CN1853489A - Antiseptic antistaling agent containing hay bacillocin - Google Patents
Antiseptic antistaling agent containing hay bacillocin Download PDFInfo
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Abstract
An antiseptic and antistaling agent in the form of wettable powder, suspension, effervescent particle, tablet, or mixture with chemical bactericide features that it contains Bacillus subtilis (CCTCC M204085). Its advantage is high effect.
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 edibility, and these are the main causes that cause the back loss of fruit and vegetable results.According to statistics, whole world average year loss late is 25%, and the rotten loss late 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, carbendazim or the like are some fruits and vegetables chemical preservation antistaling 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 and side effect, wherein most of preparations are successively announced to ban use of by Environmental Protection Agency USA 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 biologic product of antiseptic preservation of fruits and vegetables.Successively have and brewer's yeast, Trichoderma harzianum, pseudomonad, coccus and bacillus 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 antagonism 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, anticorrisive agent and the antistaling agent to them requires very high.Bacillus subtilis is can be from people and animal on one's body or the beneficial microbe 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 fungicide of chemical syntheses such as carbendazim, benomyl, Te Keduo, very helpful to guaranteeing food security.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 bacillus subtilis (Bacillus subtilis) zymotic fluid of CCTCCM204085 and antisepsis antistaling agent of living spores of containing.Bacillus 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 bacillus subtilis CCTCC M204085.
The inventor separates the bacillus subtilis CCTCC M204085 bacterial strain that screening obtains in more than 2,000 bacterial isolates that obtains from soil.Contrast test finds that it is better than control strain in the effect of antiseptic preservation of fruits and vegetables.The inventor is with this bacterial strain its biological characteristics that has been basic research, zymotechnique, pharmaceutical formulation, application 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 bacillus subtilis CCTCC M204085 fermented liquid or gemma.
The invention provides a kind of antisepsis antistaling agent, wherein contain bacillus subtilis CCTCCM204085.
Antisepsis antistaling agent of the present invention, it is wettable powder, aqueous suspension agent, effervescence granular, tablet.
The percentage by weight of each component of wettable powder is:
Bacterial cell content 5 * 10
8～1 * 10
10Individual CFU/g
Neopelex 0.2%～20%
Sodium lignosulfonate/calcium 0.1%～20%
Odium stearate 0.1%～3%
White carbon black (expanded) 2%～10%
The sericite in powder surplus
pH 4.0～7.0。
The percentage by weight of each component of aqueous suspension agent is:
Bacterial cell content 5 * 10
8～3 * 10
9Individual CFU/g
Neopelex 0.2%～20%
Chitosan 0.1％～10％
Sodium carboxymethylcellulose 0%～10%
Xanthans 0%～10%
Sodium Benzoate 0.5%～2%
Water surplus
pH 2.0～7.0。
The percentage by weight of effervescence granular, each component of tablet is:
Bacterial cell content 5 * 10
8～3 * 10
10Individual CFU/g
Surfactant 0.5%～20%
Disintegrant (acid/alkali compounds) 3%～30%
Adhesive 2%～25%
Help and collapse agent 2%～15%
The filler surplus.
Antisepsis antistaling agent of the present invention, wherein surfactant is selected from one or more in dodecyl sodium sulfate, neopelex, calcium dodecyl benzene sulfonate, sodium lignin sulfonate, calcium lignosulfonate, sodium cellulose xanghate, the withered powder of tea, the spaonin powder; The disintegrant acid compound is selected from one or more in citric acid, tartaric acid, malic acid, adipic acid, fumaric acid, lactic acid, the boric acid; Alkali compounds is selected from one or more in sodium carbonate, sodium acid carbonate, potash, the saleratus; Adhesive is selected from one or more in polyethylene glycol, polyvinylpyrrolidone, the polyvinyl alcohol (aggregate number 12000～20000); Help and collapse agent and be selected from soluble starch, dextrin, the sodium carboxymethylcellulose one or more; Filler is selected from one or more in diatomite, precipitated calcium carbonate, the sericite in powder.
Antisepsis antistaling agent of the present invention is characterized in that, it is the complex preparation that also contains chemical bactericide.Wherein chemical bactericide is selected from one or more of Bravo, Te Keduo, the bright amine of rice, iprodione, thiophanate methyl, vinclozolin, tricyclazole, carbendazim.
