CN1505680A - Biological compositions for solid waste treatment - Google Patents

Biological compositions for solid waste treatment Download PDF

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Publication number
CN1505680A
CN1505680A CNA028090608A CN02809060A CN1505680A CN 1505680 A CN1505680 A CN 1505680A CN A028090608 A CNA028090608 A CN A028090608A CN 02809060 A CN02809060 A CN 02809060A CN 1505680 A CN1505680 A CN 1505680A
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China
Prior art keywords
yeast cell
yeast
hours
electromagnetic field
cell
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
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CNA028090608A
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Chinese (zh)
Inventor
张令玉
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Ultra Biotech Ltd
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Ultra Biotech Ltd
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Filing date
Publication date
Priority claimed from US09/797,371 external-priority patent/US6391617B1/en
Priority claimed from US09/797,372 external-priority patent/US20020123127A1/en
Priority claimed from US09/797,378 external-priority patent/US6391618B1/en
Priority claimed from US09/797,437 external-priority patent/US6391619B1/en
Priority claimed from US09/797,382 external-priority patent/US20020123129A1/en
Priority claimed from US09/797,381 external-priority patent/US6436695B1/en
Priority claimed from US09/797,493 external-priority patent/US6440713B1/en
Priority claimed from US09/797,377 external-priority patent/US20020123130A1/en
Application filed by Ultra Biotech Ltd filed Critical Ultra Biotech Ltd
Publication of CN1505680A publication Critical patent/CN1505680A/en
Pending legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • A01N63/32—Yeast
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00—Treatment of water, waste water, or sewage
    • C02F1/50—Treatment of water, waste water, or sewage by addition or application of a germicide or by oligodynamic treatment
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/26—Processes using, or culture media containing, hydrocarbons
    • C12N1/28—Processes using, or culture media containing, hydrocarbons aliphatic
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds

Abstract

The present invention relates to biological compositions useful for the treatment of solid waste. The biological compositions of the invention comprises a plurality of yeast cells which is capable of suppression of growth of pathogenic microorganisms, breakdown of undesirable chemicals, such as antibiotics, insecticides and waste chemicals, and reducing the odor of organic waste matters. The yeast cells of the invention are produced by culturing the plurality of yeast cells under activation conditions in the presence of a series of electromagnetic fields. The invention also relates to methods for manufacturing the treatment composition.

Description

Handle the biological composition of solid waste
1. invention field
What the present invention relates to be used for solid waste disposal contains the zymic biological composition.Can be exercised multiple function thereby the yeast in the present composition has stimulated, comprise the degraded chemical, reduce smell and suppress microorganism.The invention still further relates to the method for these biological compositions of preparation, and the method for using these biological composition processing of waste.
2. background of invention
Every day, industrial production and agriculture production activity and city produced a large amount of solid waste.If these refuses are not correctly handled, can cause destruction serious and that continue to environment.In the period of 1995-1996, the U.S. has produced 20,800 ten thousand tons of municipal solid waste.In the municipal solid waste that is produced, 5,600 ten thousand tons (27%) by utilizing or compost is reclaimed, 3,350 ten thousand tons (16%) by high temperature incineration, 11,850 ten thousand tons (57%) is buried again.
Municipal waste can be processed, replaces burying.A kind of Municipal waste treatment process is included in high temperature incineration refuse in the incinerator.The burning Municipal waste has reduced its volume significantly like this.Municipal waste burns the ashes produced and must quilt deals carefully with to prevent the infringement of any possible unwanted component to environment.And, must be in the controlled levels that allows from the flue dust that the incinerator chimney is discharged.
At agriculture field, although mineral manure is providing very important aspect the sufficient agricultural prods to the mankind, it also is familiar with the harm of environment in recent years.Mineral manure can damage soil.For example, most of nitrogenous fertilizer can make soil acidification, therefore are unfavorable for the growth of plant and other soil organisms.Widely-used chemical nitrogen fertilizer also can suppress the activity of natural nitrogen-fixing microorganism, has reduced the natural fertilizer of soil thus.Life-time service mineral manure can also cause the serious environmental pollution.For example, because losing of nitrogenous fertilizer that leaching and soil erosion cause and phosphate fertilizer causes the eutrophication of soil and phreatic pollution and surface water.
Another kind of agricultural waste are muck, if properly do not store or remove, can cause the threat of healthy and environment.For example, it can cause environmental pollution, i.e. smell and dust; Superfluous nutrition, organism, salt and pathogen contamination surface water.For example, contain pathogenic microorganism such as intestinal bacteria, Salmonellas and Shigellae in the muck.
All things considered, the pollution of removing as improper waste treatment Policy Result is a complexity and large order.The cost of this task also is huge.Therefore, need cheap and effectively method handle the refuse that countless mankind's activity produces.
Advise in many cases in polluting control, using biological composition.Utilize the bio-feritlizer of microorganism to be suggested substitute as mineral manure.Naturally occurring nitrogen-fixing microorganism comprises bacterium, for example root nodule bacterium, vinelandii and azospirillum No. 5,071,462, United States Patent (USP) (for example see) and fungi, and for example flavus/aspergillus oryzae No. 4,670,037, United States Patent (USP) (for example see) uses in bio-feritlizer.The naturally occurring microorganism that can to make phosphate rock ore or other insoluble phosphoric acid salt solubilising be soluble phosphate is also used in bio-feritlizer, use (for example United States Patent (USP) 5,912, No. 398) respectively with nitrogen-fixing microorganism or (for example United States Patent (USP) 5 to unite use, 484, No. 464).Developed and a kind of method based on recombinant DNA technology, produce be used for bio-feritlizer can more effective fixed nitrogen, divide phosphorus decomposing and divide the bacterial isolates of potassium decomposing, for example see United States Patent (USP) 5,578, No. 486; The open WO95/09814 of PCT; Chinese patent is open: CN1081662A; CN1082016A; CN1082017A; CN1103060A; And CN1109595A.
Here citing document and do not mean that any document of admitting here to be quoted all is the prior art of being correlated with, or admit that the document quoted has substantive relation to the patentability of the application's claim.These documents are based on the obtainable information of applicant all about the statement on date with about the statement of content, do not constitute any admitting about these document dates and content exactness.
3. summary of the invention
The present invention relates to be used for the biological composition of solid waste disposal.Biological composition of the present invention comprises can suppress pathogenic microorganism growth, decompose adverse chemical product for example microbiotic, sterilant and waste chemicals and a plurality of yeast cell of reducing the organic waste smell.
In various embodiments, the yeast that the present invention uses is that the on sale and/or public can obtain on the market, such as but not limited to yeast saccharomyces cerevisiae (Saccharomyces cerevisiae).Yeast cell of the present invention prepares by cultivating a plurality of yeast cell under the activation condition that exists one group of electromagnetic field, thereby makes some metabolic function of yeast cell become very effective.Therefore, the invention still further relates to the manufacture method of this treatment compositions, be included in culturing yeast cell under the activation condition, mix the various yeast cell cultures of the present invention, the subsequent drying yeast cell is also packed the finished product.
Particularly, the present invention includes a kind of method of handling solid waste, comprise and in solid waste, add a plurality of yeast cell, and to make microbiotic in the yeast cell degradable solid refuse, wherein said yeast cell be to prepare by culturing yeast cell in the electromagnetic field that has specified range frequency and a field intensity at or a group.Can be included but not limited to penicillin, duomycin, terramycin, doxycycline, tsiklomitsin, Streptomycin sulphate, kantlex, erythromycin, Spiramycin Base, bacitracin, Polymyxin E, paraxin, cefoxitin, Xin Meisu and Vulkamycin. PA-93 by the microbiotic that yeast cell of the present invention is degraded.
The present invention also comprises a kind of method of handling solid waste, comprise and in solid waste, add a plurality of yeast cell, make the adverse chemical product in the yeast cell degradable solid refuse, wherein said yeast cell is to prepare by culturing yeast cell in the electromagnetic field that has specified range frequency and a field intensity at or a group.Can be included but not limited to toluene, ethylbenzene, Trichlorophenol, dimethylbenzene, phenyl aldehyde, propionic aldehyde, enanthaldehyde, dichlorobenzene, methyl phenyl ketone, arsanilic acid, roxarsone, Nifurazolidone, M B 15497, Trichlorphon, dinitolmide, SD-1750, monocrotophos, Rogor, dichlorodiphenyl trichloroethane (DDT) and toxaphene by the adverse chemical product that yeast cell of the present invention is degraded.The adverse chemical product also comprise organic and inorganic salts, for example ammonium compound, nitrate or nitrite and phosphoric acid salt.
The present invention further comprises the method that reduces the solid waste smell, comprise and in solid waste, add a plurality of yeast cell, make yeast cell reduce the amount of scent of molecule in the solid waste, wherein said yeast cell is to prepare by culturing yeast cell in the electromagnetic field that has specified range frequency and a field intensity at or a group.Molecule odorous includes but not limited to hydrogen sulfide, ammonia, indoles, skatole, acetate, methylamine and p-cresol.
The present invention further comprises the method that suppresses malignant bacteria growth in the solid waste, comprise and in solid waste, add a plurality of yeast cell, make yeast cell suppress the growth of malignant bacteria in the solid waste, wherein said yeast cell is to prepare by culturing yeast cell in the electromagnetic field that has specified range frequency and a field intensity at or a group.Malignant bacteria is selected from following: streptococcus aureus, pneumococcus, anthrax bacillus, Mycobacterium tuberculosis, Salmonellas, intestinal bacteria, vibrios, shigella, Clostridium botulinum and Clostridium perfringens.
Method of the present invention can use yeast cell to carry out by uniting in solid waste disposal.Biological composition of the present invention is added in the solid waste, and described biological composition comprises a kind of in the following at least yeast cell component: (a) first yeast cell component comprises antibiotic yeast cell in a plurality of degradable solid refuses; (b) second yeast cell component comprises the yeast cell of adverse chemical product in a plurality of degradable solid refuses; (c) the 3rd yeast cell component comprises the yeast cell of the amount of scent of molecule in a plurality of minimizing solid waste; (d) the 4th yeast cell component comprises the yeast cell that malignant bacteria grows in a plurality of inhibition solid waste.Handling the required time can determine by rule of thumb by the level of microbiotic, adverse chemical product, malignant bacteria and scent of molecule in the monitoring solid waste, can change in the scope in several hrs, several days and two or more weeks.
The present invention further is included in processing, storage, processing, utilizes or remove and use biological composition of the present invention in the solid waste again.
4. accompanying drawing summary
Fig. 1: the activation of yeast cell.1 yeast cell culture; 2 containers; 3 electromagnetism field sources.
Fig. 2: yeast cell adapted soil kind.4 input electrodes; 5 containers; 6 electrodes; 7 yeast cell cultures; 8 electromagnetism field sources; 9 temperature regulators.
5. detailed description of the Invention
The invention provides the biological composition that comprises yeast cell.The present invention also provides method for preparing biological composition and the method for using biological composition.
Biological composition of the present invention is used for solid waste disposal, makes usually with its storage, transportation, processing, utilizes and/or remove the health risk that is associated again and to the influence minimizing of environment.Use these compositions can reduce the total cost that solid waste is handled on community, enterprise or farm, and make the utilization again of some kind solid waste become feasible.Solid waste disposal used herein refers to the process of the physics, chemistry or the biology characteristics that change solid waste, compare with untreated solid waste, make it store, transport, utilize again, so not offensive or the threat of health or environment reduced when operating.Handle the harm that makes refuse usually and reduce, or make transportation, storage, the operation of solid waste or utilize safer again.
According to the present invention, biological composition comprises a plurality of yeast cell components.Each yeast cell component is that a group comprises and can carry out a plurality of yeast cell that one or more belong to following kind required function: (1) suppresses the growth of pathogenic agent, (2) degraded adverse chemical product, or (3) reduce organic smell.
In one embodiment, a biological composition of the present invention comprises at least one and can carry out a kind of yeast cell component in three kinds of functions.In preferred embodiments, biological composition of the present invention comprises can provide the yeast cell of whole three kinds of functions component.Therefore, preferred Biofertiliser composition comprises at least three kinds of different strains of Yeast components.The difference of yeast cell component substitutes prescription also among considering.
The material that term used herein " solid waste " general reference any kind of has been dropped because realized its purpose or useless byproduct comprises human and animal's excretory physiology refuse.The source of solid waste comprises inhabitation, commerce, agricultural and industrial activity.Refuse or rubbish that non-industry and non-agricultural solid waste are for example collected from the urban district, the material that uses when containing the food that abandons or making food, and other multiple dry substance such as paper, textiles or plastics.Particularly in residential area that restaurant and restaurant are arranged and commercial zone, the main kind of solid waste is called " rubbish " here, mainly contains decomposable food waste.Support that pathogenic organisms is grown, the malodorous rubbish that becomes owing to rotting can effectively be handled by biological composition of the present invention.The characteristics that are fit to the solid waste kind of biological composition processing of the present invention are organic content height.
The another kind of solid waste that can be handled by the present composition is mud.Term used herein " mud " extensively comprises any solid matter that precipitates from suspension in sewage storage and/or treating processes, such as but not limited to the residue or the sewage enriched material of the residue in the effluent sewage pond, municipal treatment plant.Term " mud " also comprises semi-solid thing, and sewage and sedimentary mixture.Therefore, this term comprise have extensive viscosity, the mud of density and water-content scope, and by partially disposed or stable mud.According to the source, mud can comprise multiple adverse chemical product, can not caused disadvantageous effect if do not deal carefully with to environment.Mud foul smelling, and the growth of support pathogenic organisms.
Biological composition of the present invention can also be handled the refuse that rural activity produces.Typically, animal produces refuse in following activity, such as but not limited to farm, farmland, slaughterhouse and market.The continuous generation of a large amount of animal excrements and gather and formed malodorous environment, and the health to people and domestic animal produces threat because there being pathogenic microorganism.Agricultural waste also can contain the adverse chemical product, for example antibiotic feed additive, chemical fertilizer, agricultural chemicals and weedicide, if refuse is not dealt carefully with, they can contaminate environment.
Term used herein " animal muck " extensively comprises the ight soil that contains the feedlot animal and the organic materials of urine, with or do not accompany rubbish such as for example straw that is used for traditionally fertilizing the land, hay, pad grass.Brid guano fertilizer includes but not limited to the muck that the birds tamed such as chicken, duck, turkey, goose, quail, young dove, ostrich etc. produce.Brid guano fertilizer comprises movement or the birds droppings that unacclimated birds produce.Cattle manure used herein comprises the chewing mammalian of domestication such as the refuse of cow or beef cattle generation.Term used herein " cattle manure " is not limited only to ox, also comprises other herbivore, mainly raises for their milk, meat, skin, hair and fur.Cattle manure includes but not limited to the muck that buffalo, wild ox, yak, horse, donkey, mule, sheep, goat, camel etc. produce.Cattle manure comprises the movement that is produced by unacclimated livestock.Term used herein " pig manure " includes but not limited to the muck that wild boar, dog, tame pig etc. produce.Other agricultural waste comprise the refuse that crop residues, bagasse, fruits and vegetables packaging facilities produce, the refuse that the animal product packaging facilities produces, and comprise the corpse behind the animal slaughtering.
In various embodiments, biological composition of the present invention is effective especially aspect treating refuse, mud and muck.
