CN105441370B - A kind of microbial bacterial agent and its production method - Google Patents
A kind of microbial bacterial agent and its production method Download PDFInfo
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- CN105441370B CN105441370B CN201610061792.2A CN201610061792A CN105441370B CN 105441370 B CN105441370 B CN 105441370B CN 201610061792 A CN201610061792 A CN 201610061792A CN 105441370 B CN105441370 B CN 105441370B
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- 230000001580 bacterial effect Effects 0.000 title claims abstract description 48
- 230000000813 microbial effect Effects 0.000 title claims abstract description 45
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 11
- 238000000855 fermentation Methods 0.000 claims abstract description 117
- 230000004151 fermentation Effects 0.000 claims abstract description 117
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 45
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 34
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 33
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 28
- 241000894006 Bacteria Species 0.000 claims abstract description 26
- 239000002609 medium Substances 0.000 claims description 77
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical group [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 30
- NUJOXMJBOLGQSY-UHFFFAOYSA-N manganese dioxide Chemical compound O=[Mn]=O NUJOXMJBOLGQSY-UHFFFAOYSA-N 0.000 claims description 28
- 239000002054 inoculum Substances 0.000 claims description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 239000008367 deionised water Substances 0.000 claims description 22
- 229910021641 deionized water Inorganic materials 0.000 claims description 22
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 16
- 230000004913 activation Effects 0.000 claims description 15
- 239000001963 growth medium Substances 0.000 claims description 15
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- 239000000284 extract Substances 0.000 claims description 11
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- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 210000000941 bile Anatomy 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
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- 229920001592 potato starch Polymers 0.000 claims description 8
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims description 8
- 235000008390 olive oil Nutrition 0.000 claims description 7
- 239000004006 olive oil Substances 0.000 claims description 7
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- 235000009582 asparagine Nutrition 0.000 claims description 6
- 229960001230 asparagine Drugs 0.000 claims description 6
- 235000003704 aspartic acid Nutrition 0.000 claims description 6
- 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 6
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 6
- 239000004327 boric acid Substances 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 6
- 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 group 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
- 229920000881 Modified starch Polymers 0.000 claims description 5
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- 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
- 239000008101 lactose Substances 0.000 claims description 5
- 235000019426 modified starch 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
- 229940088594 vitamin Drugs 0.000 claims description 5
- 235000013343 vitamin Nutrition 0.000 claims description 5
- 239000011782 vitamin Substances 0.000 claims description 5
- 229930003231 vitamin Natural products 0.000 claims description 5
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- 229910052785 arsenic Inorganic materials 0.000 claims description 3
- 230000008859 change Effects 0.000 claims description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical group OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 3
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 claims description 3
- 229910052753 mercury Inorganic materials 0.000 claims description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 2
- 241000881860 Paenibacillus mucilaginosus Species 0.000 claims description 2
- 239000007633 bacillus mucilaginosus Substances 0.000 claims description 2
- 235000013922 glutamic acid Nutrition 0.000 claims description 2
- 239000004220 glutamic acid Substances 0.000 claims description 2
- 239000002689 soil Substances 0.000 abstract description 9
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- 239000002068 microbial inoculum Substances 0.000 description 5
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 4
- 235000005979 Citrus limon Nutrition 0.000 description 4
- 244000131522 Citrus pyriformis Species 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 229930195725 Mannitol Natural products 0.000 description 4
- 239000000594 mannitol Substances 0.000 description 4
- 235000010355 mannitol Nutrition 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
- 229920001817 Agar Polymers 0.000 description 3
- 229920002261 Corn starch Polymers 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
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- 244000061458 Solanum melongena Species 0.000 description 3
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- 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
- 229930003451 Vitamin B1 Natural products 0.000 description 3
- 240000008042 Zea mays Species 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
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- 229960003495 thiamine Drugs 0.000 description 3
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 3
- 239000011691 vitamin B1 Substances 0.000 description 3
- 235000010374 vitamin B1 Nutrition 0.000 description 3
- 235000015099 wheat brans Nutrition 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000432824 Asparagus densiflorus Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 description 2
- 235000002597 Solanum melongena Nutrition 0.000 description 2
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001408 amides Chemical class 0.000 description 2
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 2
- 229910052796 boron Inorganic materials 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000009335 monocropping Methods 0.000 description 2
- 238000011017 operating method Methods 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 239000000811 xylitol Substances 0.000 description 2
- 235000010447 xylitol Nutrition 0.000 description 2
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 2
- 229960002675 xylitol Drugs 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 240000007817 Olea europaea Species 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 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
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- GOPYZMJAIPBUGX-UHFFFAOYSA-N [O-2].[O-2].[Mn+4] Chemical group [O-2].[O-2].[Mn+4] GOPYZMJAIPBUGX-UHFFFAOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000009305 arable farming Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- UBXWAYGQRZFPGU-UHFFFAOYSA-N manganese(2+) oxygen(2-) titanium(4+) Chemical compound [O--].[O--].[Ti+4].[Mn++] UBXWAYGQRZFPGU-UHFFFAOYSA-N 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000011368 organic material 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
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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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 present invention provides a kind of microbial bacterial agent and its production method, including bacillusmusilaginosiengineering, bacillus subtilis, bacillus licheniformis.In the microbial bacterial agent, bacillusmusilaginosiengineering viable count is 18~2,200,000,000 cfu/g, and bacillus subtilis viable count is 17~1,900,000,000 cfu/g, and bacillus licheniformis viable count is 12~1,300,000,000 cfu/g.The production method of microbial bacterial agent, including strain selection, actication of culture and spread cultivation, strain fermentation, finished product step.Microbial bacterial agent of the present invention, bacterial content is high, and miscellaneous bacteria rate is low, can effectively improve soil property.
