US5616162A
Expired / Lapsed
US
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Biological system for degrading nitroaromatics in water and soils
Assignee: Idaho Research Foundation Inc
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Filing: 1995-10-20
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Granted: 1997-04-01
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Expires: 2015-10-20
Abstract
Abstract Novel methods for biodegrading nitroaromatic compounds present as contaminants in soil or water using microorganisms are disclosed. Water is treatable directly; dry soil is first converted into a fluid medium by addition of water. The preferred method comprises two stages, each employing microorganisms: a fermentative stage, followed by an anaerobic stage. The fermentative stage is rapid, wherein an inoculum of aerobic and/or facultative microorganisms ferments a carbohydrate added to the fluid medium, exhausting the oxygen in the fluid medium and thereby inhibiting oxidative polymerization of amino by-products of the nitroaromatics. In the subsequent anaerobic stage, an inoculum of a mixed population of anaerobic microorganisms completes the mineralization of the contaminant nitroaromatics, using the remaining carbohydrate as a carbon and energy source. Preferably, the carbohydrate is a starch and the aerobic and/or facultative microorganisms are amylolytic, which cleave the starch at a moderate rate throughout both stages, ensuring a sustained supply of metabolizable carbohydrate. The microorganisms are preferably selected to be resistant to the types and concentrations of nitroaromatics present as contaminants.
Inventors
- 👤 Donald L. Crawford
- 👤 Todd O. Stevens
- 👤 Ronald L. Crawford
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 12:28
CPC Classifications
C; C02; C02F; C02F3/00; C02F3/34; A; A62; A62D; A62D3/00; A62D3/02; B; B09; B09C; B09C1/00; B09C1/10; C02F3/02; C02F3/12; C02F3/1205; C02F3/1231; C02F3/28; C02F3/2806; C02F3/286; C02F3/30; C12; C12N; C12N1/00; C12N1/20; C12N1/205; A62D2101/00; A62D2101/20; A62D2101/26; A62D2101/28; C02F2101/00; C02F2101/003; C12R; C12R2001/00; C12R2001/01; C12R2001/145; C12R2001/22; Y; Y02; Y02E; Y02E50/00; Y02E50/30; Y02W; Y02W10/00; Y02W10/10; Y10; Y10S; Y10S71/00; Y10S71/903
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
108 words
A method for biodegrading a nitroaromatic compound present as a contaminant in a sample, comprising the steps of: (a) providing a sample comprising a nitroaromatic compound; (b) adding to the sample an inoculum comprising a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (c) producing anaerobic conditions in the sample; and (d) maintaining the anaerobic conditions in the sample for a time that is sufficient for the nitroaromatic-degrading microorganisms to degrade the nitroaromatic compound.
Claim 2
Independent Claim
108 words
A method for biodegrading a nitroaromatic compound present as a contaminant in a sample, comprising the steps of: (a) providing a sample comprising a nitroaromatic compound; (b) adding to the sample an inoculum comprising a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (c) producing anaerobic conditions in the sample; and (d) maintaining the anaerobic conditions in the sample for a time that is sufficient for the nitroaromatic-degrading microorganisms to degrade the nitroaromatic compound.
Claim 3
Dependent Claim
49 words
The method of claim 1 wherein the inoculum further comprises a carbohydrate-fermenting microorganism that is resistant to the nitroaromatic compound and ferments a fermentable compound under aerobic conditions, wherein the carbohydrate-fermenting microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Lactobacillus, Bacillus, Clostridium, and Pseudomonas.
Claim 4
Dependent Claim
59 words
The method of claim 2 wherein step (c) comprises the steps of: adding to the sample an amount of the fermentable compound; and maintaining the sample under conditions suitable for fermentation by the carbohydrate-fermenting microorganism of at least a portion of the amount of the fermentable compound, thereby consuming dissolved oxygen and producing anaerobic conditions in the fluid medium.
Claim 5
Dependent Claim
12 words
The method of claim 3 wherein the fermentable compound is a carbohydrate.
Claim 6
Dependent Claim
11 words
The method of claim 4 wherein the carbohydrate is a starch.
Claim 7
Dependent Claim
13 words
The method of claim 1 wherein the inoculum is a consortium of microorganisms.
Claim 8
Dependent Claim
29 words
The method of claim 6 wherein the inoculum is an enriched consortium of microorganisms produced by culturing a naturally-occurring population of microorganisms in a medium comprising the nitroaromatic compound.
