CN100351371C - Antiseptic antistaling agent containing hay bacillocin - Google Patents
Antiseptic antistaling agent containing hay bacillocin Download PDFInfo
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Abstract
The present invention relates to an antiseptic antistaling agent with bacillus subtilis CCTCCM204085 and an application thereof. Identification proves that bacillus subtilis CCTCCM204085 belongs to typical bacillus subtilis. Compared with other kinds of bacillus subtilis, the fermentation liquor and the spore of bacillus subtilis CCTCCM204085 have better antiseptic antistaling effect on vegetables and fruits which are picked and stored. The antiseptic antistaling agent with bacillus subtilis CCTCCM204085 disclosed by the present invention is prepared from single preparations of wettable powder, a water suspending agent effervescence grains and tablets and a compounding preparation of the spore of bacillus subtilis CCTCCM204085 and chemical bactericide.
Description
Background technology
Fruit and vegetable preserves after harvesting in the process of transportation, and owing to microbial infection rots, perhaps owing to adopt the aging of back nature, quality descends, even finally loses edibleness, and these are the major causes that cause the back loss of fruit and vegetable results.According to statistics, whole world average year rate of loss is 25%, and the rotten rate of loss of developing country is then higher, reaches 40%～50%.At present, the anti-rot and fresh-keeping method that adopt countries in the world is a lot, is divided into physical fresh-keeping method, chemical fresh-keeping method, film fresh-keeping method three major types, and wherein the history with the fresh-keeping application of chemical fresh-keeping method is the longest, and effect is best.Thiophanate, benomyl, Te Keduo, derosal or the like are some fruits and vegetables Chemical Preservation preservation agents commonly used.But, enter after nineteen nineties, because these chemicals are paid attention to by the people more and more to people's toxic side effect, wherein most of preparations are successively announced to ban use of by USEPA and European Union's corresponding mechanism.
Since 1980, the numerous and confused research and development of the scientist of countries in the world can be used in the biotechnological formulation of antiseptic preservation of fruits and vegetables.Successively have and cereuisiae fermentum, trichoderma harziarum, pseudomonas, coccus and subtilis are used for the report of fruit vegetables storing disease control and the product registration is arranged.Domesticly since nineteen ninety, successively reported that several strains such as D-67-2, Bs-98, Bs-208 have the bacterial isolates of antagonistic action to plant epiphyte cause of disease bacterium, they mainly are used to the gray mold of tomato, strawberry, rice sheath blight disease, the control of asparagus stem wilt or the like field disease.
Fruit and vegetable is the food of people's immediate access often, and every day, intake was very big, and may be to eat up together with pericarp.Therefore for the sake of security, sanitas and the preservation agent to them requires very high.Subtilis is can be from people and animal on one's body or the beneficial microorganism that is separated in the body.A lot of evidences its healthy helpful to the human and animal.Just there is the people that its production is oral for the tablet confession as far back as twentieth century, to improve digestive functions.With bacillus subtilis formulation as the fruit and vegetables corrosion protection agent, thereby substitute the mycocide of chemosynthesis such as derosal, benomyl, Te Keduo, very helpful to guaranteeing food safety.The domestic bacillus subtilis strain that has business development to be worth does not see that report is arranged aspect antiseptic preservation of fruits and vegetables.
Summary of the invention
The purpose of this invention is to provide a kind of subtilis (Bacillus subtilis) fermented liquid of CCTCCM204085 and antisepsis antistaling agent of living spores of containing.Subtilis CCTCC M204085 is deposited in Chinese typical culture collection center (CCTCC) (address is in Wuhan University) on December 2nd, 2004, and preserving number is CCTCC M204085.
Another object of the present invention provides the purposes of described antisepsis antistaling agent, is used for the fruit and vegetable preservation and antisepsis of storage period.
Further purpose of the present invention provides a kind of subtilis CCTCC M204085.
The contriver separates the subtilis CCTCC M204085 bacterial strain that screening obtains in more than 2,000 bacterial isolates that obtains from soil.Simultaneous test finds that it is better than control strain in the effect of antiseptic preservation of fruits and vegetables.The contriver is with this bacterial strain its biological characteristics that has been fundamental research, zymotechnique, pharmaceutical formulation, utilisation technology and method, finally finished the present invention, having formed a class is the fruit and vegetable preserving antistaling agent of main active ingredient with subtilis CCTCC M204085 fermented liquid or gemma.
The invention provides a kind of antisepsis antistaling agent, wherein contain subtilis CCTCCM204085.
