WO2015000812A1 - Process for cleaning surfaces - Google Patents

Abstract

Process for cleaning a surface wherein said surface is treated with at least one culture of at least one organism or a formulation comprising at least one culture of at least one microorganism.

Description

Process for cleaning surfaces

The present invention relates to processes for cleaning a surface wherein said surface is treated with at least one culture of at least one organism or a formulation comprising at least one culture of at least one organism.

The present invention further relates to novel cleaning formulations.

The cleaning of surfaces is an omnipresent challenge. There is a constant need for cleaning methods for cleaning the most diverse surfaces from multiple different stains and impurities with many different application requirements and under many different application conditions.

One field that has recently gained enormous importance is the cleaning of membranes.

Membranes can be used in many applications. For example membranes can be used for water treatment or in applications involving an aqueous medium. In such applications, fouling and the growth of algae and microorganisms and biofouling resulting therefrom can be a severe prob- lem.

Approaches to reduce the problem of fouling and biofouling normally involve (back)washing such membranes. Previous approaches often made use of rather aggressive cleaning agents like acids, bases or oxidative agents like peroxides. Using such aggressive cleaning agents, however, may damage the membrane and is a potential environmental risk.

EP1928586 discloses a method for cleaning membranes using persulfates.

EP1600204 discloses a method for cleaning membranes using cleaning agents based on poly- ethers.

EP 2 252 683 discloses a method for cleaning membranes using solutions of active oxygen source.

EP1567635 discloses a method for cleaning membranes using acidic compounds. Espinasse et al. compare various cleaning agents with respect to their efficiency for cleaning various foulants (Espinasse et al., Desalination, 2012, 296, 1 -6).

The object of the present invention was therefore to provide a process for the cleaning of surfaces that can be carried out under mild conditions with a high effectivity. In particular it was an object of the present invention to provide a process for increasing the water flux through a membrane and for reducing fouling and biofouling on a membrane.

This object has been solved by a process for cleaning a surface wherein said surface is treated with at least one culture of at least one organism or a formulation comprising at least one cul- ture of at least one organism.

Processes according to the invention are in principle applicable for the cleaning of all kinds of surfaces. They are especially suitable or the cleaning of surfaces that are in constant or sporad- ic contact with water. Examples are surfaces that are exposed to sea water, rain, fluvial water, body fluids, sewage water or industrial process water. Due to this exposure to water, such surfaces often face problems with respect to fouling, in particular to biofouling due to the deposition of organic material on such surfaces.

Processes according to the invention are for example suitable for cleaning metal surfaces, surfaces of inorganic materials like ceramics, or surfaces of biological materials like skin, leather or wood. Processes according to the invention are particularly suitable for the cleaning of surfaces of organic polymers.

In one embodiment, said surface is the skin of animals or humans. In another embodiment, said surface is a wound, an injury or an irregularity of the skin of animals or humans.

In one embodiment, said surface is a porous surface.

In one especially preferred embodiment of the invention processes according to the invention are used for the cleaning of membranes. In the context of this application a membrane shall be understood to be a thin, semipermeable structure capable of separating two fluids or separating molecular and/or ionic components or particles from a liquid. A membrane acts as a selective barrier, allowing some particles, substances or chemicals to pass through, while retaining others.

Processes according to the invention are for example suitable for cleaning membranes made of or comprising as major components in the separating layer, support layer or other components of the membrane ceramic materials, polyamide (PA), polyvinylalcohol (PVA), Cellulose Acetate (CA), Cellulose Triacetate (CTA), CA-triacetate blend, Cellulose ester, Cellulose Nitrate, regenerated Cellulose, aromatic , aromatic/aliphatic or aliphatic Polyamide, aromatic, aromatic/aliphatic or aliphatic Polyimide, Polybenzimidazole (PBI), Polybenzimidazolone (PBIL), Polyacrylonitrile (PAN), PAN-poly(vinyl chloride) copolymer (PAN-PVC), PAN-methallyl sul- fonate copolymer, Poly(dimethylphenylene oxide) (PPO), Polycarbonate, Polyester, Polytetraflu- roethylene PTFE, Poly(vinylidene fluoride) (PVDF), Polypropylene (PP), Polyelectrolyte complexes, Poly(methyl methacrylate) PMMA, Polydimethylsiloxane (PDMS), aromatic, aromatic/aliphatic or aliphatic polyimide urethanes, aromatic, aromatic/aliphatic or aliphatic polyam- idimides, crosslinked polyimides or polyarylene ether, polyethersulfones, polysulfone or poly- phenylenesulfone.

