CN105441370A - Microbial agent and preparing method thereof - Google Patents
Microbial agent and preparing method thereof Download PDFInfo
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- CN105441370A CN105441370A CN201610061792.2A CN201610061792A CN105441370A CN 105441370 A CN105441370 A CN 105441370A CN 201610061792 A CN201610061792 A CN 201610061792A CN 105441370 A CN105441370 A CN 105441370A
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- microbiobacterial agent
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- 238000000034 method Methods 0.000 title abstract description 8
- 230000000813 microbial effect Effects 0.000 title abstract description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 62
- 230000004151 fermentation Effects 0.000 claims abstract description 62
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 54
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 27
- 230000004913 activation Effects 0.000 claims abstract description 16
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 11
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 11
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 32
- 239000002054 inoculum Substances 0.000 claims description 26
- 239000008367 deionised water Substances 0.000 claims description 24
- 229910021641 deionized water Inorganic materials 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 claims description 16
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 16
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 16
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 16
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 16
- 239000000843 powder Substances 0.000 claims description 16
- 238000004519 manufacturing process Methods 0.000 claims description 15
- 230000001580 bacterial effect Effects 0.000 claims description 14
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 claims description 13
- 239000000600 sorbitol Substances 0.000 claims description 13
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 10
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 10
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 claims description 8
- 241000283690 Bos taurus Species 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims description 8
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 229920002472 Starch Polymers 0.000 claims description 8
- 235000009582 asparagine Nutrition 0.000 claims description 8
- 229960001230 asparagine Drugs 0.000 claims description 8
- 235000003704 aspartic acid Nutrition 0.000 claims description 8
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 8
- 210000000941 bile Anatomy 0.000 claims description 8
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 8
- 239000004327 boric acid Substances 0.000 claims description 8
- 229910052933 brochantite Inorganic materials 0.000 claims description 8
- FUFJGUQYACFECW-UHFFFAOYSA-L calcium hydrogenphosphate Chemical compound [Ca+2].OP([O-])([O-])=O FUFJGUQYACFECW-UHFFFAOYSA-L 0.000 claims description 8
- 235000009508 confectionery Nutrition 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 229960000367 inositol Drugs 0.000 claims description 8
- 229960003512 nicotinic acid Drugs 0.000 claims description 8
- 235000001968 nicotinic acid Nutrition 0.000 claims description 8
- 239000011664 nicotinic acid Substances 0.000 claims description 8
- 229920001592 potato starch Polymers 0.000 claims description 8
- 239000008107 starch Substances 0.000 claims description 8
- 235000019698 starch Nutrition 0.000 claims description 8
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 5
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 5
- 241000251468 Actinopterygii Species 0.000 claims description 5
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 5
- 244000017020 Ipomoea batatas Species 0.000 claims description 5
- 235000002678 Ipomoea batatas Nutrition 0.000 claims description 5
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 5
- 229930195725 Mannitol Natural products 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 5
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 5
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 5
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 5
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 229960002989 glutamic acid Drugs 0.000 claims description 5
- 239000008101 lactose Substances 0.000 claims description 5
- 239000000594 mannitol Substances 0.000 claims description 5
- 235000010355 mannitol Nutrition 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- 239000000661 sodium alginate Substances 0.000 claims description 5
- 235000010413 sodium alginate Nutrition 0.000 claims description 5
- 229940005550 sodium alginate Drugs 0.000 claims description 5
- 239000004575 stone Substances 0.000 claims description 5
- 239000010455 vermiculite Substances 0.000 claims description 5
- 235000019354 vermiculite Nutrition 0.000 claims description 5
- 229910052902 vermiculite Inorganic materials 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- 239000002689 soil Substances 0.000 abstract description 9
- 241000193830 Bacillus <bacterium> Species 0.000 abstract 2
- 235000015110 jellies Nutrition 0.000 abstract 2
- 239000008274 jelly Substances 0.000 abstract 2
- 208000015181 infectious disease Diseases 0.000 abstract 1
- 230000002458 infectious effect Effects 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 48
- 238000011081 inoculation Methods 0.000 description 33
- 238000003756 stirring Methods 0.000 description 27
- 239000007788 liquid Substances 0.000 description 24
- 235000015097 nutrients Nutrition 0.000 description 19
- 230000001954 sterilising effect Effects 0.000 description 19
- 239000001963 growth medium Substances 0.000 description 18