Antisepsis antistaling agent of the present invention, it is wettable powder, water suspending agent, effervescent granule, tablet.
The weight percent of each component of wettable powder is:
Mycetocyte content 5 * 10
8～1 * 10
10Individual CFU/g
Sodium dodecylbenzene sulfonate 0.2%～20%
Sodium lignosulfonate/calcium 0.1%～20%
Sodium stearate 0.1%～3%
White carbon black (expanded) 2%～10%
The sericite in powder surplus
pH 4.0～7.0。
The weight percent of each component of water suspending agent is:
Mycetocyte content 5 * 10
8～3 * 10
9Individual CFU/g
Sodium dodecylbenzene sulfonate 0.2%～20%
Chitin 0.1%～10%
Xylo-Mucine 0%～10%
Xanthan gum 0%～10%
Sodium Benzoate 0.5%～2%
Water surplus
pH 2.0～7.0。
The weight percent of effervescent granule, each component of tablet is:
Mycetocyte content 5 * 10
8～3 * 10
10Individual CFU/g
Tensio-active agent 0.5%～20%
Disintegrating agent (acid/basic cpd) 3%～30%
Tackiness agent 2%～25%
Help and collapse agent 2%～15%
The weighting material surplus.
Antisepsis antistaling agent of the present invention, wherein tensio-active agent is selected from one or more in sodium laurylsulfonate, Sodium dodecylbenzene sulfonate, calcium dodecylbenzene sulphonate, sodium lignosulfonate, calcium lignin sulphonate, sodium cellulose xanghate, the withered powder of tea, the spaonin powder; The disintegrating agent acidic cpd is selected from one or more in citric acid, tartrate, oxysuccinic acid, hexanodioic acid, fumaric acid, lactic acid, the boric acid; Basic cpd is selected from one or more in yellow soda ash, sodium bicarbonate, salt of wormwood, the saleratus; Tackiness agent is selected from one or more in polyoxyethylene glycol, polyvinylpyrrolidone, the polyvinyl alcohol (aggregate number 12000～20000); Help and collapse agent and be selected from Zulkovsky starch, dextrin, the Xylo-Mucine one or more; Weighting agent is selected from one or more in diatomite, light calcium carbonate, the sericite in powder.
Antisepsis antistaling agent of the present invention is characterized in that, it is the compound preparation that also contains chemical bactericide.Wherein chemical bactericide is selected from one or more of m-tetrachlorophthalodinitrile, Te Keduo, prochloraz, RP-26019, thiophanate methyl, vinclozolin, tricyclazole, derosal.
The present invention further provides the purposes of described antisepsis antistaling agent, described antisepsis antistaling agent has been used for the anti-corrosive fresh-keeping of fruit vegetables Lay duration of storage, prevented infecting of mould, green mold, grey mold, ring spot, black spot, wheel mildew, brown heart, sour rot.Wherein fruit and vegetable preferably apple, pears, mandarin orange, tangerine, orange, grape, Cauliflower, Caulis et Folium Brassicae capitatae, garlic shoot, leek, tomato.
The present invention also provides a kind of subtilis CCTCC M204085.
The feature of subtilis CCTCC M204085 bacterial strain among the present invention:
A, individual morphology feature: subtilis CCTCC M204085 bacterial strain nourishing body is shaft-like, ellipse garden, two ends, size 3.0 * 0.8 (μ m), peritrichous.The ellipse garden of gemma shape, size 1.7 * 0.7 (μ m), gemma is born in the middle of the thalline, and gemma does not make sporangium expand.
B, colonial morphology: when subtilis CCTCC M204085 bacterial strain was cultivated on beef broth peptone agar plate, single bacterium colony obviously swelled, the colony edge waviness, and light yellow, quality are opaque shape, not chromogenesis.Meat soup leaves standstill and forms mycoderm when cultivating, and does not form collarium, the not muddy nothing precipitation of nutrient solution.
C, physio-biochemical characteristics: subtilis CCTCC M204085 bacterial strain and the contrast of subtilis reference culture, physiological and biochemical property is identical: H
2O
2The enzyme reaction positive, the V.P. reacting positive, the nitrate reduction positive, gelatin hydrolysis, the Trisodium Citrate utilization, otan forms, the casein hydrolysis, the starch hydrolysis all is positive.D-glucose, D-wood sugar, L-arabinose, the utilization of D-N.F,USP MANNITOL are also all positive.Utilize not aerogenesis of glucose.The lecithin reaction, the Phenylalanine dehydrogenase reaction, the tyrosine decomposition all is negative.Can grow in the 7%NaCl concentration of aqueous solution.30 ℃ of optimum growh breeding temperature are not grown more than 53 ℃.After subtilis CCTCC M204085 grew in the V.P. nutrient solution, medium pH value was 7.92.