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 and vegetable duration of storage, prevented infecting of mould, green mold, grey mold, ring spot, black spot, wheel mildew, brown rot, sour rot.Wherein fruit and vegetable preferably apple, pears, mandarin orange, tangerine, orange, grape, cauliflower, broccoli, garlic shoot, leek, tomato.
The present invention also provides a kind of bacillus subtilis CCTCC M204085.
The feature of bacillus subtilis CCTCC M204085 bacterial strain among the present invention:
A, individual morphology feature: bacillus subtilis CCTCC M204085 bacterial strain trophosome 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 bacillus subtilis CCTCC M204085 bacterial strain was cultivated on beef broth peptone agar plate, single bacterium colony obviously swelled, the colony edge waveform, 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: bacillus subtilis CCTCC M204085 bacterial strain and the contrast of bacillus 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 natrium citricum utilization, dihydroxyacetone (DHA) forms, the casein hydrolysis, the starch hydrolysis all is positive.D-glucose, D-wood sugar, L-arabinose, the utilization of D-sweet mellow wine 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 bacillus subtilis CCTCC M204085 grew in the V.P. nutrient solution, medium pH value was 7.92.
D, with the difference of common bacillus subtilis: bacillus subtilis CCTCC M204085 bacterial strain belongs to bacillus subtilis from the systematic bacteriology angle.Compare with the bacillus subtilis of having reported, the characteristics of this bacterial strain are their height drug resistances to penicillin.Penicillin concn up to the filter paper of 1000 units on dibbling bacillus subtilis CCTCC M204085 bacterial strain and move and receive on the beef broth peptone agar plate, this bacterial strain can form lawn and indicator bacterias such as citrus green mold, ash arrhizus bacteria, ring spot bacterium are produced inhibition zone by very normal growth and breeding.
Bacillus subtilis CCTCC M204085 bacterial strain with screened both at home and abroad that some that obtain can be used for fruit, vegetables corrosion-resistant saccharomycete 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
Bacillus 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.
The specific embodiment
The corrosion-resistanting fresh-keeping effect of bacillus subtilis CCTCC M204085:
Bacillus subtilis CCTCC M204085 briefly is listed as follows (seeing Table 1) to the anti-corrosive fresh-keeping of part fruit and vegetable
Table 1 bacillus subtilis CCTCC M204085 bacterial strain is to part fruit and 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 | Cauliflower | Orange | Moisture content (%) | Total acid (%) | Moisture content (%) | Total acid (%) | |
| Bacillus 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 |
| Carbendazim+antistaling agent 800mg/L | - | 57.3 | - | - | 72.3 | - | - | -0.83 | -0.0918 |
*.TL-6 another bacillus subtilis strain that obtains for inventor's separation screening (inventor can provide).
*. be sick exponential quantity.
The liquid fermentation process of bacillus 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 cultivation temperature: 28～32 ℃; Incubation time: 3 days.
B, liquid fermentation: liquid fermentation medium prescription: glucose 0%～2%, soya-bean cake powder 0.3%～4.5%, dusty yeast 0.1%～1.2%, wheat bran 0%～2.5%, ground rice 0%～4%, fish meal 0%～2.5%, corn flour 0%～4%, potassium dihydrogen phosphate 0.1%～0.8%, magnesium sulfate 0.1%～0.6%, NaCl 0%～1%.Preceding pH7.0～7.5 disappear.28 ℃～32 ℃ of liquid fermentation temperature, speed of agitator 120rpm/min～350rpm/min, throughput 1: 0.6～1.5, fermentation period changes because of different formulations, 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-processing approach: zymotic fluid anhydrates through disc centrifuge is centrifugal, stays the bacterium slurry.With the bacterium slurry dry former powder of bacillus subtilis CCTCC M204085 gemma that gets of spray drying process.The bacterium amount is 1 * 1010cfu/g-3 * 1010cfu/g in the former powder.
Bacillus subtilis CCTCC M204085 solid fermentation process
A, seed preparation
The preparation method of slant strains is the same.
The solid fermentation culture medium 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.