- 241000894006 Bacteria Species 0.000 description 13
- 239000000306 component Substances 0.000 description 10
- 239000000356 contaminant Substances 0.000 description 9
- 238000010438 heat treatment Methods 0.000 description 9
- 239000012533 medium component Substances 0.000 description 9
- 235000019198 oils Nutrition 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 6
- 235000012343 cottonseed oil Nutrition 0.000 description 6
- 235000012054 meals Nutrition 0.000 description 6
- 239000006916 nutrient agar Substances 0.000 description 6
- 239000002068 microbial inoculum Substances 0.000 description 5
- 229910001385 heavy metal Inorganic materials 0.000 description 4
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 3
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 3
- 240000003183 Manihot esculenta Species 0.000 description 3
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 244000061458 Solanum melongena Species 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 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 description 3
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 3
- 235000015278 beef Nutrition 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 210000000582 semen Anatomy 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 235000015099 wheat brans Nutrition 0.000 description 3
- 239000000811 xylitol Substances 0.000 description 3
- 235000010447 xylitol Nutrition 0.000 description 3
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 3
- 229960002675 xylitol Drugs 0.000 description 3
- 235000002597 Solanum melongena Nutrition 0.000 description 2
- 229910052785 arsenic Inorganic materials 0.000 description 2
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 2
- 229910052753 mercury Inorganic materials 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000009335 monocropping Methods 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- 241000881860 Paenibacillus mucilaginosus Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000589180 Rhizobium Species 0.000 description 1
- 238000009305 arable farming Methods 0.000 description 1
- 239000007633 bacillus mucilaginosus Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- -1 pulvis Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a microbial agent and a preparing method thereof. The microbial agent is prepared from jelly type bacillus, bacillus subtilis and bacillus licheniformis. In the microbial agent, the viable count of jelly type bacillus ranges from 1.8 billon cfu/g to 2.2 billon cfu/g, the viable count of bacillus subtilis ranges from 1.7 billon cfu/g to 1.9 billon cfu/g, and the viable count of bacillus licheniformis ranges from 1.2 billon cfu/g to 1.3 billon cfu/g. The preparing method of the microbial agent comprises the steps of strain selection, strain activation and expanding culture, strain fermentation and finished product obtaining. The microbial agent is high in strain content, low in infectious microbe rate and capable of effectively improving soil properties.
Description
Technical field
The invention provides a kind of microbiobacterial agent and production method thereof, belong to technical field of microbial fermentation.
Background technology
Microbiobacterial agent refers to that a class contains the particular product of live microorganism.It is with the process of microbial life activity and product to improve crop alimentary condition, plays soil potential fertility, stimulates crop growth, the harm of opposing germ, thus improves crop yield and quality.It not as fertilizer to supply nutrients material directly to plant like that, but its effect can not be ignored.
Microbiobacterial agent is for arable farming and growing and transplanting seedling enlarged areas, and continuous cropping continuous cropping increases, and the soil issues caused further is day by day serious, and worse and worse, quality is difficult to ensure crop yielding condition, can well improve an above-mentioned difficult problem.Microbiobacterial agent product can be divided into liquid, pulvis, granule type by formulation; Rhizobium Inoculant, vinelandii microbial inoculum, phosphorus decomposing quasi-microorganism microbial inoculum, silicate microbiobacterial agent, photosynthetic bacterium microbial inoculum, organic matter decomposing inoculant, growth promoting bacteria agent, bush mycorrhiza agent, biological restoration microbial inoculum etc. can be divided into by the microbe species included or functional performance.
Existing microbiobacterial agent product has a lot of deficiency, particularly single microbial inoculum, cannot give full play of the advantage of microbiobacterial agent.
Prior art has following defect:
(1) bacterial content of microbiobacterial agent is lower; (2) living bacteria count of microbiobacterial agent is lower; (3) the miscellaneous bacteria rate of microbiobacterial agent is high; (4) heavy metal content of microbiobacterial agent is higher; (5) quality guaranteed period of microbiobacterial agent is short.
Summary of the invention
The present invention is the deficiency solving prior art existence, provides a kind of production method of microbiobacterial agent, to realize following goal of the invention:
1, the bacterial content of microbiobacterial agent is improved;
2, the living bacteria count of microbiobacterial agent is improved;
3, the miscellaneous bacteria rate of microbiobacterial agent is reduced;
4, the heavy metal content of microbiobacterial agent is reduced;
5, the quality guaranteed period of microbiobacterial agent is extended.
For solving the problems of the technologies described above, by the following technical solutions:
A kind of microbiobacterial agent, comprises bacillusmusilaginosiengineering, subtilis, bacillus licheniformis.