D, with the difference of common subtilis: subtilis CCTCC M204085 bacterial strain belongs to subtilis from the systematic bacteriology angle.Compare with the subtilis of having reported, the characteristics of this bacterial strain are their height resistances to penicillin.Penicillin concn up to the filter paper of 1000 units on dibbling subtilis CCTCC M204085 bacterial strain and move and receive on the beef broth peptone agar plate, this bacterial strain can form lawn and indicators such as citrus green mold, ash arrhizus bacteria, ring spot bacterium are produced inhibition zone by very normal growth and breeding.
Subtilis CCTCC M204085 bacterial strain with screened both at home and abroad that some that obtain can be used for fruit, vegetables rot-resistant yeast is compared with Bacillus strain, advantage such as have that result of use is good, gemma survival rate height, zymophyte number are many.
Organism preservation explanation
Subtilis (Bacillus subtilis) CCTCC M204085 is preserved in Chinese typical culture collection center (CCTCC), the address: Wuhan University, deposit number: CCTCC M204085, preservation date: on December 2nd, 2004.
Embodiment
The corrosion-resistanting fresh-keeping effect of subtilis CCTCC M204085:
Subtilis CCTCC M204085 briefly is listed as follows (seeing Table 1) to the anti-corrosive fresh-keeping of portion water fruit vegetable
Table 1 subtilis CCTCC M204085 bacterial strain is to portion water fruit vegetable corrosion-resistanting fresh-keeping effect
| Treatment dosage | Preventive effect (%) (or decayed fruit rate) | Fruit-vegetable quality changes | |||||||
| Apple (150 days) | Orange (60 days) | ||||||||
| Apple | Sand pear | Orange | Brassica oleracea var. botrytis L. | Orange | Moisture content (%) | Total acid (%) | Moisture content (%) | Total acid (%) | |
| Subtilis CCTCC M204085 1 * 10 7cfu | 86.6 | 66.1 | 51.1 | 37.5 | 65.8 | -3.17 | -0.596 | -2.90 | -0.0477 |
| TL-6 * 1×10 7cfu | 73.4 | 56.1 | 55.2 | 45.0 | 50.1 | -3.70 | -0.750 | +0.70 | -0.1146 |
| Contrast (clear water) | 12.5 | 35.1 | 27.5 | 0.8 ** | 81.8 | 89.18 | 1.326 | 85.04 | 0.8655 |
| Bs-208 1×10 8cfu | 60.0 | 21.6 | 40.9 | 25.0 | 32.0 | -8.70 | -0.765 | -6.68 | -0.0764 |
| Derosal+preservation agent 800mg/L | - | 57.3 | - | - | 72.3 | - | - | -0.83 | -0.0918 |
*.TL-6 another bacillus subtilis strain that obtains for inventor's separation screening (contriver can provide).
*. be sick index value.
The liquid fermentation process of subtilis CCTCC M204085:
A, slant strains: slant strains culture medium prescription: conventional potato sucrose agar (peeling potatoes section 200 restrains liquor, sucrose 20 grams, agar 15 grams, water 1000ml, pH nature).Slant strains culture temperature: 28～32 ℃; Incubation time: 3 days.
B, liquid fermenting: liquid fermentation medium prescription: glucose 0%～2%, soybean cake powder 0.3%～4.5%, yeast powder 0.1%～1.2%, wheat bran 0%～2.5%, ground rice 0%～4%, fish meal 0%～2.5%, Semen Maydis powder 0%～4%, potassium primary phosphate 0.1%～0.8%, sal epsom 0.1%～0.6%, NaCl 0%～1%.Preceding pH7.0～7.5 disappear.28 ℃～32 ℃ of liquid fermenting temperature, mixing speed 120rpm/min～350rpm/min, air flow 1: 0.6～1.5, fermentation period changes because of different ingredients, the shortest 30 hours, the longest 72 hours, when sporangium is completed into and 30% left and right sides bacillus exfoliation is arranged, put jar.
C, post-treating method: fermented liquid anhydrates through disc centrifuge is centrifugal, stays the bacterium slurry.With the bacterium slurry dry former powder of subtilis CCTCC M204085 gemma that gets of spray drying process.The bacterium amount is 1 * 1010cfu/g-3 * 1010cfu/g in the former powder.