For example, membranes according to the invention can be reverse osmosis (RO) membranes, forward osmosis (FO) membranes, nanofiltration (NF) membranes, ultrafiltration (UF) membranes or microfiltration (MF) membranes. These membrane types are generally known in the art. Such membranes are for example used for the treatment of water, like the desalination of sea or brackish water or the treatment of municipal or industrial waste water. FO membranes are normally suitable for treatment of seawater, brackish water, sewage or sludge streams. Thereby pure water is removed from those streams through a FO membrane into a so called draw solution on the back side of the membrane having a high osmotic pressure. In a preferred embodiment, suitable FO membranes are thin film composite (TFC) FO membranes. Preparation methods and use of thin film composite membranes are principally known and, for example described by R. J. Petersen in Journal of Membrane Science 83 (1993) 81 -150. In a particularly preferred embodiment, suitable FO membranes comprise a fabric layer, a support layer, a separation layer and optionally a protective layer. Said protective layer can be considered an additional coating to smoothen and/or hydrophilize the surface. Said fabric layer can for example have a thickness of 10 to 500 μηη. Said fabric layer can for example be a woven or nonwoven, for example a polyester nonwoven. In a preferred embodiment, FO membranes comprise a support layer comprising as the main components at least one polysulfone, polyphenylenesulfone and/or polyethersulfone. In a preferred embodiment suitable FO membranes comprise a polyamide separation layer obtained from the condensation of a polyamine and a polyfunctional acyl halide.

RO membranes are normally suitable for removing molecules and ions, in particular monovalent ions. Typically, RO membranes separate mixtures based on a solution/diffusion mechanism. In a preferred embodiment, suitable membranes are thin film composite (TFC) RO membranes. Preparation methods and use of thin film composite membranes are principally known and, for example described by R. J. Petersen in Journal of Membrane Science 83 (1993) 81 -150. In a further preferred embodiment, suitable RO membranes comprise a fabric layer, a support layer, a separation layer and optionally a protective layer. Said protective layer can be considered an additional coating to smoothen and/or hydrophilize the surface. Said fabric layer can for exam- pie be a woven or nonwoven, for example a polyester nonwoven. In a preferred embodiment, RO membranes comprise a support layer comprising as the main components at least one polysulfone, polyphenylenesulfone and/or polyethersulfone. In a preferred embodiment suitable RO membranes comprise a polyamide separation layer obtained from the condensation of a polyamine and a polyfunctional acyl halide.

NF membranes are normally especially suitable for removing multivalent ions and large monovalent ions. Typically, NF membranes function through a solution/diffusion or/and filtration- based mechanism. NF membranes are normally used in crossflow filtration processes. In one embodiment of the invention, NF membranes comprise as the main component at least one polysulfone, polyphenylenesulfone and/or polyethersulfone. In a particularly preferred embodiment, the main components of a NF membrane are positively or negatively charged. Nanofiltra- tion membranes often comprise charged polymers comprising sulfonic acid groups, carboxylic acid groups and/or ammonium groups in combination with block copolymers useful according to the invention. In another embodiment, NF membranes comprise as the main component poly- amides, polyimides or polyimide urethanes, Polyetheretherketone (PEEK) or sulfonated poly- etheretherketone (SPEEK). UF membranes are normally suitable for removing suspended solid particles and solutes of high molecular weight, for example above 100000 Da. In particular, UF membranes are normally suitable for removing bacteria and viruses. UF membranes normally have an average pore diameter of 0.5 nm to 50 nm, preferably 1 to 40 nm, more preferably 5 to 20 nm. In a preferred embodiment, UF membranes comprise as major components polysulfones, polyethersulfones, polyphenylenesulfones or polyvinylidenenfluoride and optionally further additives like polyvinylpyrrolidone or or polyalkylene oxides like polyethylene oxides. Especially preferred membranes are UF membranes comprising polyethersulfone as the main component. In one preferred embodiment, processea according to the invention are used for cleaning UF membranes comprising 99.9 to 50% by weight of a combination of polyethersulfone and 0.1 to 50 % by weight of polyvinylpyrrolidone.

In another embodiment UF membranes comprise 95 to 80% by weight of polyethersulfone and 5 to 15 % by weight of polyvinylpyrrolidone.

MF membranes are normally suitable for removing particles with a particle size of 0.1 μηη and above. MF membranes normally have an average pore diameter of 0.05 μηη to 10 μηη, preferably 1 .0 μηη to 5 μηη. Microfiltration can use a pressurized system but it does not need to include pressure. MF membranes can be hollow fibers, flat sheet, tubular, spiral wound, hollow fine fiber or track etched. They are porous and allow water, monovalent species (Na+, CI-), dissolved organic matter, small colloids and viruses through but retain particles, sediment, algae or large bacteria. Microfiltration systems are designed to remove suspended solids down to 0.1 micrometres in size, in a feed solution with up to 2-3% in concentration. In one embodiment of the invention, MF membranes comprise as the main component at least one polysulfone, polyphe- nylenesulfone and/or polyethersulfone.

In one embodiment of the invention, membranes are present as spiral wound membranes or flat sheet membranes. In another embodiment of the invention, membranes are present as tubular membranes. In another embodiment of the invention, membranes are present as hollow fiber membranes. In yet another embodiment of the invention, membranes are present as single bore hollow fiber membranes. In yet another embodiment of the invention, membranes are present as multi bore hollow fiber membranes.

For membranes used in contact with water, the problem of fouling and biofouling is particularly grave, because biofouling occurs quite often and affects the flux and thus the efficiency of such membranes significantly.

