CN102906234A

CN102906234A - Biodegradation of renewable hydrocarbon fuel blends

Info

Publication number: CN102906234A
Application number: CN2011800252242A
Authority: CN
Inventors: M.R.福斯特; J.T.加农; D.T-W.曹
Original assignee: Butamax Advanced Biofuels LLC
Current assignee: Butamax Advanced Biofuels LLC
Priority date: 2010-05-21
Filing date: 2011-05-20
Publication date: 2013-01-30
Also published as: AU2011255275A1; MX2012013379A; CA2797555A1; WO2011146849A3; WO2011146849A2; EP2571964A2; JP2013529242A; BR112012029362A2; EP2571964A4; US20110283604A1; NZ602828A; KR20130121696A

Abstract

Biologically-produced isobutanol as a component in fuel compositions provides a valuable mechanism for introducing renewable components to fuel compositions and, at the same time, provides for reduced environmental impact of the fuel composition if it were to contaminate a given environmental area.

Description

The biological degradation of reproducible hydrocarbon fuel blend

The cross reference of related application

Present patent application relates to and requires the right of priority of the U.S. Provisional Application 61/347,127 of submitting on May 21st, 2010, and this full patent texts is incorporated herein by reference.

Invention field

The present invention relates to the field that reproducible fuel composition and described composition returning in environment are become.

Background of invention

Isopropylcarbinol is noticeable as the biofuel molecule that is applicable in the gasoline, because it can be produced by reproducible raw material, and has and manyly makes it become characteristic than the more noticeable fuel dope of ethanol; It has higher energy density, lower water-intake rate, better blend ability, and it can be used in the conventional oil engine and need not to improve (Durre, 2007, Biotech.J.2:1525-1578).Ethanol returns the assessment of the impact that becomes to confirm for the environment of gasoline component, in open system, alcohol fuel preferentially was biodegradable before benzene, toluene, ethylbenzene and dimethylbenzene (BTEX) component of gasoline, and confirm that the degraded of ethanol in the waterbearing stratum promptly consumed dissolved oxygen and other nutrition (people such as Corseuil, 1998, Wat.Res.32:2065-2072; Da Silva and Alvarez, 2002, Appl.Environ.Microbiol.70:4720-4726; The people such as Capiro, 2007, Water Research, 41:656-664).Therefore, in the gasoline existence of ethanol can cause BTEX degraded to lag behind and finally cause the BTEX feathering increase (people such as Powers, 2001, Environ.Sci.Technol.35:24A-30A).The people's such as Deeb modeling (2002, J.Environ.Engin.128:868-875) predicted in the presence of ethanol, the increase of benzene feathering length 16-34%, other people propose simultaneously, if by having produced more microbial biomass in the ethanol growth, the life cycle of benzene feathering can increase along with the alcohol concn in the gasoline and reduce (Gomez and Alvarez, 2009, Water Resour.Res.45).

The research (2009, Biomass and Bioenergy, 33:1175-1181) that Mariano and colleagues carry out is used and is indirectly measured (CO ₂Generate and reducing dyes) be evaluated in the aerobic microenvironment of the combination that comprises unpolluted soil, river or unpolluted soil and river, propyl carbinol is on the biodegradable impact of gasoline.Result's demonstration of this research, propyl carbinol can improve the degraded of gasoline in soil, and enhancement coefficient can be greater than the enhancement coefficient of realizing with ethanol.The result that the people such as Garcia-Rivero obtain (2007, J.Environ.Eng.Sci.6:389-395) show that also propyl carbinol can improve the speed of the aerobic bioreactor Degradation of hydrocarbon to the adding in the hydrocarbon mixture.Carried out using research (people 1974 such as Price, the J.Water Pollut.Contr.Fed.46:63-77 that assesses the biological degradation of isopropylcarbinol based on the measurement of biological oxygen demand (BOD) at 20 century 70s; Dias and Alexander, 1971, Appl.Microbiol.22:1114-1118), and multinomial research (c.f., the people such as Deeb, 2000, Biodegradation, 11:171-185; Pruden and Suidan, 2004, Biodegradation, 15:213-227; The people such as Somsamak, 2005, Environ.Sci.Technol.39:103-109; The people such as Vainberg, 2002, J.Environ.Eng.128:842-851) assessed the biological degradation of the trimethyl carbinol (TBA), the trimethyl carbinol is the primary biodegradation product (people such as Hatzinger of gasoline oxygenation agent methyl tertiary butyl ether (MTBE), 2001, Appl.Environ Microbiol.67:5601-5607; The people such as Steffan, 1997, Appl.Environ.Microbiol.63:4216-4222).

Exist to improve the renewable component of gasoline and/or transport fuel composition, and do not cause in the situation that environmental pollution and the environment of described component return the demand of relevant negative environmental effect.

Summary of the invention

The invention provides be used to improving the environment of hydrocarbon fuel composition under the environment release conditions and return, improve simultaneously the method and composition of the recyclability of described fuel composition.

One aspect of the present invention is to return the method that becomes be used to the environment that improves hydrocarbon fuel composition, and described method is for causing the biodegradability of one or more BTEX compounds of described gasoline to improve by add isopropylcarbinol in described composition.In one aspect, described method and composition under anaerobic provides the biological degradation effect that improves.In one aspect, described method and composition provides the biological degradation that improves under aerobic conditions.In one aspect, described method and composition provides the biological degradation that improves under the nitrate reduction condition.In one aspect, the interpolation of isopropylcarbinol has improved the biological degradation of benzene in hydrocarbon fuel composition.

Another aspect of the present invention is to return the method that becomes for the environment that improvement comprises the liquid fuel combination of ethanol, and described method is for causing the biological degradation of one or more BTEX compounds of described gasoline to be improved by adding isopropylcarbinol to described composition.

Another aspect of the present invention is to work as described ethanol for (for example being discharged into the environment compartment, soil, settling, underground water) in the hydrocarbon fuel composition component time, reduce ethanol to the method for the transmission of soil matrix, described method comprises makes isopropylcarbinol mix with described fuel composition.

Another aspect of the present invention is to reduce the BTEX feathering that is formed to the release in the environment compartment by the optional compositions of hydrocarbons that comprises ethanol, described method comprises the isopropylcarbinol that adding is an amount of, wherein said isopropylcarbinol is as the hydrocarbon of described compositions of hydrocarbons and the solubility promoter of ethanol component, thereby postpones and/or partly control the BTEX feathering and reduce the possibility that it leaks out to ground water table.

Another aspect of the present invention relates to liquid fuel combination, and described composition to be being enough to improve the recyclability of described compositions of hydrocarbons, and the amount that can not improve its potential environmental influence when described composition contaminate environment compartment comprises hydrocarbon, ethanol and isopropylcarbinol.On the other hand, by adding isopropylcarbinol, the recyclability of described fuel composition improves, and its potential environmental influence reduces, and for example, compares with the expansion of the BTEX feathering of the same combination that does not have isopropylcarbinol, the expansion of BTEX feathering can reduce, especially under the aerobic environment condition.

This paper also provides and has been used for improving the hydrocarbon fuel composition that comprises isopropylcarbinol and under anaerobic adds electron acceptor(EA) in the amount that the environment of environment compartment returns the method that becomes, described method to comprise being enough to improving the biodegradation rate of one or more BTEX components in described compartment.

This paper provides and has been used for improving the recyclability of hydrocarbon fuel composition and is limited in when being polluted by described hydrocarbon fuel composition method on the impact of environment compartment, and described method comprises mixes described hydrocarbon fuel composition with an amount of isopropylcarbinol.In embodiments, described hydrocarbon fuel composition also comprises ethanol.In embodiments, ethanol was constituted to how about 10% fuel composition before adding isopropylcarbinol.In embodiments, isopropylcarbinol provides the biological degradation of at least a improvement in the BTEX component in the described hydrocarbon fuel composition.In embodiments, isopropylcarbinol provides the biological degradation of the improvement of benzene.In embodiments, described environment compartment comprises soil matrix, and wherein said isopropylcarbinol reduces the transmission of ethanol in soil matrix.In embodiments, isopropylcarbinol hinders the BTEX feathering from the expansion of described composition.In embodiments, described isopropylcarbinol exists with the amount that the biodegradability that is suitable for making described hydrocarbon fuel composition improves.In embodiments, the biological degradation of described improvement occurs under aerobic conditions.In embodiments, the biological degradation of described improvement occurs under nitrate reduction conditioned disjunction sulfate reduction condition.

