ 75.998s 229'28έ δ00-'40000) 219.0439 67.6509 214-755 66.-486 II-0195 66.659010

S7.234I 105.8616 6^.512^6 105.1525 6^.^^52 10-3654 67.013^ 7"S45 21.54547 75.42507 23.29476 75.24S9 23.29924 33.S799 19-6157 32.2245 20.2087 36-1992 23.7m 29.33706 19.59s 31.50-7 20.22718 334.337 loo* 323.*7856 loo* 322.9675 loo* 175.5492 loo* 159.4585 loo* 152.2916 loo* 153.5181loo* 155.73$ loo* 309.8924 92.68862 261,891 7S.33M3 138.22s 41.3443127 41.69333 12. Private 70 private 5 300.0509 92.669s 322.9675 100 148.i8 84.42066 135-746 84.77S9 132.4256 86.95529 125.7975 81.S313 155.73$ 100 252.0113 77.83277 1S.3232 40.2498323 45.87475 14.1S25 2S.8286 63.1-100 126.9066 39.2939225 ^5.28201 M.02061 12 private-533 70.893600 115.3182 72.31862 11 1.8377 73.4365 Private 113-^==1 73.^31^5 104.3421 S.9982 69·9988 39.874ml 21.52395 12.26092 66.354 41.6120736 76.3307 5°121έ43 63.12445 41.1185796 66·69°°1 42.8274269 21.25172 13.32-743 20-1 Private 6 13.7S6 20.12297 13.10788 21.10S6 13.5^^11 47 200925272

DeBDE (s/hl) DSDE l90) 2,2-,-3---,-6,--NOBDE (pg/μ- 2,-,-->-,5,6,--NOBDE _β_ 2.2 - ,3,3-4,5,-,6,6'Μ0ωϋΕ (pg/μ-2.2- ,3,3-,4,5,5-,6,6--zOBDE190) 2,2-,3 ,3->4-,5,6-_OCBDE (siL)2,2\3,-,4,-,5,6--OCBDE _s_ 2.2- ,3,4,-,5,5-,6 -ocwDE (Pg-L)2.2-,3,4,4-,5,5-,6-OcBDE _s_ 2,-,3,-,4,4-6,--OCBDE (Pg/ML) 2, -,3,3-,4,4-6,6'〇CBDE _β_

2.3, -,4,4-,5-6-HPBDm (SL) 2.3, -,4,4--,6-HPBDE _s___I 2,-,3,4,4--,6-HPBDE (Pg-L 2,2·,3,--,5·'κρωσΕ _s_ 2,2-3,4,4-6,--HPBDE (SL) 2,-,3,4,4-,6,6'HPBDE _r/o) 2.3, -,4,4-,5-HXBDE (Pg-L) 2.3, -,4,4-5-ffixBDE _(./0) 52.233 1 868.4709 loo* 9S.6644 loo* 322, 08°° loo* 384.3339 loo* Private 0.699 each loo* 363.S12loo* 327.95Mloo* 337-3207loo* 375.S84loo* 738-63 81.86649 754.S38 s-87727 831.5918 88.40473 S6.2029 88.85s 3 Private 6.0805 s 。 。 。 。 。 。 。 。 。 。 275 360.7S4 95.956·Μ 684.5242 75-^600 551.1765 63.46517 671.253 71.35945 188.3629 58-8181 241.391 s.807£267.27^3 60.64775 237.4342 65.^01^6 223-652 68.26128 221.63M 65.703^^ 254.3523 67.S337 έ8. 2536 private 5.25926 349.9872 40.29925 350.5729 37.26865

Is.-6S 33.89968 141.2612 36.75S2 173.3933 39.34503 114.8636 31.63926 99-041 30.21895

Is.2139 30.5S16 78.25S8 20.8186 902.0333 -00* 868.47s loo* 940.6644 loo* 322.00000 loo* 384.3339loo* έ°-95 loo* 363b412 loo* 327.953^loo* 337.3207 100* 375.i4loo* S2.S33 100 868 -709 100 940.6644 100 322.OSS 100 384.3339 100 440.6994 100 363-0412 100 327.9534 100 337.3207 100 346.9821 92.30498 83-6472 s.75125 °°18.siM 94.19S5 861.486 91.58272 287.6541 89.30916 371-531 96-67457 393.0835 89.19538 306.3316 S .3793 2S.7S66 86-326 29^.75 Private 2 ^7.677^6 281.3654 74.84S7 706.8S9 78.3652787 360.8478 41.5497-3 363.7840° 38.673172 1-8.3416 36.7420055 144.6168 37.627^00^ 147.81s 33.5417754 141.S45 3S.84S678 129.8206 39.5850797 132.8057 39.3707453 150.0441 3-9150723 237.3596 26.313s 212.83s 24.50742 206.3327 21.93478 60-9464 1 8.68888 741 19.43233 85.4s 19.37897 ^6.59s 15.59008 52.5S72 16.03542 56.2587 Private 16.67812 Private 7.12837 12.5372 48 200925272 Example 8. Mendoza Pseudomonas Evaluation of the ability of gNSYSU to degrade multi-gas biphenyl (pcBs): 1. Preparation of multi-gas biphenyl standard: 5 10 in a suitable container Using n-decane as a solvent and formulating a polychlorinated biphenyl standard containing 12 polychlorinated biphenyl compounds [provided by the Supermicro Center of Zhengxiu University of Science and Technology] according to the components listed in Table 12 below. . 12 Table 12. Composition of Polychlorinated Standards _________ Compound Name Concentration (pg/pL) _______ 3,3,,4,4,-TeCB 100 3,4,4',5-TeCB 100 __^2 ,3,3',4,4'-PeCB 100 __ 2,3,4,4',5-PeCB 100 ___2,3',4,4',5-PeCB 100 ___2',3,4,4' ,5-PeCB 100 ___3,3',4,4',5-PeCB 100 2,3,3',4,4',5-HxCB 100 _2,3,3',4,4',5'- HxCB 100 __2,3',4,4',5,5'-HxCB 100 3,3',4,4',5,5'-HxCB 100 2,3,3,,4,4,,5, 5,-HpCB 100

