BRPI0806978A2 - composition and method of improving the functionality of a surfactant system - Google Patents

Abstract

COMPOSITION AND METHOD OF IMPROVING THE FUNCTIONALITY OF A TENSIVE SYSTEM. Compositions comprising a surfactant and a protein system are disclosed, wherein the protein system comprises the proteins and stress proteins obtained by the yeast fermentation process to obtain a fermentation mixture; subjecting the fermentation mixture to stress conditions to obtain a post-fermentation mixture; and centrifuging the post fermentation mixture and obtaining the supernatant; where the protein mixture retains its functionality under extreme conditions. Also disclosed are methods of using the above compositions as enhanced oil recovery agents, cleaning agents or as surfactant enhancing agents.

Description

COMPOSITION AND METHOD OF IMPROVING THE FUNCTIONALITY OF A TENSIVE SYSTEM

FIELD OF INVENTION

The present invention relates to the field of enhanced oil recovery and recovery solutions comprising pH and heat stable proteins and peptides and methods of using them.

BACKGROUND OF THE INVENTION

There is a critical need in the marketplace to minimize the negative toxicological profiles of chemical compositions and formulations and to increase the use of renewable resource chemicals while increasing the performance of chemical solutions and achieving those goals with low cost compounds. Broader uses of chemicals, especially those that are not isolated from the environment, include cleaning and certain industrial processes. Despite the ecological benefits, the increased urgency of developing greener chemicals includes regulatory pressure such as the European REACH initiative, which will firmly regulate all bulk chemicals in Europe. Further pressure comes from new data on exposure to substance toxicity, some of which is common and used in homes and industry, and the rising costs related to the liability of companies dealing with toxic substances.

BRIEF DESCRIPTION OF THE INVENTION

Compositions comprising a surfactant and protein mixture wherein the protein mixture comprises proteins and stress proteins obtained by the yeast fermentation process to obtain a fermentation mixture are described in the present invention; subsequently subjecting the fermentation mixture to stress conditions to obtain a post-fermentation mixture; and centrifuging the post-fermentation mixture and obtaining the supernatant; wherein the protein mixture retains its functionality even under extreme conditions. Also described herein are methods of using the above compositions as enhanced oil recovery agents, cleansing agents, or as surfactant enhancing agents.

DETAILED DESCRIPTION OF ACHIEVEMENTS

A key feature that affects the rate and / or efficiency of a chemical process is the surface energy between two or more chemical surfaces, whether liquid-liquid or solid-liquid. Surface energy between two substances is measured as interfacial stress (IFT) and is a function of both substances. The lower the IFT, the easier the two surfaces can contact. Contact between the two surfaces is a prerequisite for a chemical reaction to occur across the two surfaces. Once the reagents meet, other factors, such as pH, emulsification qualities, reaction energies, temperature, critical micelle concentration, and others, begin to affect the rate of chemical reactions.

Typically, a cleaning solution is designed to lower the IFT between the cleaning solution and the "dirt" layer, usually an oily surface, to allow the cleaner inside the cleaning solution to contact various components in the " dirt "and affect cleaning. For this reason, increased oil recovery and most cleaning solutions comprise a surfactant that lowers the IFT. Some of the currently used cleaning solutions also include the enzymes that help in cleaning.

In many cases, to maximize cleaning efficiency, especially to be effective in removing oily and greasy dirt, a highly alkaline or high pH solution is useful. See, for example, U.S. Pat. 6,025,316, 6,624,132, 7,169,237 and U.S. Patent Publication No. 20030078178, which are hereby incorporated by reference in their entirety. In some industrial applications, such as tissue cleaning, sizing agents are removed by cleaning solutions that may exceed a pH of 10. In pulp and paper processing, high pH conditions are required at various stages in the process. . At the opposite end of the spectrum, it may be necessary to use solutions that have a lower pH, that is, under acidic conditions, for use in applications such as the removal of deposits from mineral layers in bathrooms, industrial equipment, refrigeration systems, and others. still.

It is also well known that the use of hot cleaning solutions, such as the use of hot water with the cleaning agent, is desirable under some conditions. Hot solutions can better solubilize oils and minerals.

However, proteins are generally not stable or functional at extreme heat or pH, whether acidic or basic. Proteins are generally functional in a limited temperature and pH range. Out of range, the protein first loses activity and then becomes denatured. A denatured protein in a cleaning solution is not active and is not useful as a cleaning agent.

Thus, in the first aspect of the present invention, a cleaning composition comprising proteins and surfactants is described, wherein the proteins retain their stability and functionality under extreme conditions. Extreme conditions may include conditions such as temperatures above about 50 ° C to up to about 110 ° C, steam, pH levels below about 3.5, and pH levels above about 9.5, such as those encountered in operations. oil recovery levels. In some embodiments, the pH levels are above 10.5. Proteins of the disclosed compositions comprise proteins, protein fragments, peptides and stress proteins that have a size of less than 30 kDa. In some embodiments, the size range ranges from approximately 0.5 kDa to approximately 30 kDa. Throughout the present description, the protein mixture used in the compositions described in the present invention is called the "protein system".

The word "peptide" includes long chain peptides such as proteins and enzymes as well as short chain peptides such as reducers, trimers, oligomers and protein fragments. In some embodiments, the words "peptide" and "protein" are interchangeable. Accordingly, the protein systems described in the present invention may contain only short chain peptides, only long chain peptides, or a combination thereof.

In some embodiments, "retention of stability and functionality" means that, after being subjected to extreme conditions, proteins retain at least approximately 80% of their functionality compared to proteins before being subjected to extreme conditions. In other embodiments, proteins retain at least about 90% of their functionality after being subjected to extreme conditions, and in other embodiments, proteins retain at least about 95% of their functionality. Functionality can be defined in terms of the catalysis rate of a chemical reaction, decoupling of biochemical processes, decreased interfacial stress or decreased critical micelle concentration.

In some embodiments, the protein systems described in the present invention are derived from an aerobic fermentation of Saccharomyces cerevisiae, which, when mixed with the active agents or surfactants, enhances multiple chemical functions under ambient conditions or during and after exposure to conditions. extreme. The protein systems described in the present invention may also be derived from the fermentation of other yeast species, for example, kluyveromyces marxianus, kluyveromyces lactis, candida utilis, zygosaccharomyces, pichia or hansanula.

After the aerobic fermentation process, a fermentation mixture is obtained which comprises the fermented yeast cells and their secreted proteins and peptides. In some embodiments, the fermentation mixture may be subjected to additional stress, such as overheating, starvation, oxidative stress, or mechanical or chemical stress, to obtain a post-fermentation mixture. The additional stress causes additional proteins to be expressed by yeast cells and released into the fermentation mixture to form a post-fermentation mixture. These additional proteins are not normally present during a simple fermentation process. Additional proteins are known as "stress proteins", and sometimes referred to as "heat shock proteins". Once the post-fermentation mixture is centrifuged, the resulting supernatant comprises the stress proteins and proteins normally obtained during fermentation. The compositions described in the present invention comprise stress proteins. Several low molecular weight proteins may be produced by Saccharomyces cerevisiae during fermentation as practiced by those skilled in the art. These proteins appear when yeast cells are placed under stress during or near the end of the fermentation process. Although termed "heat shock proteins", stress conditions may occur during periods of lean food at mass concentrations or as a result of heat shock or pH shock conditions as described in U.S. Pat. 6,033,875, to Bussineau, et al. , incorporated by reference in its entirety. In addition, chemical stress, oxidative stress, ultrasonic vibration and other stressful conditions can cause yeast to express the formation of heat shock proteins, more precisely termed "stress proteins".

In addition, it has been found that the protein systems described in the present invention, or Living Yeast Cell Derivative, which is an alcoholic derivative of Saccharomyces cerevisiae produced by the methods set forth in U.S. Pat. 2,239,345, 2,230,478 and 2,230,479 to Seperti, which are incorporated by reference herein in their entirety, when coupled with surfactants, produce effects that simulate uncoupled oxidative phosphorylation when added to aerobic mixed culture processes such as U.S. Patent Application Publication No. 2004-0180411. Further details regarding these processes and materials are described in U.S. Patent No. 6,699,391. Each of these U.S. Patent Documents is Crude Yeast Cell Live Derivative has been further refined using dialysis membranes as determined in U.S. Patent No. 5,356,874, Bentley, resulting in polypeptides having molecular weights which range from 6,000 to 17,000 daltons as determined by the SDS-PAGE (Bentley, et al., (1990).) A key difference is that for In the compositions described in the present invention, isolation of fermentation by-products is not necessary, which makes the process cheaper.