Also provide improve comprise isopropylcarbinol hydrocarbon fuel composition under anaerobic the amount of returning the method that becomes, described method to comprise being enough to improving the biodegradation rate of one or more BTEX components of the environment in the environment compartment in described compartment, add electron acceptor(EA).In embodiments, described electron acceptor(EA) is iron, vitriol or nitrate or their combination.In embodiments, described electron acceptor(EA) is Fe (OH) ₃In embodiments, described electron acceptor(EA) is NaNO ₃In embodiments, described electron acceptor(EA) is MgSO ₄In embodiments, described one or more BTEX components comprise toluene, dimethylbenzene or benzene.In embodiments, described electron acceptor(EA) is nitrate and adds with the amount that is enough to create the nitrate reduction condition.In embodiments, described electron acceptor(EA) is vitriol and exists with the amount that is enough to create the sulfate reduction condition.In embodiments, described electron acceptor(EA) is nitrate and exists with the amount that is enough to create the nitrate reduction condition, and wherein toluene with the approximately identical fate of isopropylcarbinol in carry out biological degradation.In embodiments, described electron acceptor(EA) is vitriol and exists with the amount that is enough to create the sulfate reduction condition, and wherein toluene with the approximately identical fate of isopropylcarbinol in carry out biological degradation.In embodiments, described electron acceptor(EA) is nitrate and exists with the amount that is enough to create the nitrate reduction condition, and wherein benzene with the approximately identical fate of isopropylcarbinol in carry out biological degradation.In embodiments, described electron acceptor(EA) is vitriol and exists with the amount that is enough to create the sulfate reduction condition, and wherein the biological degradation of benzene improves there not being biological degradation under the sulfate reduction condition to compare with it.In embodiments, described electron acceptor(EA) is vitriol and exists with the amount that is enough to create the sulfate reduction condition, and wherein the biological degradation of benzene with it in the situation that do not exist the biological degradation of isopropylcarbinol to compare to improve.In embodiments, described electron acceptor(EA) is vitriol and exists with the amount that is enough to create the sulfate reduction condition, and wherein the biological degradation of benzene is compared at the biological degradation in the presence of the ethanol with it and improved.

This paper also provides and has comprised gasoline, isopropylcarbinol and Fe (OH) ₃, NaNO ₃, or MgSO ₄In at least a composition.Also provide and comprised gasoline, isopropylcarbinol and Fe (OH) ₃, NaNO ₃, KNO ₃, NHNO ₃, Na ₂SO ₄, CaSO ₄, MgSO ₄, at least a composition in chelated iron, Zero-valent Iron and the nano zero valence iron.

The accompanying drawing summary

Fig. 1 a has shown at the most 10 days degraded of isopropylcarbinol.Error line represents 95% fiducial interval.

Fig. 1 b has shown at the most 50 days degraded of isopropylcarbinol.

Fig. 1 c has shown at the most 50 days degraded of ethanol.

Fig. 2 A, 2B, 2C and 2D have shown the impact of isopropylcarbinol on the biological degradation of high density BTEX.(Fig. 2 A has shown benzene; 2B has shown toluene; 2C has shown ethylbenzene; 2D has shown total dimethylbenzene.) error line represents 95% fiducial interval.

Fig. 3 A, 3B, 3C and 3D have compared isopropylcarbinol and alcohol concn with the impact of various processing horizontals on the BTEX biological degradation.(Fig. 3 A has shown benzene; 3B has shown toluene; 3C has shown ethylbenzene; 3D has shown total dimethylbenzene.) error line represents 95% fiducial interval.

Fig. 4 A, 4B, 4C and 4D have shown the impact of isopropylcarbinol on the biological degradation of lower concentration BTEX.(Fig. 4 A has shown benzene; 4B has shown toluene; 4C has shown ethylbenzene; 4D has shown total dimethylbenzene.) error line represents 95% fiducial interval.

Fig. 5 A, 5B, 5C and 5D have shown the concentration of the biological degradation of isopropylcarbinol-higher-under various anaerobic reduction conditions.(Fig. 5 A has shown that processing 2-is unmodified; 5B has shown processing 6-nitrate reduction; 5C has shown the reduction of processing 9-iron; 5D has shown processing 12-sulfate reduction.) error line represents 95% fiducial interval.The dotted line of processing 2 represents the time that microenvironment is remodified by isopropylcarbinol.

Fig. 6 A and 6B have shown respectively the biological degradation of processing 4 (unmodified) and processing ethanol in 14 (sulfate reductions).Error line represents 95% fiducial interval.Process 4 dotted line and be illustrated in after vitriol remaining in the underground water is reduced the time that microenvironment is remodified by ethanol.

Fig. 7 A, 7B, 7C and 7D have shown in the situation that under the various anaerobic reduction condition and have or do not exist isopropylcarbinol (IBA), the biological degradation of benzene and toluene (higher concentration).(7A and 7B show respectively and just process benzene and the toluene with regard to 1,2,5 and 6; 7C and 7D show respectively and just process benzene and the toluene with regard to 8,9,11 and 12.) error line represents 95% fiducial interval.

Fig. 8 A, 8B, 8C and 8D have shown the concentration of the biological degradation of BTEX-lower-in the situation that under the various anaerobic reduction condition and have or do not exist isopropylcarbinol (IBA) or ethanol.(Fig. 8 A has shown benzene; 8B has shown toluene; 8C has shown ethylbenzene; 8D has shown total dimethylbenzene.) error line represents 95% fiducial interval.

Fig. 9 A, 9B, 9C and 9D have shown the biological degradation of isopropylcarbinol-lower concentration.(9A has shown that processing 3-is unmodified; 9B has shown processing 7-nitrate reduction; 9C has shown the reduction of processing 10-iron; 9D has shown processing 13-sulfate reduction.) error line represents 95% fiducial interval.The monitoring demonstration that replenishes, in the time of the 160th day, in the time of the 48th day, isopropylformic acid concentration has been reduced to and has been lower than analyzing and testing limit (data do not show) with regard to processing 13 with regard to processing 3.

Figure 10 A, 10B, 10C and 10D have shown in the situation that under the various anaerobic reduction condition and have or do not exist isopropylcarbinol (IBA), the biological degradation of the ethylbenzene of high density and total dimethylbenzene.(10A and 10B show respectively and just process ethylbenzene with regard to 1,2,5 and 6 and total dimethylbenzene; 10C and 10D show respectively and just process ethylbenzene with regard to 8,9,11 and 12 and total dimethylbenzene.) error line represents 95% fiducial interval.

Figure 11 and 12 has showed when the water that at first adds 1.3% and 2.6% in the E10 gasoline, the behavior of isopropylcarbinol.Among Figure 11, the isopropylcarbinol content of rising causes the water volume that reduces in the phase separation.Figure 12 has shown when occuring to separate, less ethanol has been arranged in moisture part.

Figure 13 has described the result from microbiological analysis.Error line represents 95% fiducial interval.The IBA=isopropylcarbinol.

Detailed Description Of The Invention

Unless otherwise defined, otherwise that the implication of all scientific and technical terminologies used herein and those skilled in the art understand usually is the same.As conflict, be as the criterion with present patent application (comprising its definition).In addition, unless that context has in addition is required, singular references will comprise that plural number and plural term will comprise odd number.Be all purposes, other bibliography that all publications, patent and this paper mention all is incorporated herein by reference in full.

In order further to limit the present invention, this paper provides following term and definition.

