Biodegrading Recalcitrant To Biodegradation Organic Substances

Abstract

A composition for stimulating the production and excretion of a lignolytic enzyme in a microorganism for degrading harmful substances and/or in the manufacturing of easily degradable ester containing plastics or articles made of ester containing plastic. The composition mainly includes tributyrin, triolein, fish oil, 16-hydroxyhexadecanoic acid, n-aliphatic primary fatty alcohols, polycaprolactone, aliphatic polyesters, linolenic acid, linoleic acid, alpha linolenic acid, plant polyesters, cutin, cutin derivatives, cutin monomers, omega hydroxy acids, 16-hydroxy palmitic acid, 9,16-dihydroxypalmitic acid, 10,16-dihydroxypalmitic acid, C18-hydroxy oleic acid, 9,10-epoxy-18-hydroxy stearic acid, 9,10, 18-trihydroxystearate, suberin, cork, fruit skins, vegetable skins, and their constituents and derivatives, hydroxy fatty acids, 16-hydroxy palmitic acid, 18-hydroxy stearic acid, juniperic acid, hexadecanol, linseed oil, perilla oil, amides, acetamide and N-acetyl amide, zinc, zinc salts, butyrate, acetate, lactate, manganese peroxidase, and carbamide peroxide.

Claims

1. A composition selected from a group consisting of tributyrin, triolein, fish oil, 16-hydroxyhexadecanoic acid, n-aliphatic primary fatty alcohols, polycaprolactone, aliphatic polyesters, linolenic acid, linoleic acid, alpha linolenic acid, plant polyesters, cutin, cutin derivatives, cutin monomers, omega hydroxy acids, 16-hydroxy palmitic acid, 9,16-dihydroxypalmitic acid, 10,16-dihydroxypalmitic acid, C18-hydroxy oleic acid, 9,10-epoxy-18-hydroxy stearic acid, 9,10,18-trihydroxystearate, suberin, cork, fruit skins, vegetable skins and their constituents and derivatives, hydroxy fatty acids, 16-hydroxy palmitic acid, 18-hydroxy stearic acid, juniperic acid, hexadecanol, vegetable oils, linseed oil, perilla oil, amides, acetamide and N-acetyl amide, zinc, zinc salts, butyrate, acetate, lactate, and manganese peroxidase; wherein admixing the composition with the substance resistant to biodegradation, allows said carbamide peroxide to stimulate production and excretion of enzyme superoxide dismutase in the microorganism.

2. The composition as claimed in claim 1, wherein the composition comprises one or more ingredients selected from a group consisting of 16-hydroxyhexadecanoic acid and linoleic acid.

3. The composition as claimed in claim 1, wherein the composition further comprises one or more nutrient substances for the growth of microorganism, the one or more nutrient substances selected from a group consisting of a basal mineral salt mixture, a carbon source, a nitrogen source, and a vitamin; wherein, said vitamin is selected from a group consisting of thiamine, biotin and a mixture thereof; said carbon source is selected from a group consisting of fatty acids, polyols, sugars, organic acids, cellobiose and cellulose; said nitrogen source is selected from a group consisting of ammonium nitrate, sodium nitrate, ammonium sulfate, urea and ammonia; and wherein said substances feed and encourage the growth of said microorganism or microorganisms.

4. The composition of claim 1 admixed with a degradable ester containing plastic wherein the degradation of the degradable ester plastic is hastened.

5. The composition of claim 1 admixed with an easily degradable article during manufacturing, wherein the degradation of the degradable ester plastic is hastened.

Description

DETAILED DESCRIPTION

[0020] While various aspects and features of certain embodiments have been summarized above, the following detailed description illustrates a few exemplary embodiments in further detail to enable one skilled in the art to practice such embodiments. The described examples are provided for illustrative purposes and are not intended to limit the scope of the invention.

[0021] In the following description, for the purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the described embodiments. It will be apparent to one skilled in the art however that other embodiments of the present invention may be practiced without some of these specific details. Several embodiments are described herein, and while various features are ascribed to different embodiments, it should be appreciated that the features described with respect to one embodiment may be incorporated with other embodiments as well. By the same token however, no single feature or features of any described embodiment should be considered essential to every embodiment of the invention, as other embodiments of the invention may omit such features.