Below the further improvement to technique scheme:
In described microbiobacterial agent, bacillusmusilaginosiengineering viable count is 18 ~ 2,200,000,000 cfu/g, and subtilis viable count is 17 ~ 1,900,000,000 cfu/g, and bacillus licheniformis viable count is 12 ~ 1,300,000,000 cfu/g.
A production method for microbiobacterial agent, comprise bacterial classification select, actication of culture with spread cultivation, strain fermentation, obtain finished product step.
Described actication of culture with spread cultivation in step, the activation of bacillusmusilaginosiengineering with the formula of the substratum that spreads cultivation adopted that spreads cultivation is: sweet potato powder 15 ~ 18 weight part, bean cake powder 12 ~ 15 weight part, sodium alginate 0.6 ~ 0.8 weight part, yeast extract paste 4 ~ 4.5 weight part, L-glutamic acid 0.03 ~ 0.04 weight part, bovine bile 0.03 ~ 0.05 weight part, N.F,USP MANNITOL 0.02 ~ 0.04 weight part, Manganse Dioxide 0.01 ~ 0.02 weight part, dipotassium hydrogen phosphate 0.02 ~ 0.03 weight part, magnesium sulfate 0.05 ~ 0.07 weight part, deionized water 1000 weight part.
Described actication of culture with spread cultivation in step, the activation of subtilis with the formula of the substratum that spreads cultivation adopted that spreads cultivation is: potato starch 12 ~ 14 weight part, trehalose 9 ~ 10 weight part, sorbitol 6 ~ 8 weight part, extractum carnis 4 ~ 6 weight part, aspartic acid 0.06 ~ 0.08 weight part, glucose 0.07 ~ 0.08 weight part, ironic citrate 0.02 ~ 0.04 weight part, vitaminB10 .01 ~ 0.02 weight part, 2, 6-inositol 0.03 ~ 0.04 weight part, Manganse Dioxide 0.01 ~ 0.02 weight part, dipotassium hydrogen phosphate 0.03 ~ 0.05 weight part, seven brochantite 0.04 ~ 0.06 weight parts, deionized water 1000 weight part.
Described actication of culture with spread cultivation in step, the activation of bacillus licheniformis with the formula of the substratum that spreads cultivation adopted that spreads cultivation is: lactose 18 ~ 20 weight part, fish peptone 7 ~ 9 weight part, sorbitol 4 ~ 5 weight part, treated starch 8 ~ 10 weight part, asparagine 0.06 ~ 0.08 weight part, sweet oil 0.03 ~ 0.04 weight part, ironic citrate 0.02 ~ 0.03 weight part, nicotinic acid 0.02 ~ 0.03 weight part, secondary calcium phosphate 0.06 ~ 0.09 weight part, magnesium sulfate 0.04 ~ 0.06 weight part, boric acid 1 ~ 1.4 weight part, deionized water 1000 weight part.
In described strain fermentation step, the temperature of bacillusmusilaginosiengineering fermentation is 34 DEG C, and inoculum size is 6% of fermention medium quality, and fermentation time is 30 ~ 32 hours.
In described strain fermentation step, the leavening temperature of fermentation of bacillus subtilis is 36 DEG C, and inoculum size is 6% of fermention medium quality, and fermentation time is 36 ~ 40 hours.
In described strain fermentation step, the leavening temperature of bacillus licheniformis fermentation is 35 DEG C, and inoculum size is 6% of fermention medium quality, and fermentation time is 22 ~ 24 hours.
In described obtained finished product step, the component of carrier comprises medical stone, calcium carbonate, vermiculite power, and its mass ratio is medical stone: calcium carbonate: vermiculite power=5:2:1.
Compared with prior art, the invention has the beneficial effects as follows:
1, microbiobacterial agent of the present invention, bacterial content is high, and total bacterial content is 85 ~ 8,800,000,000 cfu/g;
2, microbiobacterial agent of the present invention, living bacteria count is high, and bacillusmusilaginosiengineering viable count is 18 ~ 2,200,000,000 cfu/g, and subtilis viable count is 17 ~ 1,900,000,000 cfu/g, and bacillus licheniformis viable count is 12 ~ 1,300,000,000 cfu/g;
3, microbiobacterial agent of the present invention, miscellaneous bacteria rate is low, and miscellaneous bacteria rate is 4.6 ~ 5.2%;
4, microbiobacterial agent of the present invention, heavy metal content is low, and arsenic and compounds content thereof are 6 ~ 9mg/kg, and mercury and mercuric compounds content is 0.7 ~ 0.8mg/kg, plumbous and compounds content 9 ~ 11mg/kg;
5, microbiobacterial agent of the present invention, long quality-guarantee period, the quality guaranteed period is 20 ~ 24 months.