The term "culture" herein denotes a liquid that comprises substances released by at least one organism, optionally said at least one organism, and optionally the remaining nutrient broth. In the context of this application, the term culture shall be understood to include cultures of at least one organism as well as culture filtrates or culture supernatants of such cultures.

Thus, a culture may in one embodiment comprise at least one organism, substances released by said at least one organism and optionally the remaining nutrient broth.

In another embodiment of the invention, a culture is a culture filtrate or a culture supernatant comprising substances released by at least one organism, and optionally the remaining nutrient broth, whereas organisms themselves have been removed from such cultures. In one embodiment of the invention, said culture is a culture filtrate and is the aqueous liquid that is obtained from a culture by separating the biomass from the aqueous medium by filtration. In another embodiment said culture is a culture filtrate that is the aqueous liquid that is obtained from a culture by separating the biomass from the aqueous medium by centrifugation or sedimentation and subsequent decantation or filtration. In another embodiment said culture is the liquid that is obtained from a culture by extracting the biomass of the culture with water or another extracting medium like an organic solvent like an alcohol, acetonitrile, dimethylsulfoxide, N-methylpyrrolidone. If the biomass from a culture is extracted, the biomass is normally separated from the culture by filtration or centrifugation, optionally the cells lyzed by ultrasonication, high pressure homogenization or chemical lysis and then extracted with a suitable extraction medium like water, an organic solvent like an alcohol acetonitrile, dimethylsulfoxide, N- methylpyrrolidone.

Suitable cultures are in principle cultures of at least one organism, preferably at least one microorganism. Suitable cultures include for example cultures of fungi like yeasts, bacteria, pro- tists, algae, cells like organ or tissue cells from plants or animals.

Suitable cultures may for example be liquid cultures in a containment like a fermenter or surface cultures on solid growth plates or substrate, stab cultures or carrier-based cultures.

Preferred cultures are cultures of at least one microorganism like fungi or bacteria.

In one preferred embodiment, suitable cultures are cultures of at least one microorganism, especially at least one fungus, that is a plant pathogen. A plant pathogen herein shall denote a microorganism that causes a disease on plants or a microorganism that grows on or in a plant and that harms or weakens said plant.

Especially preferred cultures are of at least one fungus.

In one preferred embodiment, preferred cultures are cultures of at least one plant pathogenic fungus. In one preferred embodiment, preferred cultures are cultures of at least one fungus from the phylum Ascomycota,Basidiomycota or Zygomycota.

In an especially preferred embodiment, preferred cultures are cultures of at least one fungus from the class of Eurotiomycetes, Ascomycetes, Pleosporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes.

In an especially preferred embodiment, preferred cultures are cultures of at least one fungus from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilagi- nales.

In an especially preferred embodiment, preferred cultures are cultures of at least one fungus from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoace- ae, Polyporaceae, Ustilaginaceae.

Particularly preferred are cultures of at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalo- sporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides.

Cultures like culture filtrates or culture supernatants can be obtained using standard methods known in the art. Normally, preparing suitable culture filtrates comprises the following steps: a. growing a culture of an organism in an or on a medium, preferentially an aqueous medium

b. separating the medium in which the organism was grown and the biomass by filtration, centrifugation or any other method,

c. optionally extracting the biomass. To grow an organism in a culture, normally culture medium will be composed of nutrients and water, the culture is inoculated by microbial biomass. Said mixture is let to stand and optionally shaken for a certain amount of time, typically for some days to weeks.

The present invention requires cultivating or culturing microorganisms described herein, such that the culture may be used for cleaning of surfaces like membranes. The term "cultivating" includes maintaining and/or growing a living microorganism of the present invention (e.g., maintaining and/or growing a culture or strain). In one embodiment, a microorganism is cultured in liquid media. In another embodiment, a microorganism is cultured in solid media or semi-solid media. In a preferred embodiment, a microorganism is cultured in media (e.g., a sterile, liquid media) comprising nutrients essential or beneficial to the maintenance and/or growth of the microorganism. Carbon sources which may be used include sugars and carbohydrates, such as for example glucose, sucrose, lactose, fructose, maltose, molasses, starch and cellulose, oils and fats, such as for example soy oil, sunflower oil, peanut oil and coconut oil, fatty acids, such as for example palmitic acid, stearic acid and linoleic acid, alcohols, such as for example glycerol and ethanol, and organic acids, such as for example acetic acid. In a preferred embodiment, glucose, fructose or sucrose is used as carbon sources. These substances may be used individually or as a mixture. Nitrogen sources which may be used comprise organic compounds containing nitrogen, such as peptones, yeast extract, meat extract, malt extract, corn steep liquor, soya flour and urea or inorganic compounds, such as ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate and ammonium nitrate. The nitrogen sources may be used individually or as a mixture. Phosphorus sources which may be used are phosphoric acid, potassium dihydrogen phosphate or dipotassium hydrogen phosphate or the corresponding salts containing sodium. The culture medium must furthermore contain metal salts, such as for example magnesium sulfate or iron sulfate, which are necessary for growth. Further, essential growth-promoting substances such as amino acids and vitamins may also be used in addition to the above stated substances. Suitable precursors may furthermore be added to the culture medium. The stated feed substances may be added to the culture as a single batch or be fed appropriately during cultivation. Preferably, microorganisms are cultured under controlled pH. In one embodiment, microorganisms are cultured at a pH of between 4.4 and 8.5. In one embodi- ment, microorganisms are cultured at a pH of about 5.6. The desired pH may be maintained by any number of methods known to those skilled in the art. For example, basic compounds such as sodium hydroxide, potassium hydroxide, ammonia, or ammonia water, or acidic compounds, such as phosphoric acid or sulfuric acid, are used to appropriately control the pH of the culture. Also preferably, microorganisms are cultured under controlled aeration. In one embodiment, aeration is controlled by regulating oxygen levels in the culture, for example, by regulating the amount of oxygen dissolved in culture media. Preferably, aeration of the culture is controlled by agitating the culture. Agitation may be provided by a propeller or similar mechanical agitation equipment, by revolving or shaking the growth vessel (e.g. a fermenter) or by various pumping equipment. Aeration may be further controlled by the passage of sterile air or oxygen through the medium (e.g., through the fermentation mixture). Also preferably, microorganisms are cultured without excess foaming (e.g., via addition of antifoaming agents such as fatty acid poly- glycolesters).