[0022] In this application the use of the singular includes the plural unless specifically stated otherwise and use of the terms and and or is equivalent to and/or, also referred to as non-exclusive or unless otherwise indicated. Moreover, the use of the term including, as well as other forms, such as includes and included, should be considered non-exclusive. Also, terms such as element or component encompass both elements and components including one unit and elements and components that include more than one unit, unless specifically stated otherwise.

[0023] Lastly, the terms or and and/or as used herein are to be interpreted as inclusive or meaning any one or any combination. Therefore, A, B or C or A, B and/or C mean any of the following: A; B; C; A and B; A and C; B and C; A, B and C. An exception to this definition will occur only when a combination of elements, functions, steps or acts are in some way inherently mutually exclusive.

[0024] As this invention is susceptible to embodiments of many different forms, it is intended that the present disclosure be considered as an example of the principles of the invention and not intended to limit the invention to the specific embodiments shown and described.

[0025] The admixture of certain commercially available substances, as well as agricultural byproducts, will cause ligninolytic microorganisms to excrete the ligninolytic systems, resulting in the biodegradation of many unwanted organic substances, including but limited to, hydrocarbon plastics and many petroleum and natural gas based substances, including but not limited to, synthetic plastics, petroleum, gasoline, diesel, polyaromatic hydrocarbons, lubricating oils, aniline dyes, PCBs, BTEX toxins, bisphenols, phthalates, creosote, olive oil mill wastes, herbicides, pesticides, antibiotics, plant pitch, phenolic substances, explosives and their derivatives, lignin, and lignin derivatives and metabolites, including but not limited to, kraft lignins and black liquor.

[0026] Substances which stimulate the production and excretion of, or which increase the ligninolytic system production and excretion include, but are not limited to: Explosives and their metabolites, including but not limited to trinitrotoluene, polysorbate 80, ethanol, salts of transition metals, including but not limited to iron, copper, and manganese, and many organic substances which include a phenolic moiety, including but not limited to, phenolic compounds containing one or more methoxy phenolic moieties, including but not limited to, a phenol in which there are two methoxy groups attached, including but not limited to, one methoxy moiety one each side of the hydroxyl group attached to the benzene ring, not including acetosyringone, including but not limited to, kraft lignins, guaiacol, syringaldehyde, vanillin, acetovanillone, p-coumaric acid, ferulic acid, sinapic acid, coumarin, catechol, orcinol, resorcinol, eugenol, pyrogallol, acetaminophen, tannic acid, 2,5-xylidine, and many more similar compounds.

[0027] Phenolic substances, (substances in which a benzene ring has at least one hydroxy group attached to the benzene ring,) contribute to a chain of radical production, a chain reaction of radicalization by being themselves subject to being radicalized, typically first into phenoxy radicals, then in turn creating more powerful oxygen radicals, in a chain of reactions in which radicals impact and radicalize other oxygen containing substances. Linoleic acid can also contribute to the creation of radicals, due to being prone to radicalization at its unsaturated bonds, by hydrogen abstraction. The drying oils are especially prone to this form of radical creation by hydrogen abstraction at the unsaturated carbon to carbon bonds, even being able to auto-oxidize, thus initiating a cascade of oxygen radical production.

[0028] The best way to biodegrade these substances is to admix the ligninolytic system stimulating chemicals in a moist environment containing ligninolytic microorganisms, by stimulating wild microorganisms or cultured ligninolytic microorganisms with chemical stimulators of the ligninolytic systems' production and excretion. The process would be further enhanced by mixing in basal salt mixtures, including but not limited to, Murishige and Skoog's, and readily assimilated carbon sources, including but not limited to fatty acids, cellulose, polyols, starches, sugars and polymers of sugars, and nitrogen containing substances, including but not limited to, ammonium nitrate, ammonium tartrate, ammonium sulfate, urea, ammonia, and B vitamins, including but not limited to, thiamine, biotin, or thiamine and biotin containing substances, to nourish the microorganisms.

