DE3909324C1 - Microorganism and its use for microbial breakdown of petroleum hydrocarbons and mineral oil in contaminated soils, water and aerosols - Google Patents

Microorganism and its use for microbial breakdown of petroleum hydrocarbons and mineral oil in contaminated soils, water and aerosols

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Publication number
DE3909324C1
DE3909324C1 DE3909324A DE3909324A DE3909324C1 DE 3909324 C1 DE3909324 C1 DE 3909324C1 DE 3909324 A DE3909324 A DE 3909324A DE 3909324 A DE3909324 A DE 3909324A DE 3909324 C1 DE3909324 C1 DE 3909324C1
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Prior art keywords
petroleum hydrocarbons
water
microorganism
bacterial
aerosols
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Expired - Fee Related
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DE3909324A
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German (de)
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Des Erfinders Auf Nennung Verzicht
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Mikro - Bak Biotechnik 3338 Schoeningen De GmbH
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Mikro - Bak Biotechnik 3338 Schoeningen De GmbH
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    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/38—Pseudomonas
    • C12R2001/385—Pseudomonas aeruginosa

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

Abstract

The cleaning of soils, water and aerosols containing petroleum hydrocarbons is achieved by continuous addition of the bacterial mixed culture of two bacterial strains Rhodococcus erythropolis DSM 4670 and Pseudomonas aeruginosa DSM 4692. The aerobic breakdown takes place due to the formation of microorganism-specific surface-active biosurfactants which differ in their mode of action and which are produced by Rhodococcus erythropolis and Pseudomonas aeruginosa in the presence of petroleum hydrocarbons. The use of the mixed culture, in the case of substrate containing petroleum hydrocarbons, can take place in the on-site leasing process, or in in situ processes with the addition of air, pure oxygen or oxygen donors or in fixed bed, fluidised bed or submerged processes in controllable reactors with gas introduction in the presence of growth-maintaining nutrient salts. The amount of the mixed bacterial culture to be added depends on the amount of petroleum hydrocarbons in the soil, water and aerosol.

Description

Die Erfindung betrifft ein Verfahren zur Reinigung von mit Erdölkohlen­ wasserstoffen kontaminiertem Trägermaterial, Böden, Wasser und Aerosolen durch mikrobiellen Abbau mittels einer bakteriellen Mischkultur und einen bestimmten Mikroorganismus (vgl. Patentansprüche).The invention relates to a method for cleaning petroleum coal Hydrogen contaminated carrier material, soil, water and aerosols through microbial degradation using a bacterial mixed culture and a certain microorganism (see patent claims).

Mit dem vorgegebenen Einsatz einer Kohlenwasserstoff verwertenden bakteri­ ellen Mischkultur soll eine im Abbaugrad und -geschwindkeit steuer­ bare Reinigung der kontaminierten o. g. Bereiche ermöglicht werden, so daß ein sofortiger Einbau/Wiederverwendung des gereinigten Materials erfolgen kann.With the given use of a hydrocarbon utilizing bacteri The mixed culture should control the degree and speed of degradation bare cleaning of the contaminated o. g. Areas are made possible so that an immediate installation / reuse of the cleaned material can be done.

Bei den bisher angewandten Verfahren wie Bodendeponierung, Boden­ waschung, Verbrennung und Verklappung wird das kontaminierte Material lediglich verlagert. Bei derzeitig durchgeführten biologischen Ver­ fahren werden chemisch erzeugte grenzflächenaktive Verbindungen in Verbindung mit Bakterien eingesetzt. Diese Fremdstoffe sind nicht mikroorganismenspezifisch und daher uneffektiv und möglicherweise nicht vollständig metabolisierbar mit Auswirkungen auf die autochtone Fauna.In the previously used methods such as landfilling, soil Washing, incineration and dumping becomes the contaminated material just relocated. In the current biological ver drive chemically generated surfactants in Connection with bacteria used. These foreign substances are not microorganism specific and therefore ineffective and possibly not fully metabolizable with an impact on autochthonous Fauna.

