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Variovarax strains capable of degrading methyl tert-butyl either and their use

A technology of methyl tert-butyl ether and phage strains, applied in the direction of microorganism-based methods, bacteria, methods using bacteria, etc., can solve the problems of no description of bacterial strains, low efficiency and slow biodegradation methods, etc.

Inactive Publication Date: 2005-01-26
克莱沃尔有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no bacterial strains capable of utilizing MTBE as their sole carbon and energy source have been described
[0011] Thus, because pollutants produced by fuels and their additives, such as fuel oxide ethers in soil and groundwater, cannot be completely avoided, and biodegradation methods are still rather inefficient and slow, there is still a need for more efficient and faster methods of degrading these pollutants. biological method

Method used

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  • Variovarax strains capable of degrading methyl tert-butyl either and their use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Isolation and identification of bacterial strains

[0029] The strain of the present invention was isolated from activated sludge by selective enrichment using MTBE as the sole carbon source and energy source. To 1 liter of CLM medium (1g K 2 HPO 4 ·3H 2 O, 0.25g NaH 2 PO 4 .2H 2 O, 0.1g (NH 4 ) 2 SO 4 , 0.05gMgSO 4 ·7H 2 O and Ca(NO 3 ) 2 4H 2 O dissolved in 1 liter of distilled or deionized water) was added to an airtight flask with MTBE (10 μl) and 50 ml of sludge and incubated statically at 22°C. After the substrate disappeared (4 months) more MTBE was added. After 6 months, the mixed enrichment culture was suspended in 60 ml of CLM medium containing yeast extract, trace elements and MTBE (10 μl) and incubated statically at 22°C (1 month). (10 μl / 100 ml), trace elements and vitamins in CLM medium were diluted 1:5 (v / v) and incubated until turbidity was visible. A maintenance culture (called CL-EMC-1 ) was inoculated at intervals of 1.5-2 m...

Embodiment 2

[0080] Example 2 Degradation of MTBE and TBA on Laboratory Scale

[0081] Growth of strain JV-1 on MTBE and CL-3 on TBA was studied under laboratory conditions in 2 liters of airtight flasks with Teflon-wrapped stoppers containing 0.2 liters of medium. The composition that is used for enrichment and the basic salt culture medium of cultivating bacterium of the present invention is as follows (g / l distilled water or deionized water): K 2 HPO 4 ·3H 2 O, 1; NaH 2 PO 4 2H 2 O, 0.25; (NH 4 ) 2 SO 4 , 0.1; MgSO 4 ·7H 2 O, 0.05; Ca(NO 3 ) 2 4H 2 O, 0.02; FeCl 3 ·6H 2 O, 0.002, pH7.0-7.3. The medium also contains the following elements (mg / l): H 3 BO 3 , 2; FeSO 4 ·7H 2 O, 2; Na 2 SeO 3 ·5H 2 O, 1; Na 2 MoO 4 2H 2 O, 1; CoCl 2 ·6H 2 O, 1; MnSO 4 2H 2 O, 0.5; ZnSO 4 ·7H 2 O, 0.5; AlCl 3 ·6H 2 O, 0.05; NiCl 2 ·6H 2 O, 0.02; CuSO 4 ·7H 2 O, 0.01, pH 7.0-7.3. The medium was sterilized at 121°C for 20 minutes. Vitamin stock solution as follows (mg / l d...

Embodiment 3

[0082] Embodiment 3 studies on the gas chromatography mass spectrometry of new way

[0083] Use gas chromatography-mass spectrometry (GC-MS) (with the methylsiloxane capillary column (50m * 0.2mm; 0.5μm film thickness, AgilentTechnologies, U.S.A) equipped with HP5973 mass selective detector and PONA crosslinking) analysis from the embodiment 2 MTBE, TBF and TBA in the obtained culture fluid samples. The oven temperature was held at 35°C for 15 minutes, then ramped at 10°C / min to 70°C, held at that temperature for 3 minutes then ramped at 20°C / min to 250°C and held at that temperature for 5 minutes. The carrier gas (helium) was maintained at a constant column flow rate of 0.5 ml / min. Samples (m / z=25 to 200) were analyzed by electron ionization (70 eV) with full scan monitoring. Mass spectral interpretation for compound identification was performed by official standards and a database in GC-MS software (Wiley275).

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Abstract

Water-soluble ethers used as octane number enhancers in gasoline are disturbing components in soils and groundwater even in small amounts due to their strong taste and odour. This invention relates to biological purification of gasoline-contaminated soils and groundwater, more specifically to Variovorax strains being able to degrade ethers and their degradation products and to a mixed bacterial population comprising said bacteria. Further this invention relates to a process for bacterial degradation of ethers and their degradation products and to the use of one or more Variovorax strains of the invention in purifying contaminated soil or water.

Description

field of invention [0001] The present invention relates to bacteria capable of degrading ethers and their degradation products. More precisely, the invention relates to polyphage (Variovorax) strains, to mixed bacterial flora, and also to methods for the bacterial degradation of ethers and their degradation products. The invention also relates to the use of one or more of said strains for decontamination of contaminated soil and water. Background technique [0002] Ethers such as ethyl tert-butyl ether (ETBE), tert-amyl methyl ether (TAME) and methyl tert-butyl ether (MTBE) are widely used as octane boosters in unleaded gasoline. In Finland, MTBE is blended into gasoline at 9 to 13 volume percent. MTBE (which replaced lead in Europe in the 1980s) is more effective as an octane booster (fuel oxidizer) than aromatics and is safer to use from an environmental point of view. Some tertiary alcohols, such as tert-butyl alcohol (TBA) and tert-amyl alcohol (TAA), are also potenti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B09C1/00B09C1/10C02F3/34
CPCC02F2101/34C02F3/34B09C1/10B09C1/002C12R1/05C12N1/205C12R2001/05
Inventor J·沃蒂拉G·扎伊采夫
Owner 克莱沃尔有限公司
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