[0029] These nutrient mixtures can nourish ligninolytic bacteria, ligninolytic yeast, and ligninolytic fungi. The process would be further improved by supplying additional (that is moderately more than is usually found in basal salt mixtures,) but still vert minute amounts of bioassimilable mineral salts of transition metals, including but not limited to, salts of iron, copper and manganese, including but not limited to, sulfur salts thereof.

[0030] Below is an effective embodiment of the invention for biodegrading substances subject to biodegradation by ligninolytic systems, when admixed with these substances in an environment that is moist or wet, and slightly aerated, and containing colonies of ligninolytic microbes, fungi, or blue green algae, sufficient to biodegrade one metric ton of the substance to be biodegraded. The amount of the invention needed to cause effective biodegradation of recalcitrant to biodegradation substances is relatively small, less than 3% of the weight of the substance to be biodegraded. The figures are in kilograms per ton of substrate. [0031] cellulose 15 [0032] polysorbate 80 1.65 [0033] raw linseed oil 1.008 [0034] guaiacol 1.5 [0035] vanillin 1.5 [0036] refined glycerol 1.5 potassium sulfate, anhydrous 1.4944 [0037] calcium chloride anhydrous 0.30496 [0038] potassium phosphate monobasic 0.16684 magnesium sulfate 0.1584 [0039] calcium carbonate 0.09296 ammonium tartrate 0.09296 [0040] manganese sulfate 0. [0041] zinc sulfate 0.00772 boric acid 0.00548 [0042] potassium iodide 0.00076 thiamine hydrochloride 0.001 [0043] molybdenum disulfide 0.0002234 [0044] ferrous sulfate 0.00003488 [0045] cobalt chloride 0.00000222 [0046] copper sulfate pentahydrate 0.00000222 [0047] sodium benzoate 0.00000222

[0048] In one embodiment of the invention for use in conventional synthetic plastic products, an effective way of admixing the above formula in plastics which do not contain oxygen or nitrogen in their molecular structure is to make pellets of the plastics with compatibilizers that make non polar plastics compatible with polar ingredients, in which the embodiment is mixed with compatibilized plastics in higher concentrations than is intended in the end product. Such compatibilizers are available commercially from a variety of vendors. These compatibilizers are commonly available in pellets, and they usually consist of nonpolar plastics that have been modified by covalently bonding the plastic with oxygen containing groups, including but not limited to, maleic anhydride, glycidyl methacrylate, and acrylic esters. These pellets, containing higher concentrations of the embodiment than intended to be used in finished products, are meant to be later mixed with plastics lacking oxygen containing groups and otherwise lacking the embodiment.

[0049] The embodiment of the invention is effectively combined with the compatibilizer in a machine, including but not limited to, a twin screw extruder, which melts, mixes, and pelletizes the compatibilizer and the embodiment of the invention. Subsequently, the plastic to be biodegraded is admixed with the pellets consisting of a compatibilizer and the embodiment of the invention, in a machine which melts and mixes the embodiment pellets and the plastic pellets. The resulting pellets are then turned into end products by methods known to those skilled in the art of thermoplastic manufacturing. Such end products are everyday products meant to have a useful lifespan of less than one year, including but not limited to, plastic shopping bags, disposable eating utensils, packaging for food and other commercial products, bottles, straws, cups, cup lids, stirrers, stretch wrap, etc.

Biodegrading Ester Group Containing Substances

[0050] Plastics containing ester moieties are somewhat resistant to biodegradation by ligninolytic systems, probably because of hydrogen bonding within ester-containing plastics, mutually reinforcing their long molecules, making them resistant to biodegradation by oxygen radicals.

[0051] Oxygen containing plastics, containing ester moieties, including but not limited to polyethylene terephthalate, epoxy resins, polyethylene furanoate, polytrimethylene furandicarboxylate, acrylate plastics, methyl methacrylate, sodium polyacrylate, and polylactic acid can be biodegraded by esterases, including but not limited to, cutinase.