Description
Technical field
The present invention provides a kind of microbial bacterial agent and its production method, belongs to technical field of microbial fermentation.
Background technique
Microbial bacterial agent refers to a kind of particular product containing viable microbial.It is with the movable process of microbial life and
Product improves crop nutritional condition, plays soil potential fertility, stimulates crop growth, germ harm is resisted, to mention
High crop yield and quality.It supplies nutrients substance not as general fertilizer directly to plant, but its effect can not be ignored.
Microbial bacterial agent expands for arable farming and growing and transplanting seedling area, and continuous cropping continuous cropping increases, the soil further resulted in
Earth problem is got worse, and worse and worse, quality is difficult to ensure crop yielding condition, can improve above-mentioned problem well.
Microbial bacterial agent product can be divided into liquid, pulvis, granular pattern by dosage form；It can by the microbe species or functional characteristic that include
It is divided into Rhizobium Inoculant, fixed nitrogen bacteria agent, phosphorus decomposing quasi-microorganism microbial inoculum, silicate microbial bacterial agent, photosynthetic bacteria microbial inoculum, organic
Material decomposing agent, growth promoting bacteria agent, bush mycorrhiza agent, biological prosthetic microbial inoculum etc..
Existing microbial bacterial agent product has many deficiencies, and especially single microbial inoculum is unable to give full play out microorganism
The advantage of microbial inoculum.
The prior art has the disadvantage that
(1) bacterial content of microbial bacterial agent is lower；(2) living bacteria count of microbial bacterial agent is lower；(3) microbial bacterial agent
Miscellaneous bacteria rate it is high；(4) content of beary metal of microbial bacterial agent is higher；(5) shelf-life of microbial bacterial agent is short.
Summary of the invention
The present invention be solve the shortcomings of the prior art, a kind of production method of microbial bacterial agent is provided, with realize with
Lower goal of the invention:
1, the bacterial content of microbial bacterial agent is improved；
2, the living bacteria count of microbial bacterial agent is improved；
3, the miscellaneous bacteria rate of microbial bacterial agent is reduced；
4, the content of beary metal of microbial bacterial agent is reduced；
5, extend the shelf-life of microbial bacterial agent.
In order to solve the above technical problems, using following technical scheme:
A kind of microbial bacterial agent, including bacillusmusilaginosiengineering, bacillus subtilis, bacillus licheniformis.
It is further improvement to above-mentioned technical proposal below:
In the microbial bacterial agent, bacillusmusilaginosiengineering viable count is 18~2,200,000,000 cfu/g, bacillus subtilis viable bacteria
Number is 17~1,900,000,000 cfu/g, and bacillus licheniformis viable count is 12~1,300,000,000 cfu/g.
A kind of production method of microbial bacterial agent, including strain selection, actication of culture and spread cultivation, strain fermentation, be made into
Product step.
In the actication of culture and the step that spreads cultivation, the culture medium that spreads cultivation of the activation and the use that spreads cultivation of bacillusmusilaginosiengineering
Formula are as follows: 15~18 parts by weight of sweet potato powder, 12~15 parts by weight of bean cake powder, 0.6~0.8 parts by weight of sodium alginate, yeast extract 4
~4.5 parts by weight, 0.03~0.04 parts by weight of glutamic acid, 0.03~0.05 parts by weight of bovine bile, 0.02~0.04 weight of mannitol
Measure part, 0.01~0.02 parts by weight of manganese dioxide, 0.02~0.03 parts by weight of dipotassium hydrogen phosphate, 0.05~0.07 weight of magnesium sulfate
Part, 1000 parts by weight of deionized water.