As used herein, term " comprises ", " comprising ", " having ", " containing " or their any other modification will be understood to mean the integer that comprises appointment or integer group but do not get rid of any other integer or integer group.For example, the composition, mixture, technique, method, goods or the equipment that comprise series of elements needn't only limit to those elements, and can comprise the element that other is not clearly listed, or the intrinsic element of such composition, mixture, technique, method, goods or equipment.In addition, unless specify in addition, otherwise "or" refer to inclusive or, rather than refer to exclusiveness or.For example, any one all represent to satisfy condition A or B:A below are that genuine (or existence) and B are that false (or non-existent), A are that false (or non-existent) and B are that genuine (or existence) and A and B are genuine (or existence).

As used herein, as employed in whole specification sheets and the claim, term " by forming " or show such as the modification of the different tenses of " by forming " and to comprise any integer of enumerating or integer group, but can join in method, structure or the composition of appointment without additional integer or integer group.

As used herein, as employed in whole specification sheets and the claim, term " basically by forming " or show such as the modification of the different tenses of " basically by forming " and to comprise any integer of enumerating or integer group, and randomly comprise any integer of enumerating or the integer group of the basic or novel characteristics of the method, structure or the composition that significantly do not change appointment.Referring to M.P.E.P. § 2111.03.

Equally, the number that relates to element or component example (being number of times) before element of the present invention or component indefinite article " " or " a kind of " to be intended to be nonrestrictive.Therefore, " one " or " a kind of " should be interpreted as to comprise one or at least one, and the word singulative of element or component comprises that also plural number refers to, unless obviously expression odd number of numeral is arranged.

As used herein, term " invention " or " the present invention " are non-limiting term, and are not intended to mean any independent embodiment of the present invention, but contain such as the described all possible embodiment of patent application.

As used herein, the term " about " that the amount of composition of the present invention or reactant of modifying is used refers to the variation of the umerical amount that can occur by for example following mode: in fact for generation of general measure and the liquid treatment operation of enriched material or solution; By unintentional error in these operations; For the preparation of difference in manufacturing, source or the purity of the composition of composition or manner of execution etc.Term " about " also comprises the different amount owing to producing from the different equilibrium conditionss of the composition of specific starting mixt.No matter whether modify the equal parts of the claim amount of comprising by term " about ".In one embodiment, term " about " refers in 10% scope of record numerical value, preferably in 5% scope of record numerical value.

As described herein, the hydrocarbon fuel composition that comprises isopropylcarbinol provides the fuel composition of the recyclability with raising, and described recyclability is owing to equally can biodegradable reproducible component in environment.Isopropylcarbinol is reproducible component, and is as shown here, is degraded rapidly in the time of in adding the waterbearing stratum microenvironment.In addition, result's demonstration provided herein, isopropylcarbinol not only self is biodegradable, and it also provides additional biological degradation beneficial effect to gasoline under various envrionment conditionss.As describing in this area, isopropylcarbinol can be converted into by the carbon substrate that will derive from reproducible raw material such as biomass the microorganism biological ground production of isopropylcarbinol.Therefore, the isopropylcarbinol of producing provides the valuable mechanism that is used for reproducible component is introduced fuel in adding fuel composition the time biologically, and simultaneously, in the situation of the given environmental area of described fuel composition pollution, provide the environmental influence that reduces.

As everyone knows, the benzene of gasoline, toluene, ethylbenzene and total dimethylbenzene (BTEX) component are worthless environmental pollutant.In gasoline, add the recyclability that ethanol can improve gasoline, yet such interpolation also can be strengthened the infringement that BTEX causes, because ethanol can cause the expansion of BTEX feathering.In contrast, compare as additive with adding ethanol, add isopropylcarbinol in the gasoline mechanism that improves the renewable component of fuel and can not improve the BTEX feathering when described fuel blends is released to environmental area or compartment (for example, soil, settling, underground water) is provided.In addition, as shown in this paper, the biological degradation that the interpolation of isopropylcarbinol does not hinder BTEX is to for ethanol viewed degree under certain conditions.

As used herein, " biological degradation " or " degraded " refers to that the compound paid close attention to is to the elementary conversion of byproduct.

As used herein, the amount of one or more components that " environment of improvement returns " refers to reduce the hydrocarbon fuel blend in the environment compartment, the degradation rate of one or more components in the environment compartment that improves the hydrocarbon fuel blend, the size that reduces the environment compartment that contacts with one or more components of hydrocarbon fuel blend or their combination.

As used herein, " environment compartment " refers to the zone that contacts with fuel composition, and can comprise, for example, and soil, settling, underground water or their combination.

As used herein, " BTEX feathering " refers to dissolve the phase feathering.

When being used for the reference fuel composition, " recyclability of raising " refers to that the part of the described composition that improves is by being considered to and non-renewable resource, and the reproducible resource that for example fossil oil is relative with oil is produced such as biomass.

Provided hereinly be by in fuel composition, comprising isopropylcarbinol, to improve simultaneously the amount of reproducible component in the fuel composition and reduce the method for the environmental influence of fuel composition.Be applicable to the hydrocarbon part of the fuel composition of method disclosed herein, formed the blended into gasoline thing reserve that can be used for preparing gasoline, described gasoline is used for consuming at explosive motor, and described explosive motor includes but not limited to spark ignition engine.Blended into gasoline thing reserve includes but not limited to meet ASTM 4814, the blend reserve of the gasoline of EU specification EN228, and the blend reserve of reformulated gasoline.With regard to method disclosed herein, in the hydrocarbon fuel composition amount of isopropylcarbinol (by volume) comprise at least about 2%, at least about 5%, at least about 7%, at least about 10% or at least about 15% amount.In some respects, the amount of isopropylcarbinol (by volume) comprises approximately 2% to about 20% amount, and approximately 10% to about 16% amount.The amount that should be appreciated that isopropylcarbinol can be depending on vehicle technology.So, in embodiments, the amount of isopropylcarbinol can be by volume at the most approximately 85%.Isopropylcarbinol can use the hydrocarbon of any method known in the art and described fuel composition partially mixed.

In some embodiments, described hydrocarbon fuel composition also comprises ethanol, and in some embodiments, ethanol before adding isopropylcarbinol, consist of compositions of hydrocarbons at the most approximately 10%, at the most approximately 15%, at the most approximately 20% or at the most approximately 50%.In some embodiments, isopropylcarbinol has substituted ethanol in fuel composition.In some embodiments, the isopropylcarbinol adding is comprised in the fuel composition of ethanol.

Method provided herein comprises by comprise isopropylcarbinol in described composition, postpones the BTEX feathering from the method for hydrocarbon fuel composition expansion.Hydrocarbon fuel composition potential environmental influence when occuring to discharge can be assessed by the size of BTEX feathering and/or the speed of expansion and/or the concentration of BTEX feathering in the measurement involved area.The size of BTEX feathering and spreading rate can be by method as known in the art as directly to underground water sampling, with such as the method assessment of Environmental Protection Agency method SW846.

In some embodiments, the degraded of fuel composition that comprises isopropylcarbinol than comprising ethanol but the fuel composition that does not contain isopropylcarbinol occur sooner.The degraded of fuel element can be used method as known in the art such as vapor-phase chromatography (GC), for example uses U.S. EPA method 8015, or the GC-mass spectroscopy, and for example the U.S. EPA method 8260, measure in environmental sample.As shown in this paper, isopropylcarbinol is degraded soon equally with ethanol at least, and one or more BTEX components in the presence of isopropylcarbinol than in the situation that do not exist ethanol degraded faster.In some embodiments, isopropylcarbinol is with at least about 0.081d ^-1First order rate constant degraded.In some embodiments, isopropylcarbinol is with at least about 0.28d ^-1First order rate constant degraded.In some embodiments, isopropylcarbinol is with greater than about 0.074d ^-1First order rate constant degraded.In some cases, comprise the fuel composition of isopropylcarbinol and compare the BTEX biodegradation rate with raising with the fuel composition that does not contain renewable component.In some cases, the fuel composition that comprises isopropylcarbinol is compared the BTEX biodegradation rate with raising with the fuel composition that comprises ethanol but do not contain isopropylcarbinol.