[0052] Cutinase is a natural poleyesterase enzyme, the natural function of which is to biodegrade the natural polyester coating of leaves and bark. Because leaves and bark are ubiquitous, microorganisms which can biodegrade natural polyesters are ubiquitous. Once the leaves and bark have been penetrated, the microorganisms which excrete the cutinase have access to the cellulose, hemicellulose, and sugars found within the plant, which are food sources for the fungi and bacteria. The microorganisms which excrete cutinase, include but are not limited to, fungal plant pathogens, which include but are not limited to, Fusarium species, Magnaportha species, Colletotrichum species, and a few bacteria species.

[0053] Plant pathogens are ubiquitous, and they may be stimulated into excreting esterases, including but not limited to, cutinase, by substances which are constituents of plant cutin, or which resemble constituents of plant cutin. Examples of this are 16-hydroxyhexadecanoic acid, polysorbate 80, and linoleic acid, which stimulate this excretion, resulting in the biodegradation of ester group containing substances, including but not limited to, plastics, by means of severing the ester groups from the carbon atoms to which it is bonded. The resulting severed moieties are readily biodegraded by many microorganisms.

[0054] The process would be further enhanced by mixing in basal salts and readily assimilated carbon and nitrogen sources. These nutrient mixtures can nourish lipase and cutinase excreting organisms and microorganisms. The process would be further improved by supplying additional mineral salts of transition metals, including but not limited to salts of zinc, which is a constituent of cutinase, and of nonionic surfactants, including but not limited to, polysorbate 80 and 20, which increase the permeability of cell walls.

[0055] Here is an effective embodiment of the invention for biodegrading substances subject to biodegradation by cutinase or lipases, when admixed with these substances in an environment that is moist or wet, and containing colonies of cutinase or lipase excreting fungi or bacteria, including but not limited to plant pathogens, sufficient to biodegrade one metric ton of the substance to be biodegraded. The figures are in kilograms per ton of substrate.

[0056] Cellulose 15 polysorbate 80 1.65 Refined glycerol 1.5 flaxseed oil 5 [0057] 16-hydroxyhexadecanoic acid 0.8 [0058] potassium sulfate, anhydrous 1.4944 [0059] calcium chloride anhydrous 0.30496 [0060] potassium phosphate monobasic 0.1584 [0061] magnesium sulfate 0.1506 [0062] calcium carbonate 0.09296 [0063] ammonium tartrate 0.08936 [0064] manganese sulfate 0.01544 [0065] zinc sulfate 0.00772 [0066] boric acid 0.00548 [0067] potassium iodide 0.00076 [0068] thiamine hydrochloride 0.001 [0069] molybdenum disulfide 0.0002234 [0070] ferrous sulfate 0.00003488 [0071] cobalt chloride 0.00000222 [0072] copper sulfate pentahydrate 0.00000222 [0073] sodium benzoate 0.00000222

[0074] In an ester biodegrading embodiment of the invention, an optional method of admixing the above formula in plastics is to admix it with the kind of plastic containing oxygen or nitrogen to be biodegraded, in which the embodiment is mixed with plastics in higher concentrations than is intended in the end product, while the plastic is in a melted state. The embodiment of the invention is effectively combined with the oxygen or nitrogen containing synthetic plastic in a machine, including but limited to, a twin screw extruder or Brabender mixer, which melts, mixes, and pelletizes the compatibilizer and the embodiment of the invention. Subsequently, the plastic to be biodegraded is admixed with the embodiment containing pellets with more melted pellets of the plastic to be biodegraded, and the embodiment of the invention. The resulting pellets are then turned into end products by methods known to those skilled in the art of thermoplastic manufacturing, the end products including but not limited to, PET bottles.

[0075] The formula above can be used in many embodiments for the biodegradation of various substances, but substituting the correct stimulators for the flaxseed oil and 16-hydroxyhexadecanoic acid listed above in similar amounts, the different stimulators as delineated in the claims, according to the specific substance to be biodegraded, as per those in the claims attached to this patent application, those independent claims being numbered 12 and 15.