Embodiment
The production method of embodiment 1 one kinds of microbiobacterial agents
Step 1, bacterial classification are selected
The bacterial classification adopted comprises:
Bacillusmusilaginosiengineering (Bacillusmucilaginosus) preserving number is ACCC10013;
Subtilis (Bacillussubtilis) preserving number is ACCC01175;
Bacillus licheniformis (Bacilluslicheniformis) preserving number is ACCC11091;
Above-mentioned bacterial classification is all from commercially available.
Step 2, actication of culture and spread cultivation
(1) bacillusmusilaginosiengineering activation with spread cultivation
By bacillusmusilaginosiengineering strain inoculation on plate culture medium, cultivate 24 hours, obtain activated spawn for 30 DEG C; Described plate culture medium is nutrient agar;
The activated spawn of bacillusmusilaginosiengineering be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 5% of substratum quality, is placed in shaking table incubator, and rotating speed is 120 revs/min, and air flow is 1:0.4, cultivates 20 hours, obtains the seed liquor that spreads cultivation for 34 DEG C;
The formula of described liquid nutrient medium is: sweet potato powder 15g, bean cake powder 12g, sodium alginate 0.6g, yeast extract paste 4g, L-glutamic acid 0.03g, bovine bile 0.03g, N.F,USP MANNITOL 0.02g, Manganse Dioxide 0.01g, dipotassium hydrogen phosphate 0.02g, magnesium sulfate 0.05g, deionized water 1kg; PH is adjusted to be 7.
(2) subtilis activation with spread cultivation
Bacillus subtilis strain is inoculated on plate culture medium, cultivates 36 hours, obtain activated spawn for 32 DEG C; Described plate culture medium is W-Gum substratum;
The activated spawn of subtilis be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 4% of substratum quality, is placed in shaking table incubator, and rotating speed is 180 revs/min, and air flow is 1:0.6, cultivates 30 hours, obtains the seed liquor that spreads cultivation for 33 DEG C;
The formula of described liquid nutrient medium is: potato starch 12g, trehalose 9g, sorbitol 6g, extractum carnis 4g, aspartic acid 0.06g, glucose 0.07g, ironic citrate 0.02g, vitaminB10 .01g, 2,6-inositol 0.03g, Manganse Dioxide 0.01g, dipotassium hydrogen phosphate 0.03g, seven brochantite 0.04g, deionized water 1kg; PH is adjusted to be 7.2.
(3) bacillus licheniformis activation with spread cultivation
By bacillus licheniformis strain inoculation on plate culture medium, cultivate 16 hours, obtain activated spawn for 35 DEG C; Described plate culture medium is beef extract-peptone nutrient agar;
The activated spawn of bacillus licheniformis be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 4% of substratum quality, is placed in shaking table incubator, and rotating speed is 200 revs/min, and air flow is 1:0.6, cultivates 22 hours, obtains the seed liquor that spreads cultivation for 33 DEG C;
The formula of described liquid nutrient medium is: lactose 18g, fish peptone 7g, sorbitol 4g, treated starch 8g, asparagine 0.06g, sweet oil 0.03g, ironic citrate 0.02g, nicotinic acid 0.02g, secondary calcium phosphate 0.06g, magnesium sulfate 0.04g, boric acid 1g, deionized water 1kg; PH is adjusted to be 6.8.
Step 3, strain fermentation
(1) bacillusmusilaginosiengineering fermentation
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Wheat bran 20 parts, cottonseed meal 18 parts, maltose 4 parts, soyflour 4 parts, extractum carnis 1 part, bovine bile 0.5 part, Xylitol 0.2 part, Manganse Dioxide 0.1 part, dipotassium hydrogen phosphate 0.06 part, 0.1 part, magnesium sulfate, deionized water 2000 parts; PH is adjusted to be 7.
Inoculation: fermentation jar temperature is down to 30 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of bacillusmusilaginosiengineering from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 34 DEG C, and 80rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 30 hours.
(2) fermentation of bacillus subtilis
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Tapioca Starch 18 parts, sucrose 4 parts, sorbitol 1 part, extractum carnis 2 parts, aspartic acid 0.06 part, Semen Maydis powder 8 parts, ironic citrate 0.05 part, vitaminB10 .03 part, 2,6-inositol 0.05 part, Manganse Dioxide 0.06 part, dipotassium hydrogen phosphate 0.1 part, seven brochantite 0.05 part, deionized water 2000 parts; PH is adjusted to be 7.2.
