CN105802749A - Oil stain cleaner containing microbiological strain and preparation method - Google Patents
Oil stain cleaner containing microbiological strain and preparation method Download PDFInfo
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- CN105802749A CN105802749A CN201610154169.1A CN201610154169A CN105802749A CN 105802749 A CN105802749 A CN 105802749A CN 201610154169 A CN201610154169 A CN 201610154169A CN 105802749 A CN105802749 A CN 105802749A
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- fermentation
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- greasy dirt
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- 238000002360 preparation method Methods 0.000 title claims abstract description 41
- 230000002906 microbiologic effect Effects 0.000 title abstract 2
- 238000000855 fermentation Methods 0.000 claims abstract description 138
- 230000004151 fermentation Effects 0.000 claims abstract description 138
- 241000194103 Bacillus pumilus Species 0.000 claims abstract description 60
- 239000007788 liquid Substances 0.000 claims abstract description 36
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 34
- 230000004913 activation Effects 0.000 claims abstract description 23
- 239000004094 surface-active agent Substances 0.000 claims abstract description 18
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims abstract description 17
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 17
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 17
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 17
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 13
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 13
- 238000004321 preservation Methods 0.000 claims abstract description 9
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 7
- 241000193755 Bacillus cereus Species 0.000 claims description 53
- 239000003599 detergent Substances 0.000 claims description 29
- 239000012530 fluid Substances 0.000 claims description 29
- 239000012452 mother liquor Substances 0.000 claims description 29
- 230000000813 microbial effect Effects 0.000 claims description 27
- 239000000203 mixture Substances 0.000 claims description 18
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 16
- 244000061456 Solanum tuberosum Species 0.000 claims description 15
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 230000000284 resting effect Effects 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 235000012424 soybean oil Nutrition 0.000 claims description 4
- 239000003549 soybean oil Substances 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 3
- 230000007613 environmental effect Effects 0.000 claims 1
- 241000894006 Bacteria Species 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 7
- 238000004140 cleaning Methods 0.000 abstract description 3
- 239000004480 active ingredient Substances 0.000 abstract 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 abstract 1
- 235000019797 dipotassium phosphate Nutrition 0.000 abstract 1
- 229960001484 edetic acid Drugs 0.000 abstract 1
- 238000004945 emulsification Methods 0.000 abstract 1
- 239000004615 ingredient Substances 0.000 abstract 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 abstract 1
- 230000007774 longterm Effects 0.000 abstract 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 20
- 230000001954 sterilising effect Effects 0.000 description 12
- 244000005700 microbiome Species 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 238000009395 breeding Methods 0.000 description 6
- 230000001488 breeding effect Effects 0.000 description 6
- 239000008157 edible vegetable oil Substances 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 239000000706 filtrate Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 235000019198 oils Nutrition 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000004659 sterilization and disinfection Methods 0.000 description 6
- 230000001804 emulsifying effect Effects 0.000 description 5
- 150000002632 lipids Chemical class 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000006731 degradation reaction Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000002699 waste material Substances 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 239000007795 chemical reaction product Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 125000005456 glyceride group Chemical group 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 235000012015 potatoes Nutrition 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005686 eating Nutrition 0.000 description 1
- 235000006694 eating habits Nutrition 0.000 description 1
- 238000012851 eutrophication Methods 0.000 description 1
- 230000003311 flocculating effect Effects 0.000 description 1
- 239000010794 food waste Substances 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000033764 rhythmic process Effects 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D1/00—Detergent compositions based essentially on surface-active compounds; Use of these compounds as a detergent
- C11D1/66—Non-ionic compounds
- C11D1/667—Neutral esters, e.g. sorbitan esters
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/02—Inorganic compounds ; Elemental compounds
- C11D3/04—Water-soluble compounds
- C11D3/046—Salts
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/02—Inorganic compounds ; Elemental compounds
- C11D3/04—Water-soluble compounds
- C11D3/06—Phosphates, including polyphosphates
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/26—Organic compounds containing nitrogen
- C11D3/33—Amino carboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/381—Microorganisms
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D2111/00—Cleaning compositions characterised by the objects to be cleaned; Cleaning compositions characterised by non-standard cleaning or washing processes
- C11D2111/10—Objects to be cleaned
- C11D2111/14—Hard surfaces
- C11D2111/20—Industrial or commercial equipment, e.g. reactors, tubes or engines
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Inorganic Chemistry (AREA)
- Microbiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses an oil stain cleaner containing a microbiological strain and a preparation method.The cleaner comprises, by weight, 0.050-0.500 part of a surface active agent, 0.030-0.400 part of dipotassium phosphate, 0.005-0.050 part of ferrous sulfate, 0.005-0.050 part of magnesium sulfate, 0.001-0.005 part of edetic acid and 99.0-99.9 parts of fermentation liquor; the number of viable bacteria of MoHaiWei bacillus accounts for 60-70% the total number of viable bacteria in active ingredients of the fermentation liquor, and the number of viable bacteria of bacillus pumilus accounts for 30-40% the total number of viable bacteria.The preparation method includes the steps of activation of the strain, preparation of strain liquid, fermentation of the strain and preservation of the strain, and a finished product of the cleaner is obtained.The prepared cleaner has an emulsification function on oil stain ingredients in municipal pipes and can achieve a long-term oil cleaning effect and prevent the municipal pipes form being blocked.