Under the multiple situation, Municipal waste temporarily is stored in the refuse transhipment station.At transhipment station, refuse is collected line and is unloaded down from the locality, store according to kind sometimes.Then refuse is contained in and is transported to the Municipal waste processing on bigger truck or the railcar or removes the place.Usually, according to the source of solid waste, with sort operation glass, metal, inorganic with other or the nondegradable material of wood are separated from the refuse of high organic content by selection.These operations can be carried out with the known method of the industry of utilization/refuse cleaning again, for example utilize the physical characteristics of solid waste to operate as the mechanicalness of size and density.Pulverizing or grinding the size that can reduce refuse becomes particulate, and easier being operated of gained material of uniform size for example mixed and transported.Because the difference that muck, mud or rubbish are formed is analyzed a collection of waste materials sampling, detect that the amount and the kind of pathogenic organisms and adverse chemical product may be and meaning in this batch.
Though following term is considered to have in this area clear and definite implication, following still they being illustrated is beneficial to explain the present invention.
The phrase used herein growth of pathogenic agent " suppress " refers to owing to have yeast cell of the present invention in the solid waste sample, through making the pathogenic microorganism reduced number in the sample after a while or not increasing.Need be appreciated that there are not these yeast cell, the number of pathogenic agent can increase by nature in the sample.Many these microorganisms cause the disease of humans and animals, can comprise bacterium, for example, and the kind of Escherichia, salmonella, Shigella, Mycobacterium, Staphylococcus, bacillus, streptococcus and Diplococcus.
Phrase used herein " degraded adverse chemical product " instigates the environmental toxin in bad compound such as the solid waste to be converted into the biology or the biological process of inactive form, for example is the compound than small molecular weight with these compound decomposition.Antibiosis usually is present in the muck, do not wish that these compounds occur in the fertilizer of being made by muck, because the potentially dangerous of being taken in by the people is arranged, for example contain the vegetables that pollute organic fertilizer growth by edible the use, and may the propagating of antibiotics resistance in the environment.A lot of microbiotic can be added in the animal-feed to protect various domestic animals, and for example chicken, turkey and pig avoid bacterium and parasitic disease, and promote growth.A large amount of antibiotic feed additives are drained by animal, thereby gather in muck and mud.Many kinds of microbiotic use in animal surgery, such as but not limited to aminoglycoside, tetracyclines, β-Nei acyl Ammonia, glycopeptide class and Macrolide.The microbiotic example that approval is used on U.S. farm comprises, but be not limited to bacitracin, methylene-bis salicylate, Zinc-bacitracin, Moenomycin. Flavophospholopol, terramycin, duomycin, penicillin, Tylosin/sulphamethazine, roxarsone, nitrasone, monensin, X-537A, Carbadox, tiamulin, hygromycin B, nystatin, Vulkamycin. PA-93, sulfadimethoxine, ormetoprim, lincomycin, fenbendazole and virginiamycin.These antibiotic existence and amount can detect by any means known in the art in the composition, for example high performance liquid chromatography (HPLC).
The process that phrase used herein " reduces organic smell " instigates one or more scent of compound concentrations in the solid waste organism to reduce.The scent of compound, such as but not limited to hydrogen sulfide, ammonia, indoles, skatole (being the 3-Methyl-1H-indole), p-cresol and organic acid, known all is the factor of facilitating of solid waste stench character.These malodorous compounds can detect by any method well known in the art with concentration in the air sample that contacts muck at poultry manure for example, include but not limited to vapor-phase chromatography or mass spectroscopy.Smell is biological taste by the olfactory organ perception.The minimizing of the odor intensity relevant with solid waste can subjectively detect.Known several different methods and technology can be measured the intensity of smell.A kind of method of subjective measurement odor intensity is that measurement smell concerning human or animal trier can't be discovered or uncertain needed extent of dilution.In addition, can also use recognition threshold, be the higher concentration that the smell characteristics can be identified.The method of any objective or subjective detection odor intensity or technology can be used for monitoring the effect of the present composition and method.
The present inventor finds that under certain culture condition, yeast can be activated, and it is very effective that some metabolic function becomes, and makes the activatory yeast have the ability of inhibition pathogenic growth, degraded adverse chemical product or minimizing organism smell.
According to the present invention, the yeast cell component of Biofertiliser composition prepares by a plurality of yeast cell are cultivated for some time in the suitable culture base in the presence of one or continuous a plurality of alternating electromagnetic field.Culturing process makes yeast spore germination, yeast cell growth and division, can carry out in batches or carry out continuously.Term used herein " alternating electromagnetic field ", " electromagnetic field " or " EM field " are synonyms.The used electromagnetic field of the present invention can produce by several different methods well known in the art.Fig. 1 has described the sketch of exemplary device respectively.Electromagnetic wave source (3) produces the electromagnetic field of required frequency and field intensity, and this electromagnetic wave source comprises one or more signal generators that can generate electromagnetic waves, and preferred sinusoidal wave, the optimized frequency scope is 30MHz-3000MHz.Such signal generator is known in the art.Also can use the signal generator that can produce narrower range of frequency signal.If desired, can also use signal amplifier to export, thereby increase the intensity of electromagnetic field to increase signal.
Electromagnetic field can act on culture in several ways, comprises yeast cell is placed near the signal projector that links to each other with electromagnetic wave source.In one embodiment, apply electromagnetic field by the signal projector that is immersed in the electrode form in the yeast cell culture (1).In preferred embodiments, an electrode is a metal sheet, and another electrode comprises that one group is installed on the inner metal wire of container (2), can be uniformly distributed in the culture electromagnetic field energy.The number of electrode used therein line depends on the volume of culture and the diameter of electrode wires.For example, the culture of 5000ml volume, every 100ml culture can use the electrode wires of a diameter between 0.1 to 1.2mm; For the culture of volume above 1000L, every 1000L culture can use the electrode wires of a diameter between 3 to 30mm.See Fig. 1.
Though be not subjected to the constraint of any theory or mechanism, the present inventor thinks that these culture condition can activate or strengthen one or one group of expression of gene of yeast cell, thereby make some Metabolic activity of this yeast cell become more effective, produce corresponding required result.
In various embodiments, yeast belong (Saccharomyces), Schizosaccharomyces (Schizosaccharomyces), Sporobolomyces (Sporobolomyces), torulopsis (Torulopsis), Trichosporon (Trichosporon), dimension Ke Shi yeast belong (Wickerhamia), ashbya, Blastomyces (Blastomyces), mycocandida (Candida), Citeromycesbaodingensis belongs to (Citeromyces), Crebrothecium, Cryptococcus (Cryptococcus), Debaryomyces (Debaryomyces), Endomycopsis (Endomycopsis), Geotrichum (Geotrichum), Hansenula (Hansenula), Kloeckera (Kloeckera), saccharomyces oleaginosus belongs to (Lipomyces), Pichia (Pichia), the yeast of Rhodosporidium (Rhodosporidium) and Rhodotorula (Rhodotorula) can be used for the present invention.
Non-limiting examples of yeast include Saccharomyces cerevisiae strains (Saccharomyces cerevisiae Hansen) ACCC2034, ACCC2035, ACCC2036, ACCC2037, ACCC2038, ACCC2039, ACCC2040, ACCC2041, ACCC2042, AS2.1, AS2.4, AS2.11, AS2.14, AS2.16, AS2.56, AS2.69, AS2.70, AS2.93, AS2.98, AS2.101, AS2.109, AS2.110, AS2.112, AS2.139, AS2.173, AS2.174, AS2.182, AS2.196, AS2.242, AS2.336, AS2.346, AS2.369, AS2.374, AS2.375, AS2.379, AS2.380, AS2.382, AS2.390, AS2.393, AS2.395, AS2.396, AS2.397, AS2.398, AS2.399, AS2.400, AS2.406, AS2.408, AS2.409, AS2.413, AS2.414, AS2.415, AS2.416, AS2.422, AS2.423, AS2.430, AS2.431, AS2.432, AS2.451, AS2.452, AS2.453, AS2.458, AS2.460, AS2.463, AS2.467, AS2.486, AS2.501, AS2.502, AS2.503, AS2.504, AS2.516, AS2.535, AS2.536, AS2.558, AS2.560, AS2.561, AS2.562, AS2.576, AS2.593, AS2.594, AS2.614, AS2.620, AS2.628, AS2.631, AS2.666, AS2.982, AS2.1190, AS2.1364, AS2.1396, IFFI1001, IFFI1002, IFFI1005, IFFI1006, IFFI1008, IFFI1009, IFFI1010, IFFI1012, IFFI1021, IFFI1027, IFFI1037, IFFI1042, IFFI1043, IFFI1045, IFFI1048, IFFI1049, IFFI1050, IFFI1052, IFFI1059, IFFI1060, IFFI1063, IFFI1202, IFFI1203, IFFI1206, IFFI1209, IFFI1210, IFFI1211, IFFI1212, IFFI1213, IFFI1215, IFFI1220, IFFI1221, IFFI1224, IFFI1247, IFFI1248, IFFI1251, IFFI1270, IFFI1277, IFFI1287, IFFI1289, IFFI1290, IFFI1291, IFFI1292, IFFI1293, IFFI1297, IFFI1300, IFFI1301, IFFI1302, IFFI1307, IFFI1308, IFFI1309, IFFI1310, IFFI1311, IFFI1331, IFFI1335, IFFI1336, IFFI1337, IFFI1338, IFFI1339, IFFI1340, IFFI1345, IFFI1348, IFFI1396, IFFI1397, IFFI1399, IFFI1411, IFFI1413; Saccharomyces cerevisiae ellipse (Sacharomyces cerevisiae Hansen Var. ellipsoideus (Hansen) Dekker) ACCC2043, AS2.2, AS2.3, AS2.8, AS2.53, AS2.163, AS2.168, AS2.483, AS2.541, AS2.559, AS2.606, AS2.607, AS2.611, AS2.612; Xue watts yeast (Saccharomyces chevalieri Guillermond) AS2.131, AS2.213; Deshi yeast (Saccharomyces delbrueckii) AS2.285; Saccharomyces delbrueckii Lindner var.mongolicus Lodder et van Rij AS2.209, AS2.1157; less spore yeast (Saccharomyces exiguus Hansen) AS2.349, AS2.1158; fermenting yeast (Saccharomyces fermentati (Saito) Lodder et van Rij) AS2.286, AS2.343; Saccharomyces logos van laer et Denamur ex Jorgensen AS2.156, AS2.327, AS2.335; honey and yeast (Saccharomyces mellis Lodder et Kreger Van Rij) AS2.195; Saccharomyces microellipsoides Osterwalder AS2.699; ovoid yeast (Saccharomyces oviformis Osterwalder) AS2.100; Roche yeast (Saccharomyces rosei (Guilliermond) Lodder et kreger van Rij) AS2.287; Lu's yeast (Saccharomyces rouxii Boutroux) AS2.178, AS2.180, AS2.370, AS2.371; sake yeast (Saccharomyces sake Yabe) ACCC2045; A Baoli yeast (Candida arborea) AS2.566; Candida Krusei (Castellani) Berkhout AS2.1045; rum beer Candida (Candida lambica (Lindner et Genoud) van.Uden et Buckley) AS2.1182; Candida lipolytica Yeast (Candida lipolytica (Harrison) Diddens et Lodder) AS2.1207, AS2.1216, AS2.1220, AS2.1379, AS2.1398, AS2.1399, AS2.1400; Nearly smooth Candida (Candida parapsilosis (Ashford) Langeron et Talice) AS2.590; nearly smooth Candida (Candida parapsilosis (Ashford) et Talice Var.intermedia Van Rij et Verona) AS2.491; Maggi Candida (Candida pulcherriman (Lindner) Windisch) AS2.492; Candida rugousa (Anderson) Diddens et Loddeer AS2.511, AS2.1367, AS2.1369, AS2.1372, AS2.1373, AS2.1377, AS2.1378, AS2.1384; Candida tropicalis (Candida tropicalis (Castellani) Berkout) ACCC2004, ACCC2005, ACCC2006, AS2.164, AS2.402, AS2.564, AS2.565, AS2.567, AS2.568, AS2.617, AS2.1387; produce abdominal Candida (Candida utilis Henneberg Lodder et Kreger Van Rij) AS2.120, AS2.281, AS2.1180; Crebrothecium ashbyii (Guillermond) Routein AS2.481, AS2.482, AS2.1197; Geotrichum (Geotrichum candidum Link) ACCC2016, AS2.361, AS2.498, AS2.616, AS2.1035, AS2.1062, AS2.1080, AS2.1132, AS2.1175, AS2.1183; Hansenula anomala (Hansenula anomala (Hansen) Het P sydow) ACCC2018, AS2.294, AS2.295, AS2.296, AS2.297, AS2.298, AS2.299, AS2.300, AS2.302, AS2.338, AS2.339, AS2.340, AS2.341, AS2.470, AS2.592, AS2.641, AS2.642, AS2.635, AS2.782, AS2.794; Hansenula arabitolgens Fang AS2.887; Hansenula jadinii Wickerham ACCC2019; Saturn Hansenula (Hansenula saturnus (Klocker) H et P sydow) ACCC2020; Hansenula schneggii (Weber) Deker AS2.304; Hansenula subpelliculosa Bedford AS2.738, AS2.740, AS2.760, AS2.761, AS2.770, AS2.783, AS2.790, AS2.798, AS2.866; lemon-shaped Klerk yeast (Kloeckera apiculata (Reess emend.Klocker) Janke) ACCC2021, ACCC2022, ACCC2023, AS2.197, AS2.496, AS2.711, AS2.714; StarTech's fat yeast (Lipomyces starkeyi Lodder et van Rij) ACCC2024, AS2.1390; powdered Pichia (Pichia farinose (Lindner) Hansen) ACCC2025, ACCC2026, AS2.86, AS2.87, AS2.705, AS2.803; Film Bu Pichia (Pichia membranaefaciens Hansen) ACCC2027, AS2.89, AS2.661, AS2.1039; Rhodosporidium toruloides Banno ACCC2028; Rhodotorula (Rhodotorula glutinis (Fresenius) Harrison) ACCC2029, AS2.280, ACCC2030, AS2.102, AS2.107, AS2.278, AS2.499, AS2.694, AS2.703, AS2.704, AS2.1146; red yeast (Rhodotorula minuta (Saito) Harrison) AS2.277; dark red yeast (Rhodotorula rubar (Demme) Lodder) ACCC2031, AS2.21, AS2.22, AS2.103, AS2.105, AS2.108, AS2.140, AS2.166, AS2.167, AS2.272, AS2.279, AS2.282; Karnofsky yeast (Saccharomyces carlsbergensis Hansen) ACCC2032, ACCC2033, AS2.113, AS2.116, AS2.118, AS2.121, AS2.132, AS2.162, AS2.189, AS2.200, AS2.216, AS2.265, AS2.377, AS2.417, AS2.420, AS2.440, AS2.441, AS2.443, AS2.444, AS2.459, AS2.595, AS2.605, AS2.638, AS2.742, AS2.745, AS2.748, AS2.1042; grape yeast (Saccharomyces uvarum Beijer) IFFI1023, 1FFI1032, IFFI1036, IFFI1044, IFFI1072, IFFI1205, IFFI1207; Westergren yeast (Saccharomyces willianus Saccardo) AS2.5, AS2.7, AS2.119, AS2.152, AS2.293, AS2.381, AS2.392, AS2.434, AS2.614, AS2.1189; yeast (Saccharomyce sp.) AS2.311; Lu's yeast (Saccharomyces ludwigii Hansen) ACCC2044, AS2.243, AS2.508; Saccharomyces sinenses Yue AS2.1395; eight fission yeast spores (Schizosaccharomyces octosporus Beijerinck) ACCC2046, AS2.1148; S. pombe (Schizosaccharomyces pombe Linder) ACCC2047, ACCC2048, AS2.248, AS2.249, AS2.255, AS2.257, AS2.259, AS2.260, AS2.274, AS2.994, AS2.1043, AS2.1149, AS2.1178, IFFI1056; Rose Rose-colored throw spore yeast (Sporobolomyces roseus Kluyver et van Niel) ACCC2049, ACCC2050, AS2.619, AS2.962, AS2.1036, ACCC2051, AS2.261, AS2.262; white Torulopsis (Torulopsis candida (Saito) Lodder) ACCC2052, AS2.270; Torulopsis famta (Harrison) Lodder et van Rij ACCC2053, AS2.685; Torulopsis globosa (Olson et Hammer) Lodder et van Rij ACCC2054, AS2.202; Torulopsis inconspicua Lodder et van Rij AS2.75; Trichosporon behrendii Loddcr et Kreger van Rij ACCC2055, AS2.1193; heads Trichosporon yeast (Trichosporon capitatum Diddens et Lodder) ACCC2056, AS2.1385; Trichosporon cutaneum (de Beurm et al.) Ota ACCC2057, AS2.25, AS2.570, AS2.571, AS2.1374; Vickers yeast (Wickerhamia fluoresens (Soneda) Soneda) ACCC2058, AS2.1388. ...