The use of microbial agents for the degradation of substances includes U.S. Pat. 4,132,638 (the liquid is pretreated with enzymes, followed by microbial thermophilic pulp degradation), 4,666,606 (elimination of fat and odors using xeronin, which acts by stimulating metabolism of anaerobic and aerobic bacteria), 4,746. 435 (use of aerobic and anaerobic microorganisms to purify nitrogen water, etc.), 5,484,524 (use of biofilm to digest organic matter and pollutants in an aeration chamber for wastewater treatment), 5,599,451 ( the use of anaerobic and aerobic biotreatment of liquid toxic waste such as pulp and wastewater), 6.342.386 (use of a polymer and microorganism that has the ability to produce proteolytic enzymes as a surface coating to remove biofilm), which are incorporated herein by reference in their entirety.

The compositions described in the present invention include a stress protein component used in combination with a surfactant-containing composition, for example, an enhanced oil recovery composition or a cleaning composition, to increase and enhance surface active properties, decoupling. biological processes during and after exposure to extreme pH and / or elevated temperatures and to increase, increase and intensify the thermal stability of sulfated alcohol and sulfonate surfactants and other low temperature stable surfactants contained in the EOR composition. Extreme pH is defined as being below 3.5 and above 9.5. The elevated temperature is defined as up to approximately 10 ° C.

The "aerobic yeast fermentation process described in the present invention" is defined as the standard propagation conditions used in the production of commercially available bread yeast as described by Tilak Nagodawithana in "Baker's Yeast Production" and also described below. "Living Yeast Cell Derivative (LYCD) described in the present invention" is defined as an alcoholic extract obtained from yeast prepared as described below.

"Low molecular weight proteins described in the present invention" are defined as the biologically active fraction of the polypeptide comprising a size of less than 30,000 daltons, which are obtained from aerobic fermentation processes and LYCD.

"Surfactants described in the present invention" are defined as anionic sulfated alcohols, more particularly branched alcohol propoxylate sulfate surfactants, sulfonate surfactants and other surfactants as described below.

"Cleansing compositions" are defined as surfactant-containing compositions used as detergents, cleaners, degreasers, dispersants, emulsifiers, wetting agents, solubilizers or any other surfactant-containing compositions for the purpose of reducing surface stress, interfacial stress solubilize fats, grease or oil and other naturally occurring, synthetic and organic compounds.

The author of the present invention has isolated low molecular weight protein factor from aerobic yeast fermentation processes which, when coupled with surfactants, reduce critical micelle concentration, surface stress and interfacial stress of surfactants, with further reductions. critical micelle concentration, surface stress and interfacial stress observed after exposure to grease and oil. This factor was found to comprise four polypeptide fractions ranging in molecular weight from approximately 6,000 to 17,000 daltons by the results of polyacrylamide gel electrophoresis.

The compositions described in the present invention comprise an aerobic yeast fermentation supernatant, surfactants and stabilizing agents. Saccharomyces cerevisiae grows under aerobic conditions familiar to those skilled in the art using a sugar source such as molasses or soy or maize as the primary nutrient source. Alternative types of yeast that may be used in the fermentation process may include: Kluyeromyces marxianus, Kluyeromyces lactis, Candida utilis (Torula yeast), Zygosaccharomyces, Pichia and Hansanula. Those skilled in the art will recognize that other additional strains and yeast strains are potentially useful in the fermentation and production of low molecular weight proteins, the "protein system". It should be understood that these yeasts and the yeast classes described above are identified as preferred materials only and that this list is neither exclusive nor limiting to the compositions and methods described in the present invention. Other nutrients may include diastatic malt, phosphate of diammonium, magnesium sulphate, ammonium sulphate, zinc sulphate and ammonia. The yeast is propagated under continuous aeration and agitation at 30 ° C to 35 ° C and a pH range of 5.2 to 5.6 until the yeast reaches a minimum level of 4% based on dry weight. At the conclusion of the fermentation process, the yeast fermentation product is centrifuged to remove the yeast cells and the supernatant is then mixed with the surfactants and stabilizing agents, and the pH is adjusted from 4.0 to 4.6 to long-term stability.

In an alternative embodiment, the yeast fermentation process is carried on until the desired level of yeast is produced. Prior to centrifugation, yeast in the fermentation product is autolised by increasing heat to 40 ° C to 60 ° C from 2 hours to 24 hours, followed by refrigeration to below 25 ° C and centrifugation.

In another embodiment, the fermentation process is carried on until the desired level of yeast is produced. Prior to centrifugation, the yeast in the fermentation product is subjected to physical disruption of the yeast cell walls by the use of a French press, a ball mill or high pressure homogenization or another mechanical or chemical device familiar to the elements discussed in the art. technique to assist in the release of low molecular weight intracellular polypeptides. It is preferable to terminate the cell disruption process after a heating or autolysis stage, since the presence of the targeted proteins is induced by a response to thermal shock. The fermentation is then centrifuged to remove debris from the yeast cell and the supernatant is recovered.

In a third alternative embodiment, the fermentation process is carried on until the desired level of yeast is produced. After the fermentation process, the yeast cells are separated by centrifugation. Yeast cells are then partially lysed by the addition of 2.5% to 10% of a surfactant to the separated yeast cell suspension (10% -20% solids). In order to decrease protease activity in yeast cells, 1 mM EDTA is added to the mixture. The cell suspension and surfactants are gently shaken at a temperature of approximately 25 ° C to approximately 35 ° C for approximately one hour to cause partial yeast cell lysis. Cell lysis leads to increased release of intracellular proteins and other intracellular materials. After partial lysis, the partially lysed cell suspension is mixed back into the yeast and the cell solids are removed again by centrifugation. The supernatant containing the protein component is then recovered.

In another embodiment, fresh live Saccharomyces cerevisiae is added to a coated reaction vessel containing denatured methanol-alcohol. The mixture is stirred gently and heated for two hours at 60 ° C. The hot slurry is filtered and the filtered material is treated with charcoal and stirred for one hour at room temperature and filtered. The alcohol is removed under vacuum and the filtered material is further concentrated to yield an aqueous solution containing the stress proteins. This LYCD composition is then mixed with water, surfactants and stabilizing agents, and the pH is adjusted from 4.0 to 4.6 for long term stability.

In another embodiment, the heat shock process in the preceding embodiment includes various stages of agitation and heating, cooling and cycle repetition to increase the output of heat shock proteins.

In another embodiment, the LYCD is further refined to preferentially isolate active proteins having a molecular weight between 500 and 30,000 daltons using crude LYCD Anion Exchange Chromatography, followed by Molecular Sieve Chromatography. Refined LYCD is then mixed with water, surfactants and stabilizing agents, and the pH of the composition is then adjusted to between 4.0 and 4.6 to impart long term stability to the compositions.

The foregoing descriptions provide examples of a protein component suitable for use in the compositions and methods described in the present invention. These examples are not exclusive. For example, those skilled in the art will recognize that the protein component can be obtained by isolating the appropriate proteins from an alternative protein source, by protein synthesis or by other appropriate methods. The foregoing description does not lend itself to limiting the term "protein component" only to the examples included in the present invention.

Further details regarding fermentation processes and other aspects of the protein component are described in U.S. 101799.529, filed March 11, 2004, entitled "Altering Metabolism in Biological Processes", which is assigned to the assignee of this application and is hereby incorporated by reference in its entirety.