Description
Technical field
The present invention relates to greasy dirt cleaning treatment technical field, be specifically related to a kind of greasy dirt detergent containing microbial strains and preparation method thereof.
Background technology
Along with expanding economy, the material life of people is more and more abundanter, and rhythm of life is increasingly faster, and Household diet not only becomes abundant, and becomes more greasy.Edible oil use and waste becomes more, simultaneously as the reason of work and the change of dietary habit, the chance of people's outside eating also gets more and more.
And at home due to but without perfect refuse classification system and regulation, lubricant component significant portion in changing food waste is to enter in city planting ductwork by sewer pipe network, other materials mixing easily and in city planting ductwork, form oils mixture, along with temperature and seasonal variations, easily become solid kind material, block pipeline, produce the harmful gass such as stink.The cleaning of the city planting ductwork that domestic and international market is sold and dredging agent, composition is most to be made up of alkalescence or acid class chemical substance, having danger, and pipeline has corrosivity, persistency is poor, continuous discontinuity is needed to add, bringing very big inconvenience to using, and pipe network system is caused potential safety hazard, therefore it does not only reach the purpose efficiently removing oils and fats, and there is many drawbacks in its use procedure, is not suitable for widespread adoption.
Summary of the invention
It is an object of the invention to provide a kind of greasy dirt detergent containing microbial strains and preparation method thereof, for solve existing greasy dirt detergent can not effective oils composition in emulsifying and degraded pipe network, and cannot for a long time, the problem that effectively achieves the effect of edible vegetable oil and dredging pipeline.
To achieve these goals, the present invention provides following technical scheme: a kind of greasy dirt detergent containing microbial strains, it is characterized in that, the described greasy dirt detergent containing microbial strains includes following components: surfactant 0.050～0.500 weight portion, dipotassium hydrogen phosphate 0.030～0.400 weight portion, ferrous sulfate 0.005～0.050 weight portion, magnesium sulfate 0.005～0.050 weight portion, ethylenediaminetetraacetic acid 0.001～0.005 weight portion and fermentation liquid 99.0～99.9 weight portion；
The active component of described fermentation liquid can include Mo Haiwei bacillus cereus and Bacillus pumilus, wherein, it is 60～70% that the viable count of described Mo Haiwei bacillus cereus accounts for the percentage ratio of total viable count, and it is 30～40% that the viable count of described Bacillus pumilus accounts for the percentage ratio of total viable count.
Compared to prior art, greasy dirt detergent containing microbial strains of the present invention has the advantage that in the greasy dirt detergent containing microbial strains, active component is the Mo Haiwei bacillus cereus in fermentation liquid and Bacillus pumilus, both strains are all the natural function microorganism species screened from the environment that greasy dirt exists, therefore raw material natural environmental-protective, without follow-up pollution.Process via the later stage and allotment, obtain final greasy dirt detergent, it can produce natural biological metabolic product in breeding and degradation process, such as functional components such as sugar lipid surfactant, protease, cellulase, greasy dirt composition in city planting ductwork can be played the effect of emulsifying by it, the greasy dirt class solids absorbed and degrade in city planting ductwork, effectively facilitates the degraded of organic species, it is prevented that pipe network blocks；Meanwhile, it also can promote the formation of beneficial microbe colony, it is suppressed that the growth of harmful bacteria, reduces the generation of the harmful gass such as stink.Owing to natural function microorganism species relies on the metabolism of himself and degraded to carry out growing and breeding in city planting ductwork, so the effect of edible vegetable oil and dredging pipeline can be kept for a long time, persistency is strong.
nullIn addition，In order to make functional microorganism flora to play a role better，The fermentation liquid of its formation may further be enriched with appropriate surfactant、Dipotassium hydrogen phosphate、Ferrous sulfate、Magnesium sulfate and ethylenediaminetetraacetic acid，Surfactant has emulsifying、Solubilising、Washing、Anticorrosions etc. act on，Dipotassium hydrogen phosphate is microorganism、The culture medium of mushroom，Therefore nutrition can be further provided for for functional microorganism flora，Ferrous sulfate can be used for the flocculating and purifying of water，Simultaneously also can the eutrophication of anti-water-stop body and affect the growth of flora，Magnesium sulfate also can provide nutritional labeling，Ethylenediaminetetraacetic acid is a kind of chelating agen being combined with metal ion，Therefore its be added in the greasy dirt detergent that the present invention contains microbial strains can the heavy-metal ion removal inhibitory action to enzyme，Promote that the enzyme that Mo Haiwei bacillus cereus and two kinds of functional microorganism floras of Bacillus pumilus produce plays a role.