Prepare behind the spore suspension inoculation with slant strains and shake bottle or seeding tank.28～30 ℃ of fermentation temperatures, shaking speed 200rpm/min (or fermentation tank rotating speed 120rpm/min～350rpm/min, throughput 0.6%～1.2%).Ferment to logarithmic proliferation and be advisable mid-term.
B, solid fermentation culture medium
The solid fermentation culture medium prescription is: wheat bran 2%～20%, and soya-bean cake powder 1%～10%, corn flour 1%～10%,
Glucose 1%～10%, potassium dihydrogen phosphate 0.1%～2.0%, precipitated 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 culture medium preparation finishing back, 121 ℃, keeps 30 minutes.
C, fermentation parameter
Inoculum concentration 1%～10% (v/w).After the inoculation, ferment by ventilation Quchi method or plate rail method.28 ℃～32 ℃ of fermentation temperatures (control material temperature), fermentation period 48～120 hours.With the sporangium full maturity, and the standard of 30% left and right sides bacillus exfoliation for judging that fermentation stops arranged.
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 airslide disintegrating 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.
Bacillus subtilis CCTCC M204085 prepares fruit and vegetable preserving antistaling agent
A, wettable powder:
The wettable powder agent prescription
Neopelex 0.2%～20% (w/w, down together)
Sodium lignosulfonate 0.1%～20%
Expanded white carbon black 2%～20%
The former powder 5 * 10 of bacillus 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 injecting type airslide disintegrating mill, is crushed to 325 orders and sieves behind the abundant mixing, packing.
B, aqueous suspension
The former powder 5 * 10 of bacillus subtilis CCTCC M204085
8～1 * 10
10CFU/ml
Dimethylbenzene 0%～10%
Neopelex 0.2%～20%
Chitosan 0.1％～10％
Sodium carboxymethylcellulose 0%～10%
Xanthans 0%～10%
Sodium Benzoate 0.5%～2%
Water is supplemented to 100%
PH 2.0～7.0 (transferring) with watery hydrochloric acid
After above-mentioned raw materials takes by weighing in proportion, drop into the dosing chamber mixing, be milled to 95% through colloid mill and bottle behind 325 mesh sieves excessively.
C, effervescent agent (granule or tablet)
The former powder 5 * 10 of bacillus subtilis CCTCC M204085
8～3 * 10
10Individual CFU/g
Calcium dodecyl benzene sulfonate/sodium 0.2%～20%
Disintegrant (acid/alkali compounds) 3%～30%
Adhesive 2%～25%
Help and collapse agent 2%～15%
The filler surplus
Above-mentioned raw materials mixes the back and uses the tablet press machine compressing tablet or use the Squeezinggranulator granulation.
Embodiment 1: bacillus subtilis CCTCC M204085 liquid fermentation also prepares the technology of the former powder of gemma
A, bacterial classification are prepared
The culture medium 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 bacillus 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 soya-bean cake powder 1.5%, dusty yeast 0.8%, wheat bran 1.0%, rice meal 2%, corn flour 1.0%, potassium dihydrogen phosphate 0.3%, magnesium sulfate 0.3%, NaCl 0.5%.PH 8.0 before disappearing.
60L feeds intake in the 100L fermentation tank.Sterilized 30 minutes for 121 ℃.Inoculate above-mentioned bacteria suspension when being cooled to 30 ℃.29 ℃ ± 1 ℃ of liquid fermentation temperature, speed of agitator 300rpm, throughput 1: 1.2.Fermented about 32 hours, and, and put jar when having 30% left and right sides sporangium to come off to sporangium formation rate 〉=95%.Recording fermenation raw liquid living spores number is 42 * 108cfu/ml.