Surfactants

For enhanced oil cleaning and recovery applications, which include dirt removal from hard and soft surfaces, odor control, biofilm control, etc., the range of surfactants is not limiting. Anionic, nonionic and cationic surfactants may be combined with the optimized protein system based on the interaction of the surfactants and the protein system / surfactant composition / protein in the application. These include:

Anionic: Linear sodium alkylbenzene sulfonate (LABS) / sodium lauryl sulfate; sodium lauryl ether sulfates; petroleum sulfonates; linosulfonates; naphthalene sulfonates, branched alkylbenzene sulfonates;

linear alkylbenzene sulfonates; fatty acid alkylolamide sulfosuccinate; alcohol sulphates.

Cationics: Stearalconium chloride; benzalkonium chloride; quaternary ammonium compounds; amine compounds; ethosulfate compounds.

Nonionic: Dodecyl dimethylamine oxide; diethanolamide coconut alcohol ethoxylates; linear primary alcohol polyethoxylate; alkyl phenol ethoxylates; alcohol ethoxylates; EO / PO polyol block polymers; polyethylene glycol esters; fatty acid alkanolamides.

Amphoterics: Coconut Amphocarboxy Glycinate; propyl betaine cocamido; betaine derivatives; imidazolines.

In addition to the surfactants listed above, suitable nonionic surfactants include alkanolamides, amine oxides, block polymers, ethoxylated primary and secondary alcohols, ethoxylated alkyl phenols, ethoxylated fatty esters, glycerol esters, ethoxylated fatty acids and propoxylated fatty acids. , alcohols and alkyl phenols, alkyl glycoside glycol esters, polymeric polysaccharides, ethoxylated alkyl phenol sulfates and polymeric surfactants. Suitable anionic surfactants include ethoxylated amines and / or amides, sulfosuccinates and derivatives, ethoxylated alcohol sulfates, alcohol sulfates, sulfonates and sulfonic acid derivatives, phosphate esters and polymeric surfactants. Suitable amphoteric surfactants include betaine derivatives. Suitable cationic surfactants include amine surfactants. Those skilled in the art will recognize that other additional surfactants and surfactants are potentially useful in the compositions, depending on the particular cleaning application.

The EOR compositions described in the present invention include one or more sulfated alcohol and sulfonate surfactants over a wide range of concentration levels. These surfactants are anionic in nature and can be ethoxylated or propoxylated. Preferred anionic surfactants used in EOR applications include branched alcohols, preferably with an alkyl chain length of C12 to C17 and with the average propoxy groups in the molecule between 3 and 8.

Those skilled in the art will recognize that interfacial stress is highly specific with respect to the hydrophobic substrate and therefore no specific surfactant will be superior to all others when working with crude oil of different compositions. For example, West Texas light crude should better utilize a short chain surfactant length, while Indonesian light crude should require a longer chain surfactant length for more favorable results. However, for demonstration purposes, a single refined oil composition was used for the illustrations in this application.

While it may be advantageous to combine branched alcohol propoxylate sulfate surfactants with nonionic or amphoteric surfactants, the benefits of using these classes may be limited due to the low mist points. These classes of surfactants include: Nonionic: Dodecyl dimethylamine oxide; diethanolamide coconut alcohol ethoxylates; linear primary alcohol polyethoxylate; alkyl phenol ethoxylates; alcohol ethoxylates; EO / PO polyol block polymers; polyethylene glycol esters; fatty acid alkanolamides.

Amphoterics: Coconut Amphocarboxy Glycinate; propyl betaine cocamido; betaine derivatives; imidazolines.

It should be understood that these surfactants and the surfactant classes described above are identified as preferred materials only and that this list is neither exclusive nor limiting to the compositions and methods described in the present invention.

In another aspect of the present invention there is described a composition comprising stress proteins, surfactants, stabilizers and an acid. In some embodiments, the acid is selected from the group consisting of phosphoric acid, citric acid, lactic acid and hydrochloric acid.

In another aspect of the present invention, a composition comprising stress proteins, surfactants, stabilizers and a base is described. In some embodiments, the base is selected from the group consisting of sodium hydroxide, sodium metasilicate, sodium tripolyphosphate, triethanolamine, monoethanolamine and morpholine.

In another aspect of the present invention, a surface cleaning method comprising applying to the surface a composition as described in the present invention under extreme conditions is disclosed. Extreme conditions may include high temperatures and low or high pH as described above.

In the context of the present description, "cleanliness" is defined by its most fundamental characteristics: the chemical removal or surface lifting or neutralization of organic, inorganic and biologically based compounds or entities, which create or lead to: (b) unpleasant aesthetics, such as stains and dirt, (c) odors, (d) biopecia, (e) impedance or interruption of mechanical, chemical and biochemical processes or (f) capture of crude oil in underground mineral deposits.

The surfactant / protein system compositions described in the present invention enhance functionality, that is, increase cleaning efficiency under normal or environmental circumstances, or during or after exposure to extreme conditions. Increased functionality is measured in terms of one or more of: (a) removal of inorganic dirt, (b) chemical decomposition and solubilization of fats, oils and greases; (c) the creation of additional surfactants during the breakdown of fats and oils resulting in a self-catalytic cleaning synergy; (d) the removal of odors caused by urine, feces, vomiting, other biological liquids, rotten food, biofilm sludge and other sources; (e) the removal and control of biofilms; (f) the increased biodegradability of waste products associated with cleaning processes; (g) stabilized sulfated alcohols and sulfonate surfactants and other surfactants to enhance their functionality in EOR applications; or (h) uncoupled biological processes, for example oxidative phosphorylation to: (i) continue to break biofilms after active mechanical devices have stopped and / or industrial processes have been terminated or (ii) intensify EOR by enhancing microbial metabolism ( MEOR) utilizing the increase in carbon dioxide output, the by-product of the decoupling mechanism. All examples and descriptions of the efficacy of protein systems described in the present invention relate to maintaining the performance and stability of the compounds during or after exposure to extreme conditions.

Similarly, the "cleaning1" function can be summarized as the removal and / or neutralization of unwanted dirt from surfaces. The term "surfaces" may refer to hard surfaces such as floors, equipment, shelves, automobiles, minerals in the earth. (oil recovery), and still others or on soft surfaces such as fabrics and textile materials or water cleaning Unpleasant soils include such entities as oils and greases, mineral deposits, bacterial and viral substances and their secretions, compounds organic, naturally and synthetically derived compounds, smelly compounds, and combinations thereof Briefly, dust or dirt of any kind ultimately inhibits the aesthetic, visual and olfactory, mechanical or chemical processes. a breeding ground for microbial growth, whether bacterial, viral, fungal, algal, etc., or their secretions As an example, floor and equipment cleaning may include biofilm control such as biofilm growth on porous surfaces, paper processing, cleaning of cross-flow membranes such as reverse osmosis, microfiltration and ultrafiltration, industrial tank cleaning and sanitation, cooling system cleaning including cooling towers and condensers.

In some embodiments, during or after exposure to extreme environmental conditions, the protein systems and surfactant / protein system compositions described in the present invention remain stable, maintain functionality and provide improved performance compared to cleaning alone surfactants. , odor control, biofilm control, industrial chemical processes and enhanced oil recovery (EOR). In further embodiments, the protein system enhances functionality and stabilizes sulfated alcohols and sulfonate surfactants at temperatures exceeding 160 ° F. In some additional embodiments, the protein system and surfactant / protein system compositions may provide slight biological enhancements, specifically the coupling of biological processes that greatly improve cleanliness, odor control, biofilm control, oil recovery. microbial function (MEOR) and the range of functionality is maintained during and after exposure to said extreme conditions.

In some aspects, the present invention provides compositions based on surfactant binding protein systems which during and after exposure to extreme conditions exhibit the multiple functionality of the surfactants. In other aspects, methods are provided by which the production and yields of Saccharomyces cerevisiae protein systems can be increased.

The compositions and methods described in the present invention are advantageous for several reasons. Many common surfactants are stable under extreme pH conditions and temperatures up to 110 ° C and therefore these are not always distinct characteristics to choose from surfactants. Second, the control of biological contaminants, such as biofilms, has traditionally been approached as independent of associated cleanliness and chemical processes, although they are in most cases integral and interdependent. Thirdly, odor control, especially on hard surfaces, in ordinary households, in institutional and industrial products, was generally addressed by the addition of biocidal agents to cleaners or by other chemical formulations. Odor control and cleaning using traditional approaches require at least two chemical applications, one for each problem. The compositions described in the present invention are chemical formulations comprising a protein system that brings together biological and non-biological aspects of surfactant chemistry and associated processes as defined in the present invention.