Durch die nachfolgend beschriebene Erfindung werden eine Schadstoff­ verlagerung vermieden und die Bildung und Anhäufung von Nebenprodukten bei der Anwendung von nicht-mikroorganismusspezifischen Stoffen ausgeschlossen.By the invention described below, a pollutant relocation avoided and the formation and accumulation of by-products when using non-microorganism-specific substances locked out.

Erdölkohlenwasserstoffe sind ein Gemisch von Kohlenwasserstoffen unter­ schiedlicher chemischer Struktur und Eigenschaft. Sie sind nur sehr gering wasserlöslich und mit Wasser nicht mischbar.Petroleum hydrocarbons are a mixture of hydrocarbons different chemical structure and property. You are just very slightly water-soluble and immiscible with water.

Der mikrobielle Abbau der Kohlenwasserstoffe als Kohlenstoff- und Energiequelle ist wesentlich abhängig von der Löslichkeit der Kohlen­ wasserstoffe im Wasser und von der Art der enzymatischen Abbauwege. The microbial degradation of hydrocarbons as carbon and The energy source is largely dependent on the solubility of the coals Hydrogens in water and the type of enzymatic degradation pathways.  

In Gegenwart von Erdölkohlenwasserstoffen kommt es zur Bildung mikroorganismeneigener, spezifischer grenzflächenaktiver Tenside, die den mikrobiellen Abbau der Erdölkohlenwasserstoffe beschleunigen.Formation occurs in the presence of petroleum hydrocarbons microorganism-specific, specific surface-active surfactants, that accelerate the microbial degradation of petroleum hydrocarbons.

Die Erfindung macht Gebrauch von der Besonderheit der in der Misch­ kultur zum Abbau von Erdölkohlenwasserstoffen enthaltenden Bakterien­ stämme Rhodococcus erythropolis und Pseudomonas aeruginosa.The invention makes use of the peculiarity of mixing Culture for the degradation of bacteria containing petroleum hydrocarbons strains Rhodococcus erythropolis and Pseudomonas aeruginosa.

Der Bakterienstamm Rhodococcus erythropolis DSM 4670 besitzt eine lipophile Zelloberfläche. Die dadurch verursachte Affinität zum Kohlen­ wasserstoff führt in der Folge zu einem direkten Kontakt zwischen Bakterienzelle und Kohlenwasserstoff. Das Wachstum der Bakterien findet an der Grenzfläche zwischen Kohlenwasserstoff und Wasser im Kohlenwasserstoff statt.The bacterial strain Rhodococcus erythropolis DSM 4670 has one lipophilic cell surface. The affinity to coal caused by this As a result, hydrogen leads to direct contact between Bacterial cell and hydrocarbon. The growth of the bacteria takes place at the interface between hydrocarbon and water in the Hydrocarbon instead.

Während dieses Wachstumsprozesses bewirkt die Bildung eines lipophilen grenzflächenaktiven Agenz zusätzlich eine Emulgierung bzw. Mizellierung des Kohlenwasserstoffs.During this growth process causes the formation of a lipophilic surface-active agent additionally an emulsification or micellization of the hydrocarbon.

Die dadurch bewirkte Oberflächenvergrößerung des lipophilen Anteils in der Wasserphase beschleunigt den mikrobiellen Abbau.The resulting surface enlargement of the lipophilic portion in the water phase accelerates microbial degradation.

Der Stamm Pseudomonas aeruginosa DSM 4672 besitzt eine hydrophile Zelloberfläche.The Pseudomonas aeruginosa DSM 4672 strain is hydrophilic Cell surface.

In Gegenwart von Kohlenwasserstoffen werden von ihnen hydrophile grenzflächenaktive Substanzen gebildet, die zu einer Emulgierung des Kohlenwasserstoffs in der Wasserphase führen.In the presence of hydrocarbons they become hydrophilic surface-active substances are formed, which lead to an emulsification of the Lead hydrocarbon in the water phase.