Moreover, microorganisms can be cultured under controlled temperatures. In one embodiment, controlled temperatures include temperatures between 15°C and 95°C. In another embodiment, controlled temperatures include temperatures between 15°C and 70°C. Preferred temperatures are between 15°C and 55°C, more preferably between 20°C and 30°C.

Microorganisms can be cultured (e.g., maintained and/or grown) in liquid media and preferably are cultured, either continuously or intermittently, by conventional culturing methods such as standing culture, test tube culture, shaking culture (e.g., rotary shaking culture, shake flask cul- ture, etc.), aeration spinner culture, or fermentation. In a preferred embodiment, the microorganisms are cultured in shake flasks. In a more preferred embodiment, the microorganisms are cultured in a fermentor (e.g., a fermentation process). Fermentation processes of the present invention include, but are not limited to, batch, fed-batch and continuous methods of fermentation. The phrase "batch process" or "batch fermentation" refers to a closed system in which the composition of media, nutrients, supplemental additives and the like is set at the beginning of the fermentation and not subject to alteration during the fermentation, however, attempts may be made to control such factors as pH and oxygen concentration to prevent excess media acidification and/or microorganism death. The phrase "fed-batch process" or "fed-batch" fermentation refers to a batch fermentation with the exception that one or more substrates or supplements are added (e. g., added in increments or continuously) as the fermentation progresses. The phrase "continuous process" or "continuous fermentation" refers to a system in which a defined fermentation media is added continuously to a fermentor and an equal amount of used or "conditioned" media is simultaneously removed, preferably for recovery of the desired beta- lysine.

Processes according to the invention comprise treating a surface with at least one culture like a culture filtrate or culture supernatant of at least one organism or a formulation comprising at least one culture like a culture filtrate or culture supernatant of at least one organism.

Especially for cleaning membranes, processes according to the invention can be carried out under a pressure of atmospheric pressure to 100 bar, preferably 1.03 bar to 80 bar, more preferably 1 .05 bar to 50 bar, especially preferably 1.2 bar to 20 bar, especially preferably 1 .5 to 5 bar. Processes according to the invention can in principle be carried out at any temperature, as long as the cleaning medium, in most cases water, is present in liquid state. Normally processes according to the invention are carried out at temperatures between 5 and 90 °C, preferably 15 to 60 and especially preferably 20 to 45 °C. The temperatures and pressures may also be varied while processes according to the invention are being carried out.

In one embodiment of the invention, cultures can be formulated with at least one further cleaning agent or cleaning formulation. Such further cleaning agents or formulations can be applied prior to, during or after the application of said at least one culture. Suitable further cleaning agents include for example chelating agents, oxidative agents, reducing agents, surfactants, disinfectants, acids or bases.

Preferred chelating agents are for example methylglycinediacetic acid and its salts (MGDA) , ethylenediamine tetraacetic acid (EDTA), ortho- or meta phosphoric acids, diethylenetriamine- pentaacetic acid (DTPA), nitrilotriacetic acid (NTA), N-(2-yydroxyethyl)-ethylenediamine- Ν,Ν,Ν'- triacetic acid (HEDTA), diethanolglycine (DEG), 2-hydroxyethyliminodiacetic acid (EDG), citric acid, ascorbic acid or the salts of all the chelating agents listed above like their sodium salts.

Preferred oxidative agents are for example peroxides like hydrogenperoxide, persulfates, perborates, percarbonates. Preferred reducing agents are for examples sulfites, bisulfites, nitrites, phosphites, phosphon- ites.

Preferred surfactants are for example nonionic, cationic or anionic surfactants.