In the actication of culture and the step that spreads cultivation, activation and the culture medium that spreads cultivation for the use that spreads cultivation of bacillus subtilis
Formula are as follows: 12~14 parts by weight of potato starch, 9~10 parts by weight of trehalose, 6~8 parts by weight of sorbitol, 4~6 weight of beef extract
Measure part, 0.06~0.08 parts by weight of aspartic acid, 0.07~0.08 parts by weight of glucose, 0.02~0.04 weight of ironic citrate
Part, 0.01~0.02 parts by weight of vitamin B1,0.03~0.04 parts by weight of 2,6- inositol, 0.01~0.02 weight of manganese dioxide
Part, 0.03~0.05 parts by weight of dipotassium hydrogen phosphate, seven brochanites, 0.04~0.06 parts by weight, 1000 parts by weight of deionized water.
In the actication of culture and the step that spreads cultivation, activation and the culture medium that spreads cultivation for the use that spreads cultivation of bacillus licheniformis
Formula are as follows: 18~20 parts by weight of lactose, 7~9 parts by weight of fish peptone, 4~5 parts by weight of sorbitol, 8~10 weight of modified starch
Measure part, 0.06~0.08 parts by weight of asparagine, 0.03~0.04 parts by weight of olive oil, 0.02~0.03 weight of ironic citrate
Part, 0.02~0.03 parts by weight of niacin, 0.06~0.09 parts by weight of calcium monohydrogen phosphate, 0.04~0.06 parts by weight of magnesium sulfate, boric acid
1~1.4 parts by weight, 1000 parts by weight of deionized water.
In the strain fermentation step, the temperature of bacillusmusilaginosiengineering fermentation is 34 DEG C, and inoculum concentration is fermented and cultured
The 6% of matrix amount, fermentation time are 30~32 hours.
In the strain fermentation step, the fermentation temperature of fermentation of bacillus subtilis is 36 DEG C, and inoculum concentration is fermentation training
The 6% of matrix amount is supported, fermentation time is 36~40 hours.
In the strain fermentation step, the fermentation temperature of bacillus licheniformis fermentation is 35 DEG C, and inoculum concentration is fermentation training
The 6% of matrix amount is supported, fermentation time is 22~24 hours.
In the finished product step, the component of carrier includes medical stone, calcium carbonate, vermiculite power, and mass ratio is wheat
Meal stone: calcium carbonate: vermiculite power=5:2:1.
Compared with prior art, the beneficial effects of the present invention are:
1, microbial bacterial agent of the present invention, bacterial content is high, and total bacterial content is 85~8,800,000,000 cfu/g；
2, microbial bacterial agent of the present invention, living bacteria count is high, and bacillusmusilaginosiengineering viable count is 18~2,200,000,000 cfu/g,
Bacillus 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, microbial bacterial agent of the present invention, miscellaneous bacteria rate is low, and miscellaneous bacteria rate is 4.6~5.2%；
4, microbial bacterial agent of the present invention, content of beary metal is low, and arsenic and its compounds content are 6~9 mg/kg, mercury and its change
Conjunction object content is 0.7~0.8 mg/kg, 9~11 mg/kg of lead and its compounds content；
5, microbial bacterial agent of the present invention, long shelf-life, shelf-life are 20~24 months.
Specific embodiment
A kind of production method of the microbial bacterial agent of embodiment 1
Step 1, strain selection
The strain of use includes:
Bacillusmusilaginosiengineering (Bacillus mucilaginosus) deposit number is ACCC10013；
Bacillus subtilis (Bacillus subtilis) deposit number be ACCC01175；
Bacillus licheniformis (Bacillus licheniformis) deposit number is ACCC11091；
Above-mentioned strain is all from commercially available.