The preparation method that the present invention also provides for the above-mentioned greasy dirt detergent containing microbial strains, this preparation method comprises the steps:
Step S1: the activation of strain
Will be equipped with two slant tubes of Mo Haiwei Bacillus and Bacillus pumilus strain respectively to be placed in constant incubator and cultivate, incubation time is 110～130h, cultivation temperature in constant incubator is 30～35 DEG C, respectively obtains the Mo Haiwei Bacillus after activation and Bacillus pumilus；
Step S2: the preparation of seed liquor
To be inoculated in two fluid mediums respectively each carry out shaking table cultivation by the Bacillus pumilus after the step S1 Mo Haiwei bacillus cereus activated prepared and activation, to a fluid medium, the viable count of Mo Haiwei bacillus cereus is not less than 1.0 × 108Individual/ml, then obtain Mo Haiwei bacillus cereus seed liquor；To another fluid medium, the viable count of Bacillus pumilus is not less than 1.0 × 108Individual/ml, then obtain Bacillus pumilus seed liquor；
Step S3: the fermentation of strain
Carrying out first time fermentation culture by accessing in equipped with the fermentation tank of fermentation mother liquor through the step S2 Bacillus pumilus seed liquor prepared, the first time temperature of fermentation culture is 30～32 DEG C, and the time of fermentation culture first time is 8～10h；Wherein, volume is fermentation mother liquor volume 0.025～0.05% of the Bacillus pumilus seed liquor in addition fermentation tank；
Then carrying out second time fermentation culture by continuing to access in fermentation tank through the step S2 Mo Haiwei bacillus cereus seed liquor prepared, to fermentation tank, total viable count of Mo Haiwei bacillus cereus and Bacillus pumilus is not less than 5.0 × 108Individual/ml, the temperature of second time fermentation culture is 30～32 DEG C, and the time of second time fermentation culture is 36～48h, obtains strain primary fermentation liquid；Wherein, volume is fermentation mother liquor volume 0.025～0.05% of the Mo Haiwei bacillus cereus seed liquor in addition fermentation tank；
Step S4: the preservation of strain
To preserve by the step S3 strain primary fermentation liquid prepared, the fermentation liquid that must complete；
Step S5: abluent finished product
Surfactant, dipotassium hydrogen phosphate, ferrous sulfate, magnesium sulfate and ethylenediaminetetraacetic acid is added in the fermentation liquid completed through step S4, and parts by weight respectively surfactant 0.050～0.500 weight portion of each component, dipotassium hydrogen phosphate 0.030～0.400 weight portion, ferrous sulfate 0.005～0.050 weight portion, magnesium sulfate 0.005～0.050 weight portion, ethylenediaminetetraacetic acid 0.001～0.005 weight portion and fermentation liquid 99.0～99.9 weight portion, each component of mix homogeneously prepares abluent finished product.
Compared to prior art, greasy dirt detergent containing microbial strains of the present invention and preparation method thereof has the advantage that and adopts the mode of fermentation to process Mo Haiwei bacillus cereus and Bacillus pumilus both functional microorganism floras so that it is produce sugar lipid surfactant during the fermentation and multiple enzyme enters in fermentation liquid.So that when the greasy dirt detergent adopting the present invention to contain microbial strains, the oils composition that can directly exist in city planting ductwork is primary carbon source, by the sugared lipid surfactant produced and enzyme, oils composition in emulsifying and degraded city planting ductwork, the breeding of the harmful bacteria in suppression environment, avoid the generation of the problems such as stink, growth and the breeding of endogenous profitable strain in environment can be promoted again, such that it is able to reach the effect of edible vegetable oil long, effective and dredging pipeline simultaneously.And the cost of simple, easily controllable, the overall raw material of above-mentioned fermentation technology and preparation method is all relatively low, has good economic and social benefit.
Accompanying drawing explanation
By reading hereafter detailed description of the preferred embodiment, various other advantage and benefit those of ordinary skill in the art be will be clear from understanding.Accompanying drawing is only for illustrating the purpose of preferred implementation, and is not considered as limitation of the present invention.In the accompanying drawings:
Fig. 1 illustrates the flow chart of the preparation method of a kind of greasy dirt detergent containing microbial strains of the present invention.
Detailed description of the invention
The invention provides many applicable creative concepts, this creativeness concept can be reflected in a large number of in concrete context.It is only used as the exemplary illustration of the specific embodiment of the present invention in the specific embodiment described in following embodiments of the present invention, and is not meant to limit the scope of the invention.
Below in conjunction with accompanying drawing and specific embodiment, the invention will be further described.