Preferred anionic surfactants are for example polymers comprising carboxylate and/or sulfonate groups. Examples of suitable anionic surfactants include homopolymers or copolymers of (meth)acrylic acid or alkylbenzene sulfonic acids. Such anionic surfactants are for example available from BASF SE under the tradename Sokalan<^R) or Lutensit<^R). Preferred cationic surfactants are for example polymers comprising amino/ammonium groups like polyvinylamines or polyethyleneimines. Such cationic surfactants are for example available from BASF SE under the tradename Lupasol<^R>.

Preferred nonionic surfactants are for example polyalkyleneoxides like polyethyleneoxides. Suitable nonionic surfactants are for example obtainable by reacting alkyleneoxides like eth- yleneoxide and/or propyleneoxides with suitable starter alcohols. Such nonionic surfactants are for example available from BASF SE under the tradename Lutensol<^R>.

Preferred acids are for example methanesulfonic acid, phosphoric acid, sulfuric acid, hydrochlo- ric acid, hydrobromic acid, nitric acid, acetic acid, amidosulfuric acid. Especially preferred acids are methanesulfonic acid and hydrochloric acid.

Preferred bases are for example hydroxides or carbonates, like sodium or potassium hydroxide. In one embodiment, surfaces like membrane surfaces are first treated with a culture of at least one fungus and a then with a chelating agent like MGDA.

The process according to the invention allows for the cleaning of surfaces, particularly of mem- branes, under mild conditions and with high effectivity and efficiency. The process according to the invention is particularly effective for the removal of fouling and/or biofouling resulting from microorganism such as algae, bacteria, fungi or eukaryotic unicellular organism like protists.

Processes according to the invention are also easy and economical to carry out.

The process according to the invention provides a method for improving the flux through membranes, in particular the flux of water in water treatment application. Cultures useful according to the invention can thus act as flux enhancers for membranes in water treatment applications.

Another aspect of the invention are cultures of at least one organism, especially of at least one type of cells or microorganisms like fungi, bacteria, algae, especially of (pant pathogenic) fungi, especially of fungi of from the phylum Ascomycota, phylum Basidiomycota or phylum Zygomyco- ta, especially from the class of Eurotiomycetes, Ascomycetes, Pleosporomycetidae, Sordari- omycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinc- tum, Gliocladium roseum and Fusarium sporotrichoides for cleaning wounds. Another aspect of the invention are cultures of at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides for medical applications.

Another aspect of the invention is the use of cultures of at least one organism, especially of at least one type of cells or microorganisms like fungi, bacteria, yeasts, algae, especially of (plant pathogenic) fungi, especially of fungi of from the phylum Ascomycota, phylum Basidiomycota or phylum Zygomycota, especially from the class of Eurotiomycetes, Ascomycetes, Pleosporomy- cetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides for cosmetic applications like cleaning the skin.

Another aspect of the present invention relates to novel cleaning formulations comprising cultures of at least one organism as described above and at least one further cleaning agent as described above.

In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism, especially of fungi of from the phylum Ascomycota, phylum Basid- iomycota or phylum Zygomycota, especially from the class of Eurotiomycetes, Ascomycetes, Pleosporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides and at least one chelating agent.

In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one fungus, especially of fungi of from the phylum Ascomycota, phylum Basidi- omycota or phylum Zygomycota, especially from the class of Eurotiomycetes, Ascomycetes, Ple- osporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides and MGDA. In one embodiment cleaning formulations according to the invention comprise at least one culture filtrate of at least one fungus, especially of fungi of from the phylum Ascomyco- ta or phylum Basidiomycota, especially from the class of Eurotiomycetes, Ascomycetes, Pleo- sporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides and EDTA. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one acid. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism, especially of fungi of from the phylum Ascomycota, phylum Basidiomycota or phylum Zygomycota, especially from the class of Eurotiomycetes, Ascomycetes, Pleosporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides and methane sulfonic acid.

In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one surfactant. In one embodiment cleaning formula- tions according to the invention comprise at least one culture of at least one organism and one nonionic surfactant. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one polyalkylene oxide. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism, especially of fungi of from the phylum Ascomycota, phylum Basidiomycota or phylum Zygomycota, especially from the class of Eurotiomycetes, Ascomycetes, Pleosporo- mycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilagi- nales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteri- aceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides and at least one polyalkylene oxide and at least one chelator like MGDA.

In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one anionic surfactant. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one surfactant comprising sulfonate groups. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one surfactant comprising carboxylate groups. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism, especially of fungi of from the phylum Ascomycota, phylum Basidiomycota or phylum Zygomycota, especially from the class of Eurotiomycetes, Ascomycetes, Pleosporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporace- ae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides and at least one anionic surfactant and at least one chelator like MGDA.

In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one cationic surfactant. In one embodiment cleaning formulations according to the invention comprise at least one culture of at least one organism and at least one surfactant comprising ammonium groups. In one embodiment cleaning formu- lations according to the invention comprise at least one culture of at least one organism, especially of fungi of from the phylum Ascomycota, phylum Basidiomycota or phylum Zygomycota, especially from the class of Eurotiomycetes, Ascomycetes, Pleosporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes, Ustilaginomycetes, especially from the order of Eurotiales, Pleosporales, Hypocreales, Myriangiales, Polyporales, Ustilaginales, especially from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae, Ustilaginaceae, and especially selected from at least one fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Usti- lago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinc- tum, Gliodadium roseum and Fusarium sporotrichoides and at least one cationic surfactant and at least one chelator like MGDA.