The rates of chemical reactions and processes, and therefore the cleaning rate as defined above, can generally be greatly improved at elevated temperatures. In addition, chemicals in many processes require extreme pH conditions. The most naturally produced peptide compounds, such as enzymes, which have been developed for industrial and cleaning processes, have limitations on their stability and / or functionality under extreme pH conditions and at elevated temperatures. Specialized high temperature enzymes have been developed, but are even more expensive than the most common enzymes used in laundry detergents.

The protein systems described in the present invention surprisingly show not only stability, but also functionality in terms of surfactant synergies and biological and surfactant effects following exposure to extreme pH and / or elevated temperatures. Thermal stability was tested with exposure for up to 96 hours at approximately 100 ° C.

The ability of the protein system to reduce IFT and to retain this capacity during and after exposure to extreme pH levels and / or temperature elevation above 50 ° C is the common denominator that follows the other characteristics of composition. The unique characteristics of protein / surfactant system formulations that are enhanced by the reduction in IFT are as follows: reduction of critical micelle concentration; converting a portion of exposed oils and greases into additional surfactants under sterile and non-sterile conditions, providing a self-catalytic effect; devolatilization of smelly compounds; penetration of the protein system and further breakdown of biofilms after mechanical application has been discontinued by stimulating resident microflora, which enhances cleanliness, odor control and reduces total surfactant utilization; the reduction of the IFT of sulfated alcohol and sulfonate surfactants, among others; stabilization of sulfated alcohol, sulfonate surfactants and other surfactants at temperatures above 160 ° F, where typically a large increase in IFT is seen due to thermal degradation, a critical feature within EOR applications; The decoupling of oxidative phosphorylation as a fundamental mechanism affecting microflora continues after pH conditions and temperatures drop to levels that can sustain the growth of microorganisms.

An example that takes advantage of the many characteristics of surfactant / protein system compositions during and after the temperature rise above 50 ° C is floor cleaning in restaurants and food processing plants where food oils are a major floor contaminant. Floors typically comprise porous concrete, porous ceramic tiles and / or porous mortar lines. The pores are filled with oils and other organic contaminants that are the nutrients that naturally present microflora, which then create viscous biofilms within the pores. Surfactants that are used in commercially available floor cleaners do not break the oils. Surfactants generally act to emulsify oils. Therefore, when a clean floor is wet, the oil trapped in the pores rises to the top of the water film, creating a slippery layer on the floor surface. Hot water is recommended in many cases as it provides better cleaning performance. The protein system continues to work during the hot drying step as well as after drying is completed by decoupling microbial metabolism in the natural microflora. Organic nutrients continue to break down at accelerated rates, eliminating particular oils, which increases slip resistance and overall cleanliness. The pores retain much less contaminants and result in a safer floor even when wet. Finally, many odors emanate from biofilms, typically due to the presence of anaerobic bacteria. Odor reduction using the protein-based composition is a by-product of the chemical process, especially as in cleaning, using a dual mode method of action. While the protein system is not a sanitizer, by helping to remove organic materials that must remain as nutrients that could help support microbial growth, nutrient elimination leads to a more hygienic environment and can potentially reduce the amount of sanitizers, many of which have toxic elements because their purpose is to kill living cells.

An additional benefit of the broad functionality of the protein systems described in the present invention is the simplification of the chemical formulation and chemicals used in a particular process. Oils and other difficult to remove dirt were traditionally targeted by the addition of inorganic builders and caustics to surfactant mixtures. The protein system helps reduce the need for builders. At the same time, the protein system provides a formulation roadmap to reduce the toxicological and environmental traces of chemicals, especially those that people come into contact with every day. Due to the low toxicity of the protein system, the compositions can be used in a wide range of application areas such as health care, food processing, pharmaceuticals, making increased oil recovery and many more environmentally friendly industrial uses. environment.

The options for the surfactants described in the present invention are not limiting and both botanically derived and synthetic petroleum derived surfactant systems are enhanced by the protein fraction. Surfactant classes include anionic, nonionic, cationic, amphoteric surfactants. Although surfactants based on. oil are not ideal from an environmental sustainability standpoint, the addition of the protein system to the surfactant formulation still has the benefits that are not available without the protein system. These include: (a) the reduction in the amount of surfactant required for a particular process; (b) the decomposition of organic elements at the point of use and the further work on breaking down the organic elements in the discharge stream. In addition, synergism allows less surfactant to be used to achieve the same results as indicated by lower interfacial stress and critical micelle concentrations.

PH

Effects at higher alkalinity or higher pH are observed using the protein systems described in the present invention. The synergies provided by the protein system to the mixed surfactant system are evident at high pH levels. Although increasing the pH makes the product more dangerous to the user and perhaps to the discharge current, in many cases the need for cleaning outweighs the safety issue and proper usage protocols must be followed to maintain safety. A high pH cleaner may cause the need for solvents and pH neutralization may cause less environmental concern or wastewater treatment than solvents, many of which are volatile organic compounds (VOCs) and many of which have toxic effects. .

Again, the synergies provided by the protein systems described in the present invention to the surfactant systems used are maintained at lower pH levels. In the case of EOR, a wide range of pH values can be found, and the most favorable recovery is obtained from acidic conditions at alkaline pH values as high as 12-13. Near neutral pH, for more user-friendly safety, the protein systems described in the present invention have been superior to the performance of alkaline cleaners and solvent-enhanced alkaline cleaners in many cases.

Since the protein systems described in the present invention are stable upon exposure to extreme pH conditions, they continue to exert their effect on natural microflora in areas such as sewage pipes, plumbing and septic systems, where levels of pH tend to be greatly neutralized due to dilution. After mechanical application procedures such as cleaning and drying are completed, functionality is maintained and protein systems continue to work as under the other conditions described in the present invention. Without being bound by any particular theory, it is assumed that functionality will mostly occur due to the decoupling where natural microflora works to break down organic compounds, including biofilms. Without the protein system, the rate of organic degradation is not sufficient to prevent accumulation.

Enzymes

In addition to efforts to reduce the hazards of use and environmental problems caused by traditional caustic and / or solvent based cleaners such as terpenes and glycol ethers and other chemical compositions, there is much research being done on enzymes. One target area is the development of pH neutral cleaners. Since surfactants alone or with minimal builder use are not very effective at relatively neutral pHs, enzymes have been added to increase performance at neutral pHs, although performance does not address that of traditional caustic or solvent-based products. . Enzymes that have been used to enhance cleaning efficacy, particularly in the development of pH neutral cleaners, include lipase, protease and amylase. Enzymes have the distinct disadvantage of being relatively unstable, especially with regard to temperature and long-term "shelf life" stability. This greatly limits the range of applications in which enzymes can be used. See, for example, U.S. Pat. No. 6,624,132, incorporated herein by reference, which discloses that its enzyme formulation retains approximately 50% of its initial activity at 120 ° F for at least approximately 25 days after formation of the composition. The present protein systems are stable under conditions where enzymes generally are not. An alternative to enzymes is the addition of live bacteria, typically in the form of spores, stabilized in cleansing formulations until diluted in use. The main purpose of cleaning should be to reduce biology in one area so that the addition of bacteria is contraindicated. And there is the limitation of IFT, which is not increased by enzymes or bacterial spores.

The development of extremozymes, the enzymes that are extracted from naturally high temperature environments, were designed to counteract the temperature stability of lipases, proteases and amylases. The limitation is that they react slowly at low temperature. In addition, a key feature of enzymes is that they are very narrow in the range of dirt in which they break down. Enzymes can be expensive and are therefore used at low levels. Finally, there are problems related to allergenicity with enzymes in general.

In any of the above matters, the enzymes do not react synergistically with the surfactants or uncouple metabolic processes such as the protein systems described in the present invention.

Botanically Derived Compounds

In the interest of developing chemical compounds from renewable resources to reduce the confidence of petroleum-based raw materials, chemical companies have used a number of botanically derived compounds. Petroleum-based chemicals are generally hazardous and are based on crude oil, which is not a sustainable raw material.