2_ The Pseudomonas Mendocs NSYSU strain culture solution used in this experiment was prepared by the method described in Example 4 above. EXPERIMENTAL METHOD: Add 20 μM of lenticular biphenyl standard to 5 mL of 1 〇 mL of covered glass § type of official'. Then add 5 mL of Mendoza, 15 strains of NSYSU. Culture medium. The 5 tubes were placed in a constant temperature incubator (30 ° C, 120 rpm) for culture, and 1 tube was taken at the 3rd, 3rd, 6th, 9th, and 12th days of 49 200925272 to achieve high resolution. Gas Chromatograph (HRGC, HP6970) / High Resolution Mass Spectrometer (HRMS,

Micromass Autospec

Ultmiate) for concentration analysis of poly-biphenyl compounds. The experiment was repeated twice. 5 Results: 〇10 As can be seen from Table 13, the concentration of pCBs gradually decreased with the increase of culture time. In particular, when the day 12 was cultured, the concentration of certain PeCBs, HxCBs and Kunca could be reduced to about the bribe. Up to 4%, of which 2,3''4,4',5,5,_HxCB(10) solution is the best, and its concentration is reduced to about 2.6% of the original wave. This result shows that Pseudomonas mendocs NSYSU has a good ability to degrade pCBs, and the degradation effect will become more and more obvious as the action time increases. ❹ 50 200925272 Table 13. Effect of Pseudomonas Mendocs NSYSU of the present invention on the degradation of PCBs with the time of use The first experiment The second experiment ^ Days Day 0 Day 3 Day 6 Day 9 12th Day 0 Day 3 Day 6 Day 9 Day 12 Day 3,3',4,4'-TeCB (Pg/pL) 99.971 76.086 60.947 62.071 62.008 99.971 75.617 66.547 60.829 60.148 3,3_,4,4_- TeCB (%) 100* 76.108 60.965 62.089 62.026 100* 75.638 66.566 60.847 60.165 3,4,4',5-TeCB (Pg/pL) 3,4,4',5-TeCB (%) 98.580 67.794 60.203 20.539 45.828 98.580 76.418 58.397 61.617 25.534 100* 68.771 61.070 20.835 46.488 100* 77.519 59.238 62.505 25.902 - 2,3,3_,4,4'-PeCB (pg/μί) 2,3,3,,4,4,-PeCB (%) 100.679 73.686 67.565 16.225 24.587 100.679 73.953 65.538 15.901 9.840 100* 73.189 67.110 16.116 24.421 100* 73.455 65.096 15.794 9.774 · 2,3,4,4',5-PeCB (pg/μί) 2,3,4,4',5 -PeCB (%) 99.655 66.879 66.475 6.766 3.245 99.655 81.225 64.480 6.630 0.902 100* 67.111 66.705 6.789 3.257 100* 81.507 64.704 6.653 0.905 〇2,3',4,4',5-PeCB (Pg/pL) 99.679 80.260 73.750 5.287 3.051 99.679 75.063 71.537 5.181 0.871 2,3',4,4',5-PeCB (%) 100* 80.519 73.987 5.304 3.061 100* 75.305 71.768 5.198 0.874 2',3,4,4,,5-PeCB (Pg^L> 2',3,4,4,,5-PeCB (%) 99.981 67.854 67.046 4.366 2.394 99.981 86.708 65.034 4.279 0.717 100* 67.867 67.058 4.367 2.394 100* 86.725 65.047 4.279 0.717 3,3',4, 4'5-PeCB (Pg^L) 99.417 78.583 66.443 22.098 45.483 99.417 74.515 64.450 55.245 25.861 3,3',4,4',5-PeCB (%) 100* 79.044 66.833 22.227 45.750 100* 74.952 64.828 55.569 26.013 2, 3,3',4,4',5-HxCB (pg/μί) 2,3,3',4,4,,5-HxCB (%) 100.997 76.760 75.248 