It step 2, actication of culture and spreads cultivation
(1) activation of bacillusmusilaginosiengineering with spread cultivation
Bacillusmusilaginosiengineering strain is inoculated on plating medium, 30 DEG C are cultivated 24 hours, and activated spawn is obtained；Institute
Stating plating medium is nutrient agar；
The activated spawn of bacillusmusilaginosiengineering is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture substrate
The 5% of amount, is placed in shaking table culture case, and revolving speed is 120 revs/min, and ventilatory capacity 1:0.4,34 DEG C are cultivated 20 hours, are spread cultivation
Seed liquor；
The formula of the fluid nutrient medium are as follows: sweet potato powder 15g, bean cake powder 12g, sodium alginate 0.6g, yeast extract 4g, paddy ammonia
Sour 0.03g, it bovine bile 0.03g, mannitol 0.02g, manganese dioxide 0.01g, dipotassium hydrogen phosphate 0.02g, magnesium sulfate 0.05g, goes
Ionized water 1kg；Adjusting pH is 7.
(2) activation of bacillus subtilis with spread cultivation
Bacillus subtilis strain is inoculated on plating medium, 32 DEG C are cultivated 36 hours, and activated spawn is obtained；It is described
Plating medium is cornstarch culture medium；
The activated spawn of bacillus subtilis is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture medium quality
4%, be placed in shaking table culture case, revolving speed is 180 revs/min, ventilatory capacity 1:0.6,33 DEG C and cultivates 30 hours, obtains the kind that spreads cultivation
Sub- liquid；
The formula of the fluid nutrient medium are as follows: potato starch 12g, trehalose 9g, sorbitol 6g, beef extract 4g, asparagus fern
Propylhomoserin 0.06g, glucose 0.07g, ironic citrate 0.02g, vitamin B1 0.01g, 2,6- inositol 0.03g, manganese dioxide
0.01g, dipotassium hydrogen phosphate 0.03g, seven brochanite 0.04g, deionized water 1kg；Adjusting pH is 7.2.
(3) activation of bacillus licheniformis with spread cultivation
Bacillus licheniformis strain is inoculated on plating medium, 35 DEG C are cultivated 16 hours, and activated spawn is obtained；It is described
Plating medium is beef extract-peptone agar medium；
The activated spawn of bacillus licheniformis is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture medium quality
4%, be placed in shaking table culture case, revolving speed is 200 revs/min, ventilatory capacity 1:0.6,33 DEG C and cultivates 22 hours, obtains the kind that spreads cultivation
Sub- liquid；
The formula of the fluid nutrient medium are as follows: lactose 18g, fish peptone 7g, sorbitol 4g, modified starch 8g, Tianmen
Winter amide 0.06g, olive oil 0.03g, ironic citrate 0.02g, niacin 0.02g, calcium monohydrogen phosphate 0.06g, magnesium sulfate 0.04g, boron
Sour 1g, deionized water 1kg；Adjusting pH is 6.8.
Step 3, strain fermentation
(1) bacillusmusilaginosiengineering ferments
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
20 parts of wheat bran, 18 parts of cottonseed meal, 4 parts of maltose, 4 parts of soy meal, 1 part of beef extract, 0.5 part of bovine bile, wood
0.2 part of sugar alcohol, 0.1 part of manganese dioxide, 0.06 part of dipotassium hydrogen phosphate, 0.1 part of magnesium sulfate, 2000 parts of deionized water；Adjusting pH is 7.
Inoculation: fermentation jar temperature is down to 30 DEG C, under aseptic condition, and the kind that spreads cultivation of bacillusmusilaginosiengineering is added from inoculation mouth
Sub- liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 34 DEG C of fermentation jar temperature, and 80rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 30 hours.
(2) fermentation of bacillus subtilis
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
18 parts of tapioca starch, 4 parts of sucrose, 1 part of sorbitol, 2 parts of beef extract, 0.06 part of aspartic acid, 8 parts of corn flour, lemon
0.05 part of lemon acid iron, 0.03 part of vitamin B1,0.05 part of 2,6- inositol, 0.06 part of manganese dioxide, 0.1 part of dipotassium hydrogen phosphate,
Seven 0.05 part of brochanites, 2000 parts of deionized water；Adjusting pH is 7.2.
Inoculation: fermentation jar temperature is down to 30 DEG C, under aseptic condition, and the seed that spreads cultivation of bacillus subtilis is added from inoculation mouth
Liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 36 DEG C of fermentation jar temperature, and 90rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 36 hours.
(3) bacillus licheniformis ferments
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25min；
The fermentation medium comprises the following components in parts by weight:
30 parts of potato starch, 1 part of yeast powder, 10 parts of cottonseed meal, 4 parts of glucose, 0.06 part of asparagine, olive oil
0.1 part, 0.1 part of ironic citrate, 0.05 part of niacin, 0.1 part of calcium monohydrogen phosphate, 0.05 part of magnesium sulfate, 1 part of boric acid, deionized water
2000 parts；Adjusting pH is 6.8.