In addition, such as (referring to the Figure 11 and 12) as shown in this paper, in comprising the gasoline of ethanol, sneak into butanols and reduced at the volume of water when occuring that is separated, and limited the weight percent of the ethanol that described aqueous phase comprises.Be not bound by theory, it is believed that butanols by ethanol being remained in the water-soluble lower hydrocarbon fraction, can limit the amount of the ethanol that is leached to underground water.The transmission of ethanol in soil matrix reduces and can therefore postpone BTEX to the expansion of ground water table.

Therefore, will recognize that method provided herein can advantageously reduce the required time of the contaminated place of recovery, can limit the size of BTEX feathering or this two kinds of advantages can be provided, thereby the environment that improves hydrocarbon fuel composition returns.

Method provided herein can improve comprise isopropylcarbinol hydrocarbon fuel composition under anaerobic the environment in the environment compartment return.In embodiments, the described method amount that comprises being enough to improving the biodegradation rate of one or more BTEX components adds electron acceptor(EA) in described compartment.The electron acceptor(EA) that is fit to comprises nitrate, includes but not limited to NaNO ₃, NH ₄NO ₃, KNO ₃And vitriol, include but not limited to MgSO ₄And CaSO ₄, Na ₂SO ₄, and iron cpd, include but not limited to Fe (OH) ₃, chelated iron, Zero-valent Iron and nano zero valence iron.Electron acceptor(EA) can use include but not limited to the slurries form inject, lay, method by monitor well injection, gravity injection and/or vasopressing injection or their combination, to be enough to realizing that the amount of nitrate reduction, iron reduction and/or sulfate reduction condition adds in the environment compartment.The injection of vitriol and/or nitrate and/or iron cpd can be used to biostimulation sulfate-reducing bacteria and/or nitrate-reducing bacteria (if existence), pollutes at the BTEX that underground release causes because of the gasoline that comprises isopropylcarbinol with biological degradation.Such biostimulation can cause biological activity, colony and/or the metabolism of bacterium to be improved.

Aerobic conditions

To the isopropylcarbinol that adopts first approximation and the relatively demonstration of alcohol biological degradation rate, some isopropylcarbinols are processed and have been caused 0.081 ± 0.0044d of observing ^-1First order rate constant, (R ²=0.80, use from processing 4 and process 5 data, embodiment 1), and Ethanol Treatment has caused 0.074 ± 0.0023d of observing ^-1First order rate constant (R ²=0.90).Therefore, under the processing of higher concentration, isopropylcarbinol is biodegradable with the speed higher than the speed of ethanol.Isopropylcarbinol in the processing (processing 3) of low concentration has obtained 0.28 ± 0.054d ^-1Rate constant (R ²=0.69), this is than approximately large 3.5 times of the biodegradation rate constants of the isopropylcarbinol in the processing of higher concentration or ethanol.Above-mentioned difference is not only that difference by bacterial growth causes.Not bound by theory, a kind of explanation that it is believed that this difference is that the biological degradation (and growth of the bacterium of degraded isopropylcarbinol) of isopropylcarbinol is the nutrient substance restriction within initial two weeks of research, and most isopropylcarbinol biological degradation is occuring during this period of time.Isopropylcarbinol for low concentration is processed, and the nutrient substance restriction may be not so serious, and causes lower isopropylcarbinol to process the rate constant that is improved.Perhaps, the difference of isopropylcarbinol rate constant may be because the isopropylcarbinol substrate under higher concentration suppresses.In the research to the propyl carbinol biological degradation, observed substrate inhibition (Alagappan and Cowan, 2001, Biotechnol and Boengin, 75:393-405) in the concentration that is low to moderate about 3,000 μ M.

Result's demonstration provided herein, ethanol has reduced the speed of BTEX biological degradation inadvisablely more than isopropylcarbinol.What is interesting is, when relatively isopropylcarbinol and ethanol during on the affecting of BTEX biological degradation, benzene shows the most significant difference, the aerobic bioreactor Degradation of benzene in the presence of ethanol than in the presence of isopropylcarbinol, having slowed down about 12 times more.Benzene is often from the adjusting driver of the groundwater pollution in gasoline contamination place.

Anaerobic condition

Result's displaying provided herein, isopropylcarbinol easily is biodegradable under nitrate reduction, iron reduction, sulfate reduction and methanogenic condition.With regard to the condition of this research, the biological degradation of isopropylcarbinol or the BTEX that do not slow down, perhaps isopropylcarbinol degree that the BTEX biological degradation the is slowed down degree that the BTEX biological degradation slowed down less than ethanol.In some cases, the interpolation of isopropylcarbinol has improved the speed of the BTEX biological degradation of observing.Therefore, the existence of isopropylcarbinol and it are considered to more favourable than ethanol to the impact of BTEX biological degradation under the condition of the waterbearing stratum of anaerobism.

Embodiment

Embodiment 1: the aerobic test

Materials and methods

The soil and groundwater sample

The soil and groundwater collection that is used for laboratory microenvironment test is positioned at Vandenberg Air Force Base certainly, in the place 60 of CA.This place has the history of gasoline contamination, but has passed through a large amount of liquidation proceduress.The underground water of collecting is loaded in the aseptic stainless steel soda bucket (18.5L), and headspace fills nitrogen.Use is with acetic ester core sleeve

6620DT has collected locate below ground level (bgs) about 8 to 12 feet soil.Core sample in the acetic ester sleeve is exposed to air by end-blocking and sealing then and there to minimize, and spending the night on ice is transported to the laboratory, and 4 ℃ of lower storages.

In anaerobic chamber (Coy Laboratory Products, Inc., Grass Lake, MI), soil is shifted out from the acetic ester sleeve, can the initial 10cm core that once was exposed in the oxygen is terminal discarded.The soil of collecting is by consisting of with some rubbles with than the silty sand of ratchel.Make soil pass through the 0.95cm sieve, homogenize, then under 4 ℃, be stored in

In the amber glass bottle of lid, until the microenvironment setting is finished.Baseline soil and groundwater data are shown in Table 1.(NA=does not analyze; ^*SVOC (semi-volatile organic compounds) detects and comprises 0.008mgL ^-1Phenol and 0.003mgL ^-1Two (2-ethylhexyl) esters of phthalic acid; ^*Standard unit).

Table 1: the soil and groundwater data of baseline

Parameter	Underground water (mg/L)	Soil (mg/kg)
			Total organic carbon	22	1,700
The gasoline-range organism	5.7	NA
			Total SVOC	0.011*	＜0.084
Total iron	490	NA
			The iron of dissolving	＜0.1	NA
Nitrate (in N)	1.5	NA
			Vitriol is (with SO ₄ ^2-Meter)	105	NA
Basicity is (with CaCo ₃Meter)	391	NA
			Methane	0.005	NA
pH	7.2**	NA
			The oxygen of dissolving	0.8	NA

The microenvironment experiment

The group method of microenvironment experiment is the biological degradation of assessing the isopropylcarbinol of BTEX (benzene, toluene, ethylbenzene and total dimethylbenzene) and " height " and " low " concentration in soil-underground water slurries.Final converted product is not determined.In order to compare, use ethanol to replace isopropylcarbinol to prepare a kind of processing.List under the BTEX hurdle in the table 2 for the measuring object BTEX concentration of every kind of BTEX compound, table 2 has shown the experiment processing array.Contrast is revised with mercury chloride and formaldehyde.The concentration of BTEX and alcohol represents in coming source region and the possible underground water concentration that will observe in the suitable gradient feathering in its vicinity through selecting with (approx).The alcohol concn higher with respect to isopropylcarbinol concentration that uses in this research is intended to reflect isopropylcarbinol and the ethanol effective solubleness in underground water.Ethanol has the water solubility of 10 times of water solubilities that are approximately isopropylcarbinol, and the octanol-water partition coefficient of isopropylcarbinol approximately is 10 times of (Organization for Economic Cooperation and Development (Organization tor Economic Co-operation and Development) of the octanol-water partition coefficient of ethanol, 2004.SIDS Assessment Report for SIAM 19-Ethanol (CAS No.64-17-5) .Berlin, Germany; The Organization for Economic Cooperation and Development (Organization for Economic Co-operation and Development), 2004.SIDS Assessment Report for SIAM 19-Isobutanol (CAS No.78-83-1), Berlin, Germany).Therefore for the test in this research, conservatively selected the approximately ethanol volumetric molar concentration higher 3 times than isopropylcarbinol.Process behind sample collection for the 1st group and prepare in 4 days, and the 2nd group of test prepared after soil and groundwater has stored about 2 months.Experiment is processed matrix display in table 2A and 2B.