Known some yeast kind that can be activated or induce and be included in according to the present invention in the present invention has pathogenic to people and/or other live organism.Ashbya gossypii for example, Blastomyces dermatitidis (Blastomyces dermatitidis), Candida albicans (Candida albicans), Candida parakrusei, candida tropicalis (Candida tropicalis), Ma Telitan Citeromycesbaodingensis (Citeromyces matritensis), Crebrothecium ashbyii, Lauren cryptococcus (Cryptococcus laurentii), novel Cryptococcus (Cryptococcua neoformans), the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii), Debaryomyces kloeckeri, Dbaly yeast bacterium (Debaryomyces sp.) and Endomycopsis Fibnligera (Endomycopsisfibuligera).In some cases, preferably do not use such disease yeasts in the biological composition of the present invention, for example, if this use is in open place, the health of the entail dangers to mankind and/or live organism.
The yeast that common preferred yeast belongs to.Hansen yeast saccharomyces cerevisiae in the bacterial strain of yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) (Saccharomyces cerevisiae Hansen) is a preferred strain.Most preferred yeast strain is to be deposited in Chinese common micro-organisms DSMZ (CGMCC), Wine brewing yeast strain with following preserving number: AS2.504, AS2.558, AS2.413, AS2.397, AS2.69, AS2.109, AS2.607, AS2.516, AS2.561, AS2.422, AS2.393, AS2.631, AS2.982, AS2.560, AS2.467, AS2.415, AS2.375, AS2.628, AS2.1190, AS2.562, AS2.463, AS2.409, AS2.379, AS2.666, AS2.631, AS2.182, AS2.431, AS2.606, AS2.53, AS2.611, AS2.414, AS2.576, AS2.483, IFFI1211,1FF11293, IFFI1308, IFFI1210, IFFI1213, IFFI1307, IFFI1206, IFFI1052, IFFI1301, IFFI1291, IFFI1202, IFFI1021, IFFI1059, IFFI1052, IFFI1441, IFFI1008, IFFI1220, IFFI1302 and IFFI1023.
Usually, the yeast strain that the present invention uses can obtain from private or public laboratory culture thing, perhaps obtain from public DSMZ, and as American type culture collection, Manassas, No. 10801, the big ways for education, Virginia 20110-2209; And Institute of Microorganism, Academia Sinica China microbial preservation management committee China common micro-organisms DSMZ (CGMCC), Haidian District, BeiJing, China 2714 mailbox, postcode 100080.
Though preferably use pure yeast strain to begin to prepare yeast cell component of the present invention, be not limited to this.Every primary yeast cellular component can obtain by the yeast cell mixed culture of not of the same race or bacterial strain.The composition of yeast cell component can be determined with standard yeast authenticate technology well known in the art.
The ability and the efficient of activation yeast performance required function can easily detect by method known in the art before and after cultivating under condition of the present invention.For example, HPLC or mass spectroscopy can be used for detecting and analyze multiple organic molecule in the solid waste sample.Micro-biological process well known in the art can be used for detecting and the counting sample in live microorganism number and microbial count.
When handling the high relatively organic manure of bacterial count, biological composition can be prepared into the yeast cell that mainly contains bacteria growing inhibiting.When containing the solid waste of adverse chemical product, biological composition can be prepared into the yeast cell that mainly contains degraded adverse chemical product with the biological composition processing.Therefore, this biological composition can be used for the multiple situation that city, commerce, agricultural and industrial mechanism facility are run into.The present invention can also be for household use, especially in the Rural areas.
Biological composition of the present invention can directly be applied to solid waste.Known to various equivalent modifications, several different methods and equipment can be used for these yeast are mixed with solid waste.In one embodiment, zymic nutrient solution of the present invention is introduced directly in the solid waste that needs to handle.In another embodiment, zymic dry powder of the present invention mixes with solid waste, and then adds entry.This biological composition can by dispenser, atomizer and other mechanical method be used and mix with solid waste, and these methods can be automatizations.The amount of used biological composition depend in part at that time situation and the kind of solid waste, can also determine by rule of thumb.But in order to obtain effective processing, ideal situation is that every cubic metre of solid waste uses about 300 biological compositions to 600g dry weight (humidity is less than 10%).At first with yeast cell and water according to the about 30 liters mixed of every 1000g yeast (dry weight), before being applied to solid waste, cultivated 12 to 24 hours then.The effect of handling for example reduces smell or bacterial count, produces after using 24 to 72 hours.Though be not necessary, biological composition of the present invention can also unite with reodorant, sterilizing agent and the toxinicide of other kind or be used alternatingly.
5.1-5.4 joint is described respectively be used for degrading microbiotic, suppress the yeast cell component of pathogenic agent, degraded adverse chemical product and minimizing smell.The preparation method of every primary yeast cellular component has been described.5.6 joint has been described the preparation of biological composition of the present invention.In the various embodiments of the present invention, used the standard technique of yeast operation, transfer and storage.When carrying out preparation process of the present invention,, favourable though aseptic condition and clean environment are not necessarily.
The antibiotic yeast cell 5.1 degrade
The invention provides common antibiotic yeast cell in to degrade muck and the mud.
According to the present invention, the yeast cell antibiotic ability of degrading is by yeast cell is activated or strengthens cultivating in the suitable culture base in the presence of the electromagnetic field.The gained yeast cell can be used as a component of biosolids waste treatment composition of the present invention and uses.
The electromagnetic field frequency of activation or reinforcement yeast degradation microbiotic ability is usually in 70MHz arrives the scope of 600MHz.After the yeast cell growth sufficiently long time, can detect yeast cell enhanced one or more antibiotic abilities of degrading by method well known in the art.Can be included but not limited to the molecule of acyl Ammonia in the β, tetracyclines, polypeptide class, glycopeptide class, aminoglycoside and Macrolide family by the microbiotic of yeast degradation of the present invention.
It is to carry out in the liquid medium within that the present invention prepares degraded microbiotic yeast method.Contain the absorbable nutrition of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually between substratum weight about 0.1% to 5% between, preferably between about 0.5% to 2%, most preferably from about 0.8%.These carbon sources can separately or be united use in substratum.
Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has (NH 4) 2HPO 4, CaCO 3, MgSO 4, NaCl and CaSO 4
Table 1: the antibiotic zymic substratum of degrading is formed
Substratum is formed Content
Muck or mud 8.0g, dry weight,＞120 orders
????NaCl ????0.2g
????MgSO 4·7H 2O ????0.2g
????CaCO 3·5H 2O ????0.5g
????CaSO 4·2H 2O ????0.2g
Peptone ????1.5g
????K 2HPO 4 ????0.5g
Contain antibiotic extract (〉=100 μ g/ml) ????600ml
Autoclaving water ????400ml
Contain antibiotic extract by the fresh refuse of 500g such as muck, mud being scattered in about 600ml warm water (35-40 ℃) and cultivating 24 hours, filter liquide at 30-37 ℃ and remove particulate matter and prepare.Only contain certain micro-microbiotic as berry extract, can in extract, add an amount of this kind microbiotic.
It should be noted that it is not restrictive that the substratum that provides in the table 1 is formed.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.
Culturing process can be at first with cell density 10 2-10 5The yeast strain inoculum that cell/ml selects 1ml is inoculated in the 100ml substratum, and preferred inoculum density is 3 * 10 2-10 4Cell/ml.This process can increase and decrease as required in proportion.Yeast culture can be grown in the presence of an electromagnetism (EM) field or one group of electromagnetic field.If apply one group of electromagnetic field, when when an electromagnetic field switches to another electromagnetic field, yeast cell can be in same container, use same set of electromagnetic wave generator and projector.
Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives the 100.000MHz scope 70.000, and preferred 410.000 to 620.000MHz.The field intensity of electromagnetic field arrives the 250mV/cm scope 40.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or different frequency and different field intensity in the above-mentioned scope.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7,8,9 or 10 different electromagnetic field.
Though yeast cell is in the presence of electromagnetic field even cultivate several hrs and just can be activated, but yeast cell can continue to cultivate for some time (for example 2 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 144-384 hour.
For example, use exemplary device shown in Figure 1, electromagnetic output amplitude is in the 8.5-85mV/cm scope, about usually 50mV/cm.Period 1 further cultivates yeast cell another cycle under the essentially identical condition that increases to higher level in the 150-250mV scope (about usually 200mV) except amplitude after cultivating.Method of the present invention is carried out in about 25 to 30 ℃ of scopes; But, preferably carry out at 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably 3Condition under carry out preferred 0.4mol/m 3Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.
When culturing process finished, yeast cell can reclaim from culture by several different methods known in the art, and stored being lower than under about 0 ℃ to 4 ℃ temperature.The yeast cell that reclaims can store with form of powder in the drying processing.
In order to detect the activity of activatory yeast cell, can measure under different time points and the different culture condition amount of Antibiotique composition in the sample with method well known in the art such as HPLC to Antibiotique composition.For example, preparation contains the microbiotic sample (every liter of 100mg) of concentration known.Then, with 0.1ml activatory and non-activated yeast (at least 10 7Cell/ml) add 100 liters to contain in the antibiotic sample was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, by measuring sample with HPLC, residual antibiotic amount in detection and the comparison extract.
This method is applicable to the microbiotic of numerous species usually.In specific embodiment, described at the antibiotic best approach of particular types.
The yeast cell component of decomposing penicillin
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of penicillin such as penicillin G and cloxacillin.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 100.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,98 and 100MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.399 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 77MHz, 48mV/cm 15 hours; 83MHz, 48mV/cm 15 hours; 90MHz, 48mV/cm 15 hours; 96MHz, 48mV/cm 15 hours; 77MHz, 200mV/cm 30 hours; 83MHz, 200mV/cm30 hour; 90MHz, 200mV/cm 30 hours; 96MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the penicillin that the activatory yeast cell can degrade to the activity of penicillin.Prepare two 100 liters sample, each contains penicillin concn is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, by detect with HPLC and comparative sample in residual antibiotic amount.Compare with containing not the sample of activated yeast cell, the amount that contains penicillin in the sample of activated yeast cell has reduced more than 56.5%.
The yeast cell component of decomposing duomycin
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of duomycin such as duomycin and Isphamycin.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.748 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 70MHz, 48mV/cm 15 hours; 73MHz, 48mV/cm 15 hours; 88MHz, 48mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 73MHz, 200mV/cm30 hour; 88MHz, 200mV/cm 30 hours; 98MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the duomycin that the activatory yeast cell can degrade to the activity of duomycin.Prepare two 100 liters sample, each concentration that contains duomycin is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains duomycin in the sample of activated yeast cell has reduced more than 62.3%.
The yeast cell component of decomposing terramycin
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of terramycin example hydrochloric acid terramycin.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.101 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 70MHz, 48mV/cm 15 hours; 74MHz, 48mV/cm15 hour; 88MHz, 44mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 74MHz, 200mV/cm 30 hours; 88MHz, 200mV/cm 30 hours; 98MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the terramycin that the activatory yeast cell can degrade to the activity of terramycin.Prepare two 100 liters sample, each concentration that contains terramycin is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains terramycin in the sample of activated yeast cell has reduced more than 65.5%.
The yeast cell component of decomposing doxycycline
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of doxycycline.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.417 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 71MHz, 48mV/cm 15 hours; 73MHz, 48mV/cm 15 hours; 77MHz, 48mV/cm 15 hours; 88MHz, 48mV/cm 15 hours; 71MHz, 200mV/cm 30 hours; 73MHz, 200mV/cm 30 hours; 77MHz, 200mV/cm30 hour; 88MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the doxycycline that the activatory yeast cell can degrade to the activity of doxycycline.Prepare two 100 liters sample, each contains doxycycline concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains doxycycline in the sample of activated yeast cell has reduced more than 54.9%.
The yeast cell component of decomposing tsiklomitsin
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of tsiklomitsin.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.70 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 70MHz, 48mV/cm 15 hours; 75MHz, 48mV/cm 15 hours; 82MHz, 48mV/cm 15 hours; 85MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 75MHz, 200mV/cm 30 hours; 82MHz, 200mV/cm30 hour; 85MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the tsiklomitsin that the activatory yeast cell can degrade to the activity of tsiklomitsin.Prepare two 100 liters sample, each concentration that contains tsiklomitsin is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains tsiklomitsin in the sample of activated yeast cell has reduced more than 67.6%.
The yeast cell component of decomposing Streptomycin sulphate
In a specific embodiments, provide a kind of yeast cell preparation method who decomposes Streptomycin sulphate.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.441 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 70MHz, 48mV/cm 15 hours; 73MHz, 48mV/cm 15 hours; 80MHz, 48mV/cm 15 hours; 96MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 73MHz, 200mV/cm 30 hours; 80MHz, 200mV/cm30 hour; 96MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the Streptomycin sulphate that the activatory yeast cell can degrade to the activity of Streptomycin sulphate.Prepare two 100 liters sample, each concentration that contains Streptomycin sulphate is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Streptomycin sulphate in the sample of activated yeast cell has reduced more than 77.8%.
The yeast cell component of decomposing kantlex
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of kantlex.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.336 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 71MHz, 48mV/cm 15 hours; 78MHz, 48mV/cm 15 hours; 86MHz, 48mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 71MHz, 200mV/cm 30 hours; 78MHz, 200mV/cm 30 hours; 86MHz, 200mV/cm30 hour; 98MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the kantlex that the activatory yeast cell can degrade to the activity of kantlex.Prepare two 100 liters sample, each contains kantlex concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains kantlex in the sample of activated yeast cell has reduced more than 68.7%.