Cleaning formulations are very effective and efficient for cleaning surfaces, particularly membranes. They are capable of removing biofouling and of biological materials from surfaces. They are easy and economical to prepare.

Examples

Abbreviations

BF: biofilm

MEP: malt extract peptone

KGA: potato glucose

TSBY: tryptic soy broth yeast

MGDA: methylglycinediacetic acid

Cfu colony forming units

Rpm: revolutions per minute

MDF: medium density fiber

BWRO: brackish water reverse osmosis

Examples 1 to 1 1 : Production of fungal culture filtrate

In each of experiments 1 to 1 1 , the respective fungus was grown on solid or in liquid MEP medium. Liquid MEP culture medium was prepared by mixing 30 g/L malt extract and 3 g/L soya peptone in distilled water, the pH was adjusted to 5.6 and the medium sterilization by autoclav- ing at 1 15°C for 20 min. For solid MEP medium 15 g/L agar was added before autoclavation and the medium poured before cooling to room temperature. Liquid cultures of the fungi were inoculated with an agar plug of 1 cm x 1 cm covered with fungal mycelium in 50 ml of liquid MEP medium. The cultures were shaken at 100 rpm for 5 days at room temperature, for production of larger amounts this culture was transferred to 1 liter medium after 2 days.

For the production of larger amounts of culture filtrates the microorganism were grown in stirred fermenters with a working volume of 20 liters. The medium was composed of 7 g/L (NH₄)₂S0₄; 2,6 g/L MgS0₄; 5 g/L K₂HP0₄; 0,18 g/L CaCI₂; 1 ,5 g/L citric acid; 30 g/L malt extract; 3 g/L soya peptone and 17,5 ml of the trance element solution (40 g/L citric acid; 1 1 g/L ZnS0₄ ^* 7 H₂0; 8,5 g/L (NH₄)₂Fe(S0₄)₂ ^*6H₂0; 3 g/L MnS0₄ ^* H₂0; 0,8 g/L CuS0₄

^* 5 H₂0; 0,25 g/L CoS0₄ ^* 7 H₂0).

The inoculation volume was 2 % of the total fermentation broth. A pH of 5.6 and the temperature of 24°C were kept constant. Reactor was stirred at 600 rpm and run for 1 1 days.

For harvesting of the culture filtrates the culture or fermentation broth was filtered through a 0.22 μηη filter after 5 to 1 1 days of microbial growth.

Experiments 12 to 13: Production of fungal culture filtrate

Fusarium fujikuroi or Sphaceloma manihoticola fungi of experiments 12 and 13 were grown in liquid or solid potato glucose medium. For medium preparation 500 g potatoes were sliced into thin pieces, added to distilled water and steamed for 20 minutes. Subsequently the mixture was filtered through cheesecloth. The processed potatoes were mixed with 20 g/L glucose and distilled water added to a final volume of 1 .0 L. After that the medium was autoclaved at 1 15°C for 20 minutes.

Liquid cultures of the fungi were inoculated with an agar plug of 1 cm x 1 cm covered with fungal mycelium in 50 ml of liquid KGA medium. The cultures were shaken at 100 rpm for 5 days at room temperature, for production of larger amounts this culture was transferred to 1 liter medium after 2 days.

For the production of larger amounts of culture filtrates the microorganism were grown in stirred fermenters with a working volume of 20 liters. The medium was composed of 7 g/L (NH₄)₂S0₄; 2,6 g/L MgS0₄; 5 g/L K₂HP0₄; 0,18 g/L CaCI₂; 1 ,5 g/L citric acid; 30 g/L malt extract; 3 g/L soya peptone and 17,5 ml of the trance element solution (40 g/L citric acid;

1 1 g/L ZnS0₄ ^* 7 H₂0; 8,5 g/L (NH₄)₂Fe(S0₄)₂ ^*6H₂0; 3 g/L MnS0₄ ^* H₂0; 0,8 g/L CuS0₄ ^* 5 H₂0; 0,25 g/L CoS0₄ ^* 7 H₂0).

The inoculation volume was 2 % of the total fermentation broth. A pH of 5.6 and the temperature of 24°C were kept constant. Reactor was stirred at 600 rpm and run for 1 1 days. For harvesting of the culture filtrates the culture or fermentation broth containing the organism was filtered through a 0,22 μηη filter after 5 to 1 1 days of growth.

Example 14: Identification of microorganism cultures detaching biofilms

Using a Biofilm Detachment Assay (BDA-assay) cultures of microorganism were screened for their activity to detach biofilms. For that purpose the biomass was removed from culture by filtration and the culture filtrate applied in the BDA-assay.