7.529 3.413 100.997 73.215 72.991 7.379 0.979 100* 76.003 74.506 7.455 3.379 100* 72.492 72.270 7.306 0.969 2,3,3,,4,4,,5_-HxCB (Pg^L) 2,3,3,,4,4,,5,-HxCB (%) 100.993 68.022 74.678 9.823 4.625 100.993 78.143 72.437 9.626 1.169 100* 67.353 73.943 9.726 4.580 100* 77.375 71.725 9.532 1.157 Ι^ΑΑ'^,δ'-ΗχΟΒ (Pg^L) 2,3',4,4,,5,5,-HxCB (%) 99.978 71.159 73.375 2.196 1.651 99.978 78.824 71.174 2.152 0.627 100* 71.174 73.391 2.196 1.651 100* 78.841 71.189 2.152 0.627 (pg/μί) 3,3,,4,4_,5,5_-HxCB (%) 99.316 84.620 75.106 15.194 13.553 99.316 79.251 72.853 14.890 3.851 〇 100^ 85.203 75.624 15.298 13.647 100* 79.797 73.355 14.992 3.877 2,3,3·,4,4',5,5'-Ηρ€Β (Pg^L) 100.007 74.593 80.736 3.436 2.011 100.007 89.670 78.314 3.368 0.682 2,3 , 3\4, 4\5, 5'^03 (%) 100-^ 74.588 80.731 3.436 2.010 100* 89.664 78.309 3.367 0.682 * : The measured PCBs concentration on day 0 is defined as 100%. Example 9. Evaluation of the ability of Pseudomonas Mendocs NSYSU isolates to degrade polycyclic aromatic hydrocarbons (PAHs) 5 Materials: 1. Preparation of polycyclic aromatic hydrocarbon standards: in a suitable container The use of n-decane as a solvent and according to the ingredients listed in Table 14 below to formulate a compound containing 16 polycyclic aromatic hydrocarbons 51 200925272 [they are provided by the Supermicro Center of Zhengxiu University of Science and Technology] Polycyclic aromatic hydrocarbon standards. Table 14. Composition of Polycyclic Aromatic Hydrocarbon Standards Compound Name Concentration (ng/pL) Naphthalene 1 ^ (acenaphthylene) 1 Dihydrogen (acenaphthene) 1 (fluorene) 1 Phenanthrene 1 Onion (anthracene) 1 - fluoranthene 1 pyrene 1 - benzo (a) onion [benzo(a) anthracene] 1 chess (chrysene) 1 benzo (b) propylene di- Benzo(b)fluoranthene] 1 benzo(k)propanthene benzo[k]fluoranthene 1 benzo(a)f-fbenzo(a)pyrene] 1 茚(l,2,3-cd) i £[indeno(l,2,3-cd)pyrene] 1 dibenzo(a,h) onion [dibenzo(a,h)anthracene] 1 benzo(g,h,i) [benzo(g,h, i)perylene] 1

2. The Pseudomonas Mendocs NSYSU strain culture solution used in this experiment was prepared by referring to the method described in Example 4 above. Experimental Methods: © Add 50 μί polycyclic aromatic hydrocarbon standards to 5 10 mL covered glass tubes, followed by 5 mL of Pseudomonas Mendocs NSYSU strains Culture medium. The five tubes were placed in a 10 constant temperature shaking incubator (30 ° C, 120 rpm) for culture, and one tube was taken at the third, third, sixth, ninth and 12th days to obtain a high-resolution gas phase. Chromatography (HRGC, HP6970) / High Resolution Mass Spectrometer (HRMS, Micromass Autospec Ultimate) was used for concentration analysis of polycyclic aromatic hydrocarbons. The experiment was repeated twice. 15 MA ' 52 200925272 It can be seen from Table 15 that the concentration of PAHs will gradually decrease with the increase of culture time. In particular, when cultured to the 12th day, the concentration of some PAHs can be reduced to about 5 of the original concentration. 20%, in which the degradation effect of naphthalene is the best, and its concentration is reduced to about 5 to 16% of the original concentration. This result shows that: 5 N. mongolicus NSYSU has a good ability to degrade PAHs, and the degradation effect will become more and more obvious with the increase of the writing time.