Inoculation: fermentation jar temperature is down to 30 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of subtilis from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 36 DEG C, and 90rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 36 hours.
(3) bacillus licheniformis fermentation
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25min;
Described fermention medium comprises the component of following weight part:
Potato starch 30 parts, yeast powder 1 part, cottonseed meal 10 parts, glucose 4 parts, asparagine 0.06 part, 0.1 part, sweet oil, ironic citrate 0.1 part, 0.05 part, nicotinic acid, secondary calcium phosphate 0.1 part, 0.05 part, magnesium sulfate, boric acid 1 part, deionized water 2000 parts; PH is adjusted to be 6.8.
Inoculation: fermentation jar temperature is down to 30 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of subtilis from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 35 DEG C, and 80rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 22 hours.
Step 4, obtained finished product
Medical stone, calcium carbonate, vermiculite power are pressed the weight ratio mixing of 5:2:1, after sterilizing, as carrier;
3 kinds of fermented liquids step 3 fermentation obtained, filter, obtain bacillusmusilaginosiengineering bacterium liquid, subtilis bacterium liquid, bacillus licheniformis bacterium liquid;
After bacterium liquid in above-mentioned 3 being pressed the mass ratio mixing of 1:1:1, be sprayed onto on carrier, make biodiversity percentage composition be 28%.
The production method of embodiment 2 one kinds of microbiobacterial agents
Step 1, bacterial classification are selected
The bacterial classification adopted is identical with embodiment 1.
Step 2, actication of culture and spread cultivation
(1) bacillusmusilaginosiengineering activation with spread cultivation
By bacillusmusilaginosiengineering strain inoculation on plate culture medium, cultivate 24 hours, obtain activated spawn for 32 DEG C; Described plate culture medium is nutrient agar;
The activated spawn of bacillusmusilaginosiengineering be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 5% of substratum quality, is placed in shaking table incubator, and rotating speed is 120 revs/min, and air flow is 1:0.4, cultivates 21 hours, obtains the seed liquor that spreads cultivation for 35 DEG C;
The formula of described liquid nutrient medium is: sweet potato powder 17g, bean cake powder 14g, sodium alginate 0.7g, yeast extract paste 4.2g, L-glutamic acid 0.03g, bovine bile 0.04g, N.F,USP MANNITOL 0.04g, Manganse Dioxide 0.01g, dipotassium hydrogen phosphate 0.03g, magnesium sulfate 0.06g, deionized water 1kg; PH is adjusted to be 7.
(2) subtilis activation with spread cultivation
Bacillus subtilis strain is inoculated on plate culture medium, cultivates 36 hours, obtain activated spawn for 33 DEG C; Described plate culture medium is W-Gum substratum;
The activated spawn of subtilis be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 4% of substratum quality, is placed in shaking table incubator, and rotating speed is 180 revs/min, and air flow is 1:0.6, cultivates 30 hours, obtains the seed liquor that spreads cultivation for 33 DEG C;
The formula of described liquid nutrient medium is: potato starch 13g, trehalose 9.5g, sorbitol 7g, extractum carnis 4.5g, aspartic acid 0.07g, glucose 0.07g, ironic citrate 0.03g, vitaminB10 .01g, 2,6-inositol 0.03g, Manganse Dioxide 0.02g, dipotassium hydrogen phosphate 0.04g, seven brochantite 0.05g, deionized water 1kg; PH is adjusted to be 7.2.
(3) bacillus licheniformis activation with spread cultivation
By bacillus licheniformis strain inoculation on plate culture medium, cultivate 16 hours, obtain activated spawn for 35 DEG C; Described plate culture medium is beef extract-peptone nutrient agar;
The activated spawn of bacillus licheniformis be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 4% of substratum quality, is placed in shaking table incubator, and rotating speed is 200 revs/min, and air flow is 1:0.6, cultivates 22 hours, obtains the seed liquor that spreads cultivation for 33 DEG C;
The formula of described liquid nutrient medium is: lactose 19g, fish peptone 8g, sorbitol 4.5g, treated starch 9g, asparagine 0.07g, sweet oil 0.03g, ironic citrate 0.03g, nicotinic acid 0.02g, secondary calcium phosphate 0.08g, magnesium sulfate 0.05g, boric acid 1.2g, deionized water 1kg; PH is adjusted to be 6.8.
Step 3, strain fermentation
(1) bacillusmusilaginosiengineering fermentation
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Wheat bran 22 parts, cottonseed meal 18 parts, maltose 5 parts, soyflour 4.2 parts, extractum carnis 1.5 parts, bovine bile 0.5 part, Xylitol 0.3 part, Manganse Dioxide 0.1 part, dipotassium hydrogen phosphate 0.07 part, 0.1 part, magnesium sulfate, deionized water 2000 parts; PH is adjusted to be 7.