Biotransformation of Hydrogen Sulfide, to Prevent its Transformation to H2SO4 by Microbes

[0076] Superoxide dismutase, a ubiquitous enzyme known for rendering oxygen radicals less harmful, alters hydrogen sulfide into harmless substances which do not contribute to concrete erosion.

[0077] Researchers have proposed different theories regarding the origin of hydrogen sulfide in sewers. Whatever its source, hydrogen sulfide in sewers is transformed by bacteria living in biofilms attached to the concrete pipes and structures in sewers into sulfuric acid, which degrades concrete, causing billions of dollars worth of damage to sewers every year. The addition of superoxide dismutase (SOD) to sewers will greatly reduce this hugely expensive to repair damage to the concrete. Zinc/copper SOD is widely available as a commercial product from chemical suppliers, at prices that make the use of SOD in sewers a bargain, compared to repairing sewer damage, or compared to continuous admixture of inorganic chemicals that bind to hydrogen sulfide or sulfuric acid.

[0078] Extracted SOD may be admixed to H2S containing waste water by any means, including but not limited to, the admixture of copper/zinc exo-SOD to the sewer water, or by culturing SOD excreting microorganisms in the sewer water, or by the growth of said microorganisms attached by exopolysaccharides to an attachment medium placed in sewer water. Examples of these SOD excreting microorganisms include but are not limited

to, Aspergillus species, Penicillium species, and Cryptococcus species. It would be advantageous to provide pre-cultured media for the microorganisms to cling to, including but not limited to, plastic mats, brushes, ropes, foams, and fiber bunches, and commercial products such as Zeeweed, so that they would not be swept away by the flowing sewer water.

[0079] Using SOD for this purpose would be less expensive than the currently used chemical additives, because the SOD does not bind itself to the hydrogen sulfide, but rather binds additional sulfur atoms to hydrogen sulfide repeatedly, without itself being bound to the hydrogen sulfide.

[0080] Another application for SOD removal of H2S is in removal of H2S from petroleum and natural gas refining plants and effluents. H2S removal from petroleum and natural gas is referred to as sweetening. Alkanolamines such as DEA, MEA, and MDEA are currently used for this purpose, but they have to be removed from sweetening plant waste water due to environmental protection laws. SOD biotransforms H2S via combining H2S+O2, to H2S2, H2S3, H2S4 and H2S5, which substances do not have to be removed from wastewater to legally discharge the treated water in countries with rigorous environmental protection laws.

Biodegrading Substances that Contain Amide Groups

[0081] Enzymes called amidases biodegrade substances that contain amide groups, including but not limited to, polyurethanes and polyamides. Chemical stimulators will elicit the production and excretion of amidases in many microbes. These chemical stimulators include, but are not limited to, amides, acetamide, acrylamide, butyramide and urea. Organisms that can produce and excrete amidases include, but are not limited

to, Pseudomonas species, Mycobacterium species, Rhodococcus species, Streptomyces species, Klebsiella species, Burkholderia species, Alcaligenes species, and Arthrobacter species.

Biodegrading Nitrile Moiety Containing Substances

[0082] Biodegrading substances that contain nitrile groups, including but not limited to, carboxylated nitrile butadiene rubber (XNBR) and hydrogenated nitrile butadiene rubber (HNBR)

[0083] Many millions of protective XNBR nitrile rubber gloves are manufactured and disposed of every year, and many millions of tires containing HNBR are manufactured and disposed of every year. These products are very resistant to biodegradation, so it is a huge benefit to the environment to make them biodegradable. XNBR products can have biodegradation stimulating substances incorporated in them during their manufacture. Despite being very resistant to chemical penetration, XNBR has a number of moieties that can be readily biodegraded by ligninolytic systems, ester degrading enzymes, and nitrile biodegrading enzymes.

[0084] Therefore, any substance for enhancing biodegrading XNBR would optimally include the claimed inducers of the production and excretion of these enzymes and systems, and the claimed nutrients, followed by the subsequent mixing of the treated substance with the claimed microorganisms for the biodegradation of these moieties. HNBR is a hydrocarbon containing a hydrocarbon backbone, and nitrile side groups, and so is biodegradable by ligninolytic systems in combination with nitrile degrading enzymes, assisted by nutritional substances, when combined with the claimed fungi, yeast, or microbes.