Botanically derived compounds are typically isolated from their natural state and are concentrated. These include compounds such as oranges d-limonene, pineapple juice extracts and others for the purpose of replacing petroleum based chemicals.

Botanically derived compounds, however, may be toxic. For example, d-limonene has been shown to be carcinogenic, although it is used in cleaners and personal care products that come into close contact with people. Solvents such as d-limonene, although derived from natural sources, are VOCs, which negatively affect air quality. In addition, raising the temperature of these cleaning compositions above ambient conditions increases the possibility of other hazards such as explosions. In tank cleaning, for example, the elimination of solvents can greatly reduce the possibility of explosions.

Biofilms

The biofilm is a polysaccharide complex that is created by the living microflora of many types and surrounds it, and protects cells from the surrounding environment. Cells within a biofilm matrix act together to produce the biofilm, along with other products such as occasional toxins, to maintain their survival.

Biofilm is an observable problem in many areas. For example, the development of biofilms impairs the ability of biocides to kill bacteria. Secondly, in water treatment, biofilm formation can clog filters, membranes, heat exchange surfaces, etc. In fact, there are dozens of ways that biofilms interact with synthetic mechanisms. Often biofilm formation acts to undermine what is important to modern society. Thus many methods for destroying the biofilm have been tried. A general understanding of bacteria production and decomposition of the biofilm and how it is affected by a decoupling factor requires knowledge of some basic cellular processes, including that of energy production and utilization. In addition, understanding the way in which bacterial cells generate and utilize energy requires an understanding of the underlying basic science of cellular respiration.

Decoupling of Biological Processes

Specialized yeast fermentation products containing bioactive products are described in the present invention. Bioactive products include a 'decoupling' agent, the protein system, which comprises mainly low molecular weight stress proteins. The main objective for these stress proteins is to increase the utilization of the microbial substrate, ie nutrient uptake. The nutrient of interest is the biofilm and the intention is that the biofilm be degraded. Substrate utilization or nutrient uptake is increased because the decoupler terminates the oxidative phosphorylation (OP) process, which is the most efficient method by which microorganisms synthesize ATP, the final form of cellular energy. OP occurs in any or all organisms that have the ability to use oxygen as an electron acceptor, also called aerobic organisms. The process of OP occurs due to the actions of membrane-bound molecules, enzymes, coenzymes, etc. It involves and requires the transfer of electrons and protons through an 'electron transport chain' which terminates in an oxygen molecule acting as the ultimate electron acceptor. The final and indirect effect of the electron transport chain is the formation of ATP.

With a decoupler, less efficient secondary processes are used to synthesize ATP. A decoupler simply decouples or decouples the electron-transporting process from ATP formation. Also, since the uncoupler results in the loss of a proton gradient, there is a continuous loss of energy in the form of heat. Therefore, the effect of the uncoupler is twofold. It results in a dramatic increase in substrate utilization or nutrient uptake. This is primarily because the microorganism is forced to use less efficient pathways to produce ATP for general metabolic functions in order to survive. Secondly, there is a continuous loss of energy that causes the microorganism to continually add ATP because of continuous loss of energy, and this replacement of ATP is generated by inefficient methods.

An additional key manifestation of energy loss is that there is inadequate energy remaining for the formation of complex proteins that are necessary for the construction of polysaccharides or biofilms, thereby preventing the accumulation of biofilms. Increased nutrient uptake is accelerated to the point where existing biofilms become a food source for microflora, which is the assumed mechanism of biofilm removal.

Applications in EOR Experimental results show that sulfated alcohol and sulfonate surfactants may be preferred candidates for EOR as they can be effective in creating low interfacial stress (IFT) at dilute concentrations and without requiring an alkaline or alkaline agent. a co-surfactant. In addition, some of the formulations exhibit low IFT at various percent sodium chloride concentrations and thus may be suitable for use in more saline reservoirs. Due to the broad functionality of the surfactant protein system under many conditions, it is anticipated that stabilization of alcohol sulphates should be virtually identical in other surfactant classes which may have lower thermal stability. Data are provided for sulfated alcohols.

Y. Wu et al. (A Study of Branched Alcohol Propoxylate for Improved Oil Recovery, Society of Petroleum Engineers, 2005) indicate that surfactant enhanced oil recovery (EOR) has been investigated for many years, especially beginning in the seventies and eighties when the technology was placed. on a scientific basis. Unfortunately, the economic reality of process performance as proved in the early field trials made it impossible to spread the technology widely. However, the recent surge in crude oil prices has provided new impetus for considering the use of chemical EOR.

The basic physics behind the surfactant flow EOR process is that residual oil, dispersed as micron-sized ganglia, is captured by high capillary forces within the porous medium. Increasing viscous fluid flow forces or decreasing capillary forces holding the oil in place are required before the oil can be passed through the pore necks and sent to a production well. The empirical method for a successful surfactant flow is that interfacial stress (IFT) between crude oil and aqueous phase must be reduced to ultra-low interfacial values, several orders below that of a typical brine-oil reservoir system.

In addition to the requirement for low in-situ IFT, another key factor determining the technical and economic success of a surfactant flow design is minimizing the decrease in injected surfactant, with a generally solid adsorption main sink on the reservoir bar. .

A wide variety of surfactants have been investigated for their potential efficacy for chemical EOR applications. With this renewed interest in surfactant EOR, there is now the opportunity to investigate unavailable or unvestigated surfactants during this earlier development of chemical EOR technology. Branched alcohol propoxylate sulfates have emerged as an effective type of surfactant for the removal of non-aqueous phase liquids (NAPLs) from contaminated aquifer sites near the surface. This class of surfactants is limited to near surface applications due to instability at temperatures above 160 ° F. In some deep well applications, temperatures may reach temperatures as high as 250 ° F, thereby limiting the scope of use for these highly effective surfactants.

Research into the use of yeast fermentation byproducts to verify the degree to which these compounds affect the efficiency of sulfated alcohol and sulfonate surfactants, more particularly branched alcohol propoxylate sulfates, has resulted in the discovery of a group of Low molecular weight stress proteins or "protein systems" described in the present invention, when combined with surfactants, bind to surfactants, resulting in reduced interfacial stress compared to interfacial stress achieved by using the surfactants alone. A second feature of combining the protein system with the surfactants is the increase in thermal stability as measured by the degree of displacement in the surfactant interfacial tensions. A third feature of combining the protein system with the branched alcohol propoxylate sulfate surfactants is the further enhancement of the effectiveness of these surfactants for EOR through lower IFT values. A fourth feature of combining the protein system with the branched alcohol propoxylate sulfate surfactants is that it can be used in deep well applications at high temperatures.

It has been found that the protein system comprising low molecular weight stress proteins (0.5-30 kD) in combination with surfactants results in a further increase in catabolic rates without the proportional increase in biomass, and an increase in in the amount of carbon dioxide breathed, thereby further defining the active proteins responsible for the decoupling effect observed in the evaluated biological processes. Studies show that the use of the protein system or surfactants alone has little effect on catabolic or anabolic rates. A synergistic effect is observed when surfactants are combined with low molecular weight proteins. Additional studies have shown that the protein system component, when combined with surfactants, has altered the nature of a given surfactant, reducing its surface stress, critical micelle concentrations and especially its ability to convert fat and oil to soluble materials. in water, thereby greatly enhancing the cleaning efficiency of the surfactant. Although the protein components described in the present invention are preferably obtained by prior fermentation processes, the components may also be obtained by alternative methods, including the synthesis or direct isolation of proteins from other natural sources.

The addition of the protein systems described in the present invention to a sulfated alcohol surfactant-containing composition has the effect of enhancing, increasing and enhancing the surface active properties and thermal stability of the surfactants contained in the composition. This effect has particular advantages in applications where the surface active and thermally stable properties of sulfated alcohol surfactants in the compositions are desired, including the incremental cleaning and oil recovery compositions discussed in the present invention.