53 200925272 Table 15. Effect of Pseudomonas Mendocs NSYSU on Degradation of PAHs with Time of Use

The first experiment, the second experiment, the number of days, __ Day 0, Day 3, Day 6, Day 9, Day 12, Day 0, Day 3, Day 6, Day 9, Day 12, Day 12, Naphthalene (ng/pL) 1.001 0.890 0.640 0.551 0.052 1.001 0.810 0.570 0.134 0.170 naphthalene (%) 100* 88.879 63.913 55.007 5.219 100* 80.890 56.923 13.399 16.976 苊(ng/pL) 0.991 0.660 0.520 0.536 0.089 0.991 0.630 0.440 0.390 0.293 苊(%) 100* 66.573 52.451 54.096 8.978 100* 63.546 44.382 39.357 29.525 Dihydroanthracene (ng/μί) 0.999 0.790 0.640 0.646 0.108 0.999 0.780 0.610 0.430 0.334 Dihydroanthracene (%) 100* 79.066 64.053 64.677 10.818 100* 78.065 61.051 43.052 33.444 (ng/pL> 0.987 0.770 0.620 0.626 0.190 0.987 0.750 0.630 0.525 0.401 % (%) 100* 78.004 62.808 63.367 19.295 100* 75.978 63.821 53.234 40.586 Philippine (ng/μΙΟ 0.971 0.780 0.710 0.619 0.335 0.971 0.750 0.720 0.617 0.428 Philippine (%) 100* 80.300 73.093 63.755 34.485 100 * 77.211 74.123 63.557 44.064 Onion (ng/pL) 0.993 0.830 0.716 0.610 0.453 0.993 0.840 0.669 0.620 0.455 蒽(%) 100* 83.573 72.102 61.421 45.644 100* 84.580 67.365 62.428 45.842 Propylene dichloride (ng/pL> 0.997 0.890 0.784 0.760 0.498 0.997 0.900 0.814 0.760 0.480 alkadiene (%) 100* 89.226 78.607 76.193 49.905 100* 90.228 81.561 76.193 48.143 芘( Ng/pL) 0.995 0.880 0.800 0.773 0.482 0.995 0.900 0.810 0.795 0.462 芘(%) 100* 88.429 80.390 77.628 48.416 100* 90.439 81.395 79.876 46.436 Stupid (a)g(ng~L) 0.990 0.820 0.670 0.634 0.394 0.990 0.790 0.710 0.578 0.343 Stupid (a) Onion (%) 100* 82.836 67.683 64.091 39.795 100* 79.805 71.724 58.372 34.607 Chopsticks (ng/μΙΟ 1.061 1.100 0.860 0.898 0.662 1.061 1.090 0.880 0.825 0.553 chopsticks (%) 100* 103.630 81.020 84.601 62.367 100* 102.688 82.904 77.756 52.118 Benzo(b)propadienyl ruthenium (ng/μΙΟ 0.986 0.810 0.620 0.561 0.375 0.986 0.810 0.660 0.503 0.302 benzo(b)propadienyl ruthenium (%) 100* 82.178 62.902 56.898 38.078 100* 82.178 66.960 51.046 30.596 stupid (k) propadiene (ng / μΙΟ 1.165 1.120 0.960 0.844 0.597 1.165 1.090 0.990 0.803 0.512 benzene And (k) alkadiene oxime (%) 100* 96.162 82.425 72.423 51.262 100* 93.586 85.000 68.939 43.926 Benzo(a) 芘 (ng/μί) 1.037 1.010 0.710 0.621 0.420 1.037 0.980 0.740 0.499 0.322 Benzo(a)芘(%) 100* 97.387 68.460 59.870 40.520 100* 94.494 71.353 48.155 31.084 茚(l,2,3-cd> 芘(ng/pL) 0.993 0.850 0.650 0.508 0.332 0.993 0.840 0.710 0.368 0.259 茚 and (1,2, 3-cd)芘(%) 100* 85.559 65.427 51.170 33.412 100* 84.552 71.467 37.076 26.067 Dibenzo(a,h) onion (ng/μί) Dibenzo(a,h) onion (%) 1.067 0.990 0.810 0.659 0.474 1.067 1.000 0.850 0.495 0.422 100* 92.748 75.885 61.763 44.390 100* 93.685 79.632 46.406 39.507 stupid (g,h,i)茈(ng/μΙΟ 1.099 1.130 0.860 0.778 0.544 1.099 1.130 0.890 0.626 0.464 benzo (g,h,i )茈(%) 100* 102.793 78.232 70.753 49.481 100* 102.793 80.961 56.952 42.239 * : The PAHs concentrations measured on day 0 were defined as 100%, respectively. All patents and documents cited in this specification are hereby incorporated by reference in their entirety. In case of conflict, the detailed description of the case (including 5) will prevail. Although the present invention has been described with reference to the specific embodiments described above, it is obvious that modifications and variations can be made without departing from the invention. It is therefore intended that the invention may be limited as indicated by the scope of the patent. 5 Ο 10 15 ❹ [Simple description of the diagram] Figure 1 shows Pseudomonas mendocs NSYSU (which is a bacterial isolate isolated from the knife in Example 1 and characterized by Example 2) Figure 1 shows the pricking acid sequence of Pseudomonas Mendocs NSYSU with the effect of time on the degradation of 0CDF and 0CDD in the dioxin standard, with the result of the first experiment and the second Results of the second experiment; Figure 3 shows the effect of Pseudomonas mendocs NSYSU on the removal of OCDF and OCDD in the soil samples with the application time; and Figure 4 shows the hypothesis of Pseudomonas mendocs NSYSU For the removal of 0CDF and OCDD in homemade soil samples. [Main component symbol description] (none) 55