Die genannten Bakterienstämme bauen Kohlenwasserstoffe, insbesondere aromatische Kohlenwasserstoffe, durch unterschiedliche Ringspaltungs­ mechanismen (Meta-/Ortho-/Spaltung) ab. Hierdurch wird die Wahrschein­ lichkeit der Anhäufung von "dead-end-Produkten" während des Abbaus erheblich verringert.The bacterial strains mentioned build hydrocarbons, in particular aromatic hydrocarbons, through different ring splitting mechanisms (meta / ortho / cleavage). This will make the probability Ability to accumulate dead-end products during mining significantly reduced.

Die Anzucht der Bakterien erfolgt auf einem Mineralsalzmedium, bei dem als einzige Kohlenstoff- und Energiequelle ein n-Paraffingemisch der KohlenstoffkettenlängeThe bacteria are grown on a mineral salt medium in which an n-paraffin mixture as the only carbon and energy source Carbon chain length

C13-C15 vorliegt: (g/L) : NH4NO3 : 1,0; K2HPO4 : 0,25; MgSO4 × 7 H2O : 0,1; CaCl2 × 2 H2O : 0,005; FeCl3 × 6 H2O : 0,005;
Citronensäure: 0,05; Hefeextrakt: 0,05; Norpar 13 : 2,1; Leitungs­ wasser.C 13 -C 15 is: (g / L) : NH 4 NO 3 : 1.0; K 2 HPO 4 : 0.25; MgSO 4 x 7 H 2 O: 0.1; CaCl 2 x 2 H 2 O: 0.005; FeCl 3 x 6 H 2 O: 0.005;
Citric acid: 0.05; Yeast extract: 0.05; Norpar 13: 2.1; Tap water.

Die Kultivierung erfolgt aerob bei einer Temperatur von 22-28°C und einem pH-Wert von 6,5-7,5. Die Anzuchtdauer beträgt 72 Stunden.The cultivation takes place aerobically at a temperature of 22-28 ° C and a pH of 6.5-7.5. The growing period is  72 hours.

Die beschriebene bakterielle Mischkultur (vgl. Anspruch 2), nachstehend auch MIKRO-BAK KW 03 genannt, findet Anwendung zur Dekontamination von mit Erdölkohlenwasserstoffen kontaminierten Böden, Wasser und Aerosolen.The described bacterial mixed culture (see claim 2), below also called MIKRO-BAK KW 03, is used for the decontamination of Soils, water and aerosols contaminated with petroleum hydrocarbons.

Die durch diese Erfindung erreichten Vorteile liegen in der gezielten Kombination der verwandten Bakterien aufgrund der unterschiedlichen Lipophilie der Zelloberflächen dieser Bakterien und der daraus sich ergebenden Affinität zum Erdöl-Kohlenwasserstoffgemisch. Weitere Vorteile ergeben sich aus der sich ergänzenden Wirkung der unter­ schiedlichen grenzflächenaktiven Agenzien mit der daraus resul­ tierenden Ablösung von Kohlenwasserstoffen von Partikeln und Emulsions­ eigenschaften sowie auch aus der unterschiedlichen Art der enzyma­ tischen Abbauwege bei der Verwertung von Kohlenwasserstoffgemischen als Kohlenstoff- und Energiequelle.The advantages achieved by this invention lie in the targeted Combination of related bacteria due to the different Lipophilicity of the cell surfaces of these bacteria and the resulting ones resulting affinity for the petroleum-hydrocarbon mixture. Further Advantages arise from the complementary effect of the below different surface-active agents with the resultant detachment of hydrocarbons from particles and emulsions properties as well as from the different type of enzyma table ways of dismantling when recycling hydrocarbon mixtures as a source of carbon and energy.