EXAMPLES

Incremented Oil Recovery Compositions

The incremental oil recovery compositions described in the present invention generally comprise a branched alcohol propoxylate sulfate surfactant and a low molecular weight protein component produced from the aerobic yeast fermentation process described in the Detailed Description of the Preferred Embodiments. The surfactants used are manufactured and distributed by Sasol North America under the trade name Alfoterra. Auxiliary detergent ingredients may include any of a range of additives which are advantageous to obtain a desired beneficial property. These may include, but are not limited to, sodium carbonate or sodium hydroxide to adjust pH, or EDTA to minimize surfactant adsorption on forming substrates such as kaolinite clay.

Effect on Interfacial Stress: The following Alfoterra formulations were evaluated for IFT values determined by the drop drop method using a Kruss prop Shape Analysis System DSA10 to determine the effects of the low molecular weight component on this surfactant class. Since IFT is highly specific with respect to oil composition, EOR compositions can benefit from mixtures of protein-surfactant compositions tailored to create varying alkyl chain lengths or propoxylation levels. In the empirical method, shorter chain alcohols such as those in the Alfoterra 123 series will be more effective in lower molecular weight hydrocarbons, while longer chain alcohols such as those in the Alfoterra 167 series will be more effective in hydrocarbons of lower molecular weight. higher molecular weight. The surfactants evaluated for EOR applications are as follows: Alfoterra 123-4S, Alfoterra 123-8S, Alfoterra 145-4S, Alfoterra 145-8S, Alfoterra 167-4S and Alfoterra 167-7S. All Alfoterra products contain 30% active material. The stabilized yeast is based on the following:

Compositions using the above surfactants are as follows:

Concentration% by weight Component Sample A

Sample B

Stabilized Protein Component 0.00% 12.50%

(Samples B only)

(Saccharomyces cerevisae fermentation product)

Alfoterra 123-AS 33.33% 33.33%

or Alfoterra 123-8S 33.33% 33.33%

or Alfoterra 145-4S 33.33% 33.33%

or Alfoterra 145-8S 33.33% 33.33%

or Alfoterra 167-4S 33.33% 33.33%

or Alfoterra 167-7S 33.33% 33.33%

Water 66.67% 54.17%

TOTAL 100.00% 100.00%

Initial interfacial strains for the above samples were tested versus Castrol Motor Oil 10W40 as determined by the drop-down method using a Kruss prop Shape Analysis System DSA10. "A" samples denote the surfactant alone, while "B" samples contain the protein component. All samples contained 10% surfactant based on the assets and were diluted with water at a ratio of 16: 1. The results are as follows:

Stress Test # 1 Test

# 2 Test # 3

Interfacial Average Interfacial Voltage

Tension

Tension

Sample

Interfacial Interfacial mN / m Average,

mN / m

0.353

0.303

0.125

0.111

0, 222

Average, mN / m Average, mN / m

Alfoterra 123-4S "A" 0.352

0.353 0.350 Alfoterra 123-4S "B" 0.303

0.303 0.303 Alfoterra 123-8S "A" 0.128

0.13 0.128 Alfoterra 123-8S "B" 0.112

0.112 0.113 Alfoterra 145-4S "A" 0.22 0.218

0.221Alfoterra 145-4S "B" 0.169 0.172 0.169 0.167

Alfoterra 145-8 "A" 0.093 0.093

0.092 0.093

Alfoterra 145-8 "B" 0.067 0.067

0.066 0.069

Alfoterra 167-4S "A" 0.360.36

0.363 0.356

Alfoterra 167-4S "B" 0.321 0.322 0.316 0.324

Alfoterra 167-7S "A" 1.701 1.703 1.703 1.697

Alfoterra 167-7S "B" 0.867 0.868 0.869 0.865

As can be seen, interfacial stress values were reduced by 10.8% to as much as 49.1% by using the low molecular weight component with surfactants versus surfactants alone. The most favorable surfactant for the standard Castrol Motor Oil appeared to be Alfoterra 145-8S with an IFT of 0.093 mN / m. However, the addition of the low molecular weight protein component also reduced the IFT value to 0.067 mN / m or 28%.

Since sulphated alcohols in general and Alfoterra branched alcohol propoxylated sulphate surfactants in particular are thermally stable only at temperatures equal to or below approximately 160 ° F, Alfoterra samples, with and without the protein component, were heated to the boiling point (approximately 210 ° F) for three hours, and the IFT values were measured again by the same method. The results are as follows:

Stress Test # 1 Test

# 2 Test # 3Interfacial Tension Tension Tension

Interfacial Interfacial Interfacial

Sample mN / m Average, mN / m

Average, mN / m Average, mN / m

Alfoterra 123-4S "A" 0.352 0.353 0.353 0.35

Heated 123-8S "A" 0.618 0.617

0.62 0.616

Alfoterra 123-4S "B" 0.303 0.303 0.302 0.303

Heated 123-4S "B" 0.402 0.403 0.401 0.402

Addition of the protein component to Alfoterra 123-4S, a 4 molar branched C12-C13 alcohol sulphate propoxylate, reduced the IFT by 13.92%. The degree to which the IFT will change will shift depending on the type of hydrophobic that is presented to the various surfactants. However, when the surfactant / protein and surfactant / protein composition is heated to 210 ° F +, the IFT for the surfactant variable will increase by 75.6% from 352 mN / m to 618 mN / m, and the IFT for surfactant / protein composition increased from 303 mN / m to 402 mN / m or 14.2% more than the unheated surfactant value of 352 mN / m.

Stress Test # 1 Test

# 2 Test # 3

Interfacial Tension Tension Tension

Interfacial Interfacial Interfacial

Sample mN / m Average, _mN / m

Mean, mN / m Mean, mN / mAlfoterra 123-8S "A" 0.128 0.125 0.13 0.128

Heated 123-8S "A" 0.338 0.341

0.337 0.337

Alfoterra 123-8S "B" 0.112 0.111 0.112 0.113

123-8S "B" heated 0.168 0.17

0.166 0.167

Alfoterra 123-8S is similar to 123-4S, which is an 8 mol propoxylate instead of 4 mol. Data show a modest 12.5% decrease in IFT for the surfactant / protein composition versus the surfactant alone. When both variables were heated, the IFT for the surfactant variable alone increased 164.1% from 128 mN / m to 338 mN / m. The surfactant composition, however, increased from 112 mN / m to 168 mN / m, an increase of 50%, but only 31.3% from 129 mN / m for the unheated surfactant alone.

Stress Test # 1 Test

#2

Test # 3

Interfacial Average Interfacial Voltage

Tension

Tension

Sample

Interfacial Interfacial mN / m Average,

mN / m

Average, mN / m Average, mN / m Alfoterra 145-4S "A"

0.222 0.218

Heated 145-8S "A"

0.421 0.421

Heated Alfoterra 145-4S "B" 0.169 0.167 145-4S 11B "

0, 22

0, 422

0, 221

0, 425

0. 169

0, 237

0, 172

0.280.236 0.236

Alfoterra 145-4S is a 4 mol propoxylate of branched C14-15 alcohol sulfate that exhibited a 23.2% reduction in IFT when combined with the protein component. When both variables were heated for three hours, the surfactant IFT variable alone increased 91.8% from 220 mN / m to 422 mN / m. The surfactant / protein composition increased 40.2% from 169 mN / m to 237 mN / m; however, this heated surfactant / protein composition was only 7.7% higher than the only variable unheated surfactant.

Stress Test # 1 Test

# 2 Test # 3

Interfacial Average Interfacial Voltage

Tension

Tension

Sample

Interfacial Interfacial mN / m Average,

mN / m

Average, mN / m Average, mN / m

0.093

0.093

0.241

0, 24

0.067 0.111

0.066

0, 114

Alfoterra 145-8S "A" 0.092 0.093 145-8S "A" heated 0.242 0.24

Alfoterra 145-8S "B" 0.066 0.069 145-8S "B" heated 0.11 0.11

Alfoterra 145-8S, which is an 8 mol propoxylate of branched C14-15 alcohol sulphate, has the lowest IFT at 0.093 mN / m using Castrol Motor Oil 10W40 as a substrate. When the protein component is added, however, the IFT decreases by 28% to 67 mN / m. When heated, the IFT for both samples increases with the detestant variable alone increasing 159.1% from 93 mN / m to 241 mN / m. The surfactant composition, however, increased by a modest 65.7%, but with respect to the unheated surfactant sample increased by 0.018 mN / m compared to the unheated surfactant sample, 0.093 versus 0.111 mN / m.