Definition of Basal Salts

[0085] The term basal salts as used herein means a mixture of elements in the form of water soluble compounds, said elements being necessary to organisms of every kind. Salts of nitrogen, calcium, magnesium, potassium, boron, colbalt, iron, manganese, molybdum, copper, and iodine are typically included in basal salt mixtures, in the form of water soluble compounds, often in the forms of compounds containing O, H2O, SO4 and Cl2, to render them soluble in water.

Definition of Organically Assimilable Nitrogen Sources

[0086] Organically assimilable nitrogen sources means herein that a nutrient substance contains nitrogen in its composition, and said nitrogen is capable of being assimilated by microorganisms. The group of organically assimilable nitrogen sources comprises substances which include, but are not limited to, urea, nitrates, nitrites, ammonium compounds, and amino acids.

Definition of Organically Assimilable Carbon Sources

[0087] Organically assimilable carbon sources means herein that a nutrient substance contains carbon in its composition, and said carbon is capable of being assimilated by microorganisms. The group of organically assimilable carbon sources comprises substances which include, but are not limited to, sugars, polyols, carbohydrates, fatty acids, and organic acids, and compounds which contain these substances as constituents.

Definition of B Vitamins

[0088] B vitamins means herein substances which comprise a group of substances which contain B vitamins and their precursors and derivatives, including but not limited to, thiamin, biotin, bran, and substances which contain these substances as constituents.

[0089] Definition of the Words Biodegrade, Biodegradation, Compost, Home Compost, Decompost, Mineralize, Bioremediate, and Biotransform

[0090] Used herein, all of these words shall mean to so affect a problematic, difficult to biodegrade recalcitrant to biodegradation substance by means of an enzyme, an enzymatic system, or by an organism or microorganism via the production of enzymes or enzyme systems, so as to render said substances less toxic, less hazardous, less corrosive, less harmful to the environment, or in any other way less problematic.

Definition of Organic Substances

[0091] Organic substances as used herein shall mean carbon-containing compounds, as is usual, but also means compounds resulting from the activity of microorganisms, such as hydrogen sulfide and sulfuric acid.

Limitations and Claims of the Patent

[0092] Nothing in the abstract, field of invention, background, description, embodiment formulas, specification, or other non-claim parts of this patent application shall be construed as limiting the scope of the invention. Only the claims attached to this document are meant to claim or limit any aspect of the invention.

[0093] Since many modifications, variations, and changes in detail can be made to the described embodiments of the invention, it is intended that all matters in the foregoing description and shown in the accompanying drawings be interpreted as illustrative and not in a limiting sense. Furthermore, it is understood that any of the features presented in the embodiments may be integrated into any of the other embodiments unless explicitly stated otherwise. The scope of the invention should be determined by the appended claims and their legal equivalents.

[0094] In addition, the present invention has been described with reference to embodiments, it should be noted and understood that various modifications and variations can be crafted by those skilled in the art without departing from the scope and spirit of the invention. Accordingly, the foregoing disclosure should be interpreted as illustrative only and is not to be interpreted in a limiting sense. Further it is intended that any other embodiments of the present invention that result from any changes in application or method of use or operation, method of manufacture, shape, size, or materials which are not specified within the detailed written description or illustrations contained herein are considered within the scope of the present invention.

[0095] Insofar as the description above and the accompanying drawings disclose any additional subject matter that is not within the scope of the claims below, the inventions are not dedicated to the public and the right to file one or more applications to claim such additional inventions is reserved.

[0096] Although very narrow claims are presented herein, it should be recognized that the scope of this invention is much broader than presented by the claim. It is intended that broader claims will be submitted in an application that claims the benefit of priority from this application.

[0097] While this invention has been described with respect to at least one embodiment, the present invention can be further modified within the spirit and scope of this disclosure. This application is therefore intended to cover any variations, uses, or adaptations of the invention using its general principles. Further, this application is intended to cover such departures from the present disclosure as come within known or customary practice in the art to which this invention pertains and which fall within the limits of the appended claims.