C, prepare former powder
Zymotic fluid is put jar the back and is transferred pH to pH4.0 with a HCl in souring tank, through disc centrifuge the is centrifugal (separating 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, bacillus 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 soya-bean cake powder 1.5%, dusty yeast 1%, precipitated calcium carbonate 1%, corn flour 1.0%, KH2PO4 0.5%, MgSO40-3%,
PH 8.0 before disappearing.Connect ready bacterial classification behind the high pressure steam sterilization, inoculum concentration is that per 200 liters of seed liquor connect 1 eggplant bottle bacterial classification.28 ℃-30 ℃ ventilations were cultivated 12 hours, speed of agitator 250rpm, and 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 culture medium
Corn flour 8%, soya-bean 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 steams are sterilized 1 hour (glucose adds low amounts of water sterilization, 115 ℃, 20 minutes separately).Sterilization is carried out in changeing pot, and the back of having gone out is advanced running 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 culture medium 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 filtrated air off and on and ferment, the control 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 semi-finished product 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 semi-finished product 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 injecting type airslide disintegrating 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, bacillus 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 zymotic fluid 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% neopelex (w/v), 1% Chitosan (w/v), 2% sodium carboxymethylcellulose (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 semi-finished product, the result is 110 * 108cfu/ml, meets the company standard requirement.Be packaged into the test specimen of one bottle of 1000ml.
Embodiment 4, bacillus 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 neopelex 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 mixer, mixed 1 hour,, cross 325 mesh sieves after airslide disintegrating 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, bacillus subtilis CCTCC M204085 preserving fruit and vegetable utilizing agent formulation production method (effervescent tablet)
Get 50 kilograms in former powder of bacillus subtilis CCTCC M204085 gemma (50,000,000,000 cfu/g), add 30 kilograms of dodecyl sodium sulfates, 17.5 kilograms of soluble starches, 15.75 kilograms in sodium carbonate, 13.75 kilograms of abundant mixings of sodium acid carbonate.Add 12.5 kilograms of citric acids again, 2 kilograms of fumaric acid, 15.75 kilograms in tartaric acid, 25 kilograms of sodium carboxymethylcelluloses, 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 tablet press 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, bacillus subtilis CCTCC M204085 and chlorothalonil compound agent (wettable powder)
Test has proved that female powder of bacillus subtilis CCTCC M204085 (in 50,000,000,000 cfu/g) and the following proportioning of Bravo (folding hundred content) have obvious synergistic effect: bacillus subtilis CCTCC M204085: Bravo=(5～1): (1～2) through lab screening.With bacillus subtilis CCTCCM204085: Bravo=2: 1 is an example.
Take by weighing 100 kilograms in the female powder of bacillus subtilis CCTCC M204085,50 kilograms of Bravos (folding hundred) (or 75% wettable powder 67 kilograms), 20 kilograms of neopelexes, surplus is a kaolin, polishing to 200 kilogram.Be crushed to more than 325 orders with airslide disintegrating mill after machinery fully mixes, one bag of per 100 gram are packaged as finished product again.Present embodiment obtains 194 kilograms of finished products, and bacillus subtilis CCTCC M204085 living spores content is 25,000,000,000 cfu/g in the finished product, Bravo content 25%.500～600 times of thin ups soak fruit and dried in about 3 minutes and get final product during use.
Embodiment 7, bacillus subtilis CCTCC M204085 and composite dose of iprodione (iprodione) (wettable powder)
Female powder of lab screening evidence bacillus subtilis CCTCC M204085 (50,000,000,000 cfu/g) and iprodione 50% wettable powder have obvious synergistic effect: bacillus subtilis CCTCC M204085: iprodione=(10～2): (1～5) under following conditions of mixture ratios.With bacillus subtilis CCTCCM204085: iprodione=1: 1 is an example.
Take by weighing 100 kilograms in the female powder of bacillus subtilis CCTCC M204085 of 50,000,000,000 cfu/g content, 100 kilograms of iprodione 50% wettable powders, add 40 kilograms of neopelexes, beautiful mica powder complements to 300 kilograms, is crushed to more than 325 orders with airslide disintegrating mill after fully mixed.One bag of per 100 gram.Present embodiment obtains 290 kilograms of finished products, contains bacillus 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, bacillus subtilis CCTCC M204085 gemma aqueous suspension agent are used for the apple storage
Get bacillus 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 bacillus 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, antiseptic effect 93.1%, and apple total acidity rate of descent 26.2%, the contrast chemical pesticide is a carbendazol wettable powder, dilutes 1000 times, processing method is identical with bacillus subtilis CCTCC M204085.Under the equal holding conditions, antiseptic effect 84.3%, total acidity descends 31.4%.Bacillus subtilis CCTCC M204085 handles and is better than chemical agent and clear water contrast.