Subtilis CCTCC M204085 solid fermentation process
A, seed preparation
The preparation method of slant strains is the same.
The solid fermentation substratum went out behind the bacterium with liquid shaking bottle bacterial classification (or seeding tank fermented bacterium) inoculation.Seed fermentation nutrient solution prescription is: 20% potato liquor is worked as water, adds 1%～3% glucose, 0%～0.5%NaCl, 0.1%～2% peptone, 0%～0.8% yeast extract paste.
Prepare behind the spore suspension inoculation with slant strains and shake bottle or seeding tank.28～30 ℃ of leavening temperatures, shaking speed 200rpm/min (or fermentor tank rotating speed 120rpm/min～350rpm/min, air flow 0.6%～1.2%).Ferment to logarithmic proliferation and be advisable mid-term.
B, solid fermentation substratum
The solid fermentation culture medium prescription is: wheat bran 2%～20%, and soybean cake powder 1%～10%, Semen Maydis powder 1%～10%,
Glucose 1%～10%, potassium primary phosphate 0.1%～2.0%, light calcium carbonate 0.3%～5%, rice meal 0%～15% complements to 100% (w/w) with perlite.The consumption of water is: material: water=1: 1.0～1.3 (w/w).
High pressure steam sterilization is used in solid fermentation substratum preparation finishing back, 121 ℃, keeps 30 minutes.
C, fermentation parameter
Inoculum size 1%～10% (v/w).After the inoculation, ferment by ventilation Quchi method or plate rail method.28 ℃～32 ℃ of leavening temperatures (control material temperature), fermentation period 48～120 hours.With the sporocyst fully matured, and 30% left and right sides bacillus exfoliation is arranged for judging fermentation terminated standard.
D, dry and pulverizing
With the hot blast airing materials, hot blast temperature is controlled at below 55 ℃ after the fermentation ends.Earlier be crushed to 60 orders～80 orders behind the dry materials, sieve and remove thicker impurity such as wheat bran with the lower pulverizer of temperature rise.Pulverize again with micronizer mill then, mistake 325 mesh sieves.This kind powder can be used as former powder and uses, and bacteria containing amount is 1 * 1010cfu/g-1 * 1011cfu/g in the former powder.
Subtilis CCTCC M204085 prepares fruit and vegetable preserving antistaling agent
A, wettable powder:
The wettable powder agent prescription
Sodium dodecylbenzene sulfonate 0.2%～20% (w/w, down together)
Sodium lignosulfonate 0.1%～20%
Expanded white carbon black 2%～20%
The former powder 5 * 10 of subtilis CCTCC M204085
8～1 * 10
10CFU/g
Sericite in powder is supplemented to 100%
Above-mentioned raw materials takes by weighing in proportion, drops in the mixing machinery, through the jet-type micronizer mill, is crushed to 325 orders and sieves behind the abundant mixing, packing.
B, aqueous suspension
The former powder 5 * 10 of subtilis CCTCC M204085
8～1 * 10
10CFU/ml
Dimethylbenzene 0%～10%
Sodium dodecylbenzene sulfonate 0.2%～20%
Chitin 0.1%～10%
Xylo-Mucine 0%～10%
Xanthan gum 0%～10%
Sodium Benzoate 0.5%～2%
Water replenishes 100%
PH 2.0～7.0 (transferring) with dilute hydrochloric acid
After above-mentioned raw materials takes by weighing in proportion, drop into the dosing chamber mixing, be milled to 95% through colloidal mill and bottle behind 325 mesh sieves excessively.
C, effervescent (granule or tablet)
The former powder 5 * 10 of subtilis CCTCC M204085
8～3 * 10
10Individual CFU/g
Calcium dodecylbenzene sulphonate/sodium 0.2%～20%
Disintegrating agent (acid/basic cpd) 3%～30%
Tackiness agent 2%～25%
Help and collapse agent 2%～15%
The weighting material surplus
Above-mentioned raw materials mixes the back and uses the tabletting machine compressing tablet or use the Squeezinggranulator granulation.
Embodiment 1: subtilis CCTCC M204085 liquid fermenting also prepares the technology of the former powder of gemma
A, bacterial classification are prepared
The substratum of slant strains: thin slice 200 grams that the peeling potato is cut into, add water 1000ml liquor (100 ℃, 20 minutes), remove slag and stay juice.Timely and appropriate discovery moisturizing to decoction is 1000ml in the process of boiling.With sucrose 20 grams, dissolve.Add agar 15 grams again, melt agar (also will replenish the water that evaporates in right amount in the process of thawing agar).The pH nature.