Inoculation: fermentation jar temperature is down to 33 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of bacillusmusilaginosiengineering from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 34 DEG C, and 80rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 31 hours.
(2) fermentation of bacillus subtilis
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Tapioca Starch 19 parts, sucrose 5 parts, sorbitol 1.5 parts, extractum carnis 2 parts, aspartic acid 0.07 part, Semen Maydis powder 10 parts, ironic citrate 0.06 part, vitaminB10 .04 part, 2,6-inositol 0.06 part, Manganse Dioxide 0.07 part, dipotassium hydrogen phosphate 0.15 part, seven brochantite 0.06 part, deionized water 2000 parts; PH is adjusted to be 7.2.
Inoculation: fermentation jar temperature is down to 32 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of subtilis from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 36 DEG C, and 90rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 38 hours.
(3) bacillus licheniformis fermentation
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Potato starch 32 parts, yeast powder 1.5 parts, cottonseed meal 13 parts, glucose 5 parts, asparagine 0.07 part, 0.1 part, sweet oil, ironic citrate 0.2 part, 0.05 part, nicotinic acid, secondary calcium phosphate 0.2 part, 0.07 part, magnesium sulfate, boric acid 1.2 parts, deionized water 2000 parts; PH is adjusted to be 6.8.
Inoculation: fermentation jar temperature is down to 33 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of subtilis from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 35 DEG C, and 80rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 23 hours.
Step 4, obtained finished product
Identical with the working method of embodiment 1.
The production method of embodiment 3 one kinds of microbiobacterial agents
Step 1, bacterial classification are selected
The bacterial classification adopted is identical with embodiment 1.
Step 2, actication of culture and spread cultivation
(1) bacillusmusilaginosiengineering activation with spread cultivation
By bacillusmusilaginosiengineering strain inoculation on plate culture medium, cultivate 24 hours, obtain activated spawn for 34 DEG C; Described plate culture medium is nutrient agar;
The activated spawn of bacillusmusilaginosiengineering be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 5% of substratum quality, is placed in shaking table incubator, and rotating speed is 120 revs/min, and air flow is 1:0.4, cultivates 22 hours, obtains the seed liquor that spreads cultivation for 36 DEG C;
The formula of described liquid nutrient medium is: sweet potato powder 18g, bean cake powder 15g, sodium alginate 0.8g, yeast extract paste 4.5g, L-glutamic acid 0.04g, bovine bile 0.05g, N.F,USP MANNITOL 0.04g, Manganse Dioxide 0.02g, dipotassium hydrogen phosphate 0.03g, magnesium sulfate 0.07g, deionized water 1kg; PH is adjusted to be 7.
(2) subtilis activation with spread cultivation
Bacillus subtilis strain is inoculated on plate culture medium, cultivates 36 hours, obtain activated spawn for 33 DEG C; Described plate culture medium is W-Gum substratum;
The activated spawn of subtilis be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 4% of substratum quality, is placed in shaking table incubator, and rotating speed is 180 revs/min, and air flow is 1:0.6, cultivates 30 hours, obtains the seed liquor that spreads cultivation for 33 DEG C;
The formula of described liquid nutrient medium is: potato starch 14g, trehalose 10g, sorbitol 8g, extractum carnis 6g, aspartic acid 0.08g, glucose 0.08g, ironic citrate 0.04g, vitaminB10 .02g, 2,6-inositol 0.04g, Manganse Dioxide 0.02g, dipotassium hydrogen phosphate 0.05g, seven brochantite 0.06g, deionized water 1kg; PH is adjusted to be 7.2.
(3) bacillus licheniformis activation with spread cultivation
By bacillus licheniformis strain inoculation on plate culture medium, cultivate 16 hours, obtain activated spawn for 35 DEG C; Described plate culture medium is beef extract-peptone nutrient agar;
The activated spawn of bacillus licheniformis be inoculated in the liquid nutrient medium in shaking flask, inoculum size is 4% of substratum quality, is placed in shaking table incubator, and rotating speed is 200 revs/min, and air flow is 1:0.6, cultivates 22 hours, obtains the seed liquor that spreads cultivation for 33 DEG C;
The formula of described liquid nutrient medium is: lactose 20g, fish peptone 9g, sorbitol 5g, treated starch 10g, asparagine 0.08g, sweet oil 0.04g, ironic citrate 0.03g, nicotinic acid 0.03g, secondary calcium phosphate 0.09g, magnesium sulfate 0.06g, boric acid 1.4g, deionized water 1kg; PH is adjusted to be 6.8.