The yeast cell component of decomposing erythromycin
In a specific embodiments, provide a kind of yeast cell preparation method who decomposes erythromycin.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.422 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 73MHz, 48mV/cm 15 hours; 79MHz, 48mV/cm 15 hours; 88MHz, 48mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 73MHz, 200mV/cm 30 hours; 79MHz, 200mV/cm 30 hours; 88MHz, 200mV/cm30 hour; 98MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the erythromycin that the activatory yeast cell can degrade to the activity of erythromycin.Prepare two 100 liters sample, each contains erythromycin concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains erythromycin in the sample of activated yeast cell has reduced more than 72.7%.
The yeast cell component of decomposing Spiramycin Base
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of Spiramycin Base.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.620 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 70MHz, 48mV/cm 15 hours; 77MHz, 48mV/cm 15 hours; 84MHz, 48mV/cm 15 hours; 93MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 77MHz, 200mV/cm 30 hours; 84MHz, 200mV/cm30 hour; 93MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the Spiramycin Base that the activatory yeast cell can degrade to the activity of Spiramycin Base.Prepare two 100 liters sample, each contains Spiramycin Base concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Spiramycin Base in the sample of activated yeast cell has reduced more than 66.8%.
The yeast cell component of decomposing bacitracin
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of bacitracin.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 98.000MHz scope, includes but not limited to 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.486 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 75MHz, 48mV/cm 15 hours; 78MHz, 48mV/cm 15 hours; 81MHz, 48mV/cm 15 hours; 95MHz, 48mV/cm 15 hours; 75MHz, 200mV/cm 30 hours; 78MHz, 200mV/cm 30 hours; 81MHz, 200mV/cm30 hour; 95MHz, 200mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the bacitracin that the activatory yeast cell can degrade to the activity of bacitracin.Prepare two 100 liters sample, each contains bacitracin concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains bacitracin in the sample of activated yeast cell has reduced more than 71.6%.
The yeast cell component of decomposing Polymyxin E
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of Polymyxin E.The frequency of electromagnetic field that is used for activated yeast cell is 410.000 in the 470.000MHz scope, includes but not limited to 430,431,432,433,434,435,436,437,438,439,440,441,442,443,444,445,446,447,448,449,450,451,452,453,454,455,456,457,458,459 and 460MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.611 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 433MHz, 85mV/cm 12 hours; 440MHz, 85mV/cm 12 hours; 446MHz, 85mV/cm 12 hours; 457MHz, 85mV/cm 12 hours; 433MHz, 204mV/cm24 hour; 440MHz, 204mV/cm 24 hours; 446MHz, 204mV/cm 24 hours; 457MHz, 204mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the Polymyxin E that the activatory yeast cell can degrade to the activity of Polymyxin E.Prepare two 100 liters sample, each contains Polymyxin E concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Polymyxin E in the sample of activated yeast cell has reduced more than 71.6%.
The yeast cell component of decomposing paraxin
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of paraxin.The frequency of electromagnetic field that is used for activated yeast cell is 410.000 in the 470.000MHz scope, includes but not limited to 430,431,432,433,434,435,436,437,438,439,440,441,442,443,444,445,446,447,448,449,450,451,452,453,454,455,456,457,458,459 and 460MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.371 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 419MHz, 85mV/cm 12 hours; 425MHz, 85mV/cm 12 hours; 433MHz, 85mV/cm12 hour; 462MHz, 85mV/cm 12 hours; 419MHz, 183mV/cm 24 hours; 425MHz, 183mV/cm 24 hours; 433MHz, 183mV/cm 24 hours; 462MHz, 183mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the paraxin that the activatory yeast cell can degrade to the activity of paraxin.Prepare two 100 liters sample, each contains paraxin concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains paraxin in the sample of activated yeast cell has reduced more than 58.6%.
The yeast cell component of decomposing cephalosporins
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of cephalosporins such as cefoxitin, Cephaloridine, Cephaloglycin, Cephalexin Monohydrate Micro/Compacted and Cephazolin.The frequency of electromagnetic field that is used for activated yeast cell is 410.000 in the 470.000MHz scope, includes but not limited to 430,431,432,433,434,435,436,437,438,439,440,441,442,443,444,445,446,447,448,449,450,451,452,453,454,455,456,457,458,459 and 460MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.559 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 434MHz, 85mV/cm12 hour; 441MHz, 85mV/cm 12 hours; 450MHz, 85mV/cm 12 hours; 458MHz, 85mV/cm 12 hours; 434MHz, 198mV/cm 24 hours; 441MHz, 198mV/cm 24 hours; 450MHz, 198mV/cm 24 hours; 458MHz, 198mV/cm24 hour.
The activatory yeast cell is to detect by the amount of measuring the cefoxitin that the activatory yeast cell can degrade to the activity of cephalosporins.Prepare two 100 liters sample, each concentration that contains cefoxitin is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains cefoxitin in the sample of activated yeast cell has reduced more than 75.5%.
The yeast cell component of decomposing Xin Meisu
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of Xin Meisu.The frequency of electromagnetic field that is used for activated yeast cell is 550.000 in the 620.000MHz scope, includes but not limited to 555,556,557,558,559,560,561,562,563,564,565,566,567,568,569,570,571,572,573,574 and 575MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.182 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 557MHz, 85mV/cm 12 hours; 564MHz, 85mV/cm 12 hours; 568MHz, 85mV/cm 12 hours; 574MHz, 85mV/cm 12 hours; 557MHz, 231mV/cm24 hour; 564MHz, 231mV/cm 24 hours; 568MHz, 231mV/cm 24 hours; 574MHz, 231mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the Xin Meisu that the activatory yeast cell can degrade to the activity of Xin Meisu.Prepare two 100 liters sample, each concentration that contains Xin Meisu is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Xin Meisu in the sample of activated yeast cell has reduced more than 67.7%.
The yeast cell component of decomposing Vulkamycin. PA-93
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of Vulkamycin. PA-93.The frequency of electromagnetic field that is used for activated yeast cell is 550.000 in the 620.000MHz scope, includes but not limited to 590,591,592,593,594,595,596,597,598,599,600,601,602,603,604,605,606,607,608,609 and 610MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.112 8 electromagnetic field in about 25-30 ℃, the described substratum of table 1, at one group of following order: 594MHz, 85mV/cm 12 hours; 599MHz, 85mV/cm 12 hours; 602MHz, 85mV/cm 12 hours; 608MHz, 85mV/cm 12 hours; 594MHz, 231mV/cm 24 hours; 599MHz, 231mV/cm 24 hours; 602MHz, 231mV/cm24 hour; 608MHz, 231mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the Vulkamycin. PA-93 that the activatory yeast cell can degrade to the activity of Vulkamycin. PA-93.Prepare two 100 liters sample, each contains Vulkamycin. PA-93 concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with residual antibiotic amount in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Vulkamycin. PA-93 in the sample of activated yeast cell has reduced more than 69.5%.
5.2 decompose the yeast cell component of adverse chemical product
The present invention further provides can the degradable solid refuse in the yeast cell of representative chemical.
According to the present invention, the ability of yeast cell degraded adverse chemical product be by with yeast cell in the presence of the electromagnetic field in the suitable culture base cultivation activate or strengthen.The gained yeast cell can be used as a component of biosolids waste treatment composition of the present invention and uses.
The electromagnetic field frequency that is used for activating or strengthens yeast degradation adverse chemical product ability usually 30 to 280MHz, 410 to 440MHz, 660 to 690MHz, 1400 to 1435MHz and 1980 in the scope of 2210MHz.Yeast cell growth can detect the ability that the yeast cell enhanced decomposes one or more chemical by method well known in the art after the sufficiently long time.Can be included but not limited to weedicide, agricultural chemicals and the pollutent relevant by the adverse chemical product of yeast degradation of the present invention with fertilizer.
It is to carry out in the liquid medium within that the present invention prepares degraded chemical yeast method.Contain the absorbable nutrition source of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of carbohydrate source depends in part on other composition in the substratum in the substratum, but usually between substratum weight about 0.1% to 5% between, preferred about 0.5% to 2%, most preferably from about 0.8%.These carbon sources can separately or be united use in substratum.
Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has (NH 4) 2HPO 4, CaCO 3, MgSO 4, NaCl and CaSO 4
Table 2: the zymic substratum of degraded chemical is formed
Substratum is formed Content
Muck or mud 8.0g, dry weight,＞120 orders
????NaCl ????0.2g
????MgSO 4·7H 2O ????0.2g
????CaCO 3·5H 2O ????0.5g
????CaSO 4·2H 2O ????0.2g
Peptone ????1.5g
????K 2HPO 4 ????0.5g
The extract (〉=100 μ g/ml) that contains chemical ????600ml
Autoclaving water ????400ml
The used extract of substratum is by being scattered in the fresh refuse of 500g such as muck, mud and/or rubbish in the 600ml warm water (35-40 ℃) and cultivating 24 hours, filter liquide at 30-37 ℃ and remove particulate matter and make.Only contain a spot of certain chemical as berry extract, an amount of this kind chemical can be joined in the extract.
It should be noted that the medium component that provides in the table 2 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.
Culturing process can be at first with cell density 10 2-10 5The yeast strain inoculum that cell/ml selects 1ml is inoculated in the 100ml substratum, and preferred inoculum density is 3 * 10 2-10 4Cell/ml.This process can increase and decrease as required in proportion.Yeast culture can be grown in the presence of an electromagnetism (EM) field or one group of electromagnetic field.If apply one group of electromagnetic field, when an electromagnetic field switched to another electromagnetic field, yeast cell can be in same container, use same set of electromagnetic wave generator and projector.
Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives the 2210.000MHz scope 30.000 to 100.000,70.000 to 280.000,410.000 to 430.000,660.000 to 680.000 and 1980.000.The field intensity of electromagnetic field arrives the 250mV/cm scope 40.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or different frequency and different field intensity in the above-mentioned scope.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably place one group to add up to 2,3,4,5,6,7,8,9 or 10 different electromagnetic field yeast culture.
Though yeast cell is in the presence of electromagnetic field even cultivate several hrs and just can be activated, but yeast cell can continue to cultivate for some time (for example 2 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 90-480 hour.
For example, use exemplary device shown in Figure 1, electromagnetic output amplitude arrives about 300mV/cm scope about 8.Method of the present invention is carried out in about 25 to 30 ℃ of temperature ranges, but preferably carries out at 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably 3Condition under carry out preferred 0.4mol/m 3Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.
When culturing process finished, yeast cell can reclaim from culture by several different methods known in the art, and stored being lower than under about 0 ℃ to 4 ℃ temperature.The yeast cell that reclaims can store with form of powder in the drying processing.
In order to detect the activity of activatory yeast cell, can measure under different time points and the different culture condition amount of compound in the test sample with method well known in the art such as HPLC to compound.For example, preparation contains the sample (every liter of 100mg) of concentration known compound.Then, with 0.1ml activatory and non-activated yeast (at least 10 7Cell/ml) adds in 100 liters of samples that contain this compound, cultivates 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours,, detect and compare the amount of compound residual in the extract by measuring sample with HPLC.
This method can be common to the chemical of numerous species.In specific embodiment, the best approach at the particular types chemical has been described.
The yeast cell component of decomposing aromatics
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of Trichlorophenol.The frequency that is used for the electromagnetic field of activated yeast cell arrives in the 100.000MHz scope 30.000, or preferred 52 to 98MHz, includes but not limited to 52,54,56,58,60,62,64,66,68,70,72,74,76,78,80,82,84,86,88,90,92,94,96 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain IFFI1411 6 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 82MHz, 82mV/cm 25 hours; 90MHz, 82mV/cm25 hour; 98MHz, 82mV/cm 25 hours; 82MHz, 274mV/cm 32 hours; 90MHz, 274mV/cm 32 hours; 98MHz, 274mV/cm 25 hours.
The activatory yeast cell is to detect by the amount of measuring this compound that the activatory yeast cell can degrade to the activity of Trichlorophenol.Prepare two 100 liters sample, each contains this compound concentrations is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of compound residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Trichlorophenol in the sample of activated yeast cell has reduced more than 56.4%.
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of toluene or ethylbenzene.The frequency of electromagnetic field that is used for activated yeast cell is 52.000 in the 98.000MHz scope, includes but not limited to 52,54,56,58,60,62,64,66,68,70,72,74,76,78,80,82,84,86,88,90,92,94,96 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.56 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 76MHz, 89mV/cm 20 hours; 80MHz, 89mV/cm 200 hours; 86MHz, 89mV/cm 20 hours; With 96MHz, 89mV/cm 20 hours.
The activatory yeast cell is to detect by the amount of measuring this compound that the activatory yeast cell can degrade to the activity of toluene or ethylbenzene.Prepare two 100 liters sample, each contains this compound concentrations is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of compound residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains toluene in the sample of activated yeast cell has reduced more than 74.3%.
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of xylene compounds such as p-Xylol.The frequency of electromagnetic field that is used for activated yeast cell is 30.000 in 50.000MHz or 70.000 to 98.000 scopes, includes but not limited to 30,32,34,36,38,40,42,44,46,48,50,70,72,74,76,78,80,82,84,86,88,90,92,94,96,97 and 98MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.420 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 72MHz, 93mV/cm 20 hours; 80MHz, 93mV/cm 20 hours; 88MHz, 93mV/cm 20 hours; With 98MHz, 93mV/cm 20 hours.
The activity of activatory yeast cell p-Xylol compound is to detect by the amount of measuring the xylene compounds that the activatory yeast cell can degrade.Prepare two 100 liters sample, each concentration that contains xylene compounds is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of p-Xylol residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains p-Xylol in the sample of activated yeast cell has reduced more than 66.6%.
The yeast cell component of decomposing aldehyde compounds
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of phenyl aldehyde and related compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in 98.000MHz or 133-151MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,98,133,134,135,136,137,138,139,140,141,142,143,144,145,146,147,148,149,150 or 151MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.374 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 78MHz, 130mV/cm 30 hours; 86MHz, 130mV/cm 30 hours; 94MHz, 130mV/cm 30 hours; 96MHz, 130mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the phenyl aldehyde that the activatory yeast cell can degrade to the activity of phenyl aldehyde.Prepare two 100 liters sample, each concentration that contains phenyl aldehyde is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of phenyl aldehyde residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains phenyl aldehyde in the sample of activated yeast cell has reduced more than 63.6%.
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of propionic aldehyde and related compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in 98.000MHz or 145-162MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,146,147,148,149,150,151,152,153,154,155,156,157,158,159,160,161 and 162MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.414 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 76MHz, 103mV/cm 20 hours; 88MHz, 103mV/cm 20 hours; 96MHz, 103mV/cm 20 hours; 98MHz, 103mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the propionic aldehyde that the activatory yeast cell can degrade to the activity of propionic aldehyde.Prepare two 100 liters sample, each concentration that contains propionic aldehyde is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of propionic aldehyde residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains propionic aldehyde in the sample of activated yeast cell has reduced more than 73.8%.
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of enanthaldehyde compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 in the 100.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97 and 98MHz.For example, to cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.503 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 81MHz, 90mV/cm 12 hours, 85MHz, 90mV/cm 12 hours, 89MHz, 90mV/cm 12 hours, 94MHz, 90mV/cm 12 hours, 81MHz, 157mV/cm 24 hours, 85MHz, 157mV/cm24 hour, 89MHz, 157mV/cm 24 hours, 94MHz, 157mV/cm 24 hours.
Activated yeast cell is to detect by the amount of measuring the enanthaldehyde that the activatory yeast cell can degrade to the activity of enanthaldehyde.Prepare two 100 liters sample, each contains enanthaldehyde concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of enanthaldehyde residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains enanthaldehyde in the sample of activated yeast cell reduced more than 81.3% in 24 hours.