Claims (1)

200925272 X. Patent application scope · · 5 10 15 φ 20 h — a microbial reagent for removing dioxin, dioxin and/or polycyclic aromatic hydrocarbons present in a contaminated medium. There is a storage system called BCRC 910356, which is deposited at the Center for Bioresource Conservation and Research (BCRC of FIRDI) of the Food Industry Development Research Institute. 2. The microbial agent of claim 1, wherein the microbial agent degrades a dioxin having 4 to 8 gas atoms. 3. The microbial reagent according to item 2 of the patent scope, wherein the dioxin is selected from the group consisting of 2, 3, 7 8 four gas dibenzo- _ _ dioxin, 1, 2, 3, 7,8-Pentachlorodibenzopyrene-Dioxin, 2,3,4,7,8_Five gas dibenzo-corporate-Dioxin, 1,2,3,4,7,8-hexabenzodibenzo _/?_Dioxin, 1,2,3,6,7,8-hexa-dibenzo-_/?_Dioxin, 2,3,4,6,7,8_6-dibenzo-dioxin, 1, 2,3,7,8,9-six-gas dibenzo-_/?_Dioxin, (3), 4,6,7,8·seven gas two stupid and _Bu Dioxin, 1, 2, 3, 4, 7, 8,9· Heptachloro-bismuth and Dioxin, eight gas dibenzo-^^Dioxin, 2,3,7,8_ four gas two Feng and Xiachuan, (2) from five gas dibenzopyrene ^... six gas ζ Benzofuran, 2,3,6,7,8-hexa-dibenzopyrene, mm six gas, benzofuran, coffee milk-seven gas dibenzofuran, and gas dibenzofuran, 4. A combination of the microbial agents of claim 4, wherein the microbial test can be degraded - selected from the group consisting of dioxins in the following group: evolution of dioxin, poly-diphenyl _, multi-gas Biphenyl, and it's 56 200925272 5. The microbial reagent of claim 4, wherein the brominated dioxin contains 4 to 8 bromine atoms. 5 ❹ 10 15 ❹ 20 such as the microbial reagent of claim 5, wherein the desertification Dioxin is selected from the group consisting of 2,3,7,8•Tetrabromo 2 stupid and _ corpse _ Dioxin, 1,2,3,7,8-Five Dibenzopyrene dioxin, Hexabromo Dibenzo-p•dioxine, ^2^...hexabromodibenzopyrene-dioxin, 2,3,7,8-tetrabromodibenzofuran, pentabromodibenzofuran, 2,3,4, 7,8-pentabromodibenzopyrene, and combinations thereof. 7. The microbial reagent of claim 4, wherein the polybrominated diphenyl contains 2 to 1 bromine atom. The microbial reagent of the seventh aspect of the invention, wherein the polybrominated diphenyl ether is selected from the group consisting of 2,4-dibromodiphenyl ether, 4,4,-di-diphenyl ether, 2, 2',4-tribromodiphenyl ether, 2,4,4,-tribromodiphenyl ether, 2,2,4,5'-tetrabromodiphenyl, 2,3',4',6-four Bromodiphenyl, 2,2,,4,4,-tetrabromobiphenyl, 2,3',4,4,-tetrabromodiphenyl, 3,3,4,4,-four Phenyl ether, 2,2',4,4',6-penta-diphenyl ether, 2,31,4,4,6-pentaBDE, 2,2',4,4',5-five Bromodiphenyl ether, 2,2,3,4,4'-pentabromodiphenyl ether, 3,3',4,4',5·pentabromodiphenyl ether, 2,2ι,4,4·, 5,6,_ hexabromodiphenyl ether, 2,2,4,4,5,5'-hexa-diphenyl ether, 2,2',3,4,4',6-hexa-diphenyl ether, 2,2|,3,4,4',6'-hexabromodiphenyl ether, 2,2,3,4,4',5,-hexabromodiphenyl ether, 2,3,3',4 , 4',5-hexabromodiphenyl ether, 2,2,3,4,4,6,6,-heptabromodiphenyl ether, 2,2',3,4,4',5·, 6-heptabromodiphenyl ether, 2,3,3,,4,4,5,6- heptabromodiphenyl ether, 2,2',3,3',4,4',6,6,- OctaBDE, 2,2',3,4,4',5,5,,6-octabromo 57 200925272 Diphenyl ether, 2,2,,3,3',4,4',5, 6,-octaBDE, 2,2',3,3',4,5,5',6,6'-nonabromodiphenyl ether, 2,2',3,3',4,4 ',5,6,6'-nonabrominated diphenyl ether, decabromodiphenyl ether, and combinations thereof. 9. The microbial reagent of claim 4, wherein the polychlorinated biphenyl (5) contains 4 to 7 chlorine atoms. 10. The microbial reagent of claim 9, wherein the polyglycol is selected from the group consisting of 3, 3, 4, 4, - 4 gas biphenyl, Q 3, 4 , 4', 5 · four gas biphenyl, 2,3,3,,4,4,- five-phase biphenyl, 2,3,4,4,,5-penta-biphenyl, 2,3',4 , 4,, 5 - five gas biphenyl, 2,, 3, 4, 4,, 5 - five gas, stupid, 1 〇 3, 3', 4, 4', 5 · pentachlorobiphenyl, 2, 3 ,3,,4,4,,5-hexabenzene, 2,3,3',4,4',5'-hexabenzene, 2,3,,4,4',5,5, - hexa-biphenyl, 3,3',4,4',5,5'-hexabenzene, 2,3,3,,4,4',5,5,-seven-biphenyl, and The combination. U. The microbial agent of claim 1, wherein the microbial test agent degrades a polycyclic aromatic hydrocarbon hydrazine having 2 to 6 benzene rings. 