Step 3, strain fermentation
(1) bacillusmusilaginosiengineering fermentation
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Wheat bran 23 parts, cottonseed meal 19 parts, maltose 6 parts, soyflour 4.5 parts, extractum carnis 2 parts, bovine bile 0.6 part, Xylitol 0.4 part, Manganse Dioxide 0.2 part, dipotassium hydrogen phosphate 0.09 part, 0.2 part, magnesium sulfate, deionized water 2000 parts; PH is adjusted to be 7.
Inoculation: fermentation jar temperature is down to 35 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of bacillusmusilaginosiengineering from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 34 DEG C, and 80rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 32 hours.
(2) fermentation of bacillus subtilis
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Tapioca Starch 20 parts, sucrose 6 parts, sorbitol 2 parts, extractum carnis 3 parts, aspartic acid 0.08 part, Semen Maydis powder 11 parts, ironic citrate 0.07 part, vitaminB10 .04 part, 2,6-inositol 0.07 part, Manganse Dioxide 0.08 part, dipotassium hydrogen phosphate 0.2 part, seven brochantite 0.07 part, deionized water 2000 parts; PH is adjusted to be 7.2.
Inoculation: fermentation jar temperature is down to 35 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of subtilis from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 36 DEG C, and 90rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 40 hours.
(3) bacillus licheniformis fermentation
Fermention medium sterilizing: each for fermention medium component is weighed, adds in fermentor tank, start stirring, 120rpm stirs 10min; Heating, supercharging, fermentor tank internal pressure reaches 0.105MPa, and temperature reaches 121 DEG C, sterilizing 25 ~ 30min;
Described fermention medium comprises the component of following weight part:
Potato starch 34 parts, yeast powder 2 parts, cottonseed meal 15 parts, glucose 6 parts, asparagine 0.08 part, 0.2 part, sweet oil, ironic citrate 0.2 part, 0.06 part, nicotinic acid, secondary calcium phosphate 0.2 part, 0.08 part, magnesium sulfate, boric acid 1.4 parts, deionized water 2000 parts; PH is adjusted to be 6.8.
Inoculation: fermentation jar temperature is down to 35 DEG C, under aseptic condition, add the seed liquor that spreads cultivation of subtilis from inoculation mouth, inoculum size is 6% of fermention medium quality;
Fermentation: after inoculation, controls fermentation jar temperature 35 DEG C, and 80rpm stirs, and continues to pass into sterile air, avoids living contaminants; Ferment 24 hours.
Step 4, obtained finished product
Identical with the working method of embodiment 1.
Microbiobacterial agent prepared by embodiment 1-3 is detected; Detected result is in table 1;
Table 1
From table 1, microbiobacterial agent of the present invention, total bacterial content is 85 ~ 8,800,000,000 cfu/g, and bacillusmusilaginosiengineering viable count is 18 ~ 2,200,000,000 cfu/g, subtilis viable count is 17 ~ 1,900,000,000 cfu/g, bacillus licheniformis viable count is 12 ~ 1,300,000,000 cfu/g, and miscellaneous bacteria rate is 4.6 ~ 5.2%, and fineness is 120 orders, arsenic and compounds content thereof are 6 ~ 9mg/kg, mercury and mercuric compounds content is 0.7 ~ 0.8mg/kg, and plumbous and compounds content 9 ~ 11mg/kg, the quality guaranteed period is 20 ~ 24 months.
Effect experimental:
Applied by the microbiobacterial agent of embodiment 1-3 in eggplant plantation plot, every mu applies microbiobacterial agent 40 ~ 50kg, mix with topsoil, plants eggplant 1 year, soil property be improved significantly, specific targets are in table 2;
Performance index after table 2 soil improvement
From table 2, adopt after microbiobacterial agent, soil property be improved significantly, soil pH is 6.8 ~ 7.2, and the soil weight is 1.36 ~ 1.41g/cm
3, ventilation voidage is 20 ~ 23%; Heavy metal in soil content obviously reduces, and content of microorganisms significantly improves.
After adopting microbiobacterial agent, eggplant yield improves 15 ~ 18%, and commodity fruit rate improves 23 ~ 25%, and malformed fruit rate reduces 9 ~ 13%.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.The above is the citing of best mode for carrying out the invention, and the part wherein do not addressed in detail is the common practise of those of ordinary skill in the art; Protection scope of the present invention is as the criterion with the content of claim, and any equivalent transformation carried out based on technology enlightenment of the present invention, also within protection scope of the present invention.
