WO1988001255A1 - Process for the degradation of coal tar constituents by white rot fungi - Google Patents

Process for the degradation of coal tar constituents by white rot fungi Download PDF

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Publication number
WO1988001255A1
WO1988001255A1 PCT/US1987/002084 US8702084W WO8801255A1 WO 1988001255 A1 WO1988001255 A1 WO 1988001255A1 US 8702084 W US8702084 W US 8702084W WO 8801255 A1 WO8801255 A1 WO 8801255A1
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WO
WIPO (PCT)
Prior art keywords
coal tar
fungus
degradation
source
carbon
Prior art date
Application number
PCT/US1987/002084
Other languages
French (fr)
Inventor
Steven D. Aust
John A. Bumpus
Original Assignee
Michigan State University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Michigan State University filed Critical Michigan State University
Priority to AT87905864T priority Critical patent/ATE100072T1/en
Priority to DE3788794T priority patent/DE3788794T2/en
Publication of WO1988001255A1 publication Critical patent/WO1988001255A1/en

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi

Definitions

  • the present invention relates to a process for the degradation of polycyclic aromatic compounds found in coal tar mixtures.
  • the present invention relates to the biodegradation of coal tar.
  • microorganisms are not able to cause complete biodegradation of polycyclic aromatic compounds.
  • a few species of bacteria and fungi are able to cause extensive degradation of relatively simple polycyclic aromatic compounds (biphenyl and naphthalene, for example) .
  • polyaromatic compounds such as benzo (a)pyrene, a compound composed of five aromatic rings is resistant to biodegradation by most, but not all microorganisms and is acknowledged to be a persistent environmental pollutant.
  • microorganisms are able to degrade certain polycyclic aromatic compounds, their usefulness in microbial waste treatment systems is often limited for the following reasons: (1) degradation is often not extensive, that is to say degradation does not always proceed to carbon dioxide as an end product of metabolism; (2) some metabolites may be even more toxic than the parent compound and as environmentally persistent and (3) many microorganisms used in waste treatment systems are not able to degrade a wide variety of organopollutants, and thus mixtures of microorganisms often have to be used instead of a single species.
  • the process comprises mixing of coal tar or polycyclic aromatic constituents found in coal tar with white rot fungi and appropriate nutrients under aerobic conditions.
  • the mixture may also be supplemented by providing additional lignin degrading oxidases and hydrogen peroxide.
  • the coal tar constituents, including polycyclic aromatic compounds, are degraded to carbon dioxide.
  • this invention relates to the ability of white rot fungi (e.g. Phanerochaete chrysosporium) to degrade normally hard-to-degrade polycyclic aromatic compounds found in coal tar.
  • white rot fungi e.g. Phanerochaete chrysosporium
  • organopollutants such as single aromatic ring compounds will also be degraded by white rot fungi.
  • 14 CO 2 has been found to be the ultimate degradation product in all cases tested, it is apparent that any potentially toxic intermediates from the degradation of the coal tar are also degraded.
  • the process is very non-specific and capable of completely degrading a wide variety of organic molecules and thus is well suited for treatment of coal tar contaminated materials.
  • treatment systems can consist of adding large amounts of these enzymes to effect initial oxidative degradation of the pollutants in the coal tar. Supplemental enzyme treatment can also take place in the presence of white rot fungi or other microorganisms.
  • Example 1 Because coal tar is a complex mixture containing well over 50 compounds, it was necessary to select representative compounds for evaluation. If these compounds are degraded, the rest will be degradable. The compounds studied and the reasons for their selection are listed below:
  • Each compound was incubated in aqueous culture as described by Bumpus, et al. in U.S. application Serial No. 702,944 filed February 19, 1985.
  • the culture medium was nutrient nitrogen deficient and was composed of 56 mM glucose, 1.2 mM ammonium tartrate, 20 mM dimethyl succinate buffer, pH 4.2-4.3.
  • Cultures (10 ml) also contained trace amounts of thiamine and minerals as described by Kirk, T.K. et al. Arch. Microbiol. 117, 277-285 (1977).
  • the initial concentration of 14 C-p-cresol, 14 C-2-methylnaphthalene and 14 C-phenanthrene was 5 nmoles/culture.
  • the initial concentration of 14 C-benzo(a)pyrene was 1.25 nmoles/culture. Evolution of 14 CO 2 from each coal tar constituent was monitored as described by Kirk et al. and in Serial No. 702,944. As can be seen from Figures 1 and 2, the representative compounds are degraded up to the percentage levels indicated. The medium can be supplemented with more nutrient medium including particularly an additional carbon source and the process reinitiated until the degradation is complete. As can be seen 2-methylnaphthalene is the most difficult to degrade and will therefore require more repeated treatments.
  • the carbon source can be glucose, cellulose or any other compound which provides energy to the white rot fungus.
  • the culture media for enabling the white rot fungus to degrade coal tar or derivatives thereof are well known to those skilled in the art.
  • the process can be performed in the field or in a reaction vessel where large amounts of liquid are used. It is preferred to use a reaction vessel to prevent leaching into the ground water.