AnwendungsbeispieleExamples of use

  • 1. 10 m3 Mineralöl kontaminierten Erdreiches (11 000-17 500 mg Mineralöl/kg Boden) werden in einer Schichthöhe von ca. 25 cm auf einer ölundurchlässigen Folie ausgebreitet (Gesamtfläche ca. 40 m2) und einmalig mit 5 l Bakterienmischkultur MIKRO-BAK KW 03 besprüht. Die Bodentemperatur muß größer oder gleich 13°C und nicht höher oder gleich 37°C sein. Die Bodenfeuchte ist während der Dekontaminations­ phase auf größer als oder gleich 45% zu halten.1. 10 m 3 mineral oil contaminated soil (11 000-17 500 mg mineral oil / kg soil) are spread out in a layer height of approx. 25 cm on an oil-impermeable film (total area approx. 40 m 2 ) and once with 5 l mixed bacteria culture MIKRO- BAK KW 03 sprayed. The floor temperature must be greater than or equal to 13 ° C and not higher than or equal to 37 ° C. The soil moisture must be kept at or above 45% during the decontamination phase.
  • 2. Auf 10 m3 mit 13,85 g Mineralöl kontaminierten Wassers werden zur Dekontamination 1,85 l Bakterienmischkultur MIKRO-BAK KW 03 (10% des Mineralölvolumens) aufgesprüht.
    Die Anwendung ist bis zur Unterschreitung der behördlichen Grenzwerte durchzuführen.    2. 1.85 l of mixed bacterial culture MIKRO-BAK KW 03 (10% of the mineral oil volume) are sprayed onto 10 m 3 of water contaminated with 13.85 g mineral oil.
    The application must be carried out until the official limits are undershot.
  • 3. Mineralölhaltige Aerosole werden an beliebigem Trägermaterial abge­ schieden und dort mit der Bakterienmischkultur MIKRO-BAK KW 03 im Verhältnis 1 : 10 zur Trägeroberfläche besprüht.3. Mineral oil-containing aerosols are removed from any carrier material divorce and there with the mixed bacteria culture MIKRO-BAK KW 03 in Sprayed ratio 1:10 to the carrier surface.

Claims (2)

1. Rhodococcus erythropolis DSM 4670.1. Rhodococcus erythropolis DSM 4670. 2. Verwendung des Stammes nach Anspruch 1 in Mischkultur mit Pseudomonas aeruginosa DSM 4672 zum mikrobiellen Abbau von Erdölkohlenwasserstoffen und Mineralölen.2. Use of the strain according to claim 1 in mixed culture with Pseudomonas aeruginosa DSM 4672 for the microbial degradation of petroleum hydrocarbons and mineral oils.
DE3909324A 1989-03-17 1989-03-17 Microorganism and its use for microbial breakdown of petroleum hydrocarbons and mineral oil in contaminated soils, water and aerosols Expired - Fee Related DE3909324C1 (en)

Priority Applications (1)

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DE3909324A DE3909324C1 (en) 1989-03-17 1989-03-17 Microorganism and its use for microbial breakdown of petroleum hydrocarbons and mineral oil in contaminated soils, water and aerosols

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DE3909324A DE3909324C1 (en) 1989-03-17 1989-03-17 Microorganism and its use for microbial breakdown of petroleum hydrocarbons and mineral oil in contaminated soils, water and aerosols

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4102405A1 (en) * 1991-01-28 1992-08-20 Heinrich Weseloh Fa Procedure for establishing load-fill dump for biologically treatable waste - by placing waste in ground depression in layers each provided with drainage pipes and addn. of nutrients
EP2051784A2 (en) * 2006-07-31 2009-04-29 INDIAN OIL CORPORATION Ltd. Method for bio-assisted treatment of hydrocarbon contaminated soil

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ATCC, Catalogue of Strains I, 1982, S. 168,182 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4102405A1 (en) * 1991-01-28 1992-08-20 Heinrich Weseloh Fa Procedure for establishing load-fill dump for biologically treatable waste - by placing waste in ground depression in layers each provided with drainage pipes and addn. of nutrients
EP2051784A2 (en) * 2006-07-31 2009-04-29 INDIAN OIL CORPORATION Ltd. Method for bio-assisted treatment of hydrocarbon contaminated soil
EP2051784A4 (en) * 2006-07-31 2011-07-06 Indian Oil Corp Ltd Method for bio-assisted treatment of hydrocarbon contaminated soil

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