In order to detect the edible vegetable oil effect of the greasy dirt detergent of the microbial strains contained in following each embodiment, intercepting same section of greasy dirt discharge pipeline section, the greasy dirt discharge pipeline section total length of intercepting is 150cm, and internal diameter is 4cm.This greasy dirt being discharged pipeline section and is equally divided into three sections, every segment length is 50cm, respectively every section of greasy dirt is discharged pipeline section and is numbered 1,2 and 3.Every section of greasy dirt discharge pipeline section before treatment is weighed respectively, wherein,
The quality m1 of the greasy dirt discharge pipeline section being numbered 1 is 856.45g；
The quality m1 of the greasy dirt discharge pipeline section being numbered 2 is 834.63g；
The quality m1 of the greasy dirt discharge pipeline section being numbered 3 is 803.82g.
The present invention provides a kind of greasy dirt detergent containing microbial strains, and the consumption that makes of the greasy dirt detergent containing microbial strains in following each embodiment is the 0.05% of every section of greasy dirt discharge pipeline section total measurement (volume).By calculating, the volume of above-mentioned every section of greasy dirt discharge pipeline section is 628cm3, then in each embodiment, the consumption that makes of the greasy dirt detergent containing microbial strains of every section of greasy dirt discharge pipeline section use is 0.314ml.
Preparation method and the composition thereof of the greasy dirt detergent containing microbial strains is further described hereafter by specific embodiment.
Embodiment one
As it is shown in figure 1, the present embodiment adopts following step to prepare an abluent successively.
Step S1: the activation of strain
Two slant tubes being respectively provided with Mo Haiwei Bacillus and Bacillus pumilus strain are placed in the constant incubator of 32 DEG C and cultivate 120h, respectively obtain the Mo Haiwei bacillus cereus after activation and the Bacillus pumilus after activation.
Step S2: the preparation of seed liquor
The preparation of fluid medium: take peeled potatoes 200.0g, it is cut into the fritter of 1 centimeter square, after the 800ml that adds water boils, little fire keeps boiling 30 minutes, use three layers filtered through gauze, can repeatedly produce, make filtrate reach 1000ml, the filtrate obtained is the potato juice (being left out impurity) that potato quality mark is 20%, takes sucrose 20g and is dissolved in potato juice completing the preparation of fluid medium.Fluid medium after preparation adopts steam sterilization mode sterilizing 20 minutes under the environment of 121 DEG C, obtains the fluid medium produced.Two fluid mediums are produced in the manner described above.
The preparation of Mo Haiwei bacillus cereus seed liquor: carry out shaking table cultivation by being inoculated in a fluid medium by the Mo Haiwei bacillus cereus after the step S1 activation prepared, the rotating speed that shaking table is cultivated is 150r/min, cultivation temperature is 32 DEG C, after cultivating 48 hours, the viable count of the Mo Haiwei bacillus cereus after cultivation is 1.2 × 108CFU/ml, obtains Mo Haiwei bacillus cereus seed liquor；
The preparation of Bacillus pumilus seed liquor: will be inoculated in another fluid medium carry out shaking table cultivation by the Bacillus pumilus after the step S1 activation prepared, the rotating speed that shaking table is cultivated is 150r/min, cultivation temperature is 32 DEG C, after cultivating 48 hours, after cultivation, the viable count of Bacillus pumilus is 1.4 × 108CFU/ml, obtains Bacillus pumilus seed liquor.
Wherein, shaking table is cultivated and is referred to and be positioned in shaking table by thalline to carry out the process cultivated, and in the process that shaking table is cultivated, thalline and culture medium are fully contacted, and meet the needs of thalli growth, increase dissolved oxygen amount simultaneously, are beneficial to the breathing of thalline.
Step S3: the fermentation of strain
Producing of fermentation mother liquor: measure ammonium nitrate 7.5g, sodium chloride 3.0g, dipotassium hydrogen phosphate 2.0g, magnesium sulfate 0.2g, green vitriol 0.2g, yeast extract 0.5g, one-level soybean oil 4.0g, distilled water 1000ml mix homogeneously make solution, and to regulate pH be 8.0, fermentation mother liquor after being configured, fermentation mother liquor after configuration is accessed in fermentation tank, at the temperature of 121 DEG C, adopt steam sterilization mode sterilizing 60min, obtain the fermentation mother liquor produced.
To access equipped with the fermentation tank of fermentation mother liquor carries out first time fermentation culture by the seed liquor of the step S2 Bacillus pumilus prepared, and the volume of Bacillus pumilus seed liquor is fermentation mother liquor volume 0.03%, first time fermentation culture is cultivation 9h under the condition of culture of constant temperature 31 DEG C；Then will continue to access in the fermentation tank after first time fermentation culture by the step S2 Mo Haiwei bacillus cereus seed liquor prepared, proceed second time fermentation culture, and volume is fermentation mother liquor volume the 0.03% of Mo Haiwei bacillus cereus seed liquor, second time fermentation culture is cultivation 40h under the condition of culture of constant temperature 31 DEG C.Above-mentioned first time fermentation culture and second time fermentation culture all adopt the mode of aerobic cultivation.Aerobic cultivation refers to cultivating microorganism under the environment of aerobic, makes microorganism and air be fully contacted and be bred in such circumstances and metabolism.Second time fermentation culture terminates, and Mo Haiwei bacillus cereus and total viable count of Bacillus pumilus in fermentation tank are 6.4 × 108Individual/ml, obtains strain primary fermentation liquid.