The yeast cell component of decompoing halogenated benzene compound
In a specific embodiments, provide the preparation method of the yeast cell of a kind of decompoing halogenated benzene compound such as Meta Dichlorobenzene.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 163.000 in the 183.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,163,164,165,166,167,168,169,170,171,172,173,174,175,176,177,178,179,180,181,182 and 183MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.483 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 72MHz, 107mV/cm20 hour; 80MHz, 107mV/cm 10 hours; 90MHz, 107mV/cm 30 hours; 94MHz, 107mV/cm 40 hours.
The activity of activatory yeast cell santochlor is to detect by the amount of measuring the dichlorobenzene that the activatory yeast cell can degrade.Prepare two 100 liters sample, each concentration that contains dichlorobenzene is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of dichlorobenzene residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains dichlorobenzene in the sample of activated yeast cell has reduced more than 64.6%.
The yeast cell component of decomposing methyl phenyl ketone and related compound
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of methyl phenyl ketone and related compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 175.000 in the 191.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,175,176,177,178,179,180,181,182,183,184,185,186,187,188,189,190 and 191MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.256 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 76MHz, 124mV/cm 20 hours; 82MHz, 124mV/cm30 hour; 90MHz, 124mV/cm 40 hours; 98MHz, 124mV/cm 20 hours.
The activatory yeast cell is to detect by the amount of measuring the methyl phenyl ketone that the activatory yeast cell can degrade to the activity of methyl phenyl ketone.Prepare two 100 liters sample, each contains methyl phenyl ketone concentration is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of methyl phenyl ketone residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains methyl phenyl ketone in the sample of activated yeast cell has reduced more than 75.5%.
The yeast cell component of decomposing arsanilic acid and related compound
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of arsanilic acid and related compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 183.000 in the 205.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,183,184,185,186,187,188,189,190,191,192,193,194,195,196,197,198,199,200,201,202,203,204 and 205MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.745 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 78MHz, 133mV/cm 30 hours; 88MHz, 133mV/cm 40 hours; 92MHz, 133mV/cm30 hour; 96MHz, 133mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the arsanilic acid that the activatory yeast cell can degrade to the activity of arsanilic acid.Prepare two 100 liters sample, each concentration that contains arsanilic acid is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the residual antibiotic amount of arsanilic acid in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains arsanilic acid in the sample of activated yeast cell has reduced more than 75.5%.
The yeast cell component of decomposing roxarsone and related compound
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of roxarsone and related compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 114.000 in the 128.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,114,115,116,117,118,119,120,121,122,123,124,125,126,127 and 128MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.173 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 78MHz, 110mV/cm 10 hours; 92MHz, 110mV/cm 10 hours; 78MHz, 213mV/cm 30 hours; 92MHz, 213mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the roxarsone that the activatory yeast cell can degrade to the activity of roxarsone.Prepare two 100 liters sample, each concentration that contains roxarsone is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of roxarsone residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains roxarsone in the sample of activated yeast cell has reduced more than 67.9%.
The yeast cell component of decomposing the Nifurazolidone compound
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of Nifurazolidone and related compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 200.000 in the 220.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,200,201,202,203,204,205,206,207,208,209,210,211,212,213,214,215,216,217,218,219 and 220MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.397 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 74MHz, 98mV/cm30 hour; 76MHz, 98mV/cm 20 hours; 86MHz, 98mV/cm 30 hours; 94MHz, 98mV/cm 30 hours.
The activatory yeast cell is to detect by the amount of measuring the Nifurazolidone that the activatory yeast cell can degrade to the activity of Nifurazolidone.Prepare two 100 liters sample, the concentration of each Furanzolidon-containing is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of Nifurazolidone residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Nifurazolidone in the sample of activated yeast cell has reduced more than 81.4%.
The yeast cell component of decomposing M B 15497
In another embodiment, provide a kind of preparation method who decomposes the yeast cell of M B 15497 and related compound.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 213.000 in the 229.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,210,211,212,213,214,215,216,217,218,219,220,221,222,223,224,225,226,227,228 and 229MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.452 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 78MHz, 112mV/cm 30 hours; 82MHz, 112mV/cm 30 hours; 86MHz, 112mV/cm 30 hours; 94MHz, 112mV/cm 20 hours.
The activatory yeast cell is to detect by the amount of measuring the M B 15497 that the activatory yeast cell can degrade to the activity of M B 15497.Prepare two 100 liters sample, each concentration that contains M B 15497 is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of M B 15497 residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains M B 15497 in the sample of activated yeast cell has reduced more than 67.9%.
The yeast cell component of decomposing the trichlorophonum compound
The preparation method of the yeast cell of a kind of trichlorophonum of decomposition and related compound is provided in another embodiment.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 220.000 in the 250.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,220,222,224,226,228,230,232,234,236,238,240,242,244,245,248 and 250MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.100 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 74MHz, 219mV/cm 30 hours; 86MHz, 219mV/cm20 hour; 96MHz, 219mV/cm 30 hours; 98MHz, 219mV/cm 20 hours.
The activatory yeast cell is to detect by the amount of measuring the trichlorophonum that the activatory yeast cell can degrade to the activity of trichlorophonum.Prepare two 100 liters sample, each concentration that contains trichlorophonum is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of trichlorophonum residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains trichlorophonum in the sample of activated yeast cell has reduced more than 72 4%.
The yeast cell component of decomposing dinitolmide
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of dinitolmide and related compound.Dinitolmide is a 2-methyl-3, and the 5-dinitrobenzamide is also named 3,5-dinitro o toluamide.The frequency of electromagnetic field that is used for activated yeast cell is 70.000 to 98.000MHz or 220.000 in the 250.000MHz scope, includes but not limited to 70,72,74,76,78,80,82,84,86,88,90,92,94,96,97,98,220,222,224,226,228,230,232,234,236,238,240,242,244,246,248 and 250MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.189 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 76MHz, 202mV/cm 30 hours; 82MHz, 202mV/cm30 hour; 90MHz, 202mV/cm 20 hours; 96MHz, 202mV/cm 20 hours.
The activatory yeast cell is to detect by the amount of measuring 3, the 5 dinitro o toluamides that the activatory yeast cell can degrade to the activity of dinitolmide.Prepare two 100 liters sample, each concentration that contains dinitolmide is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of dinitolmide residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains dinitolmide in the sample of activated yeast cell has reduced more than 72.4%.
Remove ammonium compound (NH 4) the yeast cell component
In a specific embodiments, provide a kind of preparation method who removes or reduce the yeast cell of ammonium compound level in the solid waste.The frequency of electromagnetic field that is used for activated yeast cell is 660 to 680MHz or 2160 in the 2190MHz scope, preferred 661,662,663,664,665,666,667,668,669,670,671,672,673,674,675,676,677,678,679,680,2170,2171,2172,2173,2174,2175,2176,2177,2178,2179,2180,2181,2182,2183,2184,2185MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.614 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 662MHz, 152mV/cm 18 hours; 666MHz, 152mV/cm 18 hours; 672MHz, 152mV/cm18 hour; 678MHz, 152mV/cm 18 hours; 662MHz, 310mV/cm 25 hours; 666MHz, 310mV/cm 25 hours; 672MHz, 310mV/cm 35 hours; 678MHz, 310mV/cm 35 hours.
Activatory zymic activity is to detect by the amount of measuring activated yeast cell removal ammonium compound.Compare with containing not the sample of activated yeast cell, the amount that contains ammonium compound in the sample of activated yeast cell obviously reduces (＞93.6%).
Remove the yeast cell component of nitrate and nitrite
In a specific embodiments, provide a kind of preparation method who removes or reduce the yeast cell of nitrate and nitrite level in the solid waste.The frequency of electromagnetic field that is used for activated yeast cell is 661.000 in the 680.000MHz scope, includes but not limited to 661,662,663,664,665,666,667,668,669,670,671,672,673,674,675,676,677,678,679 and 680MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.14 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 661MHz, 126mV/cm 25 hours; 665MHz, 126mV/cm 25 hours; 672MHz, 126mV/cm 25 hours; 676MHz, 126mV/cm25 hour; 661MHz, 196mV/cm 25 hours; 665MHz, 196mV/cm 25 hours; 672MHz, 196mV/cm 38 hours; 676MHz, 196mV/cm 38 hours.
The activatory yeast cell is to detect by the amount of measuring activated yeast cell removal nitrate to the activity of nitrate.Prepare two 100 liters sample, each concentration that contains nitrate is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of residual nitric acid salt in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains nitrate in the sample of activated yeast cell has reduced more than 69.7%.
Remove the biological yeast cell component that can utilize phosphorus
In a specific embodiments, provide that biology can utilize phosphorus such as HPO in a kind of removal solid waste 4 2-, H 2PO 4 -Deng the preparation method of yeast cell.In a specific embodiments, provide a kind of preparation method who removes or reduce the yeast cell of nitrate and nitrite in the solid waste.The frequency that is used for the electromagnetic field of activated yeast cell arrives in the 440.000MHz scope 80.000, preferred 86.000 to 120.000MHz or 410.000 to 440.000MHz, includes but not limited to 86,88,90,92,94,96,98,100,102,104,106,108,110,112,114,116,118,120,410,411,412,413,414,415,416,417,418,419,420,421,422,423,424,425,426,427,428,429 and 430MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.620 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 98MHz, 68mV/cm 24 hours; 112MHz, 68mV/cm24 hour; 108MHz, 68mV/cm 24 hours; 118MHz, 68mV/cm 24 hours; 98MHz, 240mV/cm 24 hours; 112MHz, 240mV/cm 24 hours; 108MHz, 240mV/cm 42 hours; 118MHz, 240mV/cm 42 hours.
The activatory yeast cell is to detect by the amount of measuring the utilized phosphorus that the activatory yeast cell can remove to the activity that can utilize phosphorus.Prepare two 100 liters sample, each contains biology and can utilize the concentration of phosphorus to be 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, can utilize the amount of phosphorus with residual biology in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, contain in the sample of activated yeast cell and can utilize the amount of phosphorus to reduce more than 65.8%.
The yeast cell component of decomposing Trichlorphon
In a specific embodiments, provide a kind of preparation method who decomposes the Trichlorphon and the yeast cell of relevant organophosphorus pesticide compound.The frequency of electromagnetic field that is used for activated yeast cell is 1980.000 to 2020.000, preferred 2000.000 in the 2020.000MHz scope, includes but not limited to 2000,2001,2002,2003,2004,2005,2006,2007,2008,2009,2010,2011,2012,2013,2014,2015,2016,2017,2018,2019 and 2020MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.440 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 2000MHz, 125mV/cm 10 hours; 2004MHz, 125mV/cm 10 hours; 2009MHz, 125mV/cm 24 hours; 2018MHz, 125mV/cm 24 hours; 2000MHz, 168mV/cm 10 hours; 2004MHz, 168mV/cm 10 hours; 2009MHz, 168mV/cm 56 hours; 2018MHz, 168mV/cm 56 hours.
The activatory yeast cell is to detect by the amount of measuring the Trichlorphon that the activatory yeast cell can degrade to the activity of Trichlorphon.Prepare two 100 liters sample, each concentration that contains Trichlorphon is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of Trichlorphon residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, after 48 hours, the amount that contains Trichlorphon in the sample of activated yeast cell has reduced more than 10%.
The yeast cell component of decomposing SD-1750
In a specific embodiments, provide the preparation method of a kind of decomposition SD-1750 (DDVP) and the yeast cell of relevant organophosphorus pesticide compound.The frequency of electromagnetic field that is used for activated yeast cell is 1983.000 in the 2118.000MHz scope, includes but not limited to 1983,1988,1993,1998,2003,2008,2013,2018,2023,2028,2033,2038,2043,2048,2053,2058,2063,2068,2073,2078,2083,2088,2093,2098,2103,2108,2113 and 2118MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.443 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 1993MHz, 140mV/cm 24 hours; 2023MHz, 140mV/cm 24 hours; 2083MHz, 140mV/cm 24 hours; 2103MHz, 140mV/cm 24 hours; 1993MHz, 190mV/cm 24 hours; 2023MHz, 190mV/cm 24 hours; 2083MHz, 190mV/cm 56 hours; 2103MHz, 190mV/cm 56 hours.
The activatory yeast cell is to detect by the amount of measuring the SD-1750 that the activatory yeast cell can degrade to the activity of SD-1750.Prepare two 100 liters sample, each concentration that contains SD-1750 is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of SD-1750 residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains SD-1750 in the sample of activated yeast cell has reduced more than 67.5%.
The yeast cell component of decomposing monocrotophos
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of monocrotophos and associated pesticide.The frequency of electromagnetic field that is used for activated yeast cell is 1983.000 in the 2118.000MHz scope, includes but not limited to 1983,1988,1993,1998,2003,2008,2013,2018,2023,2028,2033,2038,2043,2048,2053,2058,2063,2068,2073,2078,2083,2088,2093,2098,2103,2108,2113 and 2118MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.93 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 2998MHz, 165mV/cm 24 hours; 2033MHz, 165mV/cm 24 hours; 2058MHz, 165mV/cm 24 hours; 2113MHz, 165mV/cm 24 hours; 2998MHz, 202mV/cm 56 hours; 2033MHz, 202mV/cm 56 hours; 2058MHz, 202mV/cm 24 hours; 2113MHz, 202mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the monocrotophos that the activatory yeast cell can degrade to the activity of monocrotophos.Prepare two 100 liters sample, each concentration that contains monocrotophos is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of monocrotophos residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains monocrotophos in the sample of activated yeast cell has reduced more than 73.4%.
The yeast cell component of decomposing Rogor
In a specific embodiments, provide a kind of preparation method who decomposes the yeast cell of Rogor and relevant Pesticidal compound.The frequency of electromagnetic field that is used for activated yeast cell is 1983.000 in the 2118.000MHz scope, includes but not limited to 1983,1988,1993,1998,2003,2008,2013,2018,2023,2028,2033,2038,2043,2048,2053,2058,2063,2068,2073,2078,2083,2088,2093,2098,2103,2108,2113 and 2118MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.379 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 1988MHz, 195mV/cm 24 hours; 2023MHz, 195mV/cm 24 hours; 2088MHz, 195mV/cm 24 hours; 2108MHz, 195mV/cm 24 hours; 1988MHz, 277mV/cm 56 hours; 2023MHz, 277mV/cm 56 hours; 2088MHz, 277mV/cm 24 hours; 2108MHz, 277mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the Rogor that the activatory yeast cell can degrade to the activity of Rogor.Prepare two 100 liters sample, each concentration that contains Rogor is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of Rogor residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains Rogor in the sample of activated yeast cell has reduced more than 69.6%.
The yeast cell component of decomposing dichlorodiphenyl trichloroethane (DDT)
In a specific embodiments, provide a kind of decomposing D DT preparation method with the yeast cell of the organic Pesticidal compound of relevant dichloro.The frequency of electromagnetic field that is used for activated yeast cell is 1420.000 in the 1435.000MHz scope, includes but not limited to 1420,1421,1422,1423,1424,1425,1426,1427,1428,1429,1430,1431,1432,1433,1434,1435MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.415 8 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 1423MHz, 75mV/cm 24 hours; 1426MHz, 75mV/cm24 hour; 1433MHz, 75mV/cm 24 hours; 1435MHz, 75mV/cm 24 hours; 1423MHz, 146mV/cm 56 hours; 1426MHz, 146mV/cm 56 hours; 1433MHz, 146mV/cm 24 hours; 1435MHz, 146mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the DDT that the activatory yeast cell can degrade to the activity of DDT.Prepare two 100 liters sample, each concentration that contains DDT is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of DDT residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains DDT in the sample of activated yeast cell has reduced more than 78.5%.