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Environmental & Geological Engineering (AREA)
  • Botany (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Water Supply & Treatment (AREA)
  • Hydrology & Water Resources (AREA)
  • Soil Sciences (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

A process for degrading coal tar or polycyclic aromatic constituents thereof using lignin degrading oxygenase fungal enzymes and hydrogen peroxide. The fungus is preferably living and is most preferably a white rot fungus such as Phanerochaete chrysosporium. The process provides a means for the treatment of coal tar wastes.

Description

PROCESS FOR THE DEGRADATION OF COAL TAR CONSTITUENTS BY WHITE ROT FUNGI
Cross-Reference to Related Application
This application is a continuation-in-part of application Serial No. 702,944 filed February 19, 1985. Background of the Invention (1) Summary of the Invention
The present invention relates to a process for the degradation of polycyclic aromatic compounds found in coal tar mixtures. In particular the present invention relates to the biodegradation of coal tar. (2) Prior Art
Most microorganisms are not able to cause complete biodegradation of polycyclic aromatic compounds. A few species of bacteria and fungi are able to cause extensive degradation of relatively simple polycyclic aromatic compounds (biphenyl and naphthalene, for example) . On the other hand, polyaromatic compounds such as benzo (a)pyrene, a compound composed of five aromatic rings is resistant to biodegradation by most, but not all microorganisms and is acknowledged to be a persistent environmental pollutant. Although some microorganisms are able to degrade certain polycyclic aromatic compounds, their usefulness in microbial waste treatment systems is often limited for the following reasons: (1) degradation is often not extensive, that is to say degradation does not always proceed to carbon dioxide as an end product of metabolism; (2) some metabolites may be even more toxic than the parent compound and as environmentally persistent and (3) many microorganisms used in waste treatment systems are not able to degrade a wide variety of organopollutants, and thus mixtures of microorganisms often have to be used instead of a single species. The ability of a microorganism to degrade a broad spectrum of organopollutants is important in the treatment of sites contaminated with coal tar, a complex mixture of aromatic and polycyclic aromatic compounds that are by-products of the coking and coal gasification industries. For years coal tar wastes have been deposited in unlined dump sites where they have contaminated soils and ground water. The problem is to use microorganism to degrade the constituents found in coal tar. Objectives
It is therefore an objective of the present invention to provide a process for the microbial degradation of the constituents of coal tar and polycyclic aromatic constituents thereof. Further, it is an objective of the present invention to provide a process which is simple and economical. These and other objectives will become increasingly apparent by reference to the following description and to the drawings. In The Drawings Figures 1 and 2 are graphs showing the ability of a white rot fungus (i.e. Phanerochaete chrysosporium) to mineralize representative 14C-labeled compounds which are found in coal tar. General Description The present invention relates to a process for biodegrading coal tar or constituents thereof. The process comprises mixing of coal tar or polycyclic aromatic constituents found in coal tar with white rot fungi and appropriate nutrients under aerobic conditions. The mixture may also be supplemented by providing additional lignin degrading oxidases and hydrogen peroxide. The coal tar constituents, including polycyclic aromatic compounds, are degraded to carbon dioxide.
Specifically this invention relates to the ability of white rot fungi (e.g. Phanerochaete chrysosporium) to degrade normally hard-to-degrade polycyclic aromatic compounds found in coal tar. At the same time, easier-to-degrade organopollutants such as single aromatic ring compounds will also be degraded by white rot fungi. Furthermore, because 14CO2 has been found to be the ultimate degradation product in all cases tested, it is apparent that any potentially toxic intermediates from the degradation of the coal tar are also degraded. The process is very non-specific and capable of completely degrading a wide variety of organic molecules and thus is well suited for treatment of coal tar contaminated materials. Furthermore, because degradative enzymes are secreted by white rot fungi, treatment systems can consist of adding large amounts of these enzymes to effect initial oxidative degradation of the pollutants in the coal tar. Supplemental enzyme treatment can also take place in the presence of white rot fungi or other microorganisms. Specific Description