Sterilization according to a conventional method, beveling, cultivate 2 days in advance after, the subtilis CCTCC M204085 original seed that activated of inoculation, 30 ℃ ± 1 ℃ constant temperature culture 3 days.The thalline of microscopy more than 95% stops when all having formed gemma cultivating.
Above-mentioned cultured test tube slant bacterial classification is washed with sterilized water, break up, make it into uniform spore suspending liquid, handled 30 minutes through 75 ℃ of waters bath with thermostatic control.This gemma suspension is chilled to switching eggplant bottle inclined-plane after the room temperature, and every test tube slant bacterial classification connects 2-3 eggplant bottle inclined-plane.30 ℃ ± 1 ℃ constant temperature culture 12 hours washes with sterilized water again, and jolting is dispersed as bacteria suspension.With this bacterial suspension inoculation 100L liquid fermentation tank.
B, fermentation
Liquid fermentation medium is: glucose 2%, and soybean cake powder 1.5%, yeast powder 0.8%, wheat bran 1.0%, rice meal 2%, Semen Maydis powder 1.0%, potassium primary phosphate 0.3%, sal epsom 0.3%, NaCl 0.5%.PH 8.0 before disappearing.
60L feeds intake in the 100L fermentor tank.Sterilized 30 minutes for 121 ℃.Inoculate above-mentioned bacteria suspension when cooling off 30 ℃.29 ℃ ± 1 ℃ of liquid fermenting temperature, mixing speed 300rpm, air flow 1: 1.2.Fermented about 32 hours, and, and put jar when having 30% left and right sides sporangium to come off to sporangium rate of formation 95%.Recording fermenation raw liquid living spores number is 42 * 108cfu/ml.
C, prepare former powder
Fermented liquid is put jar the back and is transferred pH to pH4.0 with a HCl in souring tank, through disc centrifuge the is centrifugal (separation factor 〉=8000g), stay bacterium to starch that anhydrates.The beautiful mica powder (fineness 325 orders) of 2% (w/v), making beating will by volume be added in the bacterium slurry.Use spray-drying tower, become former powder with bacterium slurry spray is dried.The import wind-warm syndrome was 180 ℃ when spray was done, 70 ℃ of outlet wind-warm syndrome.After testing, the several 5 * 1010cfu/g of living spores in the former powder.
Embodiment 2, subtilis CCTCC M204085 solid fermentation also prepare the technology of former powder
A, bacterial classification are prepared: with embodiment 1.
B, solid fermentation seed are prepared:
The seed of solid fermentation prepares in liquid fermentation tank.Its seed culture based formulas is: glucose 2%, and soybean cake powder 1.5%, yeast powder 1%, light calcium carbonate 1%, Semen Maydis powder 1.0%, KH2PO4 0.5%, MgSO40.3%,
PH 8.0 before disappearing.Connect ready bacterial classification behind the high pressure steam sterilization, inoculum size is that per 200 liters of seed liquor connect 1 eggplant bottle bacterial classification.28 ℃ of-30 ℃ of aerated culture 12 hours, mixing speed 250rpm, air quantity progressively is increased to 1: 10 at 1: 0.6.In this enforcement, fermentation obtains 200 liters of seed liquor.
C, solid fermentation substratum
Semen Maydis powder 8%, soybean cake powder 10%, wheat bran 20%, ground rice 10%, glucose 2%, perlite 50%.Water is 1: 1 with the ratio of material.121 ℃ of high pressure steam are sterilized 1 hour (glucose adds less water sterilization, 115 ℃, 20 minutes separately).Sterilization is carried out in changeing pot, and the back of having gone out is advanced tap water by interlayer and is cooled to 28 ℃.
D, inoculation and fermentation
After 1 ton of (after adding water) sterilization of above solid medium and the cooling, evenly add in the substratum material that changes in the pot by the G/W of aseptic pipeline with 200 liters of seed liquor and sterilization separately.Pot is rotated on edge joint kind limit, makes seed liquor and mixing of materials even.With the material that the connects kind fermentation bed of packing into, material thickness 10cm.Feed sterile air off and on and ferment, controlled temperature is 28 ℃～30 ℃.Fermentation period is 72 hours.Sampling microscopy in the time of 72 hours, all ripe and stuck fermentation when 30% left and right sides bacillus exfoliation is arranged sporangium.Present embodiment fermentation sync rates 〉=96%.