Inoculation: fermentation jar temperature is down to 30 DEG C, under aseptic condition, and the seed that spreads cultivation of bacillus subtilis is added from inoculation mouth
Liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 35 DEG C of fermentation jar temperature, and 80rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 22 hours.
Step 4, finished product
Medical stone, calcium carbonate, vermiculite power are mixed by the weight ratio of 5:2:1, after sterilizing, as carrier；
Step 3 is fermented 3 kinds of fermentation liquids obtained, filtering obtains bacillusmusilaginosiengineering bacterium solution, bacillus subtilis bacterium
Liquid, bacillus licheniformis bacterium solution；
After bacterium solution in above-mentioned 3 is mixed by the mass ratio of 1:1:1, it is sprayed on carrier, makes biodiversity percentage composition
28%。
A kind of production method of the microbial bacterial agent of embodiment 2
Step 1, strain selection
The strain of use is same as Example 1.
It step 2, actication of culture and spreads cultivation
(1) activation of bacillusmusilaginosiengineering with spread cultivation
Bacillusmusilaginosiengineering strain is inoculated on plating medium, 32 DEG C are cultivated 24 hours, and activated spawn is obtained；Institute
Stating plating medium is nutrient agar；
The activated spawn of bacillusmusilaginosiengineering is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture substrate
The 5% of amount, is placed in shaking table culture case, and revolving speed is 120 revs/min, and ventilatory capacity 1:0.4,35 DEG C are cultivated 21 hours, are spread cultivation
Seed liquor；
The formula of the fluid nutrient medium are as follows: sweet potato powder 17g, bean cake powder 14g, sodium alginate 0.7g, yeast extract 4.2g, paddy
Propylhomoserin 0.03g, bovine bile 0.04g, mannitol 0.04g, manganese dioxide 0.01g, dipotassium hydrogen phosphate 0.03g, magnesium sulfate 0.06g,
Deionized water 1kg；Adjusting pH is 7.
(2) activation of bacillus subtilis with spread cultivation
Bacillus subtilis strain is inoculated on plating medium, 33 DEG C are cultivated 36 hours, and activated spawn is obtained；It is described
Plating medium is cornstarch culture medium；
The activated spawn of bacillus subtilis is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture medium quality
4%, be placed in shaking table culture case, revolving speed is 180 revs/min, ventilatory capacity 1:0.6,33 DEG C and cultivates 30 hours, obtains the kind that spreads cultivation
Sub- liquid；
The formula of the fluid nutrient medium are as follows: potato starch 13g, trehalose 9.5g, sorbitol 7g, beef extract 4.5g,
Aspartic acid 0.07g, glucose 0.07g, ironic citrate 0.03g, vitamin B1 0.01g, 2,6- inositol 0.03g, titanium dioxide
Manganese 0.02g, dipotassium hydrogen phosphate 0.04g, seven brochanite 0.05g, deionized water 1kg；Adjusting pH is 7.2.
(3) activation of bacillus licheniformis with spread cultivation
Bacillus licheniformis strain is inoculated on plating medium, 35 DEG C are cultivated 16 hours, and activated spawn is obtained；It is described
Plating medium is beef extract-peptone agar medium；
The activated spawn of bacillus licheniformis is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture medium quality
4%, be placed in shaking table culture case, revolving speed is 200 revs/min, ventilatory capacity 1:0.6,33 DEG C and cultivates 22 hours, obtains the kind that spreads cultivation
Sub- liquid；
The formula of the fluid nutrient medium are as follows: lactose 19g, fish peptone 8g, sorbitol 4.5g, modified starch 9g, day
Asparagine 0.07g, olive oil 0.03g, ironic citrate 0.03g, niacin 0.02g, calcium monohydrogen phosphate 0.08g, magnesium sulfate 0.05g,
Boric acid 1.2g, deionized water 1kg；Adjusting pH is 6.8.
Step 3, strain fermentation
(1) bacillusmusilaginosiengineering ferments
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
22 parts of wheat bran, 18 parts of cottonseed meal, 5 parts of maltose, 4.2 parts of soy meal, 1.5 parts of beef extract, bovine bile 0.5
Part, 0.3 part of xylitol, 0.1 part of manganese dioxide, 0.07 part of dipotassium hydrogen phosphate, 0.1 part of magnesium sulfate, 2000 parts of deionized water；Adjust pH
It is 7.