For the biofilm detachment assay (BDA-Assay), Staphylococcus epidermidis cells were scraped from an agar plate and suspended in 1 ml TSBY medium (30 g/L tryptic soy broth; 3 g/L yeast extract; pH 7) to a density of 5 x 10⁵ cfu/ml. 200 μΙ of the diluted culture were added to the wells of a 96 well microtiter plate and incubated over night at 37°C on a shaker (50 rpm). During the incubation the bacteria adhere on the well surface and formed a biofilm. The supernatant was carefully removed afterwards and the well washed with H₂0.

To test the biofilm dispersal activity 200 μΙ of the microorganism culture filtrate or control solution , i.e. sterile medium were added to the Staphylococcus epidermidis biofilm and incubated for 2 hours. After the incubation time, the supernatants were carefully removed, the well washed with buffer and the biofilm stained with crystal violet. To this end, 250 μΙ of 0.5 % crystal violet solution was added to each well and the biofilm stained for 10 min. Unbound crystal violet was removed by pipetting and the well rinsed again with water. For quantification the biofilm was dissolved in 33 % acetic acid and the O.D. at 570 nm measured.

The filtrates of 13 strains of microorganism led to biofilm removal in the range of 71 - 91 % (table 1 ). 5 of the 13 hits belong to the genus Fusarium. For the exclusion of media-based biofilm detaching effects, the media used for microorganism cultures were tested in the BDA-Assay (table 1 , row 14, 15). These control samples didn^'t show any or only very weak biofilm degradation.

Table 1 : S. epidermidis biofilm dispersal by different culture filtrates of different biofilms

Example 15: Detachment of natural water biofilm by treatment with culture filtrates

The identified culture filtrates showed very good removal of the artificial biofilm produced in the multi titer plate. Of interest was to study the effect on natural biofilms. Therefore, glass slides were placed in a creek for 7 weeks. The biofilm formed on the slides (see figure 2, buffer controls) strongly stick to its surface. Each slide showed two areas along its longitudinal axle: the rough top region to which the biofilm stuck much stronger, and the lower region were the glass surface was evenly smooth.

The slides were incubated over night with culture filtrates of Experiments No. 1 , No. 3 No. 4, No. 5, No. 6, and intensively washed with tap water. Subsequently, the central region of each slide was gently wiped with soft tissue.

The biofilm of the slides exposed to the buffer controls did not show any biofilm removal (figure 1 ). Even in the central region, which had been softly rubbed, only minor biofilm detachment could be observed.

Culture filtrate No. 4 and No. 5 effectively removed the natural biofilm at which No. 4 performed best. These results showed that the fungal culture filtrates were also effective against natural biofilms.

Figure 1 shows the effects of cleaning of biofilm-covered slides with fungal culture filtrates. Buffer controls: pH 4; pH 5 ; pH 8; slides were incubated with 50 ml of culture filtrates of the fungi No. 3, 4, 5, 6. All the slides were incubated on a shaker (50 rpm) at 37°C. The culture filtrates were prepared from lab cultures grown in MEP medium.

Example 16: Detachment of natural water biofilm by treatment with formulations of culture filtrates with further cleaning agents

It was tested if a pretreatment of a natural biofilm with the MGDA and/or a nonionic surfactant composed of alkyl endcapped polyethylene glycol could improve the cleaning efficiency of the candidate filtrates. First the biofilm covered slides were incubated with 10 mM of the nonionic surfactant and 100 mM of MGDA for two hours, subsequently the bio- film was incubated with the culture filtrates for 24 hours. The slides treated with these additional chemicals were brighter than the non-preincubated slides. Figure 2 shows the effects of cleaning of MGDA / a alkyl endcapped polyethylene glycol surfactant preincubat- ed biofilm-covered slides with fungal culture filtrates. This showed that the additional chemical treatment enhanced the cleaning efficiency of the culture filtrates.

Example 17: Cleaning of fresh/process water from a MDF manufacturing plant using RO membrane

The cleaning effect of the filtrates on reverse osmosis membranes comprising a polyam- ide separating layer on a carrier layer of polyethersulfone was analyzed. Experiment 17 was conducted with a fresh water reverse osmosis membrane. Membrane pieces of 6.5 x 23.5 cm were embedded into a flux measuring cell and exposed to synthetic salt solution (0.1 % NaCI) at 20 bar. For each experiment, three measuring cells were used in parallel and the values averaged.

For cleaning of the membranes, the membrane pieces were incubated with the culture filtrates for 12 hours. After incubation with the cleaning solution, the membranes were washed and fixed in the measuring cell.

After cleaning with the culture filtrates No. 4, No. 5 and No. 6, the flux increased significantly (table 3). If a treatment of 1 h with 0,5 % MGDA was applied in addition the flux was further increased. Best result was achieved by cleaning with No. 5 culture filtrate, followed by treatment with MGDA (flux increase of 146.8 %). NaCI retention was not affected by the cleaning procedure, showing that no deterioration of the membrane occurred (table 3).

Table 3: Flux and NaCI retention of none (control) and the culture filtrate cleaned membranes. After cleaning with the culture filtrates the indicated (+MGDA) samples were incubated with 10 mM MGDA for 1 h. Best cleaning effect was achieved with No. 5 + 10 mM MGDA.