E, fermentation work in-process drying
Blow material on the fermentation bed with 40～45 ℃ of big air quantity of aseptic hot blast, wind direction from bottom to top.Through 12 hours, the fermentation work in-process were air-dry, water content 4.9%.
F, crushing screening are made former powder
With beater disintegrating machine precomminution one time, fineness reaches 80 orders.Advance the jet-type micronizer mill again and pulverize second time, fineness reaches 325 orders.It is 6 * 1010cfu/g that the survey living spores is counted the result.Detection through indexs such as fineness, living spores number, water contents all meets the company standard requirement.
Embodiment 3, subtilis CCTCC M204085 preserving fruit and vegetable utilizing agent formulation production method (production of living spores aqueous suspension)
Ferment by embodiment 1 method, 1000L feeds intake.After the fermentation ends fermented liquid is acidified to pH4.0 through HCl, centrifugal with disc centrifuge, obtain 300 liters in dense thick bacterium slurry.By the volume ratio of bacterium slurry, add 6% Sodium dodecylbenzene sulfonate (w/v), 1% Chitosan (w/v), 2% Xylo-Mucine (w/v), 1% Sodium Benzoate (w/v), dimethylbenzene 5% (v/v) weighs 345 kilograms altogether.Fully stir the living spores number in the back detection preparation work in-process, the result is 110 * 108cfu/ml, meets the company standard requirement.Be packaged into the test sample of one bottle of 1000ml.
Embodiment 4, subtilis CCTCC M204085 preserving fruit and vegetable utilizing agent formulation production method (wettable powder)
Claim to press former powder (500 * 108cfu/g) 100 kilograms add Sodium dodecylbenzene sulfonate 45kg, sodium lignosulfonate 22.5kg, ammonium sulfate 45.0kg, white carbon black (expanded) 90kg, sericite in powder 147.5kg, the heavy 450kg of total that embodiment 1 produces.In mixing machine, mixed 1 hour,, cross 325 mesh sieves after micronizer mill.After testing, the living spores number is 110 * 108cfu/g in the product, meets company standard.Be packaged as 200 gram/bags.
Embodiment 5, subtilis CCTCC M204085 preserving fruit and vegetable utilizing agent formulation production method (effervescent tablet)
Get 50 kilograms in former powder of subtilis CCTCC M204085 gemma (50,000,000,000 cfu/g), add 30 kilograms of sodium laurylsulfonates, 17.5 kilograms of Zulkovsky starches, 15.75 kilograms in yellow soda ash, 13.75 kilograms of abundant mixings of sodium bicarbonate.Add 12.5 kilograms of citric acids again, 2 kilograms of fumaric acid, 15.75 kilograms in tartrate, 25 kilograms of Xylo-Mucines, fully mixing.Complementing to the material gross weight with sericite in powder is 250 kilograms, abundant again mixing.With mixed material compressing tablet, become the effervescent tablet of every heavily about 2 grams with tabletting machine.Present embodiment obtains 249.2 kilograms in tablet at last, and every gram contains living spores 98.7 hundred million in the tablet.
Embodiment 6, subtilis CCTCC M204085 and chlorothalonil compound agent (wettable powder)
Test has proved that female powder of subtilis CCTCC M204085 (in 50,000,000,000 cfu/g) and the following proportioning of m-tetrachlorophthalodinitrile (folding hundred content) have obvious synergistic effect: subtilis CCTCC M204085: m-tetrachlorophthalodinitrile=(5～1): (1～2) through lab screening.With subtilis CCTCCM204085: m-tetrachlorophthalodinitrile=2: 1 is an example.
Take by weighing 100 kilograms in the female powder of subtilis CCTCC M204085,50 kilograms of m-tetrachlorophthalodinitriles (folding hundred) (or 75% wettable powder 67 kilograms), 20 kilograms of Sodium dodecylbenzene sulfonatees, surplus is a kaolin, polishing to 200 kilogram.Be crushed to micronizer mill more than 325 orders after mechanical thorough mixing is even, one bag of per 100 gram are packaged as finished product again.Present embodiment obtains 194 kilograms of finished products, and subtilis CCTCC M204085 living spores content is 25,000,000,000 cfu/g in the finished product, m-tetrachlorophthalodinitrile content 25%.500～600 times of thin ups soak fruit and dried in about 3 minutes and get final product during use.