Claim 9
Dependent Claim
23 words
The method of claim 6 wherein the inoculum comprises an amount of a sample previously subjected to a nitroaromatic biodegradation treatment utilizing microorganisms.
Claim 10
Dependent Claim
35 words
The method of claim 1 further comprising the step of adding to the sample a member of the group consisting of a nutritional source of nitrogen and a nutritional source of phosphorous for the microorganisms.
Claim 11
Dependent Claim
15 words
The method of claim 1 further comprising the step of adding water to the sample.
Claim 12
Dependent Claim
28 words
The method of claim 10 wherein the sample is a member of the group consisting of a water and an aqueous slurry of soil or other particulate matter.
Claim 13
Dependent Claim
21 words
The method of claim 1, wherein the sample is maintained at a pH within a range of about 6 to 8.
Claim 14
Dependent Claim
24 words
The method of claim 1 wherein the sample is maintained at a temperature within a range of about 10° C. to about 40° C.
Claim 15
Dependent Claim
22 words
The method of claim 1 wherein the sample, under the anaerobic conditions, has a redox potential of less than about -200 mV.
Claim 16
Dependent Claim
16 words
The method of claim 1 further comprising the step of sequestering the sample in a vessel.
Claim 17
Dependent Claim
25 words
The method of claim 15 further comprising the step of covering the vessel so as to decrease gas exchange between the sample and the atmosphere.
Claim 18
Independent Claim
100 words
A method for degrading a nitroaromatic compound present as a contaminant in a sample comprising the steps of: (a) providing a sample comprising a nitroaromatic compound and a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (b) producing anaerobic conditions in the sample; and (c) maintaining the anaerobic conditions in the sample for a time that is sufficient for the microorganisms to degrade the nitroaromatic compound.
Claim 19
Independent Claim
100 words
A method for degrading a nitroaromatic compound present as a contaminant in a sample comprising the steps of: (a) providing a sample comprising a nitroaromatic compound and a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (b) producing anaerobic conditions in the sample; and (c) maintaining the anaerobic conditions in the sample for a time that is sufficient for the microorganisms to degrade the nitroaromatic compound.
Claim 20
Dependent Claim
49 words
The method of claim 17 wherein the sample further comprises a carbohydrate-fermenting microorganism that is resistant to the nitroaromatic compound and ferments a fermentable compound under aerobic conditions, wherein the carbohydrate-fermenting microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Lactobacillus, Bacillus, Clostridium, and Pseudomonas.
Claim 21
Dependent Claim
63 words
The method of claim 18 wherein step (b) comprises the steps of: adding to the sample an amount of the fermentable compound; and maintaining the sample containing the added carbon source under conditions suitable for fermentation of at least a portion of the amount of the carbon source by the first microorganisms, thereby consuming dissolved oxygen and producing anaerobic conditions in the sample.
Claim 22
Dependent Claim
12 words
The method of claim 19 wherein the fermentable compound is a carbohydrate.
Claim 23
Dependent Claim
11 words
The method of claim 20 wherein the carbohydrate is a starch.
Claim 24
Dependent Claim
18 words
The method of claim 17 wherein the sample comprises a consortium of microorganisms that includes the nitroaromatic-degrading microorganism.
Claim 25
Dependent Claim
35 words
The method of claim 17 further comprising the step of adding to the sample a member of the group consisting of a nutritional source of nitrogen and a nutritional source of phosphorous for the microorganisms.
Claim 26
Dependent Claim
21 words
The method of claim 17 wherein the sample is maintained at a pH within a range of about 6 to 8.
Claim 27
Dependent Claim
24 words
The method of claim 17 wherein the sample is maintained at a temperature within a range of about 10° C. to about 40° C.
Claim 28
Dependent Claim
22 words
The method of claim 17 wherein the sample, under the anaerobic conditions, has a redox potential of less than about -200 mV.
Claim 29
Dependent Claim
15 words
The method of claim 17 further comprising the step of adding water to the sample.
Claim 30
Dependent Claim
27 words
The method of claim 27 wherein the sample is a member of the group consisting of water and an aqueous slurry of soil or other particulate matter.
Claim 31
Dependent Claim
16 words
The method of claim 17 further comprising the step of sequestering the sample in a vessel.
Claim 32
Dependent Claim
25 words
The method of claim 29 further comprising the step of covering the vessel so as to decrease gas exchange between the sample and the atmosphere.