Inoculation: fermentation jar temperature is down to 33 DEG C, under aseptic condition, and the kind that spreads cultivation of bacillusmusilaginosiengineering is added from inoculation mouth
Sub- liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 34 DEG C of fermentation jar temperature, and 80rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 31 hours.
(2) fermentation of bacillus subtilis
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
19 parts of tapioca starch, 5 parts of sucrose, 1.5 parts of sorbitol, 2 parts of beef extract, 0.07 part of aspartic acid, 10 parts of corn flour,
0.06 part of ironic citrate, 0.04 part of vitamin B1,0.06 part of 2,6- inositol, 0.07 part of manganese dioxide, dipotassium hydrogen phosphate 0.15
Part, 0.06 part of seven brochanite, 2000 parts of deionized water；Adjusting pH is 7.2.
Inoculation: fermentation jar temperature is down to 32 DEG C, under aseptic condition, and the seed that spreads cultivation of bacillus subtilis is added from inoculation mouth
Liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 36 DEG C of fermentation jar temperature, and 90rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 38 hours.
(3) bacillus licheniformis ferments
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
32 parts of potato starch, 1.5 parts of yeast powder, 13 parts of cottonseed meal, 5 parts of glucose, 0.07 part of asparagine, olive
Oily 0.1 part, 0.2 part of ironic citrate, 0.05 part of niacin, 0.2 part of calcium monohydrogen phosphate, 0.07 part of magnesium sulfate, 1.2 parts of boric acid, deionization
2000 parts of water；Adjusting pH is 6.8.
Inoculation: fermentation jar temperature is down to 33 DEG C, under aseptic condition, and the seed that spreads cultivation of bacillus subtilis is added from inoculation mouth
Liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 35 DEG C of fermentation jar temperature, and 80rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 23 hours.
Step 4, finished product
It is identical as the operating method of embodiment 1.
A kind of production method of the microbial bacterial agent of embodiment 3
Step 1, strain selection
The strain of use is same as Example 1.
It step 2, actication of culture and spreads cultivation
(1) activation of bacillusmusilaginosiengineering with spread cultivation
Bacillusmusilaginosiengineering strain is inoculated on plating medium, 34 DEG C are cultivated 24 hours, and activated spawn is obtained；Institute
Stating plating medium is nutrient agar；
The activated spawn of bacillusmusilaginosiengineering is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture substrate
The 5% of amount, is placed in shaking table culture case, and revolving speed is 120 revs/min, and ventilatory capacity 1:0.4,36 DEG C are cultivated 22 hours, are spread cultivation
Seed liquor；
The formula of the fluid nutrient medium are as follows: sweet potato powder 18g, bean cake powder 15g, sodium alginate 0.8g, yeast extract 4.5g, paddy
Propylhomoserin 0.04g, bovine bile 0.05g, mannitol 0.04g, manganese dioxide 0.02g, dipotassium hydrogen phosphate 0.03g, magnesium sulfate 0.07g,
Deionized water 1kg；Adjusting pH is 7.
(2) activation of bacillus subtilis with spread cultivation
Bacillus subtilis strain is inoculated on plating medium, 33 DEG C are cultivated 36 hours, and activated spawn is obtained；It is described
Plating medium is cornstarch culture medium；
The activated spawn of bacillus subtilis is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture medium quality
4%, be placed in shaking table culture case, revolving speed is 180 revs/min, ventilatory capacity 1:0.6,33 DEG C and cultivates 30 hours, obtains the kind that spreads cultivation
Sub- liquid；
The formula of the fluid nutrient medium are as follows: potato starch 14g, trehalose 10g, sorbitol 8g, beef extract 6g, asparagus fern
Propylhomoserin 0.08g, glucose 0.08g, ironic citrate 0.04g, vitamin B1 0.02g, 2,6- inositol 0.04g, manganese dioxide
0.02g, dipotassium hydrogen phosphate 0.05g, seven brochanite 0.06g, deionized water 1kg；Adjusting pH is 7.2.
(3) activation of bacillus licheniformis with spread cultivation
Bacillus licheniformis strain is inoculated on plating medium, 35 DEG C are cultivated 16 hours, and activated spawn is obtained；It is described
Plating medium is beef extract-peptone agar medium；
The activated spawn of bacillus licheniformis is inoculated in the fluid nutrient medium in shaking flask, inoculum concentration is culture medium quality
4%, be placed in shaking table culture case, revolving speed is 200 revs/min, ventilatory capacity 1:0.6,33 DEG C and cultivates 22 hours, obtains the kind that spreads cultivation
Sub- liquid；
The formula of the fluid nutrient medium are as follows: lactose 20g, fish peptone 9g, sorbitol 5g, modified starch 10g, Tianmen
Winter amide 0.08g, olive oil 0.04g, ironic citrate 0.03g, niacin 0.03g, calcium monohydrogen phosphate 0.09g, magnesium sulfate 0.06g, boron
Sour 1.4g, deionized water 1kg；Adjusting pH is 6.8.