Embodiment 9, bacillus 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 bacillus subtilis CCTCC M204085 wettable powder, add water 25kg, be diluted to the bacillus subtilis CCTCCM204085 spore suspending liquid of 2 * 108cfu/ml, to only adopt down from tree, three days fresh orange 200kg immerses the bacillus 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 carbendazim is the chemical preservative contrast, is blank with the clear water.Two kinds of processing with orange amount and pack the storage situation and bacillus subtilis CCTCCM204085 processing identical.Preserve check result after 120 days.It is 84.7% that bacillus subtilis CCTCC M204085 (wettable powder) handles antiseptic effect, and the carbendazim effect is 86.1%, and both differences are not remarkable, prove that bacillus subtilis CCTCC M204085 might replace carbendazim to be used for the orange anti-corrosive fresh-keeping.
Embodiment 10, bacillus subtilis CCTCC M204085 effervescent 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 bacillus subtilis CCTCC M204085 effervescence granular 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% carbendazim (w.p) is that the standard chemical medicament contrasts.Room temperature condition 90 days antiseptic effect bacillus subtilis CCTCC M204085 of storage down is 66.1%, and carbendazim is 60.0%, and bacillus subtilis CCTCC M204085 handles and is better than the carbendazim processing.
Embodiment 11, bacillus subtilis CCTCC M204085 gemma aqueous suspension agent are used for the cauliflower mildew-resistant
Get 500 milliliters of bacillus 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 wrapper, 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 bacillus subtilis CCTCC M204085 aqueous suspension is handled mildew-resistant reaches 78.2% (calculating by sick index).
Embodiment 12, bacillus 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 sprayer behind the mixing and go up the orange surface that is about to harvesting.The specification requirement 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 bacillus 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 bacillus subtilis CCTCC M204085 bacterium.
Claims (10)
1, a kind of antisepsis antistaling agent wherein contains bacillus subtilis (Bacillus subtilis) CCTCCM204085.
2, antisepsis antistaling agent according to claim 1 is characterized in that, it is wettable powder, aqueous suspension agent, effervescence granular or tablet.
3, antisepsis antistaling agent according to claim 2, wherein the percentage by weight of each component of wettable powder is:
Bacterial cell content 5 * 10
8～1 * 10
10Individual CFU/g
Neopelex 0.2%～20%
Sodium lignosulfonate/calcium 0.1%～20%
Odium 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 percentage by weight of each component of aqueous suspension agent is:
Bacterial cell content 5 * 10
8～3 * 10
9Individual CFU/g
Neopelex 0.2%～20%
Chitosan 0.1％～10％
Sodium carboxymethylcellulose 0%～10%
Xanthans 0%～10%
Sodium Benzoate 0.5%～2%
Water surplus
pH 2.0～7.0。
5, antisepsis antistaling agent according to claim 2, wherein the percentage by weight of effervescence granular or each component of tablet is:
Bacterial cell content 5 * 10
8～3 * 10
10Individual CFU/g
Surfactant 0.5%～20%
Disintegrant 3%～30%
Adhesive 2%～25%
Help and collapse agent 2%～15%
The filler surplus
Wherein disintegrant is acid compound or alkali compounds.
6, antisepsis antistaling agent according to claim 5, wherein surfactant is selected from one or more in dodecyl sodium sulfate, neopelex, calcium dodecyl benzene sulfonate, sodium lignin sulfonate, calcium lignosulfonate, sodium cellulose xanghate, the withered powder of tea, the spaonin powder; The disintegrant acid compound is selected from one or more in citric acid, tartaric acid, malic acid, adipic acid, fumaric acid, lactic acid, the boric acid; The disintegrant alkali compounds is selected from one or more in sodium carbonate, sodium acid carbonate, potash, the saleratus; Adhesive is selected from one or more in polyethylene glycol, polyvinylpyrrolidone, the polyvinyl alcohol; Help and collapse agent and be selected from soluble starch, dextrin, the sodium carboxymethylcellulose one or more; Filler is selected from one or more in diatomite, precipitated 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 Bravo, Te Keduo, the bright amine of rice, iprodione, thiophanate methyl, vinclozolin, tricyclazole, carbendazim.
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 bacillus subtilis (Bacillus subtilis) CCTCC M204085.
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