Claim 33
Independent Claim
179 words
A method for biodegrading a nitroaromatic compound present as a contaminant in a sample, comprising the steps of: (a) adding to a sample comprising a nitroaromatic compound an inoculum comprising (i) a carbohydrate-fermenting microorganism that is resistant to the nitroaromatic compound and ferments a carbohydrate under aerobic conditions, wherein the carbohydrate-fermenting microorganism of a genus selected from the group consisting of Klebsiella, Enterobacter, Lactobacillus, Bacillus, Clostridium, and Pseudomonas and (ii) a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (b) adding an amount of the carbohydrate to the sample; (c) maintaining the sample under conditions suitable for fermentation of at least a portion of the amount of the carbon source by the carbohydrate-fermenting microorganism, thereby producing anaerobic conditions in the sample; and (d) maintaining the anaerobic conditions in the sample for a time that is sufficient for the nitroaromatic-degrading microorganism to degrade the nitroaromatic compound.
Claim 34
Independent Claim
179 words
A method for biodegrading a nitroaromatic compound present as a contaminant in a sample, comprising the steps of: (a) adding to a sample comprising a nitroaromatic compound an inoculum comprising (i) a carbohydrate-fermenting microorganism that is resistant to the nitroaromatic compound and ferments a carbohydrate under aerobic conditions, wherein the carbohydrate-fermenting microorganism of a genus selected from the group consisting of Klebsiella, Enterobacter, Lactobacillus, Bacillus, Clostridium, and Pseudomonas and (ii) a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (b) adding an amount of the carbohydrate to the sample; (c) maintaining the sample under conditions suitable for fermentation of at least a portion of the amount of the carbon source by the carbohydrate-fermenting microorganism, thereby producing anaerobic conditions in the sample; and (d) maintaining the anaerobic conditions in the sample for a time that is sufficient for the nitroaromatic-degrading microorganism to degrade the nitroaromatic compound.
Claim 35
Independent Claim
175 words
A method for degrading a nitroaromatic compound present as a contaminant in a sample comprising the steps of: (a) providing a sample comprising a nitroaromatic compound and (i) a carbohydrate-fermenting microorganism that is resistant to the nitroaromatic compound and ferments a carbohydrate under aerobic conditions, wherein the carbohydrate-fermenting microorganism of a genus selected from the group consisting of Klebsiella, Enterobacter, Lactobacillus, Bacillus, Clostridium, and Pseudomonas, and (ii) a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (b) adding an amount of the carbohydrate to the sample; (c) maintaining the sample under conditions suitable for fermentation of at least a portion of the amount of the carbohydrate by the carbohydrate-fermenting microorganisms, thereby producing anaerobic conditions in the sample; and (d) maintaining the anaerobic conditions in the sample for a time that is sufficient for the nitroaromatic-degrading microorganisms to degrade the nitroaromatic compound.
Claim 36
Independent Claim
175 words
A method for degrading a nitroaromatic compound present as a contaminant in a sample comprising the steps of: (a) providing a sample comprising a nitroaromatic compound and (i) a carbohydrate-fermenting microorganism that is resistant to the nitroaromatic compound and ferments a carbohydrate under aerobic conditions, wherein the carbohydrate-fermenting microorganism of a genus selected from the group consisting of Klebsiella, Enterobacter, Lactobacillus, Bacillus, Clostridium, and Pseudomonas, and (ii) a nitroaromatic-degrading microorganism that degrades the nitroaromatic compound under anaerobic conditions, wherein the nitroaromatic-degrading microorganism is of a genus selected from the group consisting of Klebsiella, Enterobacter, Bacteroides, Fusobacterium, Desulfovibrio, Desulfuromonas, Clostridium, Desulfotomaculum, Sporosarcina, Lactobacillus, Bacillus, Pseudomonas, Veillonella, Acidaminococcus, Methanobacterium, Methanococcus, and Archaeoglobus; (b) adding an amount of the carbohydrate to the sample; (c) maintaining the sample under conditions suitable for fermentation of at least a portion of the amount of the carbohydrate by the carbohydrate-fermenting microorganisms, thereby producing anaerobic conditions in the sample; and (d) maintaining the anaerobic conditions in the sample for a time that is sufficient for the nitroaromatic-degrading microorganisms to degrade the nitroaromatic compound.
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