Chemistry: molecular biology and microbiology – Process of utilizing an enzyme or micro-organism to destroy... – Destruction of hazardous or toxic waste
Reexamination Certificate
1999-10-14
2001-07-31
Beisner, William H. (Department: 1744)
Chemistry: molecular biology and microbiology
Process of utilizing an enzyme or micro-organism to destroy...
Destruction of hazardous or toxic waste
C435S244000, C504S117000
Reexamination Certificate
active
06268206
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to compositions for bioremediation and detoxication of contaminated media and facilitating plant growth in contaminated media. More specifically, compositions containing microorganisms treated with compounds containing any one of: cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), adenylate cyclase, guanylate cyclase, forskolin, or the homologs, analogs and derivatives of such compounds, and methods for making and using such compositions are disclosed.
BACKGROUND
0
F THE INVENTION
Hydrocarbons, including petroleum based hydrocarbons, can be released into the environment from industrial processes through discharges and emissions, from surface spills such as open water spills, above ground storage tank overfilling and releases, from oil spills and tanker purging during ship transport, from oil drilling and exploration operations, from agricultural operations such as cropland weed and pest management associated with chemical applications, and from subsurface releases such as leaking underground storage tanks, undesirably polluting the environment. “Petroleum based hydrocarbons” is understood to include by way of example, and not as a limitation to the present invention, chemicals toxic to the environment such as crude oil, distilled or refined fuels, halogenated pesticides, halogenated herbicides, halogenated aromatic hydrocarbons, polycyclic aromatic hydrocarbons, and/or methane and the like. Release of such toxic chemicals is referred to herein as a “contaminant”. The environment into which the contaminant is released, be it liquid or solid or gaseous media, includes by way of example, and not as a limitation to the present invention, surface water, ground water, wells, rivers, estuaries, the ocean, surface sediment, subsurface soil, surface soil, foliage and/or soil containing natural plant growth, and/or air, and the like and is referred to herein as “contaminated media”.
Oil spilled at sea often reaches the littoral zone and becomes coated on or mixed with soil, or other solids, such as rocks, and vegetation. Oil adsorption and absorption on or in these materials renders the petroleum contaminants less mobile and difficult to clean up or remediate using conventional physical/mechanical remediation techniques, such as filtering, excavation or mechanical removal. Such techniques also tend to adversely affect the ecosystems in proximity to the contaminants.
Contaminants discharged from industrial processes are often mixed as an aqueous influent to treatment systems and then as an effluent to receiving waters. These chemicals are also adsorbed on suspended solids in water and can settle out and thus become incorporated into the sediment. Once settled in the sediment, the contaminants become less mobile and difficult to remediate using conventional physical/mechanical remediation techniques without causing damage to surrounding ecosystems.
Herbicides and pesticides used in cropland management are applied to plants and released in spills or as excess in over application. These applications and releases are washed into the soil and plants from irrigation practices or from atmospheric precipitation. Such herbicides and pesticides accumulate in the rhizosphere where conventional physical/mechanical remediation techniques also prove difficult or detrimental to the surrounding ecosystem.
Distilled or refined fuels, including chemical mixtures such as gasoline and diesel fuels stored in above ground or underground storage tanks released to the environment from overspills during tank filling and tank and piping failures are adsorbed or absorbed respectively into the soil and plants adjacent to such releases. There they tend to become less mobile and/or accumulate on or in the groundwater resulting in dissolved aqueous and non-aqueous phase liquids containing these chemical contaminants. Further migration of these chemicals also results in widespread aquifer contamination. Such contaminated media are also difficult to remediate and can harm human health and sensitive ecosystems.
Bioremediation of these contaminants tends to be the best technology and most cost effective means to remediate such contaminated media. What is meant by “bioremediation” or “bioremediation activity” is the microbial oxidation and/or mineralization (i.e., biodegradation) of contaminants. “Mineralization” means the bioconversion of contaminants to carbon dioxide, water and occasionally new microbial cell growth. “Detoxication” (sometimes designated “detoxification”) refers to the change in a contaminant (molecule or complex mixture) that renders it less harmful to one or more susceptible organisms (i.e. microorganisms, plants, animals or humans). See M. Alexander,
Biodegradation and Bioremediation,
pp. 41-48 (1994), incorporated herein by reference.
Many naturally occurring microorganisms are useful in microbial oxidation and mineralization of such contaminants because they generate enzymes that oxidize and mineralize the contaminants through what is known metabolically as catabolism. However a disadvantage of conventional microbial oxidation and mineralization is that the rate of biochemical oxidation (i.e., bioremediation) of the contaminated media is often undesirably slow. One reason for the slow bioremediation rate of the contaminants is that the microbes may exist only in small amounts naturally in the contaminated media. It has also been purported that useful indigenous microorganisms tend to be attacked by predatory microorganisms, thereby keeping the amount of indigenous microorganisms relatively low. In addition, naturally occurring microorganisms may only produce small amounts of the enzymes required for oxidation and mineralization thereby also causing slow oxidation and mineralization rates of the contaminants. It is believed that the rate of enzyme production occurs at low rates and can be ultimately inhibited through a cellular phenomenon known as catabolite repression.
It is known that microorganisms tend to exhibit decreased production of enzymes useful in oxidizing and mineralizing contaminants when insulted with toxic contaminants thereby decreasing or stopping the rate of biodegradation of such contaminants. These reasons, either combined or independently, render microbial biodegradation of contaminated media undesirably slow and/or incomplete to mineralization.
Genetically engineered microorganisms (GEMs) have been developed in an attempt to overcome some of the above problems with naturally occurring microorganisms. GEMs however tend to be unacceptable to environmental regulatory authorities for widespread and uncontrolled environmental application due to their uncertain effects on the environment.
There are numerous enzymes present in microorganisms known to catalyze the biodegradation of the contaminants. In bacteria, cAMP has been found to play a role in the formation of the constitutive enzymes necessary to catalyze the breakdown of secondary sugars such as galactose and arabinose in the presence of glucose. However, enzymes known to be associated with the break down of sugars, such as glucose, lactose and galactose, and not those associated with the oxidation and mineralization of toxic contaminants tend to exhibit cAMP induced activity. For example, H. V. Rickenberg,
Cyclic AMP in Prokaryotes,
pp. 353-369 (1974), (“Rickenberg”), describes the synthesis of several proteins (enzymes) controlled by cAMP. It has been demonstrated that cAMP is required for the effective synthesis of beta-galactosidase and tryptophanase in
E. Coli.
Rickenberg further describes that in “
E. Coli
exogenous cAMP overcomes both the severe transient (citation omitted) and less severe steady-state catabolite repression (citation omitted) of the synthesis of beta-galactosidase caused by the presence of glucose in the medium.” H. V. Rickenberg,
Cyclic AMP in Prokaryotes,
pp. 354-355.
It was found that inhibition of biolumenescence of Photobacterium phosphoreum is a useful measure of general toxicity associated with solids including soil
Beisner William H.
St. Onge Steward Johnston & Reens LLC
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