Example 1 Because coal tar is a complex mixture containing well over 50 compounds, it was necessary to select representative compounds for evaluation. If these compounds are degraded, the rest will be degradable. The compounds studied and the reasons for their selection are listed below:
(1) p-cresol - This monocyclic aromatic compound is an abundant component found in coal tar. It is relatively water soluble and, therefore, represents a major threat to ground water. This compound is representative of one (1) ring compounds found in coal tar and is relatively easily degraded. It is noted that degradation of p-cresol alone by P. chrysosporium has been demonstrated by others.
(2) 2-methylnaphthalene - This compound is an abundant component found in coal tar. It is slightly water soluble, and, therefore a threat to ground water. This compound, along with naphthalene makes up greater than 70% of the organopollutants in some coal tar contaminated sites. This compound is representative of two ring polycyclic compounds found in coal tar. It is noted that degradation of this compound by P. chrysosporium has not been reported by others.
(3) phenanthrene - This polycyclic compound is found in coal tar and is representative of a suspect carcinogenic compound. This compound is representative of three ring compounds found in coal tar. It is noted that degradation of this compound by P. chrysosporium has not been reported by others.
(4) benzo (a)pyrene - This polycyclic aromatic compound is found in coal tar and is representative of a carcinogenic compound. It is very difficult to degrade in the environment and is representative of five ring compounds found in coal tar. It is noted that the inventors were the first to demonstrate degradation of this compound by P. chrysosporium.
The degradation of various 14C-labeled coal tar constituents, i.e. p-cresol, phenanthrene, benzo (a)pyrene and 2-methylnaphthalene was accomplished separately in cultures of P. chrysosporium as shown in Figures 1 and 2. Degradation, as assayed by 14CO2 evolution was monitored for thirty (30) days at which time supplemental glucose was added and degradation was monitored for another 30 days (60 days total).
Each compound was incubated in aqueous culture as described by Bumpus, et al. in U.S. application Serial No. 702,944 filed February 19, 1985. The culture medium was nutrient nitrogen deficient and was composed of 56 mM glucose, 1.2 mM ammonium tartrate, 20 mM dimethyl succinate buffer, pH 4.2-4.3. Cultures (10 ml) also contained trace amounts of thiamine and minerals as described by Kirk, T.K. et al. Arch. Microbiol. 117, 277-285 (1977). The initial concentration of 14C-p-cresol, 14C-2-methylnaphthalene and 14C-phenanthrene was 5 nmoles/culture. The initial concentration of 14C-benzo(a)pyrene was 1.25 nmoles/culture. Evolution of 14CO2 from each coal tar constituent was monitored as described by Kirk et al. and in Serial No. 702,944. As can be seen from Figures 1 and 2, the representative compounds are degraded up to the percentage levels indicated. The medium can be supplemented with more nutrient medium including particularly an additional carbon source and the process reinitiated until the degradation is complete. As can be seen 2-methylnaphthalene is the most difficult to degrade and will therefore require more repeated treatments.
The carbon source can be glucose, cellulose or any other compound which provides energy to the white rot fungus. The culture media for enabling the white rot fungus to degrade coal tar or derivatives thereof are well known to those skilled in the art.
The process can be performed in the field or in a reaction vessel where large amounts of liquid are used. It is preferred to use a reaction vessel to prevent leaching into the ground water.
It is intended that the foregoing description be only illustrative and that this invention be limited only by the hereinafter appended claims.