Example 18: Cleaning of brackish water using a reverse osmosis membrane

The fouling cake and the biofilm formed on membranes depend on the nature of the feed water. It was therefore tested if the culture filtrates were effective on membranes from different water treatment plants. Therefore a reverse osmosis membrane comprising a poly- amide separating layer on a carrier layer of polyethersulfone from a brackish water desalination plant was analyzed (table 4). With cleaning solution 1 , consisting of an acid, alkaline and protease enzyme treatment the flux increased 60,9 %. No flux increment was observed on the MGDA treated membrane. Surprisingly the flux increased by 185% on the membrane cleaned with the culture filtrate No.6. This effect was enhanced, if a treatment with 10 mM MGDA was done after the cleaning procedure with the filtrate (239% flux increase). The cleaning efficiency of the 10 fold diluted filtrate was the same as the undi- luted filtrate, the cleaning efficiency dropped only slightly when the supernatant was further diluted 1 :100. Consistent with the previous experiment the cleaning with culture supernatant had no effect on the salt retention.

Table 4: Cleaning of BWRO cleaned with the culture filtrate of No. 6. The culture filtrate was diluted 1 : 10 and 1 :100 fold. Cleaning with the filtrate was done for 12 hours. Certain samples an additional treatment with 10 mM MGDA was done for 1 hours.

An additional BWRO membrane was cleaned with the culture filtrates No. 5 and No. 6. Also on this additional membrane a good cleaning efficiency with No. 6 was observed Table 5). The flux increased after cleaning with the supernatant from No. 5, showing that also this filtrate works as membrane cleaner.

In summary the filtration experiments showed that the culture filtrates identified with the BDA assay were able to clean fouled membranes. Remarkable was that membranes fed with different source waters were cleaned with the filtrates, showing that the cleaning do not depend on a specific foulant.

Table 5: Cleaning of BWRO membrane with culture filtrate No. 5 and No. 6. As control the membrane was cleaned with medium only.

Claims

Process for cleaning surfaces

Claims 1 . Process for cleaning a surface wherein said surface is treated with at least one culture of at least one organism or a formulation comprising at least one culture of at least one organism, wherein said at least one organism is selected from fungi or bacteria.

2. Process according to at least one of the preceding claims, wherein said at least one organism is a plant pathogen.

3. Process according to at least one of the preceding claims, wherein said at least one or- ganism is a fungus from the phylum Ascomycota, phylum Basidiomycota or phylum Zygomycete.

4. Process according to at least one of the preceding claims, wherein said at least one organism is a fungus from the class Eurotiomycetes, Ascomycetes, Pleosporomycetidae, Sordariomycetes, Dothideomycetes, Agaricomycetes or Ustilaginomycetes.

5. Process according to at least one of the preceding claims, wherein said at least one organism is a fungus from the order of Eurotiales, Pleosporales, Hypocreales, Myriangial- es, Polyporales or Ustilaginales.

6. Process according to at least one of the preceding claims, wherein said at least one organism is a fungus from the family of Trichocomaceae, Pleosporaceae, Nectriaceae, Beionecteriaceae, Elsinoaceae, Polyporaceae or Ustilaginaceae.

7. Process according to at least one of the preceding claims, wherein said at least one organism is a fungus selected from Verticillium agaricinum, Fusarium proliferatum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Cephalosporium sp., Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrichoides.

8. Process according to at least one of the preceding claims, wherein said culture is obtained in a process comprising the following steps:

a. growing a culture of an organism in a or on a medium,

b. separating the medium in which the organism was grown and the biomass, c. optionally extracting the biomass.

9. Process according to at least one of the preceding claims, wherein said surface is selected from an organic polymer, metal surfaces, surfaces of inorganic materials like ceramics, or surfaces of biological materials like skin, leather or wood.

10. Process according to at least one of the preceding claims, wherein said surface is a porous surface.

1 1 . Process according to at least one of the preceding claims, wherein said surface is the surface of a membrane.

12. Process according to at least one of the preceding claims, wherein said surface is treated with a culture of at least one organism under a pressure of atmospheric pressure to 100 bar.

13. Process according to at least one of the preceding claims, wherein said surface is treated with a culture of at least one fungus in combination with at least one further cleaning agent, wherein said at least one further cleaning agent is applied prior to, simultaneously or after said at least one culture of at least one organism.

14. Process according to claim 16, wherein said at least one further cleaning agent is selected from chelating agents, peroxides, surfactants, disinfectants, acids or bases.

15. Cleaning formulation comprising at least one culture of at least one organism and at least one further cleaning agent, wherein said at least one further cleaning agent is selected from chelating agents, peroxides, surfactants, disinfectants, acids or bases.

16. Culture of at least one fungus selected from Verticillium agaricinum, Fusarium prolifera- tum, Fusarium sulphureum, Fomes fomentarius, Ustilago maydis, Aspergillus giganteus, Drechslera avenae, Fusarium tricinctum, Gliocladium roseum and Fusarium sporotrich- oides for medical applications.

17. Culture of at least one organism for cleaning wounds.