12. The microbial reagent of claim 11, wherein the polycyclic aromatic hydrocarbon is selected from the group consisting of naphthalene, anthracene, indoline, phenanthrene, onion, and allene.第,芘,Benzene (Pseudo-onion, chopsticks, 2〇4 benzo(b)propadienyl fluorene, benzo(k)propadienyl fluorene, benzo(a)pyrene, hydrazine (l, 2,3-cd) hydrazine, dibenzo (a, h) onion, benzo (g, h, i) oxime, and combinations thereof. 13. The microbial reagent of claim 1, further comprising There are at least one micro-clearing agent selected from the following groups: 200925272 5 〇10 organism: microorganism of the genus Pseudomonas, microorganism of the genus Desmoococcus CDeAfl/ococcoiWa, Candida (Q "Mountain." microorganisms, microorganisms of the genus Escherichia (i?A〇i/ococcw«s), microorganisms of the genus Aeromonas (Jeromowas), microorganisms of the genus Rhizobium, Leptobacter Microorganisms of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus The microorganism and the microorganism of the genus Bacillus (5aci (7/w5·). 14. The microorganism reagent according to the πth aspect of the patent application, wherein the microorganism capable of removing environmental pollutants is selected from the group consisting of: Psemi/omo«as reszwovoraws strain CA10, Pseudomonas vonii (Piet/i/owowa·!? verom'z·) PH-03, Dehalomyces sp. CDe/ia /ococcoiWei· w.) Strain CBDB1, Pseudomonas mendoc (PseMdowowa·? we«i/oci«a) FERM P-18187, W5w<a«ai/iz7, and combinations thereof. 15. The microbial agent of claim 1, wherein the microbial agent is manufactured in a dosage form selected from the group consisting of: a culture solution, a suspension, a granule, a powder, a lozenge ( A tablet, a pill, a capsule, and a slurry. The microbial reagent of claim 1, further comprising a biocompatible carrier. 17. The microbial agent of claim 16, wherein the Pseudomonas parasiticus NSYSU is captured by the biocompatible carrier (59) 2009 25272 5 Ο 10 15 ❹ 20 18. The microbial agent of claim 17, wherein the biocompatible carrier is selected from the group consisting of silica gel, temple powder, loam, chitin, and several Chitosan, polyvinyl alcohol, alginic acid, polyacrylamide, carrageenan, agarose, gelatin, cellulose , cellulose acetate, dextran, and collagen. 19. The microbial reagent of claim 16, wherein the Pseudomonas putida NSYSU is supported on On the biocompatible carrier. 20. The microbial agent of claim 19, wherein the biocompatible carrier is selected from the group consisting of: glass, ceramic, metal oxide, activated carbon Activated carbon, kaolinite, bentonite, zeolite, alumina, anthracite, glutaraldehyde, polyacrylic acid, polyuric acid Polyethylene, polyvinyl chloride, ion exchange resin, epoxy resin (ep0Xy resin), photosetting resin, polyester (p〇iyester), and polystyrene (polystyrene). 21. The microbial agent of claim 1, wherein the microbial agent is also resistant to mercury. 22. The microbial agent of claim 1, wherein the contaminated 60 200925272 medium is selected from the group consisting of: soil (s〇il), sludge (sludge), and "child" ( Sediment), groundwater, waste 5 〇10 15 ❹ 20 23. The microbial agent of claim 22, wherein the contaminated medium is selected from the group consisting of: land, orchard land, Grazing grassland, well water, fish culture ponds, factory wastewater, domestic sewage, and sewage sludge. 24. A method for removing dioxin, dioxin, and/or polycyclic aromatic hydrocarbons present in a contaminated medium, comprising: using a microbial agent as in claim 1 Treating the contaminated medium such that dioxin, dioxin, and/or polycyclic aromatic hydrocarbons present in the contaminated medium are degraded by Pseudomonas mendocensis NSYSU in the microbial agent and disappear. 