Table 2A: the 1st group experiment processing array

Table 2B: the 2nd group experiment processing array

Prepared microenvironment by each that the 40g on-site soil is placed 54 glass serum bottles (volume of each glass serum bottle is about 160mL).BTEX and alcohol (isopropylcarbinol or ethanol) are added in the processing bottle, to reach the aimed concn that shows among table 2A and the B.Fill bottle so that stay the headspace of 10mL with underground water.Contrast is revised with microbiostatic activity with mercury chloride (being 700mg/L in the bottle).Subsequently, after 4 days, use formaldehyde (being 1%v/v in the bottle) to revise contrast active with restriction micro-organisms.Minimum 3, maximum 8 repetition have been prepared in each processing.

The microenvironment for preparing under 15 ℃ on the gyrate shaker of 100rpm operation incubation.For BTEX and oxygen the headspace in each bottle is monitored.Calculate moisture BTEX concentration by using Henry's law.Analyzed the sample of water for isopropylcarbinol and ethanol and possible isopropylcarbinol degraded product (isobutyric aldehyde and isopropylformic acid).The headspace of each bottle is periodically poured oxygen, to keep the aerobic conditions in the bottle.The headspace of each control bottle also is washed into oxygen, loses because of the possible BTEX that the qi of chong channel ascending adversely process causes with assessment.

Analytical procedure

Be equipped with the Varian CP-3900 gas chromatograph of FID and Restek QSPLOT post (30m length, 0.32mm diameter) by use, analyzed headspace gas for BTEX, injector temperature is 260 ℃, and detector temperature is 290 ℃.Be equipped with impulsive discharge helium ionization detector (PDHID) and CP-Molsiene 5A post and CP-ParaBond Q post (to be 10m length by use, 0.32mm Varian CP-3800 gas chromatograph diameter), analyzed headspace oxygen, injector temperature is 210 ℃, and detector temperature is 240 ℃.Oxygen was at 1.20 minutes wash-outs.By at first collecting 130 μ L with the sub-sample of mercury chloride preservation, measured the alcohol concn in the water (and isobutyric aldehyde and isopropylformic acid).Flame ionization detector (FID) and Stabilwax DA post (30m length is equipped with by use, 320 μ m diameters) Varian CP-3800 gas chromatograph, analyzed these samples, injector temperature is 280 ℃, and detector temperature is 280 ℃.

Microbiological analysis

In when beginning experiment, midway (generally) and collected sample (2mL) from the microenvironment of revising with BTEX and isopropylcarbinol when finishing, to assess microbial population along with the variation of flow of research.According to method SM9215C, after fetching immediately serial dilution sample and be plated on R2A agar (BDDifco) and basic salt culture medium (BSM; The people such as Hareland, 1975, J.Bacteriol.121:272-285) on the agar.In the container of the sealing that contains BTEX or isopropylcarbinol incubation BSM dull and stereotyped, with the bacterium of selecting to grow at these substrates.After 3 days, after 10 days, manually carried out enumeration for selective medium for R2A agar.Sample (～8mL) also freezing immediately under-70 ℃, and when research finishes, transport Microbial Insights in dry ice, Rockford, TN analyzes to be used for CENSUS.CENSUS is based on the counting to eubacterium 16S rRNA gene copy number, adopt quantitative polyase chain reaction (qPCR) detection method come quantitatively total eubacterium (about the more information of CENSUS method referring to Microbial Insights, 2009http: //www.microbe.com/how-census-works.html and http://microbe.com/census-applications/anaerobic-btex.html; Respectively at 07.07.2009 and 05.08.09 access).

The degraded of isopropylcarbinol and ethanol

In any contrast, all do not observe the gathering or isobutyric gathering of measurable reduction (＞10%), isobutyric aldehyde of isopropylcarbinol or ethanol.The biological degradation of processing the isopropylcarbinol in 3 and 4 is shown among Fig. 1.In the processing (processing 3) of low concentration, isopropylcarbinol was degraded in 7 days to being lower than analyzing and testing limit (3 μ M).Isopropylcarbinol degraded product isobutyric aldehyde and isopropylformic acid detect in the time of 4 days first at incubation, and isobutyric aldehyde was reduced in 5 days subsequently and is lower than detectability.Isobutyric concentration at first raises, but the sample of getting after 82 days has been confirmed isopropylformic acid further degraded (not display data) of same also quilt in microenvironment.Isobutyric biodegradable product is not determined, but research shows the preceding, and isopropylformic acid easily carries out biological degradation under aerobic conditions, and butyrates easily is converted into CO under aerobic conditions ₂(Miller, 2001, J.Anim.Sci., 79:2503-2512; Bonartseva, 2003, Appl.Biochem.Biotech.109:285-301).

In the processing (processing 4, Fig. 1 b) of higher concentration, isopropylcarbinol starting point concentration degraded from 3,400 μ M in 23 days is limit to being lower than analyzing and testing.Along with isopropylcarbinol is degraded, two kinds of isopropylcarbinol degradation productions, isobutyric aldehyde and isobutyric formation and degraded subsequently are observed.Isobutyric aldehyde reached the peak concentration of 900 μ M at the 9th day, and dropped to the analyzing and testing limit that is lower than 11 μ M after 19 days.Isopropylformic acid was increased to the peak concentration of 1,750 μ M at the 25th day, then dropped to 100 μ M at the 48th day.In the second group of microenvironment that is used for comparing isopropylcarbinol and ethanol (processing 5), isopropylcarbinol is degraded with similar timetable, and isopropylformic acid is observed with similar amount.Yet, only have the isobutyric aldehyde (78 μ M) of trace content to be observed.

In the microenvironment that ethanol is revised, alcohol concn is reduced to the analyzing and testing limit that is lower than 22 μ M from 11,000 μ M in about 40 to 45 days (Fig. 1 c).In contrast, do not observe the reduction of ethanol.Be not subjected to the restriction of available nutrient substance in order to ensure the degraded of viewed ethanol, the nutrient substance of improved basic salt culture medium people such as (, 1975) Hareland form was added whole processing at 22 days.Do not observe measurable rising of ethanol (or any other compound) speed, show that the biological degradation of pollutent is not subjected to the restriction of nutrient substance operability about 22 days in above-mentioned experimental system.

The BTEX degraded

With the microenvironment that the BTEX of higher concentration revises, contain or do not contain isopropylcarbinol, be shown among Fig. 2.Result about low concentration BTEX is provided among Fig. 4.The one-level of the retardation time of observing and recurrence represents with the sky and be shown in the table 3 effective half-life.Be the transformation period that returns to add retardation time effective half-life.(IBA=isopropylcarbinol; ± value indication 95% fiducial interval.)

Table 3: for the retardation time of BTEX and (τ effective half-life of recurrence _1/2 )

BTEX concentration in the high density contrast did not demonstrate observable decline during about 25 days, and in this time, had observed the reduction of concentration in the contrast (at the most about 20%) for some compounds.Again revised contrast with formaldehyde subsequently, with microbiostatic activity; Additional formaldehyde also prevents further reduction in the contrast.In the time of 25 days, most BTEX compound is degraded, so these losses do not affect data evaluation.Except total dimethylbenzene, in lower concentration BTEX contrast, do not observe significant (＞10%) and reduce, wherein during 10 days of this experiment, observe the reduction of total xylene concentration about 25%.