Step S4: the preservation of strain
By be the airtight resting state preservation of lucifuge 6 days when 15 DEG C by the step S3 strain primary fermentation liquid prepared, obtain the fermentation liquid completed.
Step S5: abluent finished product
Take through step S1～S4 fermentation liquid 1000g completed, adding surfactant 2.51g, dipotassium hydrogen phosphate 2.01g, ferrous sulfate 0.25g, magnesium sulfate 0.25g and ethylenediaminetetraacetic acid 0.03g in fermentation liquid, the end-product that mix homogeneously obtains is an abluent.It should be noted that surfactant, dipotassium hydrogen phosphate, ferrous sulfate, magnesium sulfate and ethylenediaminetetraacetic acid are commercially available prod in embodiment.
Surfactant refers to the material adding the interface state generation significant change that can make its solution system on a small quantity, and it has fixing hydrophilic and oleophilic group, can align on the surface of solution.Surfactant in the present embodiment is preferably fatty glyceride.
According to detection, in an abluent, it is 64% that the viable count of Mo Haiwei bacillus cereus accounts for the percentage ratio of total viable count, and it is 36% that the viable count of Bacillus pumilus accounts for the percentage ratio of total viable count.Mo Haiwei bacillus cereus concentration in deodorizer is 4.67 × 108Individual/ml, bacillus pumilus concentration in deodorizer is 2.63 × 108Individual/ml.
This abluent is applied in the greasy dirt discharge pipeline section being numbered 1, occupation mode is that the abluent taking 0.314ml directly sprays in the pipeline opening of the greasy dirt discharge pipeline section being numbered 1, after standing three days, again weigh this and be numbered the greasy dirt discharge pipeline section of 1, weight m2=263.54g.Dispel after totally it addition, this greasy dirt to be discharged the greasy dirt on pipeline section, again weigh this and be numbered greasy dirt discharge pipeline section of 1, nt wt net weight m3=245.20g.
According to formula w=(m1-m2)/(m1-m3) × 100%, obtain the oily waste degradation rate w=97.0% of an abluent.
Embodiment two
Continuing referring to Fig. 1, the present embodiment adopts following step to prepare No. two abluents successively.
Step S1: the activation of strain
Two slant tubes being respectively provided with Mo Haiwei Bacillus and Bacillus pumilus strain are placed in the constant incubator of 30 DEG C and cultivate 130h, respectively obtain the Mo Haiwei bacillus cereus after activation and the Bacillus pumilus after activation.
Step S2: the preparation of seed liquor
The preparation of fluid medium: take peeled potatoes 150.0g, it is cut into the fritter of 1 centimeter square, after the 850ml that adds water boils, little fire keeps boiling 30 minutes, use three layers filtered through gauze, can repeatedly produce, make filtrate reach 1000ml, the filtrate obtained is the potato juice (being left out impurity) that potato quality mark is 15%, takes sucrose 25g and is dissolved in potato juice completing the preparation of fluid medium.Fluid medium after preparation adopts steam sterilization mode sterilizing 20 minutes under the environment of 121 DEG C, obtains the fluid medium produced.Two fluid mediums are produced in the manner described above.
The preparation of Mo Haiwei bacillus cereus seed liquor: carry out shaking table cultivation by being inoculated in a fluid medium by the Mo Haiwei bacillus cereus after the step S1 activation prepared, the rotating speed that shaking table is cultivated is 130r/min, cultivation temperature is 35 DEG C, after cultivating 50 hours, the viable count of the Mo Haiwei bacillus cereus after cultivation is 1.3 × 108CFU/ml, obtains Mo Haiwei bacillus cereus seed liquor；
The preparation of Bacillus pumilus seed liquor: will be inoculated in another fluid medium carry out shaking table cultivation by the Bacillus pumilus after the step S1 activation prepared, the rotating speed that shaking table is cultivated is 130r/min, cultivation temperature is 35 DEG C, after cultivating 50 hours, after cultivation, the viable count of Bacillus pumilus is 1.5 × 108CFU/ml, obtains Bacillus pumilus seed liquor.
Step S3: the fermentation of strain
Producing of fermentation mother liquor: measure ammonium nitrate 4.04g, sodium chloride 2.02gg, dipotassium hydrogen phosphate 1.01g, magnesium sulfate 0.1g, green vitriol 0.1g, yeast extract 0.3g, one-level soybean oil 2.53g, distilled water 1000ml mix homogeneously make solution, and to regulate pH be 7.5, fermentation mother liquor after being configured, fermentation mother liquor after configuration is accessed in fermentation tank, at the temperature of 121 DEG C, adopt steam sterilization mode sterilizing 60min, obtain the fermentation mother liquor produced.