25. The microbial agent of claim 24, wherein the contaminated "quality" is selected from the group consisting of: soil, sludge, sediment, groundwater, wastewater, and waste gas. The method of claim 25, wherein the contaminated medium is selected from the group consisting of: land, orchard land, grazing grassland, _|+* 7, fishery pond, factory wastewater, domestic sewage, and 27. The method of applying for a patent (4), in which the dioxin present in the contaminated medium contains 4 to 8 gas atoms. 28. The method of claim 27 The dioxin present in the contaminated 61 200925272 medium is selected from the group consisting of 2,3,7,8_ tetrachlorodibenzo-Ocein, 1,2,3,7,8-five Gas dibenzo- _p_dioxine, 2,3,4,7,8-penta-dibenzo-p-dioxin, ι,2,3,4,7,8-hexa-dibenzo-Osin, 1, 2,3,6,7,8-hexa-dibenzo-p_dioxine, -5 2,3'4,6,7,8-hexa-dibenzo-$_dioxine, 1,2,3,7 ,8,9-six-gas dibenzo- corpse- Ossin, 1,2,3,4,6,7,8-seven gas dibenzo- _ _ dioxin, 1,2,3,4,7,8,9·seven gas dibenzo-household-Dioxin , eight gas diphenyl dioxin Q xin, 2,3,7,8-tetrazed dibenzofuran, 1,2,3,7,8-penta-dibenzofuran, 1,2,3,4 ,7,8·hexachlorodibenzofuran, i,2,3,6,7,8-hexaphenodiphenyl 1 〇 and astringent, l2,3,7,8,9. hexabenzodibenzopyrene a sulphur, 1, 2, 3, 4, 6, 7, 8-seven gas and a sigma-methane, an octa-benzoic benzopyrene, and combinations thereof. 29. The method of claim 24, The dioxin-like compound present in the contaminated enamel is selected from the group consisting of desertified dioxin, poly-di-dimethicone, poly-biphenyl, and a combination thereof. ❹ 3〇^ The method of claim 29, wherein the dioxin compound is brominated dioxin and contains 4 to 8 bromine atoms. 1 · The method of claim 30 of claim 4, the + dynasty is selected from 2 In the following group: 2,3,7,8-four desert dibenzo-p-dioxin, U,3,7,8-pentabromodibenzo-Ocein, hexabromodi- and dioxins, 1,2 ,3,7,8,9 - Hexabromo 2 stupid and _/?_Dioxin, 2'3,7,8·Four desert dibenzopyrene, 123,7,8_Five desert dibenzo-bite, 2,3,4,7, 8-Homobromodibenzofuran, and a combination thereof. 32. The method of claim 29, wherein the dioxin compound 62 200925272 is a polybrominated diphenyl ether and contains 2 to ι of a bromine atom. 33. The method of claim 32, wherein the polybrominated diphenyl ether is selected from the group consisting of 2,4-dibromodiphenyl ether and 4,4'-dibromodiphenyl ether. , 2,2',4-tri-diphenyl ether, 2,4,4,-tri-diphenyl ether, 2,2',4,5'-. 5 tetrabromodiphenyl ether, 2,3,, 4,6-tetrabromodiphenyl ether, 2,2,4,4'-tetrabromodiphenyl ether, 2,3,,4,4,-tetraphenylene, 3,3,,4, 4,_tetrabromodiphenyl ether, 2,2',4,4',6-pentabromodiphenyl ether, 2,3',4,4',6-pentabromodiphenyl ether, ◎ 2,2' , 4,4',5-pentabromodiphenyl ether, 2,2',3,4,4,-pentabromodiphenyl ether, 3,3',4,4',5-pentabromodiphenyl ether, 2,2,,4,41,5,6,_hexabromodiphenyl ether, 10 2,2',4,4',5,5'-hexabromodiphenyl ether, 2,2',3,4 ,4,6-hexabromodiphenyl ether, 2,2',3,4,4',6'-hexabromodiphenyl ether, 2,2',3,4,4',5,-hexabromo Diphenyl ether, 2,3,3',4,4',5-hexabromodiphenyl ether, 2,2',3,4,4',6,6'-heptabromodiphenyl ether, 2,2 ',3,4,4',5',6-heptabromodiphenyl ether, 2,3,3',4,4,5,6-heptbromodiphenyl bond, 2,2^3,3 ^4,4,,6,6^eight desert, diphenyl hydrazine 2,2,,3,4,4',5,5',6-octabromo-15 diphenyl ether, 2,2,3,3,4,4,5,6'-octaBDE Ether, ® 2,2',3,3',4,5,5',6,6'-nonabrominated diphenyl ether, 2,2,3,3,4,4,5,6, 6,-nonabrominated diphenyl ether, decabromodiphenyl ether, and combinations thereof. 34. The method of claim 29, wherein the dioxin compound is polyglycol and contains 4 to 7 gas atoms. 35. The method of claim 34, wherein the polyglycol is selected from the group consisting of 3,3', 4,4,-tetrachlorobiphenyl, 3, 4, 4, , 5-tetrahydrobiphenyl, 2,3,3',4,4'-five-biphenyl, 2,3,4,4,,5-penta-biphenyl, 2,3',4,4' , 5-penta-biphenyl, 2,3,4,4,5-pentachlorobiphenyl, 3,3,,4,4,,5_ five-phase biphenyl, 2,3,3,,4, 4,,5-hexachlorobiphenyl, 2,3,3,,4,4,,5,-six gas phase 63 200925272 Ben, 2,3,,4,4.,5,5,-six gas Benzene, 3,3,4,4,5,5,_hexa-biphenyl, 2'3'3'4'4'5'5-biphenyl, and combinations thereof. 