The transformation period (table 3) of the BTEX compound that under aerobic conditions, obtains generally within other people viewed scope (referring to summary: United States Geological Survey, 2006, " Description; properties, and degradation of selected volatile organic compounds detected in groundwater "-consult selected document.Open File Report 2006-1338)。The result of comparative microenvironment (

processing

4,5 and 6) demonstration, ethanol generally shows the disadvantageous effect to BTEX biological degradation higher than isopropylcarbinol (Fig. 3 and table 3).Unique exception is ethylbenzene, and wherein isopropylcarbinol and ethanol have similarly affected the biological degradation of ethylbenzene.In addition, in the processing (processing 1 and 3) of lower concentration, the interpolation of isopropylcarbinol caused the biological degradation of ethylbenzene and total dimethylbenzene little but measurable rising may be because the degraded person of ethylbenzene and total dimethylbenzene accidental growth on isopropylcarbinol.

Process BTEX effective half-life in 5 with about factor of two to five less than processing in 4 (table 3).Yet the biodegradation rate of isopropylcarbinol is approximately identical in these two kinds of processing.

Microorganism characterizes

What the enumeration of microorganism and qPCR analyzed the results are shown among Figure 13.These data presentation, the microorganism concn in the microenvironment raises the duration of research.The enumeration of microorganism all demonstrates similar trend with total eubacterium data, although the eubacterium concentration of measuring by analysis of molecules is higher than the microbe colony counting.Aerobic plate count and by the difference no wonder between total bacterium of analysis of molecules, because some bacteriums well-grown on whole agar plates not, then a kind of method has been measured the bacterium of educable, that can not cultivate, anaerobism and aerobic.The plate count data validation isopropylcarbinol of can either degrading have the natural bacteria of the BTEX that can degrade to be present in the on-the-spot material.As indicated in the processing of lower concentration, microbial growth becomes limited when substrate (BTEX or isopropylcarbinol) is depleted.

Embodiment 2: the anaerobism test

Materials and methods

Soil and groundwater

The collection of soil and groundwater and sample and handling procedure are for as mentioned above.The soil and groundwater data presentation of baseline is in table 4.(NA=does not analyze; * SVOC detects and comprises 0.008mgL-1 phenol and two (2-ethylhexyl) esters of 0.003mgL-1 phthalic acid; * standard unit)

Table 4: underground water and soil parameters

Parameter	Underground water (mg/L)	Soil (mg/kg)
			Total organic carbon	22	1,700
The gasoline-range organism	5.7	NA
			Total SVOC	0.011*	＜0.084
Total iron	490	NA
			The iron of dissolving	＜0.1	NA
Nitrate (in N)	1.5	NA
			Vitriol is (with SO ₄ ^2-Meter)	105	NA
Basicity is (with CaCo ₃Meter)	391	NA
			Methane	＜0.005	NA
ph	7.2**	NA
			The oxygen of dissolving	0.8	NA

The preparation of microenvironment

For microenvironment experiment, from nitrate reduction under the condition of methanogenic scope, assessed BTEX and the isopropylcarbinol biological degradation in soil-underground water slurries.BTEX and isopropylcarbinol (table 5) have been assessed in " height " and " low " concentration.Use ethanol to substitute isopropylcarbinol and prepared two kinds of processing.Electron acceptor(EA) concentration reflection adds the amount (table 5) of sample and does not comprise background election acceptor density in the on-the-spot underground water.The concentration of BTEX and alcohol represents in coming source region and the possible underground water concentration that may observe in the suitable gradient feathering in its vicinity.The alcohol concn higher with respect to isopropylcarbinol concentration that uses in this research is intended to reflect isopropylcarbinol and the ethanol effective solubleness in underground water.Ethanol has the water solubility of 10 times of water solubilities that are approximately isopropylcarbinol, and the octanol-water partition coefficient of isopropylcarbinol is the about 10 times (Organization for Economic Cooperation and Development (Organization for Economic Co-operation and Development) of the octanol-water partition coefficient of ethanol, 2004.SIDSAssessment Report for SIAM 19-Ethanol (CAS No.64-17-5) .Berlin, Germany; The Organization for Economic Cooperation and Development (Organization for Economic Co-operation and Development), 2004.SIDS Assessment Report for SIAM 19-Isobutanol (CAS No.78-83-1), Berlin, Germany).Because the biodegradation rate of expection ethanol is higher than the speed of isopropylcarbinol, has therefore conservatively selected to test than the about high 3 times ethanol volumetric molar concentration of isopropylcarbinol.

Whole microenvironment preparations are all carried out in anaerobic room.Prepared microenvironment by the 40g on-site soil being placed 160mL glass serum bottle.BTEX and alcohol are added in the processing bottle, with the aimed concn that obtains to show in the table 5.(the target BTEX concentration for each BTEX compound is listed under the BTEX hurdle.) fill bottle with underground water, stay the headspace of about 2mL.With mercury chloride (bottle in be 700mgL ^-1) and formaldehyde (bottle in for by volume 1%) revised the contrast microenvironment with the restriction micro-organisms activity.Analyze the processing of having prepared minimum 3 repetitions for pure and mild BTEX.Each processes additional monitoring electron acceptor(EA) and the methane of being recycled and reused for.

Use is with the curling bottle that sealed of isoprene-isobutylene rubber stopper of Teflon, and in 15 ℃ of incubations on the gyrate shaker of 100rpm operation.If necessary, added the additional electron acceptor to keep desired reductive condition.Contrast is revised with mercury chloride and formaldehyde.The electron acceptor(EA) concentration of revising is shown in last hurdle.The nutrient substance of improved basic salt culture medium (people such as Hareland, 1975) form was added whole processing in the time of 22 days.

Table 5: experiment processing array

Process	BTEX(μM)	Isopropylcarbinol (μ M)	Ethanol (μ M)	Electron acceptor(EA) (μ M)
					Contrast 1	180/38/38/75	-	-	-
Contrast 2	180/38/38/75	3,400	-	-
					Contrast 3	15/3.8/3.8/7.5	68	-	-
Contrast 4	15/3.8/3.8/7.5	-	11,000	-
					Process 1	180/38/38/75	-	-	-
Process 2	180/38/38/75	3,400	-	-
					Process 3	15/3.8/3.8/7.5	68	-	-
Process 4	15/3.8/3.8/7.5	-	11,000	-
					Process 5	180/38/38/75	-	-	7,100(NaNO ₃)
Process 6	180/38/38/75	3,400	-	7,100(NaNO ₃)
					Process 7	15/3.8/3.8/7.5	68	-	7,100(NaNO ₃)
Process 8	180/38/38/75	-	-	2,600(Fe(OH) ₃)
					Process 9	180/38/38/75	3,400	-	2,600(Fe(OH) ₃)
Process 10	15/3.8/3.8/7.5	68	-	2,600(Fe(OH) ₃)
					Process 11	180/38/38/75	-	-	3,200(MgSO ₄)
Process 12	180/38/38/75	3,400	-	3,200(MgSO ₄)
					Process 13	15/3.8/3.8/7.5	68	-	3,200(MgSO ₄)
Process 14		-	11,000	3,200(MgSO ₄)

Analytical procedure

The Varian CP-3900 gas chromatograph (GC) that is equipped with FID and Restek QSPLOT post by use uses the GC that is equipped with FID and Restek Rt-Alumina post for methane for BTEX, has analyzed headspace gas.By using Henry's law to calculate concentration in the water.