To access equipped with the fermentation tank of fermentation mother liquor carries out first time fermentation culture by the seed liquor of the step S2 Bacillus pumilus prepared, and the volume of Bacillus pumilus seed liquor is fermentation mother liquor volume 0.025%, first time fermentation culture is cultivation 10h under the condition of culture of constant temperature 30 DEG C；Then will continue to access in the fermentation tank after first time fermentation culture by the step S2 Mo Haiwei bacillus cereus seed liquor prepared, proceed second time fermentation culture, and volume is fermentation mother liquor volume the 0.025% of Mo Haiwei bacillus cereus seed liquor, second time fermentation culture is cultivation 48h under the condition of culture of constant temperature 30 DEG C.Above-mentioned first time fermentation culture and second time fermentation culture all adopt the mode of aerobic cultivation.Second time fermentation culture terminates, and Mo Haiwei bacillus cereus and total viable count of Bacillus pumilus in fermentation tank are 7.0 × 108Individual/ml, obtains strain primary fermentation liquid.
Step S4: the preservation of strain
By be the airtight resting state preservation of lucifuge 7 days when 18 DEG C by the step S3 strain primary fermentation liquid prepared, obtain the fermentation liquid completed.
Step S5: abluent finished product
Take through step S1～S4 fermentation liquid 1000g completed, adding fatty glyceride 0.50g, dipotassium hydrogen phosphate 0.30g, ferrous sulfate 0.50g, magnesium sulfate 0.50g and ethylenediaminetetraacetic acid 0.01g in fermentation liquid, the end-product that mix homogeneously obtains is No. two abluents.
According to detection, in No. two abluents, it is 69% that the viable count of Mo Haiwei bacillus cereus accounts for the percentage ratio of total viable count, and it is 31% that Bacillus pumilus accounts for the percentage ratio of total viable count.Mo Haiwei bacillus cereus concentration in deodorizer is 4.97 × 108Individual/ml, bacillus pumilus concentration in deodorizer is 2.23 × 108Individual/ml.
These No. two abluents are applied in the greasy dirt discharge pipeline section being numbered 2, occupation mode same abluent, after standing three days, again weigh this and are numbered the greasy dirt discharge pipeline section of 2, weight m2=267.42g.Dispel after totally it addition, this greasy dirt to be discharged the greasy dirt on pipeline section, again weigh this and be numbered greasy dirt discharge pipeline section of 2, nt wt net weight m3=241.32g.
According to formula w=(m1-m2)/(m1-m3) × 100%, obtain the oily waste degradation rate w=95.6% of No. two abluents.
Embodiment three
Continuing referring to Fig. 1, the present embodiment adopts following step to prepare No. three abluents successively.
Step S1: the activation of strain
Two slant tubes being respectively provided with Mo Haiwei Bacillus and Bacillus pumilus strain are placed in the constant incubator of 35 DEG C and cultivate 130h, respectively obtain the Mo Haiwei bacillus cereus after activation and the Bacillus pumilus after activation.
Step S2: the preparation of seed liquor
The preparation of fluid medium: take peeled potatoes 250.0g, it is cut into the fritter of 1 centimeter square, after the 750ml that adds water boils, little fire keeps boiling 30 minutes, use three layers filtered through gauze, can repeatedly produce, make filtrate reach 1000ml, the filtrate obtained is the potato juice (being left out impurity) that potato quality mark is 25%, takes sucrose 15g and is dissolved in potato juice completing the preparation of fluid medium.Fluid medium after preparation adopts steam sterilization mode sterilizing 20 minutes under the environment of 121 DEG C, obtains the fluid medium produced.Two fluid mediums are produced in the manner described above.
The preparation of Mo Haiwei bacillus cereus seed liquor: carry out shaking table cultivation by being inoculated in a fluid medium by the Mo Haiwei bacillus cereus after the step S1 activation prepared, the rotating speed that shaking table is cultivated is 180r/min, cultivation temperature is 30 DEG C, after cultivating 45 hours, the viable count of the Mo Haiwei bacillus cereus after cultivation is 1.0 × 108CFU/ml, obtains Mo Haiwei bacillus cereus seed liquor；
The preparation of Bacillus pumilus seed liquor: will be inoculated in another fluid medium carry out shaking table cultivation by the Bacillus pumilus after the step S1 activation prepared, the rotating speed that shaking table is cultivated is 180r/min, cultivation temperature is 30 DEG C, after cultivating 45 hours, after cultivation, the viable count of Bacillus pumilus is 1.2 × 108CFU/ml, obtains Bacillus pumilus seed liquor.
Step S3: the fermentation of strain
Producing of fermentation mother liquor: measure ammonium nitrate 5.10g, sodium chloride 5.10g, dipotassium hydrogen phosphate 5.10g, magnesium sulfate 0.51g, green vitriol 0.51g, yeast extract 0.51g, one-level soybean oil 3.57g, distilled water 1000ml mix homogeneously make solution, and to regulate pH be 8.5, fermentation mother liquor after being configured, fermentation mother liquor after configuration is accessed in fermentation tank, at the temperature of 121 DEG C, adopt steam sterilization mode sterilizing 60min, obtain the fermentation mother liquor produced.