Step 3, strain fermentation
(1) bacillusmusilaginosiengineering ferments
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
23 parts of wheat bran, 19 parts of cottonseed meal, 6 parts of maltose, 4.5 parts of soy meal, 2 parts of beef extract, 0.6 part of bovine bile,
0.4 part of xylitol, 0.2 part of manganese dioxide, 0.09 part of dipotassium hydrogen phosphate, 0.2 part of magnesium sulfate, 2000 parts of deionized water；Adjust pH be
7。
Inoculation: fermentation jar temperature is down to 35 DEG C, under aseptic condition, and the kind that spreads cultivation of bacillusmusilaginosiengineering is added from inoculation mouth
Sub- liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 34 DEG C of fermentation jar temperature, and 80rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 32 hours.
(2) fermentation of bacillus subtilis
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
20 parts of tapioca starch, 6 parts of sucrose, 2 parts of sorbitol, 3 parts of beef extract, 0.08 part of aspartic acid, 11 parts of corn flour, lemon
0.07 part of lemon acid iron, 0.04 part of vitamin B1,0.07 part of 2,6- inositol, 0.08 part of manganese dioxide, 0.2 part of dipotassium hydrogen phosphate,
Seven 0.07 part of brochanites, 2000 parts of deionized water；Adjusting pH is 7.2.
Inoculation: fermentation jar temperature is down to 35 DEG C, under aseptic condition, and the seed that spreads cultivation of bacillus subtilis is added from inoculation mouth
Liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 36 DEG C of fermentation jar temperature, and 90rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 40 hours.
(3) bacillus licheniformis ferments
Fermentation medium sterilizing: fermentation medium each component is weighed, and is added in fermentor, is started stirring, 120rpm
Stir 10min；Heating, pressurization, fermentation pressure inside the tank reach 0.105MPa, and temperature reaches 121 DEG C, and sterilize 25~30min；
The fermentation medium comprises the following components in parts by weight:
34 parts of potato starch, 2 parts of yeast powder, 15 parts of cottonseed meal, 6 parts of glucose, 0.08 part of asparagine, olive oil
0.2 part, 0.2 part of ironic citrate, 0.06 part of niacin, 0.2 part of calcium monohydrogen phosphate, 0.08 part of magnesium sulfate, 1.4 parts of boric acid, deionized water
2000 parts；Adjusting pH is 6.8.
Inoculation: fermentation jar temperature is down to 35 DEG C, under aseptic condition, and the seed that spreads cultivation of bacillus subtilis is added from inoculation mouth
Liquid, inoculum concentration are the 6% of fermentation medium quality；
Fermentation: after inoculation, controlling 35 DEG C of fermentation jar temperature, and 80rpm stirring is continually fed into filtrated air, avoids miscellaneous bacteria dirty
Dye；Fermentation 24 hours.
Step 4, finished product
It is identical as the operating method of embodiment 1.
The microbial bacterial agent of embodiment 1-3 preparation is detected；Testing result is shown in Table 1；
Table 1
Seen from table 1, microbial bacterial agent of the present invention, total bacterial content are 85~8,800,000,000 cfu/g, bacillusmusilaginosiengineering viable bacteria
Number be 18~2,200,000,000 cfu/g, bacillus subtilis viable count be 17~1,900,000,000 cfu/g, bacillus licheniformis viable count be 12~
1300000000 cfu/g, miscellaneous bacteria rate are 4.6~5.2%, and fineness is 120 mesh, and arsenic and its compounds content are 6~9 mg/kg, mercury and its change
Conjunction object content is 0.7~0.8 mg/kg, and 9~11 mg/kg of lead and its compounds content, the shelf-life is 20~24 months.
Effect experiment:
The microbial bacterial agent of embodiment 1-3 is applied in eggplant plantation plot, applies 40~50kg of microbial bacterial agent per acre,
With topsoil mix, plant eggplant 1 year, soil property be improved significantly, specific targets are shown in Table 2；
Performance indicator after 2 soil improvement of table
As can be seen from Table 2, after using microbial bacterial agent, soil property be improved significantly, soil pH is 6.8~7.2, soil
Bulk density is 1.36~1.41g/cm3, ventilation voidage is 20~23%；Heavy metal in soil content is substantially reduced, content of microorganisms
It significantly improves.