Be equipped with the Varian CP-3800 gas chromatograph of FID and Stabilwax DA post by use, analyzed the determining alcohol in the water (and possible isopropylcarbinol degraded product isobutyric aldehyde and isopropylformic acid).Collected moisture sample, to be used for carrying out anion analysis via ion chromatography (Dionex DX-120, Sunnyvale, CA).Nitrate also uses Quant Nitrate test strip (EMD Chemicals, Gibbstown, NJ) periodically to measure.Total uses Hach test kit (Hach, Loveland, CO) to measure according to the specification sheets of manufacturers with the iron that dissolves.

Microbiological analysis

When this experiment beginning and end, obtained microenvironment slurries sub-sample sample (approximately 8mL) from

processing

2,6,9 and 12, to determine that microbial population is through the variation of this research process.Sample is freezing immediately under-70 ℃, and (when this research finishes) be transported to Microbial Insights at dry ice, Rockford, and TN is to be used for passing through CENSUS ^TMQuantitative PCR technique (Microbial Insights, 2009http: //www.microbe.com/how-census-works.html and http://microbe.com/census-applications/anaerobic-btex.html; Respectively at 07.07.2009 and 05.08.09 access) carry out the quantitative polyase chain reaction (qPCR) to total eubacterium, denitrifying bacterium, iron and sulfate-reducing bacteria and methane-producing bacteria.

The degraded of isopropylcarbinol and ethanol

By degradable, and its degradation rate changes under different anaerobic conditions isopropylcarbinol in the processing of high density.Fig. 5 has shown isopropylcarbinol and the electron acceptor(EA) concentration in the processing (

process

2,6,9 and 12) of higher concentration.For unmodified processes and displays the background sulfate concentration.In the contrast microenvironment, do not observe the reduction of isopropylcarbinol or ethanol, or isobutyric aldehyde or isobutyric gathering.In the microenvironment that nitrate is revised, observed the most rapidly isopropylcarbinol biological degradation.In 16 days, isopropylcarbinol (is processed 6) and is degraded to being lower than detectability under the nitrate reduction condition.The degraded of nitrate and isopropylcarbinol is utilized simultaneously, and before the 14th day again modification, drops to and can not detect in the time of the 13rd day.Processing in 6, until all do not observe measurable reduction (not display data) of background vitriol in 19 days.

In the unmodified microenvironment (processing 2) of processing (the processing 12) neutralization that vitriol is revised, all observe the biological degradation of isopropylcarbinol, wherein had limited background vitriol.In order to assess isopropylcarbinol biological degradation of (that is, methanogenic condition) after vitriol exhausts, in the time of 88 days, again added isopropylcarbinol (to the final concentration of 3,400 μ M) to the microenvironment bottle of processing 2.Additional isopropylcarbinol was degraded in about 30 days.Yet the methane that trace (＜2 μ M) level only arranged is being processed in 2 after the exhausting of vitriol and is being observed, and the methane level in itself and the contrast is close.

Isopropylcarbinol in the processing that iron is revised was biodegradable in about 80 days to being lower than the analyzing and testing limit.Ferric iron by monitoring in 44 days demonstrates the concentration that scope is about 18 to 36 μ tM.Yet, only have the ferrous iron (at the most 36 μ M) of relatively low-level dissolving to be observed.Owing to the formation of Iron sulfuret in the microenvironment bottle, because observed the precipitation of black in the situation that can not exist measurable ferrous iron to gather.In addition, with the formation of Iron sulfuret consistently, the background sulfate concentration reduces in the processing that iron is revised.The reduction of isopropylcarbinol concentration is associated with the reduction of sulphate content.

In the microenvironment (processing 3,7,10 and 13) of low isopropylcarbinol concentration, in the processing that iron is revised in about 25 days, in about 15 days, isopropylcarbinol is by complete biodegradable (Fig. 9) in the processing unmodified, that nitrate is revised revises with vitriol.Outside the electron acceptor(EA) of revising, each microenvironment also comprises the background nitrate content of about 100 μ M in the underground water material, and has observed the reduction relevant with the biological degradation of isopropylcarbinol of background nitrate between incubation period.The degraded of one mole of isopropylcarbinol needs four molar nitric acid salt in theory, suppose and be reduced to nitrogen (McCarty fully, " Bioengineering issues related in situ remediation of contaminated soils and groundwater " Environmental Technology, in:Omenn, G.S., Reducing risks from environmental chemicals through biotechnology.Plenum press, New York, the 143-162 page or leaf, 1988).Enrichment culture experiment shows, the molar ratio of the nitrate of consumption and the benzene of degraded is ten, is that (Burland and Edwards, 1999, ApplEnviron Microbiol 65 (2): 529-533) for the twice of theoretical numerical value.The background nitrate of 100 μ M can be conducive to the biological degradation of about 13 μ M isopropylcarbinols, accounts for 18% of initial isopropylcarbinol in the processing of lower concentration.In the processing of lower concentration, do not observe background vitriol and reduce.

The isobutyric aldehyde of isopropylformic acid and trace is accredited as instantaneous biological degradation intermediate; In whole processing, all observed these two kinds of compounds biological degradation subsequently.Isopropylformic acid gathers near (factors with about 2) stoichiometric amount, except the processing that the nitrate of high density is revised (only observe therein 5% gather).Be difficult to explain isopropylformic acid limited generation in the nitrate of high density is processed.

In the microenvironment (processing 4 and 14) that ethanol is revised, ethanol had been degraded in about 60 days under the sulfate reduction condition to being lower than analyzing and testing limit (Fig. 6).At the 44th day, in processing 4 and 14, all detect methane (being respectively 4 and 25 μ M), but be reduced to non-detection level by the 78th day.

In order to assess the biological degradation of ethanol after vitriol exhausts, ethanol was re-filled in the time of 88 days to the bottle of processing 4.In 90 days, occured ethanol subsequently until be lower than the biological degradation of analyzing and testing limit.This result meets the ethanol research in front anaerobism, and those researchs have been reported in the situation that there is not vitriol, ethanol via the biological degradation of fermentation (people such as Laanbroek, 1982, Arch.Microbiol.133:178-184).

The BTEX degraded

At this experimental session, the decline of BTEX concentration is negligible (＜15%) in the contrast.BTEX biological degradation in higher concentration is processed is shown in Fig. 5 and 7, and the result of low concentration BTEX (with comparative Ethanol Treatment) is shown among Fig. 8.Just process with regard to 4, two last time points do not have available data, because BTEX is refilled unintentionally to this processing.

In whole high density microenvironments of revising with electron acceptor(EA), all observe the biological degradation (Fig. 7) of toluene.When in the situation that do not exist alcohol during incubation, about 38 μ M toluene were degraded to being lower than detectability under nitrate reduction, iron reduction and sulfate reduction condition respectively in 80 days.The existence of isopropylcarbinol shows slightly influential under nitrate reduction and sulfate reduction condition to the degraded of toluene, and the degraded of the toluene that in the microenvironment that iron is revised, slowed down.Yet in unmodified high density microenvironment (processing 2), measurable toluene concentration of not observing with respect to contrast reduces trend.Background vitriol in the processing 2 is at 30 days initial internal consumptions, and supposition is the biological degradation because of isopropylcarbinol, and lacks the reason that electron acceptor(EA) may be toluene sustainable existence in unmodified processing.Exist therein in the microenvironment of low concentration of sufficient electron acceptor(EA), isopropylcarbinol is at unmodified (limited vitriol) and anyly all the biological degradation of toluene is shown slight influence (Fig. 8) in the microenvironment of electron acceptor(EA) modification.The result (Figure 10) of ethylbenzene and total dimethylbenzene is similar to those results of toluene, although compare with toluene, total ethylbenzene and the biological degradation of dimethylbenzene occur slightly slow.