To access equipped with the fermentation tank of fermentation mother liquor carries out first time fermentation culture by the seed liquor of the step S2 Bacillus pumilus prepared, and the volume of Bacillus pumilus seed liquor is fermentation mother liquor volume 0.05%, first time fermentation culture is cultivation 8h under the condition of culture of constant temperature 32 DEG C；Then will continue to access in the fermentation tank after first time fermentation culture by the step S2 Mo Haiwei bacillus cereus seed liquor prepared, proceed second time fermentation culture, and volume is fermentation mother liquor volume the 0.05% of Mo Haiwei bacillus cereus seed liquor, second time fermentation culture is cultivation 36h under the condition of culture of constant temperature 32 DEG C.Above-mentioned first time fermentation culture and second time fermentation culture all adopt the mode of aerobic cultivation.Second time fermentation culture terminates, and Mo Haiwei bacillus cereus and total viable count of Bacillus pumilus in fermentation tank are 5.4 × 108Individual/ml, obtains strain primary fermentation liquid.
Step S4: the preservation of strain
By be the airtight resting state preservation of lucifuge 5 days when 19 DEG C by the step S3 strain primary fermentation liquid prepared, obtain the fermentation liquid completed.
Step S5: abluent finished product
Take through step S1～S4 fermentation liquid 1000g completed, adding fatty glyceride 5.05g, dipotassium hydrogen phosphate 4.04g, ferrous sulfate 0.05g, magnesium sulfate 0.05g and ethylenediaminetetraacetic acid 0.05g in fermentation liquid, the end-product that mix homogeneously obtains is No. three abluents.
According to detection, in No. three abluents, it is 62% that the viable count of Mo Haiwei bacillus cereus accounts for the percentage ratio of total viable count, and it is 38% that Bacillus pumilus accounts for the percentage ratio of total viable count.Mo Haiwei bacillus cereus concentration in deodorizer is 3.53 × 108Individual/ml, bacillus pumilus concentration in deodorizer is 2.17 × 108Individual/ml.
These No. three abluents are applied in the greasy dirt discharge pipeline section being numbered 3, occupation mode same abluent, after standing three days, again weigh this and are numbered the greasy dirt discharge pipeline section of 3, weight m2=258.69g.Dispel after totally it addition, this greasy dirt to be discharged the greasy dirt on pipeline section, again weigh this and be numbered greasy dirt discharge pipeline section of 1, nt wt net weight m3=243.56g.
According to formula w=(m1-m2)/(m1-m3) × 100%, obtain the oily waste degradation rate w=97.3% of an abluent.
In the preparation process in accordance with the present invention, owing to adopting the mode of fermentation to process Mo Haiwei bacillus cereus and Bacillus pumilus both functional microorganism floras so that it is produce sugar lipid surfactant during the fermentation and multiple enzyme enters in fermentation liquid.So that when the greasy dirt detergent adopting the present invention to contain microbial strains, the oils composition that can directly exist in city planting ductwork is primary carbon source, by the sugared lipid surfactant produced and enzyme, oils composition in emulsifying and degraded city planting ductwork, the breeding of the harmful bacteria in suppression environment, avoid the generation of the problems such as stink, growth and the breeding of endogenous profitable strain in environment can be promoted again, such that it is able to reach the effect of edible vegetable oil long, effective and dredging pipeline simultaneously.
The raw material simple, easily controllable, overall due to the fermentation technology of the present invention and the cost of preparation method are all relatively low, have good economic and social benefit.
It should be noted that above-described embodiment the present invention will be described rather than limits the invention, and those skilled in the art can design alternative embodiment without departing from the scope of the appended claims.In the claims, any reference marks that should not will be located between bracket is configured to limitations on claims.Word " comprises " and does not exclude the presence of the element or step not arranged in the claims.
Claims (8)
1. the greasy dirt detergent containing microbial strains, it is characterized in that, the described greasy dirt detergent containing microbial strains includes following components: surfactant 0.050～0.500 weight portion, dipotassium hydrogen phosphate 0.030～0.400 weight portion, ferrous sulfate 0.005～0.050 weight portion, magnesium sulfate 0.005～0.050 weight portion, ethylenediaminetetraacetic acid 0.001～0.005 weight portion and fermentation liquid 99.0～99.9 weight portion；
The active component of described fermentation liquid includes Mo Haiwei bacillus cereus and Bacillus pumilus, wherein, it is 60～70% that the viable count of described Mo Haiwei bacillus cereus accounts for the percentage ratio of total viable count, and it is 30～40% that the viable count of described Bacillus pumilus accounts for the percentage ratio of total viable count.