Embodiment 7, subtilis CCTCC M204085 and composite dose of RP-26019 (iprodione) (wettable powder)
Female powder of lab screening evidence subtilis CCTCC M204085 (50,000,000,000 cfu/g) and RP-26019 50% wettable powder have obvious synergistic effect: subtilis CCTCC M204085: RP-26019=(10～2): (1～5) under following conditions of mixture ratios.With subtilis CCTCCM204085: RP-26019=1: 1 is an example.
Take by weighing 100 kilograms in the female powder of subtilis CCTCC M204085 of 50,000,000,000 cfu/g content, 100 kilograms of RP-26019 50% wettable powders, add 40 kilograms of Sodium dodecylbenzene sulfonatees, beautiful mica powder complements to 300 kilograms, is crushed to more than 325 orders with micronizer mill after fully mixed.One bag of per 100 gram.Present embodiment obtains 290 kilograms of finished products, contains subtilis CCTCC M204085 living spores 16,700,000,000 cfu/g in the finished product, iprodione 16.7%.500 times of thin ups soak fruit and dried in about 3 minutes and get final product during use.
Embodiment 8, subtilis CCTCC M204085 gemma water suspending agent are used for the apple storage
Get subtilis CCTCC M204085 aqueous suspension 0.5kg, add water 25kg, be diluted to the aqueous suspensions of 2 * 108cfu/ml, fully stir (about 15 minutes) and 100kg red fuji apple (every crowd of 10kg) is immersed in the subtilis CCTCC M204085 aqueous suspensions soak time 5 minutes afterwards in batches.Pull out and dry, single fruit cover mesh bag.The vanning storage.Storage is 5 months under the room temperature, preservative effect 93.1%, and apple total acidity rate of descent 26.2%, the contrast chemical pesticide is a carbendazol wettable powder, dilutes 1000 times, treatment process is identical with subtilis CCTCC M204085.Under the equal holding conditions, preservative effect 84.3%, total acidity descends 31.4%.Subtilis CCTCC M204085 handles and is better than chemical agent and clear water contrast.
Embodiment 9, subtilis CCTCC M204085 wettable powder are used for the orange anti-corrosive fresh-keeping
(the living spores number is 110 * 108cfu/g) 0.5Kg in the product to get subtilis CCTCC M204085 wettable powder, add water 25kg, be diluted to the subtilis CCTCCM204085 spore suspending liquid of 2 * 108cfu/ml, to only adopt down from tree, three days fresh orange 200kg immerses the subtilis CCTCC M204085 suspension in batches, steep after 5 minutes the orange taking-up is dried, pack with the small plastic bag single fruit, adorn a carton for per then 10 kilograms.Preserve under the room temperature.With the derosal is the Chemical Preservative contrast, is blank with the clear water.Two kinds of processing with orange amount and pack the storage situation and subtilis CCTCCM204085 processing identical.Preserve check result after 120 days.It is 84.7% that subtilis CCTCC M204085 (wettable powder) handles preservative effect, and the derosal effect is 86.1%, and both differences are not remarkable, prove that subtilis CCTCC M204085 might replace derosal to be used for the orange anti-corrosive fresh-keeping.
Embodiment 10, subtilis CCTCC M204085 effervescent granule are used for the preservation and freshness of pear
(the living spores number is 110 * 108cfu/g) in the product, adds the spore suspension that water 24.5kg is diluted to 2.2 * 108cfu/ml to get the subtilis CCTCC M204085 effervescent granule 0.5kg of embodiment 5 preparation.The spore suspension that dilution is good evenly sprays at pear epidermis to be gathered, and begins to gather after air-dry 1-2 hour.Handle with care when gathering, do not hinder the surface of pear.Gathering, the back is timely packs with thin soft paper single fruit, divides cartoning then, 10 kilograms of pears of every case.The processing that sprays clear water is as blank, and what spray 0.1% derosal (w.p) is that the standard chemical medicament contrasts.Room temperature condition 90 days preservative effect subtilis CCTCC M204085 of storage down is 66.1%, and derosal is 60.0%, and subtilis CCTCC M204085 handles and is better than the derosal processing.