The yeast cell component of decomposing toxaphene
In a specific embodiments, provide a kind of preparation method who decomposes the toxaphene and the yeast cell of the organic Pesticidal compound of relevant chloro.The frequency of electromagnetic field that is used for activated yeast cell is 1420.000 in the 1435.000MHz scope, includes but not limited to 1420,1421,1422,1423,1424,1425,1426,1427,1428,1429,1430,1431,1432,1433,1434,1435MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.504 4 electromagnetic field in about 25-30 ℃, the described substratum of table 2, at one group of following order: 1420MHz, 120mV/cm 24 hours; 1426MHz, 120mV/cm 24 hours; 1431MHz, 120mV/cm 24 hours; 1434MHz, 120mV/cm 24 hours.
The activatory yeast cell is to detect by the amount of measuring the toxaphene that the activatory yeast cell can degrade to the activity of toxaphene.Prepare two 100 liters sample, each concentration that contains toxaphene is 100mg/L.Respectively 0.1ml activatory and non-activated yeast cell (are all contained 10 then 7Cell/ml) add in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, with the amount of toxaphene residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains toxaphene in the sample of activated yeast cell has reduced more than 70.8%.
5.3 reduce the yeast cell component of smell
The present invention also provides the yeast cell of the smell that can reduce solid waste such as muck, mud and/or rubbish.Although be not subjected to the constraint of any theory or mechanism, the present inventor thinks that yeast cell of the present invention can reduce the smell of solid waste by reducing, absorb or decomposing known and unknown malodorous compound in the solid waste.But, there is no need to confirm that these compounds are decomposed.As long as after using yeast cell of the present invention, one group of subjective smell that detects of main body has been reduced just passable.
According to the present invention, the yeast cell that preparation can reduce the solid waste smell is undertaken by culturing cell in the presence of electromagnetic field, in the suitable culture base.The electromagnetic field frequency of activation or this function of reinforcement yeast arrives in the 2380MHz scope 2160 usually.Yeast cell growth can detect the ability that yeast cell reduces the solid waste smell by method well known in the art after the sufficiently long time.
The method that the present invention prepares the yeast cell that reduces smell is to carry out in the liquid medium within.Contain the absorbable nutrition source of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of carbohydrate depends in part on other composition in the substratum in the substratum, but usually between substratum weight about 0.1% to 5% between, preferred about 0.5% to 2%.Most preferably from about 0.8%.These carbon sources can separately or be united use in substratum.
Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has (NH 4) 2HPO 4, CaCO 3, MgSO 4, NaCl and CaSO 4
It should be noted that it is not restrictive that the substratum that provides in the table 38 is formed.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.
Culturing process can be at first with cell density 10 2-10 5Individual cell/ml, preferred 3 * 10 2-10 4Individual cell/ml is inoculated in the yeast strain inoculum that 1ml selects in the 100ml substratum.This process can increase and decrease as required in proportion.Yeast culture can be grown in the presence of an electromagnetism (EM) field or one group of electromagnetic field.If apply one group of electromagnetic field, when an electromagnetic field switched to another electromagnetic field, yeast cell can be in same container, use same set of electromagnetic wave generator and projector.
Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field can be 2160 to 2380MHz, and preferred 2160.000 to 2250.000MHz or 2280.000 in the 2380.000MHz scope.The field intensity of electromagnetic field arrives the 300mV/cm scope 25.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or different frequency and different field intensity in the above-mentioned scope.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7,8,9 or 10 different electromagnetic field.
Though yeast cell is in the presence of electromagnetic field even cultivate several hrs and just can be activated, but yeast cell can continue to cultivate for some time (for example 2 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 80-320 hour.
For example, use exemplary device shown in Figure 1, electromagnetic output amplitude is in the 25-200mV/cm scope.After fs cultivates, yeast cell is further cultivated for some time under the essentially identical condition the higher level increasing to except amplitude in the 250-300mV/cm scope.Method of the present invention is carried out in about 25 to 30 ℃ of scopes, but preferably carries out at 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably 3Condition under carry out preferred 0.4mol/m 3Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.
When culturing process finished, yeast cell can reclaim from culture by several different methods known in the art, and stored being lower than under about 0 ℃ to 4 ℃ temperature.The yeast cell that reclaims can store with form of powder in the drying processing.
Can detect the ability that the yeast cell of cultivating reduces the organic materials smell with any means known in the art.Stench chemical such as hydrogen sulfide, ammonia, indoles, p-cresol, skatole and organic acid amount can detect with any means known in the art in the organic materials test sample, include but not limited to vapor-phase chromatography, olfactometry, mass spectroscopy or use the smell test panel.
For example, in order to detect the activity of activated yeast cell, can use mass spectroscopy (as VG micromass) to measure under different time points and the different breeding conditions amount of malodorous compound in the test sample to malodorous compound.For example, the malodorous compound (reaching every liter of 100mg) with known quantity adds in the aqueous extract of 10 liters of muck.Then, with 0.1ml activation and non-activated yeast (at least 10 7Cell/ml) adds in 10 liters of samples that contain this compound, cultivates 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect and compare the amount of malodorous compound remaining in the extract.
The yeast cell component of the smell that the minimizing sulfocompound causes
In one embodiment of the invention, provide a kind of preparation method who removes the hydrogen sulfide sulfur-bearing relevant or contain the yeast cell of sulfydryl (SH-) molecule with other.The yeast cell of removing the hydrogen sulfide sulfur-bearing relevant with other or containing sulfydryl (SH-) molecule can prepare by cell is cultivated in the presence of the electromagnetic field of 2160.000 to 2250.000MHz scopes, includes but not limited to 2160,2165,2170,2175,2180,2185,2190,2195,2200,2205,2210,2215,2220,2225,2230,2235,2240,2245 and 2250MHz.
For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.559 4 electromagnetic field in about 25-30 ℃, the described substratum of table 3, at one group of following order: 2165MHz, 240mV/cm 20 hours; 2175MHz, 240mV/cm 20-60 hour; 2200MHz, 240mV/cm 20 hours; 2235MHz, 240mV/cm 20 hours.
The activatory yeast cell is to have down by measuring the activatory yeast cell that the change of the amount of hydrogen sulfide detects to sulfur-bearing or the activity that contains sulfydryl (SH-) compound.Prepare two 100 liters sample, each hydrogen sulfide containing concentration is 100mg/L.Then 0.1ml activatory and the non-activated yeast cell of 0.1ml (are all contained 10 7Cell/ml) add respectively in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, the amount of residual hydrogen sulfide in detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains hydrogen sulfide in the sample of activated yeast cell has reduced more than 59.8%.
Reduce the yeast cell component contain the smell that the NH-compound causes
In another embodiment of the invention, provide a kind of preparation method who removes the ammonia and the yeast cell of the relevant NH-of containing compound.The yeast cell of removing ammonia and the relevant NH-of containing compound can prepare by cell is cultivated in the presence of the electromagnetic field of 2160.000 to 2250.000 MHz scopes, includes but not limited to 2160,2165,2170,2175,2180,2185,2190,2195,2200,2205,2210,2215,2220,2225,2230,2235,2240,2245 and 2250MHz.
For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.423 4 electromagnetic field in about 25-30 ℃, the described substratum of table 3, at one group of following order: 2160MHz, 250mV/cm 20 hours; 2175MHz, 250mV/cm 20 hours; 2210MHz, 250mV/cm 20 hours; 2245MHz, 250mV/cm 10 hours.
The activatory yeast cell is to detect by the change of measuring ammonia amount under the existence of activatory yeast cell to ammonia and the activity that contains the NH-compound.Prepare two 100 liters sample, containing the NH-compound concentrations in each sample is 100mg/L.Then 0.1ml activatory and the non-activated yeast cell of 0.1ml (are all contained 10 7Cell/ml) add respectively in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, the amount of residual ammonia in detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains ammonia in the sample of activated yeast cell has reduced more than 69.6%.
The yeast cell component of the smell that minimizing indoles and other related compound cause
Among the present invention, provide a kind of preparation method who decomposes the yeast cell of indoles and other related compound such as skatole.The yeast cell that decomposes indoles and other related compound can prepare by cell is cultivated in the presence of the electromagnetic field of 2160.000 to 2250.000MHz scopes, includes but not limited to 2160,2165,2170,2175,2180,2185,2190,2195,2200,2205,2210,2215,2220,2225,2230,2235,2240,2245 and 2250MHz.
For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.612 4 electromagnetic field in about 25-30 ℃, the described substratum of table 3, at one group of following order: 2165MHz, 240mV/cm 40 hours; 2180MHz, 240mV/cm 20 hours; 2200MHz, 240mV/cm 40 hours; 2220MHz, 240mV/cm 20 hours.
The activatory yeast cell is to detect by the amount of measuring activatory yeast cell removal indoles to the activity of indoles and other related compound.Prepare two 100 liters sample, the concentration that contains the indoles related compound in each sample is 100mg/L.Then 0.1ml activatory and the non-activated yeast cell of 0.1ml (are all contained 10 7Cell/ml) add respectively in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, by the amount of indoles residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains indoles in the sample of activated yeast cell has reduced more than 71.3%.
The yeast cell component of the smell that the minimizing organic acid causes
In another embodiment, provide a kind of preparation method who removes the yeast cell of organic acid odorous such as formic acid, acetate, propionic acid, butyric acid and other voltaile fatty acid.The yeast cell that can reduce the organic acid smell can prepare by cell is cultivated in the presence of the electromagnetic field of 2160.000 to 2250.000MHz scopes, includes but not limited to 2160,2165,2170,2175,2180,2185,2190,2195,2200,2205,2210,2215,2220,2225,2230,2235,2240,2245 and 2250MHz.
For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.53 4 electromagnetic field in about 25-30 ℃, the described substratum of table 42, at one group of following order: 2315MHz, 290mV/cm 30 hours; 2335MHz, 290mV/cm 10 hours; 2355MHz, 290mV/cm 20 hours; 2375MHz, 290mV/cm 10 hours.
The activatory yeast cell is to detect by the change of measuring acetate amount under the existence of activatory yeast cell to the organic acid activity.Prepare two 100 liters sample, containing organic acid concentration in each sample is 100mg/L.Then 0.1ml activatory and the non-activated yeast cell of 0.1ml (are all contained 10 7Cell/ml) add respectively in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, by the amount of acetate residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains acetate in the sample of activated yeast cell has reduced more than 89.4%.
The yeast cell component of the smell that minimizing aliphatics replacement amine causes
In another embodiment, provide a kind of removal or degrading aliphatic to replace amine, for example methylamine, dimethylamine or Trimethylamine 99 reduce the preparation method of the yeast cell of the smell that these compounds cause thus.Remove or the yeast cell of these amines of degrading can prepare by cell is cultivated in the presence of the electromagnetic field of 2160.000 to 2250.000MHz scopes, include but not limited to 2160,2165,2170,2175,2180,2185,2190,2195,2200,2205,2210,2215,2220,2225,2230,2235,2240,2245 and 2250MHz.
For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.541 4 electromagnetic field in about 25-30 ℃, the described substratum of table 3, at one group of following order: 2160MHz, 250mV/cm 20 hours; 2190MHz, 250mV/cm 10 hours; 2210MHz, 250mV/cm 40 hours; 2250MHz, 250mV/cm 40 hours.
The activatory yeast cell is to detect by the amount of measuring this amine under the existence of activatory yeast cell to the activity of methyl substituted amine.Prepare two 100 liters sample, the concentration that contains methyl substituted amine in each sample is 100mg/L.Then 0.1ml activatory and the non-activated yeast cell of 0.1ml (are all contained 10 7Cell/ml) add respectively in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, by the amount of methyl substituted amine residual in HPLC measurement and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains methyl substituted amine in the sample of activated yeast cell has reduced more than 82.2%.
The yeast cell component of the smell that minimizing p-cresol and related compound cause
The preparation method of the yeast cell of a kind of removal or degraded p-cresol and related compound is provided in another embodiment.Remove or the yeast cell of degraded p-cresol and other related compound can prepare by cell is cultivated in the presence of the electromagnetic field of 2160.000 to 2250.000MHz scopes, include but not limited to 2160,2165,2170,2175,2180,2185,2190,2195,2200,2205,2210,2215,2220,2225,2230,2235,2240,2245 and 2250MHz.
For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.163 4 electromagnetic field in about 25-30 ℃, the described substratum of table 3, at one group of following order: 2300MHz, 98mV/cm 20 hours; 2370MHz, 98mV/cm 15 hours; 2300MHz, 250mV/cm 20 hours; 2370MHz, 250mV/cm 30 hours.
The activatory yeast cell is to exist the change of the amount of following p-cresol and related compound to detect by measuring the activatory yeast cell to the activity of p-cresol and related compound.Prepare two 100 liters sample, the concentration that contains p-cresol in each sample is 100mg/L.Then 0.1ml activatory and the non-activated yeast cell of 0.1ml (are all contained 10 7Cell/ml) add respectively in two samples was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, by the amount of p-cresol residual in HPLC detection and the comparative sample.Compare with containing not the sample of activated yeast cell, the amount that contains p-cresol in the sample of activated yeast cell has reduced more than 92.5%.
5.4 pathogenic agent inhibition yeast cell component
The present invention also provides the yeast cell of the pathogenic microorganism growth that can suppress to exist in the solid waste.Usually, owing to there is the utilizable abundant nutrition of these pathogenic microorganisms in the solid waste, the number of pathogenic agent increases fast after after a while.But, in the presence of pathogenic agent inhibition zymic of the present invention, the constant in time or minimizing of the number of pathogenic agent in the solid waste of handling.Though be not subjected to the constraint of any theory or mechanism, the present inventor thinks that the yeast cell of the inhibition pathogenic agent that exists in the solid waste has formed the environment that is unfavorable for the pathogenic microorganism growth.
According to the present invention, the ability of yeast influence/control pathogenic agent number is activated or strengthens by yeast is cultivated in the presence of electromagnetic field.Gained pathogenic agent inhibition yeast cell uses as the component of solid waste disposal composition of the present invention.
The electromagnetic field frequency of the ability of activation or reinforcement yeast control pathogenic microorganism number arrives in the 50MHz scope 30 usually.Yeast cell growth can detect the ability of yeast cell influence/control pathogenic agent number by method well known in the art after the sufficiently long time.
The method that the present invention prepares pathogenic agent inhibition yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition source of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of carbohydrate depends in part on other composition in the substratum in the substratum, but usually between substratum weight about 0.1% to 5% between, preferred about 0.5% to 2%, most preferably from about 0.8%.These carbon sources can separately or be united use in substratum.
Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has (NH 4) 2HPO 4, CaCO 3, MgSO 4, NaCl and CaSO 4
Table 4: pathogenic agent inhibition zymic substratum is formed
Substratum is formed Content
Zulkovsky starch ????8.0g
Sucrose ????5g
????NaCl ????0.2g
????MgSO 4·7H 2O ????0.2g
????CaCO 3·5H 2O ????0.5g
????CaSO 4·2H 2O ????0.2g
Peptone ????1.5g
????K 2HPO 4 ????0.5g
Autoclaving water ????400ml
The pathogenic agent extract ????600ml
The used pathogenic agent extract of substratum is to remove the particulate matter preparation by refuse 30-37 ℃ of cultivation 24 hours down, filter liquide in about 600ml warm water (35-40 ℃) of 500g being contained pathogenic agent.It should be noted that it is not restrictive that the substratum that provides in the table 4 is formed.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.
Culturing process can be at first with cell density 10 2-10 5Cell/ml, preferred 3 * 10 2-10 4Cell/ml is inoculated in the yeast strain inoculum that 1ml selects in the 100ml substratum.This process can increase and decrease as required in proportion.Yeast culture can be grown in the presence of an electromagnetism (EM) field or one group of electromagnetic field.If apply one group of electromagnetic field, when an electromagnetic field switched to another electromagnetic field, yeast cell can be in same container, use same set of electromagnetic wave generator and projector.
Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field can be 30.000 to 50.000,500.000 to 650.000 and 1000.000 in the 1150.000MHz scope.The field intensity of electromagnetic field arrives the 200mV/cm scope 20.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7,8,9 or 10 different electromagnetic field.
Though yeast cell is in the presence of electromagnetic field even cultivate several hrs and just can be activated, but yeast cell can continue to cultivate for some time (for example 2 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 144-272 hour.
For example, use exemplary device shown in Figure 1, electromagnetic output amplitude is in the 20-180mV/cm scope.Fs further cultivates yeast cell for some time under the essentially identical condition the higher level that increases to except amplitude in the 200-350mV/cm scope after cultivating.
When culturing process finished, the yeast cell that suppresses pathogenic agent can reclaim from culture by several different methods known in the art, and in about 0 ℃ to 4 ℃ storage.The yeast cell that suppresses pathogenic agent can also store with form of powder in the drying processing.
Can be with any microorganism count method known in the art, for example optical densitometric method, the solid dielectric plating dilution method of counting or in microscopically individual cells counting process detect the ability of the yeast control pathogenic agent number that suppresses pathogenic agent.Can use that dyeing is distinguished or definite sample in different microorganism strains or kinds, or detect their viability.When wishing that pathogenic agent inhibition yeast works to one group of pathogenic microorganism, can monitor the number of more than one typical pathogenic microorganism and assess pathogenic agent inhibition zymic performance.
For example, in the presence of the pathogenic agent inhibition yeast of different concns, the solid waste sample that contains the concentration known pathogenic microorganism is cultivated the identical time under identical conditions, and the identical yeast strain that cultural method is handled according to the present invention is not as negative control.Also can detect the growth of pathogenic agent under the normal condition with not adding any zymic sample.The number of pathogenic agent before and after detecting and relatively cultivating.
Prepare 1 liter of culture, every milliliter contains 10 at least 10The pathogenic microorganism of cell.(every milliliter contains 2 * 10 to add 1ml activatory yeast cell in 1 liter of pathogenic microorganism culture 7To 5 * 10 7Individual yeast), cultivated 24 hours at 30 ℃.Comprise and contain the not contrast of activated yeast cell.Detect and the number of microorganism in the culture more separately.Be several embodiment below, wherein studied the malignant bacteria of specific kind.
The yeast cell component that suppresses streptococcus aureus
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses the yeast cell of staphylococcus aureus growth.The frequency of electromagnetic field that is used for activated yeast cell is 30.000 in the 50.000MHz scope, includes but not limited to 30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49 and 50MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.595 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 30MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 43MHz, 26mV/cm 12 hours; 47MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 43MHz, 150mV/cm 24 hours; 47MHz, 150mV/cm 24 hours.
The activatory yeast cell is to detect by the growth of measuring streptococcus aureus under the existence of activatory yeast cell to the activity of streptococcus aureus.Contained streptococcus aureus grows to cell counting and reaches 10 in the solid waste extract in substratum 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell of streptococcus aureus in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 2.6% in the sample of activated yeast cell.
The yeast cell component that suppresses pneumococcus
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses the yeast cell of pneumococcus growth.The frequency of electromagnetic field that is used for activated yeast cell is 30.000 in the 50.000MHz scope, includes but not limited to 30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49 and 50MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain IFFI1021 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 30MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 42MHz, 26mV/cm 12 hours; 49MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 42MHz, 150mV/cm 24 hours; 49MHz, 150mV/cm 24 hours.
The activatory yeast cell is to detect by the growth of measuring pneumococcus under the existence of activatory yeast cell to the activity of pneumococcus.Contained pneumococcus grows to cell counting and reaches 10 in the solid waste extract in substratum 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell counting of pneumococcus in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 3% in the sample of activated yeast cell.
The yeast cell component that suppresses anthrax bacillus
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses the yeast cell of anthrax bacillus growth.The frequency of electromagnetic field that is used for activated yeast cell is 20.000 in the 45.000MHz scope, includes but not limited to 20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44 and 45MHz.For example, will cultivate in the presence of the yeast cell of anthrax bacillus strains A S2.390 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 24MHz, 100mV/cm 24 hours; 37MHz, 100mV/cm 24 hours; 40MHz, 100mV/cm 24 hours; 45MHz, 100mV/cm 24 hours; 24MHz, 190mV/cm 24 hours; 37MHz, 190mV/cm 24 hours; 40MHz, 190mV/cm24 hour; 45MHz, 190mV/cm 24 hours.
The activatory yeast cell is to detect by the growth of measuring anthrax bacillus under the existence of activatory yeast cell to the activity of anthrax bacillus.Contained anthrax bacillus grows to cell counting and reaches 10 in the solid waste extract in substratum 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell counting of anthrax bacillus in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 2.6% in the sample of activated yeast cell.
The yeast cell component that suppresses Mycobacterium tuberculosis
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses the yeast cell of Mycobacterium tuberculosis growth.The frequency of electromagnetic field that is used for activated yeast cell is 30.000 in the 50.000MHz scope, includes but not limited to 30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49 and 50MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.431 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 33MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 45MHz, 26mV/cm 12 hours; 47MHz, 26mV/cm 12 hours; 33MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 45MHz, 150mV/cm 24 hours; 47MHz, 150mV/cm 24 hours.
The activatory yeast cell is to detect by the growth of measuring Mycobacterium tuberculosis under the existence of activatory yeast cell to the activity of Mycobacterium tuberculosis.Make Mycobacterium tuberculosis contained in the solid waste extract in substratum, grow to cell counting and reach 10 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell counting of Mycobacterium tuberculosis in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 2.9% in the sample of activated yeast cell.
Suppress colibacillary yeast cell component
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses the yeast cell of intestinal bacteria growth.The frequency of electromagnetic field that is used for activated yeast cell is 30.000 in the 50.000MHz scope, includes but not limited to 30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49 and 50MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.561 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 30MHz, 26mV/cm 12 hours; 34MHz, 26mV/cm 12 hours; 38MHz, 26mV/cm 12 hours; 49MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 3443MHz, 150mV/cm 24 hours; 38MHz, 150mV/cm 24 hours; 49MHz, 150mV/cm 24 hours.
The activatory yeast cell is that colibacillary growth detects under the existence of activatory yeast cell by measuring to colibacillary activity.Make intestinal bacteria contained in the solid waste extract in substratum, grow to cell counting and reach 10 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect colibacillary cell counting in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 48% in the sample of activated yeast cell.
The yeast cell component that suppresses Salmonellas
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses the yeast cell of Salmonella growth.The frequency of electromagnetic field that is used for activated yeast cell is 30.000 in the 50.000MHz scope, includes but not limited to 30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49 and 50MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.178 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 30MHz, 26mV/cm 12 hours; 33MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 38MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 33MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 38MHz, 150mV/cm24 hour.
The activatory yeast cell is to detect by the growth of measuring Salmonellas under the activated yeast cell existence to the activity of Salmonellas.Make Salmonellas contained in the solid waste extract in substratum, grow to cell counting and reach 10 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell counting of Salmonellas in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 62% in the sample of activated yeast cell.
The yeast cell component that suppresses vibrios
In a specific embodiment of the present invention, provide a kind of yeast cell preparation method who suppresses vibrios.The frequency of electromagnetic field that is used for activated yeast cell is 500.000 in the 550.000MHz scope, includes but not limited to 520,521,522,523,524,525,526,527,528,529,530,531,532,533,534,535,536,537,538,539 and 540MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.377 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 521MHz, 150mV/cm 24 hours; 527MHz, 150mV/cm 24 hours; 531MHz, 150mV/cm 24 hours; 538MHz, 150mV/cm 24 hours; 521MHz, 276mV/cm 24 hours; 527MHz, 276mV/cm 24 hours; 531MHz, 276mV/cm 24 hours; 538MHz, 276mV/cm 24 hours.
The activatory yeast cell is to detect by the growth of measuring vibrios under the activated yeast cell existence to the activity of vibrios.Make vibrios contained in the solid waste extract in substratum, grow to cell counting and reach 10 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell counting of vibrios in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 5.6% in the sample of activated yeast cell.
The yeast cell component that suppresses shigella
In a specific embodiment of the present invention, provide a kind of preparation method who suppresses the yeast cell of shigella.The frequency of electromagnetic field that is used for activated yeast cell is 600.000 in the 650.000MHz scope, includes but not limited to 630,631,632,633,634,635,636,637,638,639,640,641,642,643,644,645,646,647,648,649 and 650MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.395 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 630MHz, 180mV/cm 24 hours; 636MHz, 180mV/cm 24 hours; 641MHz, 180mV/cm 24 hours; 649MHz, 180mV/cm 24 hours; 630MHz, 314mV/cm 24 hours; 636MHz, 314mV/cm 24 hours; 641MHz, 314mV/cm 24 hours; 649MHz, 314mV/cm 24 hours.
The activatory yeast cell is to detect by the growth of measuring shigella under the activated yeast cell existence to the activity of shigella.Make shigella contained in the solid waste extract in substratum, grow to cell counting and reach 10 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell counting of shigella in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 4.6% in the sample of activated yeast cell.
Suppress botulinal yeast cell component
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses botulinal yeast cell.The frequency of electromagnetic field that is used for activated yeast cell is 1000.000 in the 1050.000MHz scope, includes but not limited to 1010,1011,1012,1013,1014,1015,1016,1017,1018,1019,1020,1021,1022,1023,1024,1025,1026,1027,1028,1029,1030,1031,1032,1033,1034 and 1035MHz.For example, the yeast cell of Wine brewing yeast strain AS2.432 is cultivated in the presence of about 25-30 ℃, 8 electromagnetic field in the described substratum of table 4, at one group of following order: 1012MHz, 180mV/cm 24 hours; 1018MHz, 180mV/cm 24 hours; 1024MHz, 180mV/cm 24 hours; 1033MHz, 180mV/cm 24 hours; 1012MHz, 323mV/cm 24 hours; 1018MHz, 323mV/cm 24 hours; 1024MHz, 323mV/cm 24 hours; 1033MHz, 323mV/cm 24 hours.
The activatory yeast cell is that botulinal growth detects under the existence of activatory yeast cell by measuring to botulinal activity.Make Clostridium botulinum contained in the solid waste extract in substratum, grow to cell counting and reach 10 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect botulinal cell counting in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 5.1% in the sample of activated yeast cell.
The yeast cell component that suppresses Clostridium perfringens
In a specific embodiments of the present invention, provide a kind of preparation method who suppresses the yeast cell of Clostridium perfringens.The frequency of electromagnetic field that is used for activated yeast cell is 1100.000 in the 1150.000MHz scope, includes but not limited to 1100,1101,1102,1103,1104,1105,1106,1107,1108,1109,1110,1111,1112,1113,1114,1115,1116,1117,1118,1119 and 1120MHz.For example, will cultivate in the presence of the yeast cell of Wine brewing yeast strain AS2.432 8 electromagnetic field in about 25-30 ℃, the described substratum of table 4, at one group of following order: 1102MHz, 180mV/cm24 hour; 1106MHz, 180mV/cm 24 hours; 1113MHz, 180mV/cm 24 hours; 1117MHz, 180mV/cm 24 hours; 1102MHz, 301mV/cm 24 hours; 1106MHz, 301mV/cm 24 hours; 1113MHz, 301mV/cm 24 hours; 1117MHz, 301mV/cm 24 hours.
The activatory yeast cell is to detect by the growth of measuring Clostridium perfringens under the existence of activatory yeast cell to the activity of Clostridium perfringens.Make Clostridium perfringens contained in the solid waste extract in substratum, grow to cell counting and reach 10 10Cell/ml.Then 1ml activatory and the non-activated yeast cell of 1ml (are all contained 10 7Cell/ml) adds respectively in two 1 liter the bacterial cultures sample, cultivates 24 hours at 20-37 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect the cell counting of Clostridium perfringens in the sample by traditional bacterial cell method of counting.Compare with containing not the sample of activated yeast cell, contain that cell counting has reduced 6.2% in the sample of activated yeast cell.
5.5 adapt to
In another embodiment of the invention, save any restraining activatory yeast cell fully according to 5.1-5.10 and can in the presence of pending solid waste sample, further be cultivated as mixture.Following examples have been described this and have been improved the process for selective of solid waste composition properties.
The extract of pending solid waste such as muck or mud, be by will be about the 1000g brid guano fertile mix, soak with 1000 to 3000ml water prepared in about 48 hours.Then extract and about 1000g argol fertilizer (dry weight, promptly humidity is less than 10%) are mixed forming suspension, yeast cell is added wherein.Contain 5 * 10 to the major general 7The above 1ml yeast of cell/ml adds in the suspension.According to the number of used activated yeast cell bacterial strain, can add the yeast cell of about 50ml.If only use the minority bacterial strain, each bacterial strain can add 5 to the 10ml yeast cell.This method can enlarge or reduce as required in proportion.The mixture of yeast and solid waste was cultivated about 120-280 hour in the presence of one group of electromagnetic field.According to the bacterial strain of used yeast, the frequency of each electromagnetic field meets one of frequency of 5.1-5.4 joint description.If use multiple different yeast strain, can unite and use following 5 frequency band: 20-50MHz, 60-150MHz, 400-700MHz, 1400-1600MHz, 2000-2500MHz; Each is about 24 to 56 hours.Usually, the field intensity that in this method, imposes on yeast cell be at 20mV/cm in the scope of 350mV/cm.
This culture is being cultivated under the round-robin temperature between about 5 ℃ to about 37 ℃.For example, in a typical circulation, the temperature of culture can be from about 37 ℃, kept the about 1-2 of this temperature hour, transfer to 26-30 ℃ then, kept the about 2-4 of this temperature hour, and be reduced to 5-10 ℃ and kept the about 1-2 of this temperature hour then, temperature is increased to about 37 ℃ of another circulations of beginning more then.Recirculation is up to end of processing.After last temperature cycle finished, the temperature of culture was reduced to 3-4 ℃, and keeps the about 5-6 of this temperature hour.Behind this end of processing, as filtering yeast cell is separated from substratum, reclaims with traditional method.Yeast cell after the adaptation is stored in 4 ℃.Fig. 2 has described an exemplary means of this cultural method.