2. the greasy dirt detergent containing microbial strains according to claim 1, it is characterised in that described Mo Haiwei bacillus cereus concentration in described fermentation liquid is not less than 1.0 × 108Individual/ml, described Bacillus pumilus concentration in described fermentation liquid is not less than 5.0 × 107Individual/ml.
3. the preparation method containing the greasy dirt detergent of microbial strains, it is characterised in that comprise the steps:
Step S1: the activation of strain
Will be equipped with two slant tubes of Mo Haiwei Bacillus and Bacillus pumilus strain respectively to be placed in constant incubator and cultivate, incubation time is 110～130h, cultivation temperature in constant incubator is 30～35 DEG C, respectively obtains the Mo Haiwei bacillus cereus after activation and the Bacillus pumilus after activation；
Step S2: the preparation of seed liquor
To be inoculated in two fluid mediums respectively each carry out shaking table cultivation by the Bacillus pumilus after the step S1 Mo Haiwei bacillus cereus activated prepared and activation, to a fluid medium, the viable count of Mo Haiwei bacillus cereus is not less than 1.0 × 108Individual/ml, then obtain Mo Haiwei bacillus cereus seed liquor；To another fluid medium, the viable count of Bacillus pumilus is not less than 1.0 × 108Individual/ml, then obtain Bacillus pumilus seed liquor；
Step S3: the fermentation of strain
Carrying out first time fermentation culture by accessing in equipped with the fermentation tank of fermentation mother liquor through the step S2 Bacillus pumilus seed liquor prepared, the first time temperature of fermentation culture is 30～32 DEG C, and the time of fermentation culture first time is 8～10h；Wherein, volume is fermentation mother liquor volume 0.025～0.05% of the Bacillus pumilus seed liquor in addition fermentation tank；
Then carrying out second time fermentation culture by continuing to access in fermentation tank through the step S2 Mo Haiwei bacillus cereus seed liquor prepared, to fermentation tank, total viable count of Mo Haiwei bacillus cereus and Bacillus pumilus is not less than 5.0 × 108Individual/ml, the temperature of second time fermentation culture is 30～32 DEG C, and the time of second time fermentation culture is 36～48h, obtains strain primary fermentation liquid；Wherein, volume is fermentation mother liquor volume 0.025～0.05% of the Mo Haiwei bacillus cereus seed liquor in addition fermentation tank；
Step S4: the preservation of strain
To preserve by the step S3 strain primary fermentation liquid prepared, the fermentation liquid that must complete；
Step S5: abluent finished product
Surfactant, dipotassium hydrogen phosphate, ferrous sulfate, magnesium sulfate and ethylenediaminetetraacetic acid is added in the fermentation liquid completed through step S4, and parts by weight respectively surfactant 0.050～0.500 weight portion of each component, dipotassium hydrogen phosphate 0.030～0.400 weight portion, ferrous sulfate 0.005～0.050 weight portion, magnesium sulfate 0.005～0.050 weight portion, ethylenediaminetetraacetic acid 0.001～0.005 weight portion and fermentation liquid 99.0～99.9 weight portion, each component of mix homogeneously prepares abluent finished product.
4. the preparation method of the greasy dirt detergent containing microbial strains according to claim 3, it is characterised in that in described step S2, in fluid medium, the parts by weight of each component are: potato juice 97～99 weight portion, sucrose 1～3 weight portion；
Wherein, described potato juice comprises Rhizoma Solani tuber osi and water, and in described potato juice, Rhizoma Solani tuber osi is 15～25 weight portions, and water is 75～85 weight portions.
5. the preparation method of the greasy dirt detergent containing microbial strains according to claim 3, it is characterized in that, described step S2 is inoculated with two fluid mediums of Mo Haiwei bacillus cereus and Bacillus pumilus respectively when shaking table is cultivated, the rotating speed that shaking table is cultivated is 130～180r/min, temperature is 30～35 DEG C, and incubation time is 45～50 hours.
6. the preparation method of the greasy dirt detergent containing microbial strains according to claim 3, it is characterized in that, in described step S3, each composition weight number of fermentation mother liquor is: ammonium nitrate 0.40～1.00 weight portion, sodium chloride 0.2～0.5 weight portion, dipotassium hydrogen phosphate 0.1～0.5 weight portion, magnesium sulfate 0.01～0.05 weight portion, green vitriol 0.01～0.05 weight portion, yeast extract 0.03～0.08 weight portion, one-level soybean oil 0.20～0.50 weight portion, distilled water 98.00～99.00 weight portion, the pH value of described fermentation mother liquor is adjusted to 7.5～8.5.
7. the preparation method of the greasy dirt detergent containing microbial strains according to claim 3, it is characterised in that in described step S3, first time fermentation culture and second time fermentation culture all adopt the mode of aerobic cultivation.
8. the preparation method of the greasy dirt detergent containing microbial strains according to any one in claim 3～7, it is characterised in that in described step S4, the operation that preserves of strain preserves 5～7 days for the airtight resting state of lucifuge under the environmental condition below 20 DEG C.
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