In the processing of the higher BTEX concentration that does not contain isopropylcarbinol, in whole 162 days incubation, under any anaerobic condition, do not observe the biological degradation (Fig. 7) of certain benzene.As if yet the existence of isopropylcarbinol has stimulated the biological degradation of benzene under iron reduction and sulfate reduction condition, because the benzene concentration of processing in 9 and 12 begins to reduce when experiment finishes.In the processing of the lower concentration under the sulfate reduction condition, through after about 300 days incubation, the concentration of benzene is reduced in containing the microenvironment of isopropylcarbinol (processing 13) and is lower than analyzing and testing limit (0.50 μ M), and is reduced to about 0.85 μ M in containing the microenvironment of ethanol (processing 14).The biological degradation of benzene is being processed beginning before 120 days in 14, and is processing generation between 160 days and 300 days in 13.

Microbiological analysis

The microbiological analysis result who shows in the table 6 has generally shown the rising of microorganism concn between incubation period.(value is with cell mL ^-1Expression; ± value indication 95% fiducial interval.)

Table 6: the result of microbiological analysis

In the microenvironment that comprises ferric iron (processing 9) or vitriol (processing 2 and 12), corresponding iron reduction and the colony of sulfate-reducing bacteria have increased respectively 1000 times.Because nitrate exhausts when approaching the end of detesting incubation in processing 6, so the colony of methanogen has increased 100 times.Having observed slightly lower level methanogen biomass in other are processed increases.

In the processing that nitrate is revised, do not observe the increase of denitrifying bacterium growth.A kind of explanation of this observations is, although two kinds of different nitrate reductase genes are monitored, the functioning gene of the denitrifying bacterium in the system is not by quantitatively.Perhaps, although the activity of denitrifying bacterium may be basic, but the growth of denitrifying bacterium may be low, viewed in studying such as the propyl carbinol of the people such as Dubbels under Denitrification Conditions (2009, Int.J.Syst.Evol.Microbiol.59:1576-1578).Equally, the zymogenic bacteria that growth does not rely on nitrate reduction also may relate to isopropylcarbinol degraded under Denitrification Conditions (people such as Laanbroek, 1982, Arch.Microbiol.133:178-184).

Embodiment 3: isopropylcarbinol is as the cosolvent of ethanol

Under 65 °F, isopropylcarbinol-alcohol-gasoline blend has been carried out the research of water tolerance scope and the test that is separated.The isopropylcarbinol of incremental change is mixed with E10 gasoline, and then added 1.3% or 2.6% water to whole blends.For some blend, water need to be increased to 2.6 volume %, absorbed a large amount of water because comprise the blended into gasoline thing of ethanol, and 1.3% water always is not enough to cause for the water of the separation of analyzing and the formation (that is, the water of 1.3 volume % is absorbed fully by higher isopropylcarbinol blend) of hydrocarbon phase.This is found among Figure 11, and therein, 1.3 volume % are enough to obtain two kinds of phases to the amount of isopropylcarbinol reaches 5 volume % in E10 till, but with regard to the isopropylcarbinol of 10 volume % among the E10, and the water that its level need to be increased to 2.6 volume % causes and is separated.

Data presentation among Figure 12, the amount that is extracted into the ethanol of water reduces along with the rising of isopropylcarbinol concentration, shows that isopropylcarbinol is as the cosolvent of ethanol.By GC the concentration of hydrocarbon and aqueous phase is all measured.

The description of aforementioned particular will disclose general essence of the present invention fully, so that other staff can be in the situation that do not break away from universal of the present invention, need not to use the technical knowledge in this area with carrying out undo experimentation, easily revise and/or change a plurality of application of this type of particular.Therefore, this type of changes and modification is intended to based on instruction provided herein and guidance, in the described disclosed purpose and scope that is equal to embodiment.Should understand the term of this paper or the purpose of term is that description is not restriction, therefore the term of this specification sheets or term are according to originally being description and hard-core purpose, like this, the term of this standard or term are explained according to instruction and guidance by those skilled in the art.

Claims

1. be used for to improve the recyclability of hydrocarbon fuel composition and be limited in when being polluted by described hydrocarbon fuel composition method on the impact of environment compartment, it is included in an amount of isopropylcarbinol of addings in the described hydrocarbon fuel composition with for the biodegradability that improves described hydrocarbon fuel composition.

2. the process of claim 1 wherein that described hydrocarbon fuel composition also comprises ethanol.

3. the process of claim 1 wherein that described ethanol consisted of at the most approximately 10% of described fuel composition before adding isopropylcarbinol.

4. the process of claim 1 wherein that described isopropylcarbinol provides the biological degradation of at least a improvement in the BTEX component of described hydrocarbon fuel composition.

5. the process of claim 1 wherein that described isopropylcarbinol provides the biological degradation of the improvement of benzene.

6. the method for claim 2, described environment compartment comprises soil matrix, and wherein the interpolation of isopropylcarbinol reduces the transmission of ethanol in soil matrix.

7. the process of claim 1 wherein that the interpolation of isopropylcarbinol hinders the BTEX feathering from the expansion of described composition.

8. the process of claim 1 wherein that the interpolation of isopropylcarbinol has improved the biodegradability of the various components of described hydrocarbon fuel composition.

9. claim 4 or 5 method, the biological degradation of wherein said improvement occurs under aerobic conditions.

10. claim 4 or 5 method, the biological degradation of wherein said improvement occurs under nitrate reduction or sulfate reduction condition.

11. improve comprise isopropylcarbinol hydrocarbon fuel composition under anaerobic the environment in the environment compartment return the method that becomes, it amount that comprises being enough to improving the biodegradation rate of one or more BTEX components adds electron acceptor(EA) in described compartment.

12. the method for claim 11, wherein said electron acceptor(EA) are iron, vitriol or nitrate or their combination.

13. the method for claim 11, wherein said electron acceptor(EA) are Fe (OH) ₃

14. the method for claim 11, wherein said electron acceptor(EA) is NaNO ₃

15. the method for claim 11, wherein said electron acceptor(EA) is MgSO ₄

16. the method for claim 11, wherein said one or more BTEX components comprise toluene.

17. the method for claim 11, wherein said one or more BTEX components comprise dimethylbenzene.

18. the method for claim 11, wherein said one or more BTEX components comprise benzene.

19. the method for claim 11, wherein said electron acceptor(EA) are nitrate and add with the amount that is enough to create the nitrate reduction condition.

20. the method for claim 11, wherein said electron acceptor(EA) are vitriol and exist with the amount that is enough to create the sulfate reduction condition.

21. the method for claim 11, wherein said electron acceptor(EA) be nitrate and exist with the amount that is enough to create the nitrate reduction condition, and wherein toluene with the approximately identical fate of isopropylcarbinol in carry out biological degradation.

22. the method for claim 11, wherein said electron acceptor(EA) be vitriol and exist with the amount that is enough to create the sulfate reduction condition, and wherein toluene with the approximately identical fate of isopropylcarbinol in carry out biological degradation.

23. the method for claim 11, wherein said electron acceptor(EA) be nitrate and exist with the amount that is enough to create the nitrate reduction condition, and wherein benzene with the approximately identical fate of isopropylcarbinol in carry out biological degradation.

24. the method for claim 11, wherein said electron acceptor(EA) be vitriol and exist with the amount that is enough to create the sulfate reduction condition, and wherein the biological degradation of benzene is compared with its biological degradation when not having the sulfate reduction condition and improved.

25. the method for claim 11, wherein said electron acceptor(EA) be vitriol and exist with the amount that is enough to create the sulfate reduction condition, and wherein the biological degradation of benzene with it in the situation that do not exist the biological degradation of isopropylcarbinol to compare to improve.

26. the method for claim 11, wherein said electron acceptor(EA) be vitriol and exist with the amount that is enough to create the sulfate reduction condition, and wherein the biological degradation of benzene is compared at the biological degradation in the presence of the ethanol with it and improved.

27. composition, it comprises gasoline, isopropylcarbinol and Fe (OH) ₃, NaNO ₃, or MgSO ₄In at least a.

28. composition, it comprises gasoline, isopropylcarbinol and Fe (OH) ₃, NaNO ₃, KNO ₃, NHNO ₃, Na ₂SO ₄, CaSO ₄, MgSO ₄, at least a in chelated iron, Zero-valent Iron and the nano zero valence iron.