5.6 the preparation of biological composition
Biological composition of the present invention can prepare by the yeast cell culture of yeast cell being cultivated under the suitable conditions according to 5.1 to 5.4 joints and mix aequum.Because biological composition is not just to be used to handle solid waste, the yeast of biological composition can go on foot the desiccating method dryings with two at once.In first drying step, yeast cell is being no more than in first drying machine that drying is no more than 10 minutes under 65 ℃ the temperature, makes the rapid dormancy of yeast cell.Yeast cell is being no more than in second drying machine that drying is no more than 30 minutes under 70 ℃ the temperature then, further removes moisture.After two stages, water-content should be less than 5%.Preferably, should observe temperature and time of drying in two drying step, yeast cell just can not lose activity and function like this.Then the exsiccant yeast cell is cooled to room temperature.Can also screen the exsiccant yeast cell with separator, select the particle of preferred size.Pack the exsiccant yeast cell with sack packer then.
The invention is not restricted to the category of described specific embodiments, they are indivedual examples of all respects of the present invention, and method that function is suitable and component are also in category of the present invention.Undoubtedly, except show here and the content described, according to the description and the accompanying drawing of front, multiple improvement of the present invention will be conspicuous for those those skilled in the art.These improve also within claim category subsequently.

Claims (19)

1. a processing contains the method for antibiotic solid waste, and described method comprises add a plurality of yeast cell in solid waste, makes yeast cell degraded microbiotic, and wherein said a plurality of yeast cell comprise a kind of in following at least:
(a) by with yeast cell in frequency is 70 to 100MHz scopes, field intensity is to cultivate the yeast cell for preparing in 8.5 to one of 250mV/cm or one group of electromagnetic field, it can be degraded and be selected from following microbiotic: penicillin, duomycin, terramycin, doxycycline, tsiklomitsin, Streptomycin sulphate, kantlex, erythromycin, Spiramycin Base and bacitracin;
(b) by with yeast cell in frequency is 410 to 470MHz scopes, field intensity is to cultivate the yeast cell for preparing in 8.5 to one of 250mV/cm or one group of electromagnetic field, it can be degraded and be selected from following microbiotic: Polymyxin E, paraxin and cefoxitin; Or
(c) by with yeast cell in frequency is 550 to 620MHz scopes, field intensity is to cultivate the yeast cell for preparing in 8.5 to one of 250mV/cm or one group of electromagnetic field, it can be degraded and be selected from following microbiotic: Xin Meisu and Vulkamycin. PA-93.
2. a processing contains the method for the solid waste of adverse chemical product, and described method comprises add a plurality of yeast cell in solid waste, makes yeast cell degraded adverse chemical product, and wherein said a plurality of yeast cell comprise a kind of in following at least:
(a) by with yeast cell in frequency is 52 to 98MHz scopes, field intensity is to cultivate the yeast cell for preparing, its can degrade toluene, ethylbenzene or Trichlorophenol in 8 to one of 300mV/cm or one group of electromagnetic field;
(b) by with yeast cell in frequency is 30 to 50MHz or 70 to 98MHz scopes, field intensity is to cultivate the yeast cell for preparing, its xylene compounds of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
(c) by with yeast cell in frequency is 70 to 98MHz or 133 to 151MHz scopes, field intensity is to cultivate the yeast cell for preparing, its phenyl aldehyde of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
(d) by with yeast cell in frequency is 70 to 98MHz or 145 to 162MHz scopes, field intensity is to cultivate the yeast cell for preparing, its propionic aldehyde of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
(e) by with yeast cell in frequency is 70 to 100MHz scopes, field intensity is to cultivate the yeast cell for preparing, its enanthaldehyde of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field; With
(f) by with yeast cell in frequency is 70 to 98MHz or 163 to 183MHz scopes, field intensity is to cultivate the yeast cell for preparing, its dichlorobenzene of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
3. a processing contains the method for the solid waste of adverse chemical product, and described method comprises add a plurality of yeast cell in solid waste, makes yeast cell degraded adverse chemical product, and wherein said a plurality of yeast cell comprise a kind of in following at least:
(a) by with yeast cell in frequency is 70 to 98MHz or 175 to 191MHz scopes, field intensity is to cultivate the yeast cell for preparing, its methyl phenyl ketone of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
(b) by with yeast cell in frequency is 70 to 98MHz or 183 to 205MHz scopes, field intensity is to cultivate the yeast cell for preparing, its arsanilic acid of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
(c) by with yeast cell in frequency is 70 to 98MHz or 114 to 128MHz scopes, field intensity is to cultivate the yeast cell for preparing, its roxarsone of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
(d) by with yeast cell in frequency is 70 to 98MHz or 200 to 220MHz scopes, field intensity is to cultivate the yeast cell for preparing, its Nifurazolidone of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field;
(e) by with yeast cell in frequency is 70 to 98MHz or 213 to 229MHz scopes, field intensity is to cultivate the yeast cell for preparing, its M B 15497 of can degrading in 8 to one of 250mV/cm or one group of electromagnetic field; With
(f) by with yeast cell in frequency is 70 to 98MHz or 220 to 250MHz scopes, field intensity is to cultivate the yeast cell for preparing, its can degrade trichlorophonum or dinitolmide in 8 to one of 250mV/cm or one group of electromagnetic field.
4. a processing contains the method for the solid waste of adverse chemical product, and described method comprises add a plurality of yeast cell in solid waste, makes yeast cell reduce the amount of adverse chemical product, and wherein said a plurality of yeast cell comprise a kind of in following at least:
(a) by with yeast cell in frequency is 660 to 680MHz or 2160 to 2190MHz scopes, field intensity is to cultivate the yeast cell for preparing in 25 to one of 300mV/cm or one group of electromagnetic field, it can reduce the amount of ammonium compound;
(b) by with yeast cell in frequency is 661 to 680MHz scopes, field intensity is to cultivate the yeast cell for preparing in 8 to one of 250mV/cm or one group of electromagnetic field, it can reduce the amount of nitrate or nitrite; Or
(c) by with yeast cell in frequency is 86 to 120MHz or 410 to 430MHz scopes, field intensity is to cultivate the yeast cell for preparing in 8 to one of 250mV/cm or one group of electromagnetic field, it can reduce phosphatic amount.
5. a processing contains the method for the solid waste of adverse chemical product, and described method comprises add a plurality of yeast cell in solid waste, makes yeast cell degraded adverse chemical product, and wherein said a plurality of yeast cell comprise a kind of in following at least:
(a) by with yeast cell in frequency is 1980 to 2118MHz scopes, field intensity is to cultivate the yeast cell for preparing in 25 to one of 300mV/cm or one group of electromagnetic field, it can be degraded and be selected from following adverse chemical product: Trichlorphon, SD-1750, monocrotophos and Rogor;
(b) by with yeast cell in frequency is 1420 to 1435MHz scopes, field intensity is to cultivate the yeast cell for preparing in 25 to one of 300mV/cm or one group of electromagnetic field, it can degradation of dichloro-diphenyl-trichloroethane or toxaphene.
6. method that reduces the solid waste smell, comprise and in solid waste, add a plurality of yeast cell, make yeast cell reduce the amount of scent of molecule in the solid waste, wherein said yeast cell by with yeast cell in frequency is 2160 to 2380MHz scopes, field intensity is to cultivate in 25 to 300mV/cm one or the one group of electromagnetic field to prepare, described molecule odorous is selected from following: hydrogen sulfide, ammonia, indoles, skatole, acetate, methylamine and p-cresol.
7. a processing contains the method for the solid waste of pathogenic bacterium, and described method comprises add a plurality of yeast cell in solid waste, makes yeast cell suppress the growths of pathogenic bacterium in the solid waste, and wherein said a plurality of yeast cell comprise a kind of in following at least:
(a) by with yeast cell in frequency is 20 to 50MHz scopes, field intensity is to cultivate the yeast cell for preparing in 20 to one of 350mV/cm or one group of electromagnetic field, it can suppress streptococcus aureus, pneumococcus, anthrax bacillus, Mycobacterium tuberculosis, Salmonellas or colibacillary growth;
(b) by with yeast cell in frequency is 500 to 550MHz scopes, field intensity is to cultivate the yeast cell for preparing in 20 to one of 350mV/cm or one group of electromagnetic field, it can suppress the growth of vibrios;
(c) by with yeast cell in frequency is 600 to 650MHz scopes, field intensity is to cultivate the yeast cell for preparing in 20 to one of 350mV/cm or one group of electromagnetic field, it can suppress the growth of Shigellae;
(d) by with yeast cell in frequency is 1000 to 1050MHz scopes, field intensity is to cultivate the yeast cell for preparing in 20 to one of 350mV/cm or one group of electromagnetic field, it can suppress botulinal growth; With
(e) by with yeast cell in frequency is 1100 to 1150MHz scopes, field intensity is to cultivate the yeast cell for preparing in 20 to one of 50mV/cm or one group of electromagnetic field, it can suppress the growth of Clostridium perfringens.
8. a method of handling solid waste comprises adding a kind of biological composition in solid waste, and described biological composition comprises a kind of of following yeast cell component at least:
(a) the first yeast cell component, it comprises antibiotic a plurality of yeast cell in the degradable solid refuse, the described first yeast cell component prepares by yeast cell is cultivated in one or one group of following electromagnetic field: frequency is selected from 70 to 100MHz, 410 to 470MHz and 550 to 620MHz, and field intensity is 8 to 250mV/cm;
(b) the second yeast cell component, it comprises a plurality of yeast cell of adverse chemical product in the degradable solid refuse, the described second yeast cell component prepares by yeast cell is cultivated in one or one group of following electromagnetic field: frequency is selected from 30 to 100MHz, 70 to 280MHz, 410 to 430MHz, 660 to 680MHz and 1980 to 2210MHz, and field intensity is 8 to 250mV/cm;
(c) the 3rd yeast cell component, it comprises a plurality of yeast cell that reduce the amount of scent of molecule in the solid waste, described the 3rd yeast cell component be by with yeast cell one or a class frequency 2160 to 2380MHz, field intensity cultivates in the electromagnetic field of 300mV/cm scope 25 and prepares;
(d) the 4th yeast cell component, it comprises a plurality of yeast cell that suppress pathogenic bacterium growth in the solid waste, described the 4th yeast cell component prepares by yeast cell is cultivated in one or one group of following electromagnetic field: frequency is selected from 30 to 50MHz, 500 to 550MHz, 600 to 650MHz, 1000 to 1050MHz and 1100 to 1150MHz, and field intensity is 20 to 350mV/cm; With
Make the amount of microbiotic, adverse chemical product, scent of compound and pathogenic bacterium in the yeast cell minimizing solid waste in the yeast cell component.
9. the method for claim 8, wherein biological composition comprises yeast cell component (a) and (b), (c) and (d).
10. the method for claim 8, wherein said yeast cell is the cell that is selected from following yeast kind: yeast saccharomyces cerevisiae, Xue's watt yeast, De Shi yeast, saccharomyces exiguus, saccharomyces fermentati, Saccharomyces logos, honeybee yeast, Saccharomyces microellipsoides, ellipsoideus yeast, Luo Shi yeast, Lu Shi yeast, saccharomyces sake, saccharomyces uvarum, Wei Shi yeast, saccharomyces ludwigii, Saccharomyces sinenses and saccharomyces carlsbergensis.
11. the method for claim 8, wherein said yeast cell is a brewing yeast cell.
12. the method for claim 11, wherein said biological composition comprises dry yeast cell, adds about 300 in every cubic metre of solid waste to this biological composition of 600g.
13. the method for claim 11 wherein adds in described solid waste before the described dry yeast cell, with described dry yeast cell and the water mixed according to about 30 liters of about 1000g yeast cell, and cultivates about 12 to 24 hours.
14. composition that comprises antibiotic a plurality of yeast cell in the degradable solid refuse, wherein said a plurality of yeast cell prepares yeast cell cultured method in or one group of electromagnetic field by comprising, described electromagnetic field has (i) one or more frequencies that are selected from following scope: 70 to 100MHz, 410 to 470MHz and 550 to 620MHz and (ii) field intensity 8 to 250mV/cm.
15. composition that comprises a plurality of yeast cell of adverse chemical product in the degradable solid refuse, wherein said a plurality of yeast cell prepares yeast cell cultured method in or one group of electromagnetic field by comprising, described electromagnetic field has (i) one or more frequencies that are selected from following scope: 30 to 100MHz, 70 to 280MHz, 410 to 430MHz, 660 to 680MHz and 1980 to 2210MHz and (ii) field intensity 8 to 250mV/cm.
16. composition that comprises a plurality of yeast cell that reduce the solid waste smell, wherein said a plurality of yeast cell prepares yeast cell cultured method in or one group of electromagnetic field by comprising, described electromagnetic field has (i) one or more 2160 frequency and (ii) field intensity 25 to 300mV/cm in the 2380MHz scope.
17. one kind comprises the composition that suppresses a plurality of yeast cell of pathogenic bacterium growth in the solid waste, wherein said a plurality of yeast cell prepares yeast cell cultured method in or one group of electromagnetic field by comprising, described electromagnetic field has (i) one or more frequencies that are selected from following scope: 30 to 50MHz, 500 to 550MHz, 600 to 650MHz, 1000 to 1050MHz and 1100 to 1150MHz and (ii) field intensity 20 to 350mV/cm.
18. claim 14,15,16 or 17 composition, wherein said yeast cell is the cell that is selected from following yeast kind: yeast saccharomyces cerevisiae, Xue's watt yeast, De Shi yeast, saccharomyces exiguus, saccharomyces fermentati, Saccharomyces logos, honeybee yeast, Saccharomycesmicroellipsoides, ellipsoideus yeast, Luo Shi yeast, Lu Shi yeast, saccharomyces sake, saccharomyces uvarum, Wei Shi yeast, saccharomyces ludwigii, Saccharomyces sinenses and saccharomyces carlsbergensis.
19. claim 14,15,16 or 17 composition, wherein said yeast cell is a dry yeast cell.
CNA028090608A 2001-03-01 2002-03-01 Biological compositions for solid waste treatment Pending CN1505680A (en)

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US09/797,437 2001-03-01
US09/797,371 US6391617B1 (en) 2001-03-01 2001-03-01 Yeast compositions for converting bio-available nitrogen in a culture medium to intracellular nitrogen
US09/797,493 2001-03-01
US09/797,372 US20020123127A1 (en) 2001-03-01 2001-03-01 Methods and compositions for reducing odor
US09/797,378 US6391618B1 (en) 2001-03-01 2001-03-01 Methods and compositions for degrading environmental toxins
US09/797,437 US6391619B1 (en) 2001-03-01 2001-03-01 Methods and compositions for suppressing growth of algae
US09/797,381 2001-03-01
US09/797,382 US20020123129A1 (en) 2001-03-01 2001-03-01 Methods and compositions for degrading nitrogen-containing compounds
US09/797,382 2001-03-01
US09/797,381 US6436695B1 (en) 2001-03-01 2001-03-01 Yeast compositions for converting bio-available phosphorus in a culture medium to intracellular phosphorus
US09/797,378 2001-03-01
US09/797,371 2001-03-01
US09/797,377 2001-03-01
US09/797,493 US6440713B1 (en) 2001-03-01 2001-03-01 Methods and compositions for suppressing growth of pathogenic microbes
US09/797,377 US20020123130A1 (en) 2001-03-01 2001-03-01 Methods and compositions for degrading polymeric compounds
US09/797,372 2001-03-01

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