Embodiment 11, subtilis CCTCC M204085 gemma water suspending agent are used for the Cauliflower mildew-resistant
Get 500 milliliters of subtilis CCTCC M204085 aqueous suspensions, add 24.5 liters in water, be sprayed at the cauliflower surface of the Cauliflower that is about to gather behind the mixing.Require spray cloth to get even as far as possible.The air-dry back cauliflower of gathering at once.Each cauliflower is single uses soft paper pack, leaves in the carton 5 kilograms in every case then in.If the spray clear water is a blank.Temperature is preservation in 7 ℃ the freezer, OOBA result after 30 days.The effect that subtilis CCTCC M204085 aqueous suspension is handled mildew-resistant reaches 78.2% (by sick Index for Calculation).
Embodiment 12, subtilis CCTCC M204085 chlorothalonil compound agent are used for the orange storage
Get composite dose 250 gram and add 25 liters in water, be sprayed onto tree with atomizer behind the mixing and go up the orange surface that is about to harvesting.The technical requirements that air-dry back strictness is plucked by orange is plucked.Orange single fruit under adopting is cased the back well with soft paper bag, and every case 5kg does blank with clear water.Preservation check result after 2 months in 7 ℃ of freezers.In two months storage periods, blank decayed fruit rate is 41.9%, decayed fruit rate 17.5% is handled in the single agent of subtilis CCTCC M204085, handles decayed fruit rate only 8.3% for composite dose, and the composite dose of anti-effect of rotting is better than single agent of subtilis CCTCC M204085 bacterium.
Claims (10)
1, a kind of antisepsis antistaling agent wherein contains subtilis (Bacillus subtilis) CCTCCM204085.
2, antisepsis antistaling agent according to claim 1 is characterized in that, it is wettable powder, water suspending agent, effervescent granule or tablet.
3, antisepsis antistaling agent according to claim 2, wherein the weight percent of each component of wettable powder is:
Mycetocyte content 5 * 10
8～1 * 10
10Individual CFU/g
Sodium dodecylbenzene sulfonate 0.2%～20%
Sodium lignosulfonate/calcium 0.1%～20%
Sodium stearate 0.1%～3%
Expanded white carbon black 2%～10%
The sericite in powder surplus
pH 4.0～7.0。
4, antisepsis antistaling agent according to claim 2, wherein the weight percent of each component of water suspending agent is:
Mycetocyte content 5 * 10
8～3 * 10
9Individual CFU/g
Sodium dodecylbenzene sulfonate 0.2%～20%
Chitin 0.1%～10%
Xylo-Mucine 0%～10%
Xanthan gum 0%～10%
Sodium Benzoate 0.5%～2%
Water surplus
pH 2.0～7.0。
5, antisepsis antistaling agent according to claim 2, wherein the weight percent of effervescent granule or each component of tablet is:
Mycetocyte content 5 * 10
8～3 * 10
10Individual CFU/g
Tensio-active agent 0.5%～20%
Disintegrating agent 3%～30%
Tackiness agent 2%～25%
Help and collapse agent 2%～15%
The weighting material surplus
Wherein disintegrating agent is acidic cpd or basic cpd.
6, antisepsis antistaling agent according to claim 5, wherein tensio-active agent is selected from one or more in sodium laurylsulfonate, Sodium dodecylbenzene sulfonate, calcium dodecylbenzene sulphonate, sodium lignosulfonate, calcium lignin sulphonate, sodium cellulose xanghate, the withered powder of tea, the spaonin powder; The disintegrating agent acidic cpd is selected from one or more in citric acid, tartrate, oxysuccinic acid, hexanodioic acid, fumaric acid, lactic acid, the boric acid; The disintegrating agent basic cpd is selected from one or more in yellow soda ash, sodium bicarbonate, salt of wormwood, the saleratus; Tackiness agent is selected from one or more in polyoxyethylene glycol, polyvinylpyrrolidone, the polyvinyl alcohol; Help and collapse agent and be selected from Zulkovsky starch, dextrin, the Xylo-Mucine one or more; Weighting agent is selected from one or more in diatomite, light calcium carbonate, the sericite in powder.
7, antisepsis antistaling agent according to claim 1 is characterized in that, it also contains chemical bactericide.
8, antisepsis antistaling agent according to claim 7, wherein chemical bactericide is selected from one or more of m-tetrachlorophthalodinitrile, Te Keduo, prochloraz, RP-26019, thiophanate methyl, vinclozolin, tricyclazole, derosal.
9, the purposes of arbitrary described antisepsis antistaling agent of a kind of claim 1-8 is characterized in that, described antisepsis antistaling agent is used for the anti-corrosive fresh-keeping of fruit and vegetable.
10, a kind of subtilis (Bacillus subtilis) CCTCC M204085.
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