Claims

WE CLAIM:
-1- A process for degrading a coal tar or a constituent thereof which comprises:
(a) providing the coal tar or a polycyclic aromatic constituent thereof mixed with fungal enzymes containing a lignin degrading oxygenase and with hydrogen peroxide under aerobic conditions; and
(b) degrading the coal tar or polycyclic aromatic constituents thereof to carbon dioxide.
-2-
The process of Claim 1 wherein the enzymes and hydrogen peroxide are provided by a fungus along with a carbon source enabling degradation of the coal tar or constituents thereof by the fungus.
-3-
The process of Claim 2 wherein the fungus is living.
-4- The process of Claim 3 wherein the fungus is a white rot fungus.
-5- The process of Claim 2 wherein the fungus has been grown in a culture medium containing a minimal nutrient nitrogen level.
-6- The process of Claim 7 wherein the culture media includes a source of carbon, thiamine and minerals for the fungus. -7- The process of Claim 6 wherein the source of carbon is glucose.
-8- The process of Claim 6 wherein the source of carbon is a carbohydrate.
-9-
The process of Claim 1 wherein the coal tar is mixed with soil from the environment and wherein the soil is mixed with the fungal enzymes and hydrogen peroxide.
-10The process of Claim 9 wherein the soil is mixe with a fungus as the source of the fungal enzymes and hydrogen peroxide.
-11- The process of Claim 1 performed in a reaction vessel.
PCT/US1987/002084 1986-08-22 1987-08-21 Process for the degradation of coal tar constituents by white rot fungi WO1988001255A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AT87905864T ATE100072T1 (en) 1986-08-22 1987-08-21 PROCESSES FOR DEGRADING COAL TAR COMPONENTS WITH POT FUNGI.
DE3788794T DE3788794T2 (en) 1986-08-22 1987-08-21 METHOD FOR DEGRADING CARBON SEED INGREDIENTS WITH rotten mushrooms.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US89900086A 1986-08-22 1986-08-22
US899,000 1986-08-22

Publications (1)

Publication Number Publication Date
WO1988001255A1 true WO1988001255A1 (en) 1988-02-25

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PCT/US1987/002084 WO1988001255A1 (en) 1986-08-22 1987-08-21 Process for the degradation of coal tar constituents by white rot fungi

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EP (1) EP0278973B1 (en)
CA (1) CA1325988C (en)
DE (1) DE3788794T2 (en)
WO (1) WO1988001255A1 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993019811A1 (en) * 1992-04-01 1993-10-14 Biotal Ltd. Biocatalysis
NL1000372C2 (en) * 1995-05-16 1996-11-19 Tno Aerobic removal of difficult organic cpds. from aq. waste or ground water

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101941703B1 (en) 2016-11-22 2019-04-11 한국에너지기술연구원 Production method of organic acid and high rank coal via biological low rank coal conversion
CN108941191A (en) * 2018-07-03 2018-12-07 上海松沅环境修复技术有限公司 A kind of repairing method of microorganism of oil-polluted soils

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3979283A (en) * 1974-09-25 1976-09-07 Bioteknika International, Inc. Microbial degradation of DDT

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3979283A (en) * 1974-09-25 1976-09-07 Bioteknika International, Inc. Microbial degradation of DDT

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Applied and Emvironmental Microbiology 43(5), May 1982, (Washington, DC), CERNIGLIA et al, "Glucuronide and Sulfate Conjugation in the Fungal Metabolism of Aromatic Hydrocarbons", pages 1070-1075. *
Applied and Environmental Microbiology 44(1), July 1982, (Washington, DC), COHEN et al, "Degragation of Coal by Fungi Polyporus Versicolor and Poria Monticola", pages 23-27. *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993019811A1 (en) * 1992-04-01 1993-10-14 Biotal Ltd. Biocatalysis
NL1000372C2 (en) * 1995-05-16 1996-11-19 Tno Aerobic removal of difficult organic cpds. from aq. waste or ground water

Also Published As

Publication number Publication date
CA1325988C (en) 1994-01-11
EP0278973A1 (en) 1988-08-24
EP0278973A4 (en) 1989-03-23
EP0278973B1 (en) 1994-01-12
DE3788794D1 (en) 1994-02-24
DE3788794T2 (en) 1994-05-19

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