Claims (10)
1. a microbiobacterial agent, is characterized in that: comprise bacillusmusilaginosiengineering, subtilis, bacillus licheniformis.
2. a kind of microbiobacterial agent according to claim 1, it is characterized in that: in described microbiobacterial agent, bacillusmusilaginosiengineering viable count is 18 ~ 2,200,000,000 cfu/g, and subtilis viable count is 17 ~ 1,900,000,000 cfu/g, and bacillus licheniformis viable count is 12 ~ 1,300,000,000 cfu/g.
3. a production method for microbiobacterial agent, is characterized in that: comprise bacterial classification select, actication of culture with spread cultivation, strain fermentation, obtain finished product step.
4. the production method of a kind of microbiobacterial agent according to claim 3, it is characterized in that: described actication of culture with spread cultivation in step, the activation of bacillusmusilaginosiengineering with the formula of the substratum that spreads cultivation adopted that spreads cultivation is: sweet potato powder 15 ~ 18 weight part, bean cake powder 12 ~ 15 weight part, sodium alginate 0.6 ~ 0.8 weight part, yeast extract paste 4 ~ 4.5 weight part, L-glutamic acid 0.03 ~ 0.04 weight part, bovine bile 0.03 ~ 0.05 weight part, N.F,USP MANNITOL 0.02 ~ 0.04 weight part, Manganse Dioxide 0.01 ~ 0.02 weight part, dipotassium hydrogen phosphate 0.02 ~ 0.03 weight part, magnesium sulfate 0.05 ~ 0.07 weight part, deionized water 1000 weight part.
5. the production method of a kind of microbiobacterial agent according to claim 3, it is characterized in that: described actication of culture with spread cultivation in step, the activation of subtilis with the formula of the substratum that spreads cultivation adopted that spreads cultivation is: potato starch 12 ~ 14 weight part, trehalose 9 ~ 10 weight part, sorbitol 6 ~ 8 weight part, extractum carnis 4 ~ 6 weight part, aspartic acid 0.06 ~ 0.08 weight part, glucose 0.07 ~ 0.08 weight part, ironic citrate 0.02 ~ 0.04 weight part, vitaminB10 .01 ~ 0.02 weight part, 2, 6-inositol 0.03 ~ 0.04 weight part, Manganse Dioxide 0.01 ~ 0.02 weight part, dipotassium hydrogen phosphate 0.03 ~ 0.05 weight part, seven brochantite 0.04 ~ 0.06 weight parts, deionized water 1000 weight part.
6. the production method of a kind of microbiobacterial agent according to claim 3, it is characterized in that: described actication of culture with spread cultivation in step, the activation of bacillus licheniformis with the formula of the substratum that spreads cultivation adopted that spreads cultivation is: lactose 18 ~ 20 weight part, fish peptone 7 ~ 9 weight part, sorbitol 4 ~ 5 weight part, treated starch 8 ~ 10 weight part, asparagine 0.06 ~ 0.08 weight part, sweet oil 0.03 ~ 0.04 weight part, ironic citrate 0.02 ~ 0.03 weight part, nicotinic acid 0.02 ~ 0.03 weight part, secondary calcium phosphate 0.06 ~ 0.09 weight part, magnesium sulfate 0.04 ~ 0.06 weight part, boric acid 1 ~ 1.4 weight part, deionized water 1000 weight part.
7. the production method of a kind of microbiobacterial agent according to claim 3, is characterized in that: in described strain fermentation step, and the temperature of bacillusmusilaginosiengineering fermentation is 34 DEG C, and inoculum size is 6% of fermention medium quality, and fermentation time is 30 ~ 32 hours.
8. the production method of a kind of microbiobacterial agent according to claim 3, it is characterized in that: in described strain fermentation step, the leavening temperature of fermentation of bacillus subtilis is 36 DEG C, and inoculum size is 6% of fermention medium quality, and fermentation time is 36 ~ 40 hours.
9. the production method of a kind of microbiobacterial agent according to claim 3, it is characterized in that: in described strain fermentation step, the leavening temperature of bacillus licheniformis fermentation is 35 DEG C, and inoculum size is 6% of fermention medium quality, and fermentation time is 22 ~ 24 hours.
10. the production method of a kind of microbiobacterial agent according to claim 3, is characterized in that: in described obtained finished product step, the component of carrier comprises medical stone, calcium carbonate, vermiculite power, and its mass ratio is medical stone: calcium carbonate: vermiculite power=5:2:1.
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