Stress Test # 1 Test # 2 Test # 3

Interfacial Tension Tension

Tension

Interfacial Average

Interfacial Interfacial

Sample mN / m Average, _mN / m

Average, mN / m Average, mN / m

Alfoterra 167-4S "A" 0.808 0.805

0.806 0.813

Heated 167-4S "A" 1.701 1.703

1.703 1.697

Alfoterra 167-4S "B" 0.759 0.762

0.757 0.757

Heated 167-4S "B" 0.867 0.868

0.869 0.865

Alfoterra 167-4S, which is a 4 mol propoxylate of branched C16-17 alcohol sulphate, shows that a 6.1% reduction for IFT when the protein component is added reduces the IFT from 0.808 mN / m to 0.759 mN / m. When heated, the IFT for the surfactant variable alone increases by 110.5% from 0.808 mN / m to 1.701 mN / m. The surfactant / protein composition, however, only increases by 14.2% from 0.759 mN / m to 0.868 mN / m, and the IFT is only 7.3% higher than the only variable unheated surfactant.

Stress Test # 1 Test

# 2 Test # 3

Interfacial Tension TensionTension

Interfacial Interfacial Interfacial

Sample mN / m Average, mN / m

Average, mN / m Average, mN / m

Alfoterra 167-7S "A" 0.36 0.36

0.363 0.356

Heated 167-7S "A" 0.645 0.64

0.642 0.639

Alfoterra 167-7S "B" 0.321 0.322

0.316 0.324

Heated 167-7S "B" 0.35 0.349

0.347 0.353

Alfoterra 167-7S, which is a 7 mole propoxylate of branched C16-17 alcohol sulphate, shows that a 10.8% reduction for IFT when the protein component is added reduces the IFT by 0.3 60 mN / m to 0.321 mN / m. When heated, the IFT for the surfactant variable alone increases by 79.2% from 0.360 mN / m to 0.645 mN / m. The surfactant / protein composition, however, only increases by 9.0% from 0.321 mN / m to 0.350 mN / m, and the IFT is actually 2.8% lower than the only variable unheated surfactant.

An additional feature is that of the conversion of oil and grease to surfactant materials as described in U.S. Patent Application no. 11 / 322,104 of 2,005, wherein the levels of surfactant materials, as measured by critical micelle concentrations, are significantly increased by continued contact of the protein / surfactant compositions and the grease or oil substrate. This effect is further accentuated when contact occurs under non-sterile conditions. As an example, a test of a commercially available Bilge Water Cleaner with a pH value of 12.5, available from Western Marine, was performed versus a Bilge Water Cleaner prototype using a surfactant / protein composition with a pH value. of 4.5. Tests using a Kruss prop Shape Analysis System DSA10 with diesel fuel as a substrate are as follows:

64: 1 west

64: 1

Marine System Composition of

in Pond Water Protein /

(Diesel) surfactant in

Pond water

(Diesel)

Solution Properties Grease Pre-Exposure Surface tension prior to grease drop exposure (mN / m) 33,05 27,40

CMC before grease drop exposure (ppm) 94 179 *

Properties and Effects During Grease Exposure Initial interfacial tension on grease drop exposure (mN / m) 1.86 0.50

Grease balance interfacial tension (mN / m) 0.78 0.38

Programming for interfacial tension balance (minutes) 2.000 Grease drop volume after 2.88 0 minutes = 48 hours (ul) 1.63 0.00

Programming for the drop volume balance of

grease

(minutes)> 2,880

210

Solution Properties After Grease Exposure Surface tension 5.0 ml retention after grease drop exposure (mN / m) 29.07 27.18

CMC after grease drop exposure (ppm) 55 76

Calculated Properties Based on the Data Above Converted Grease Water Retention Concentration (ppm) 674 1,000

CMC change pre-grease versus post-grease (ppm) 41 103

Percentage of, 5,0 ul of grease converted to soluble components (%) 67,4 100,0

Percentage of 5,0 ul of grease converted to "surfactant-type" materials (%) 4.1 10.3

Percentage of grease that is converted to "surfactant type" (%) 6.1 10.3 *

In this case, the surfactant / protein composition converted 10.3% of the grease into "surfactant type" materials.

Another feature of the surfactant / protein composition is that it can increase carbon dioxide production in the presence of bacteria. This characteristic is similar to the effects sought by Microbial Incremental Oil Recovery where gas production and the resulting increase in pressure will help to facilitate cleaning of oil in the field toward the oil well. In US Patent Application 20040180411, "Altering the Metabolism in Biological Processes", the surfactant / protein composition demonstrated the ability to significantly increase carbon dioxide production. In a controlled experiment using bioreactors and a method to capture and measure carbon dioxide production, the surfactant / protein composition increased carbon dioxide production by 432.9%.

Test Method Carbon mass balance studies used a sterile Tryptic Soy Broth solution that was inoculated with Polyseed, a mixture according to a proprietary mixture of aerobic bacteria normally used for five days of BOD testing. Tryptic Soy Broth was chosen as a nutrient because it is completely soluble. Therefore, any suspended solids or particulate matter that is developed during the study period is assumed to be biomass produced as a result of carbon source assimilation. Since 51% of bacteria are known to comprise carbon, it is possible to determine the rate of carbon in the nutrient substrate that is converted to biomass by analyzing unfiltered versus filtered samples for total organic carbon at the beginning of the study, followed by sample analysis at any time during the study.

Carbon mass equilibrium studies were performed to determine the ability of the compositions described in the present invention to affect changes in carbon uptake, embiomass carbon conversion rate and carbon dioxide respiration. The studies were performed using an Applikon bioreactor using air that was sprayed through a 1.5 N sodium hydroxide solution followed by spraying through deionized water twice to remove all carbon dioxide from the aeration source. Exhaust air from the bioreactor is then sprayed through a 1.5 N sodium hydroxide solution to capture all carbon dioxide created in the bioreactor during the test period.

A Tryptic Soy Broth solution is prepared by adding 72 grams of the sterile 10% Tryptic Soy Broth concentrate to 2,400 ml of deionized water twice in a 4 liter beaker. Two Polyseed inoculum capsules are added to the nutrient solution. The inoculated nutrient is heated and maintained at 30 ° C with continuous shaking using a stir bar and incubated for fourteen hours. Prior to transferring the nutrient to the bioreactor, the nutrient solution is filtered through four layers of cotton gauze to remove the grain used as a substrate for dried bacteria in Polyseed. Two liters if the nutrient solution is loaded into the bioreactor. Untreated "controls" are performed as a baseline and "treated" samples have 10 ppm of test composition added to the nutrient.

The bioreactor is then sealed and the carbon dioxide free air is sprayed at a feed rate of 0.5 liter per minute while the bioreactor temperature is maintained at 30 ° C and the turbine mixer is rotated at 500 rpm for the duration of the test. Exhaust air is sprayed through a 1.5 M sodium hydroxide solution to capture the carbon dioxide that is breathed. The nutrient is tested at time 0 and again at the conclusion of the study. Filtered and unfiltered nutrient samples are analyzed for total organic carbon.

The exhaust air from the bioreactor is sprayed through 200 ml of the 1.5 N sodium hydroxide solution. Upon completion of the test, the sodium hydroxide solution is transferred to a beaker and 20 ml of a chloride solution. 3.5 N barium solution is neutralized with 4 N hydrochloric acid using a pH meter and burette to determine the volume of hydrochloric acid solution required to neutralize the solution. This is the value for B. The standardization factor is created by neutralizing 200 ml of 1.5 N sodium hydroxide solution with 4N hydrochloric acid using a pH meter and a burette to determine the volume of hydrochloric acid solution required to neutralize the solution. This is the value for S. The amount of carbon breathed as carbon dioxide is then calculated using S and B in the following equation: C = 6N (B-S) where N = 7.5.