36. The method of claim 24, wherein the cyclarene aromatic hydrocarbon present in the contaminated material has 2 to 6 benzene rings. 5 Ο 10 15 20 37. The method of claim 4, wherein the polycyclic aromatic hydrocarbons present in the contaminated medium are selected from the group consisting of: naphthalene, dangerous, Dihydrogen, phenanthrene, phenanthrene, onion, propylidene, ratio, benzo (a) onion, chopsticks, stupid and (b) alkadiene, benzo (k) propylene, and stupid (4) You, Festival and (l, 2, 3-cd), dibenzo (a, h) onion, benzo (g, h, i) oxime, and combinations thereof. 38. The method of claim 24, wherein the microbial agent is combinable with at least one microorganism selected from the group consisting of microorganisms capable of removing environmental pollutants: microorganisms of Pseudomonas, dehalogenation Microorganisms of the genus Cocci, microorganisms of the genus Candida, microorganisms of the genus Escherichia, microorganisms of the genus Aeromonas, microorganisms of the genus Rhizobium, microorganisms of the genus Lezmomonas, microorganisms of the genus Arthrobacter, Fula Microorganisms of the genus Dictyobacter, microorganisms of the genus Flavobacterium, and microorganisms of the genus Bacillus. 39. The method of claim 38, wherein the microorganism capable of removing environmental pollutants is selected from the group consisting of Pseudomonas resin Pseudomonas strain CA10, Pseudomonas ferosus PH- 03. Dehalomyces sp. strain CBDB1, Pseudomonas mendocensis ferm P-18187, Candida viswanathii, and their paper. 64 200925272 Sequence Listing <110> National University of Sun Yat-Sen University of Science and Technology<120> Microbial Reagent 5 for removing dioxin pollutants present in a contaminated medium and its use method, <130> NSYSU <160&gt 3 <170> Patent In version 3.4 Ο 10 15 Ο <210> 1 <211> 20 <212> DNA <213> Artificial <220><223> For PCR amplification of bacterial strain The universal forward primer of the 16S rRNA gene F1 <400> 1 attgaacgct ggcggcaggc 20 <210> 2 <211> 20 <212> DNA <213> Artificial <220><223> A general reverse primer for amplifying a 16S rRNA gene of a bacterial strain R1 20 200925272 <400> 2 cccagtcatg aatcactccg 20 5 <210> 3 <211> 1453 <212> DNA <213> Mendoza pseudomons Bacteria NSYSU <400> 3 at tgaacgct ggcggcaggc t taacacatg caagtcgagc ggatgaaggg agcttgctcc 60 ctgat t tagc ggcggacggg tgagtaatgc ct aggaatet gcctggtagt gggggataac 120 gt t ccgaaag gaacgctaat accgcat acg tcctacggga gaaagcaggg gaccttcggg 180 cct tgcgcta tcagatgagc ctaggtcgga ttagctagtt ggtgaggtaa tggctcacca 240 aggcgacgat ccgtaactgg tctgagagga tgatcagtca cactggaact gagacacggt 300 ccagactcct acgggaggca gcagtgggga atat tggaca atgggcgaaa gcctgatcca 360 gccatgccgc gtgtgtgaag aaggtct teg gattgtaaag cact t taagt tgggaggaag 420 ggcat taacc taatacgt ta gtgt 11 tgac gt taccaaca gaataageae cggcaact tc 480 gtgccagcag ccgcggtaat acgaagggtg caagcgt taa teggaat tac tgggcgtaaa 540 gcgcgcgtag gtggttcgtt aagt tggatg tgaaagcccc gggctcaacc tgggaactgc 600 atccaaaact ggcgagctag agtacggtag agggtggtgg aat t tcctgt gtagcggtga 660 aatgcgtaga tataggaagg aacaccagtg gegaaggega ccacctggac tgatactgac 720 actgaggtgc gaaagcgtgg ggagcaaaca ggattagata ccctggtagt ccacgccgta 780 aacgatgtca actagccgtt ggaatccttg agattttagt ggcgcagcta acgcat taag 840 t tgaccgcct ggggagtacg gccgcaaggt taaaactcaa atgaattgac gggggcccgc 900 acaagcggtg gagcatgtgg tttaattega agcaacgcga agaaccttac ctggcct tga 960 10 15 Ο lanthanum 20 2 200925272 catgctgaga actttccaga gatggat tgg tgccttcggg aactcagaca caggtgctgc 1020 atggctgtcg tcagctcgtg tcgtgagatg tgggttaagt cccgtaacga gcgcaaccct 1080 tgtccttagt taccagcacc tcgggtgggc actctaagga gactgccggt gacaaaccgg 1140 aggaaggtgg ggatgacgtc aagtcatcat ggccct tacg gccagggcta cacacgtgct 1200 acaatggtcg gtacaaaggg t tgccaagcc gcgaggtgga gctaatccca taaaaccgat 1260 cgtagtccgg atcgcagtct gcaactcgac tgcgtgaagt cggaatcgct agtaatcgtg 1320 aatcagaatg tcacggtgaa tacgttcccg ggcct tgtac acaccgcccg tcacaccatg 1380 ggagtgggtt gctccagaag tagctagtct aacct tcggg aggacggt ta ccacggagtg 1440 at tcatgact ggg 1453 10 3