The Carbon Mass Balance can then be calculated as follows: Carbon Nutrient Consumed = Increased Carbon Biomass + Carbon Respirated as Carbon Dioxide

Results:

Surfactant Composition /% Control Protein Difference

Carbon based contaminant reduction 175.3 328.6 87.50%

Carbon breathed as carbon dioxide 49.5 263.8 432.90%

Biomass carbon as sludge 142,876.8 -46.20%

Interfacial Stress Testing in Extreme pH Conditions

Surface tensions and interfacial tensions were measured versus Castrol Motor Oil 10W40 at 32: 1 dilutions of each of five samples using a surfactant system, with and without the stabilized protein component, and adjusted to the following pH values: pH = 1, 2, 7.5 (surfactant system only), 12 and 13. The formulas are as follows:

Example 1

Example 2

Stabilized protein component 20.00% 0.00%

Ethoxylated linear alcohol (6.0 E.O.) 7.50% 7.50%

Sodium Lauryl Ether Sulphate (3.0 E.O.) 1.50% 1.50%

Water 71.00% 91.00%

Total 100.00% 100.00%

pH adjusted to 1, 2, 7.5, 12 or 13 with phosphoric acid or sodium hydroxide (50% solution).

Surface tensions were determined by the Wilhelmy plate method, and interfacial tensions were determined by the DuNouy ring method, both using a Kruss Processor Tensiometer K100. All measurements were made in triplicate. The results of surface stress studies are shown in Table 1, while the results of interfacial stress studies are shown in Table 2.

TABLE 147/49

Surface Tension Data - 32: 1 Dilutions

Stress Test # 1 Test

Test # 3

Interfacial Tension Tension Tension

Interfacial Interfacial Interfacial

Sample mN / m Average, _mN / m

Average, mN / m Average, mN / m

Example 1 at pH = 1 29.97 29.93

29.99 29.99

Example 1 at pH = 2 29.83 29.83

29.84 29.81

Example lapH = 12 29.88 29.87

29.87 29.89

Example lapH = 13 30.32 30.35

30.27 30.34

Example 2 at pH = 1 30.54 30.51

30.53 30.58

Example 2 at pH = 7.5 28.79 28.81

28.79 28.76

Example 2 at pH = 13 30.96 30.96

30.97 30.94 TABLE 2

Surface Tension Data - 32: 1 Dilutions

Stress Test # 1 Test

Test # 3

Interfacial Tension Tension Tension

Interfacial Interfacial Interfacial

Sample mN / m Average, _mN / m

Average, mN / m Average, mN / mExample 1 at pH = 1 3.42 3.40 3.42

3.44

Example 1 at pH = 2.36 3.36 3.35

3.38

Example lapH = 12 3.49 3.47 3.48

3.51

Example 1 at pH = 13 3.92 3.93 3.91

3.92

Example 2 at pH = 1.80 4.77 4.82

4.80

Example 2 at pH = 7.5 3.94 3.89

3.97 3.95

Example 2 at pH = 13 5.22 5.23 5.19

5.25

Discussion of pH Data The pH data are plotted above and appear to indicate that there is a modest surface elevation and interfacial stress as extreme pHs are approached, and that the elevation is more significant at high versus low pH. The data clearly indicate that the protein / surfactant composition is preventing a portion of the surface and interfacial stress from increasing, which can otherwise be seen for the protein / surfactant composition surfactant package alone. A bowl-shaped (or even V-shaped) curve is observed with minimum values close to the mean pH range in both data series for protein / surfactant composition as well as for surfactant samples only from existing data.

All patents, patent applications and literature references cited in this specification are hereby incorporated by reference in their entirety.

Thus, the compounds, systems and methods described in the present invention offer many benefits over the prior art. While the above description contains many specificities, they should not be construed as limitations on the scope of the invention, but instead serve as an exemplification of preferred embodiments thereof. Many other variations are possible.

Accordingly, the scope of the present invention should be determined not by the embodiments illustrated above, but by the appended claims and their legal equivalents.

Claims (33)

1. COMPOSITION, characterized in that it comprises a surfactant and a protein system, wherein the protein mixture comprises proteins and stress proteins obtained by the yeast defermentation process to obtain a fermentation mixture, subjecting the fermentation mixture to the conditions of stress to obtain a post-fermentation mixture; ecentrifugation of the post-fermentation mixture and obtaining the supernatant, wherein the protein mixture maintains its functionality under extreme conditions. Composition according to Claim 1, characterized in that the stress conditions are selected from the group consisting of overheating, starvation, oxidative stress, mechanical stress and chemical stress. COMPOSITION according to claim 1, characterized in that the extreme conditions are selected from the group consisting of high temperature, high pH and low pH. Composition according to Claim 1, characterized in that the high temperature is above 50 ° C. Composition according to Claim 1, characterized in that the high temperature is a temperature below 110 ° C. Composition according to Claim 1, characterized in that the high pH is a pH above 9.5. Composition according to Claim 1, characterized in that the low pH is a pH below Composition according to Claim 1, characterized in that the stress proteins range in size from approximately 5 kDa to approximately 30 kDa. COMPOSITION according to claim 1, characterized in that the functionality of the protein mixture under extreme conditions is at least approximately 80% of the functionality of the protein mixture before subjecting to extreme conditions. Composition according to Claim 1, characterized in that the fermentation step is an aerobic fermentation. Composition according to Claim 1, characterized in that the surfactant comprises one or more of an anionic surfactant, a nonionic surfactant, a cationic surfactant and an amphoteric surfactant. Composition according to Claim 1, characterized in that the functionality is selected from the group consisting of reduction of surface tension, reduction of critical micelle concentration, and decoupling of biochemical processes. Composition according to Claim 12, characterized in that the decoupling comprises the control of biofilms. Composition according to Claim 13, characterized in that the control of the biofilms comprises the prevention, reduction and removal of the biofilms. Composition according to Claim 12, characterized in that the decoupling comprises oxidative phosphorylation. Composition according to Claim 13, characterized in that the biofilms are on a porous surface. Composition according to Claim 16, characterized in that the porous surface is selected from the group consisting of concrete, plaster, tiles, porous media, wastewater sludge, and fabrics. Composition according to Claim 17, characterized in that the porous medium comprises crude oil. Composition according to Claim 12, characterized in that the decoupling comprises accelerated and increased CO2 production as a by-product. Composition according to Claim 19, characterized in that accelerated CO2 production is used in the recovery of recovered oil to improve the microbial recovery of improved oil (MEOR) process. Composition according to Claim 1, characterized in that the composition comprises stress proteins, surfactants, stabilizers and an acid. Composition according to Claim 21, characterized in that the acid is selected from the group consisting of phosphoric acid, citric acid, lactic acid and hydrochloric acid. Composition according to Claim 1, characterized in that the composition comprises stress proteins, surfactants, stabilizers and a base. Composition according to Claim 23, characterized in that the base is selected from the group consisting of sodium hydroxide, sodium metasilicate, sodium carbonate, sodium tripolyphosphate, triethanolamine, monoethanolamine, and morpholine. 25. METHOD OF IMPROVING THE FUNCTIONALITY OF A TENSIVE SYSTEM, characterized in that it comprises the addition of stress proteins to the surfactant system, wherein stress proteins are obtained by the yeast defermentation process to obtain a fermentation mixture; fermentation mixture under stress conditions to obtain a post fermentation mixture; and centrifugation of the post-fermentation mixture and obtaining the supernatant, wherein the stress proteins maintain their functionality under extreme conditions. Method according to claim 25, characterized in that said surfactant system comprises sulfated alcohol surfactants. Method according to claim 25, characterized in that the surfactant system comprises sulfonated surfactants. Method according to claim 25, characterized in that the functionality of the surfactant system is increased by increasing the thermal stability of the surfactant system. Method according to claim 28, characterized in that the surfactant system is stable at a temperature above 160 ° F. Method according to claim 25, characterized in that the functionality of the surfactant system is increased by increasing the acid stability of the surfactant system. Method according to claim 30, characterized in that the surfactant system is stable at a pH below 3.5. Method according to claim 25, characterized in that the functionality of the surfactant system is increased by increasing the alkaline stability of the surfactant system. Method according to claim 32, characterized in that the surfactant system is stable at a pH greater than 9.5.