After microbial bacterial agent, eggplant yield improves 15~18%, and commodity fruit rate improves 23~25%, malformed fruit rate
Reduce 9~13%.
Finally, it should be noted that the foregoing is only a preferred embodiment of the present invention, it is not intended to restrict the invention,
It, for those skilled in the art, still can be with although describing the invention in detail with reference to the foregoing embodiments
It modifies the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features.It is all
Within the spirit and principles in the present invention, any modification, equivalent replacement, improvement and so on should be included in guarantor of the invention
Within the scope of shield.The above is the citing of best mode for carrying out the invention, wherein the part that do not address in detail is this field
The common knowledge of those of ordinary skill；Protection scope of the present invention is based on the contents of the claims, any based on of the invention
Technical inspiration and the equivalent transformation carried out, also within protection scope of the present invention.
Claims (1)
1. a kind of production method of microbial bacterial agent, it is characterised in that: including strain selection, actication of culture and spread cultivation, strain hair
Ferment, finished product step；
In the actication of culture and the step that spreads cultivation, the activation of bacillusmusilaginosiengineering and matching for the culture medium that spreads cultivation for the use that spreads cultivation
Side are as follows: 15~18 parts by weight of sweet potato powder, 12~15 parts by weight of bean cake powder, 0.6~0.8 parts by weight of sodium alginate, yeast extract 4~
4.5 parts by weight, 0.03~0.04 parts by weight of glutamic acid, 0.03~0.05 parts by weight of bovine bile, 0.02~0.04 weight of mannitol
Part, 0.01~0.02 parts by weight of manganese dioxide, 0.02~0.03 parts by weight of dipotassium hydrogen phosphate, 0.05~0.07 weight of magnesium sulfate
Part, 1000 parts by weight of deionized water；
In the actication of culture and the step that spreads cultivation, the activation of bacillus subtilis and the formula of the culture medium that spreads cultivation for the use that spreads cultivation
Are as follows: 12~14 parts by weight of potato starch, 9~10 parts by weight of trehalose, 6~8 parts by weight of sorbitol, 4~6 weight of beef extract
Part, 0.06~0.08 parts by weight of aspartic acid, 0.07~0.08 parts by weight of glucose, 0.02~0.04 parts by weight of ironic citrate,
0.01~0.02 parts by weight of vitamin B1,0.03~0.04 parts by weight of 2,6- inositol, 0.01~0.02 parts by weight of manganese dioxide,
0.03~0.05 parts by weight of dipotassium hydrogen phosphate, seven brochanites, 0.04~0.06 parts by weight, 1000 parts by weight of deionized water；
In the actication of culture and the step that spreads cultivation, the activation of bacillus licheniformis and the formula of the culture medium that spreads cultivation for the use that spreads cultivation
Are as follows: 18~20 parts by weight of lactose, 7~9 parts by weight of fish peptone, 4~5 parts by weight of sorbitol, 8~10 weight of modified starch
Part, 0.06~0.08 parts by weight of asparagine, 0.03~0.04 parts by weight of olive oil, 0.02~0.03 weight of ironic citrate
Part, 0.02~0.03 parts by weight of niacin, 0.06~0.09 parts by weight of calcium monohydrogen phosphate, 0.04~0.06 parts by weight of magnesium sulfate, boric acid
1~1.4 parts by weight, 1000 parts by weight of deionized water；
In the strain fermentation step, the temperature of bacillusmusilaginosiengineering fermentation is 34 DEG C, and inoculum concentration is fermented and cultured matrix
The 6% of amount, fermentation time are 30~32 hours；
In the strain fermentation step, the fermentation temperature of fermentation of bacillus subtilis is 36 DEG C, and inoculum concentration is fermentation medium
The 6% of quality, fermentation time are 36~40 hours；
In the strain fermentation step, the fermentation temperature of bacillus licheniformis fermentation is 35 DEG C, and inoculum concentration is fermentation medium
The 6% of quality, fermentation time are 22~24 hours；
In the finished product step, the component of carrier includes medical stone, calcium carbonate, vermiculite power, and mass ratio is medical stone:
Calcium carbonate: vermiculite power=5:2:1；
In microbial bacterial agent obtained, bacillusmusilaginosiengineering viable count is 18~2,200,000,000 cfu/g, bacillus subtilis viable count
For 17~1,900,000,000 cfu/g, bacillus licheniformis viable count is 12~1,300,000,000 cfu/g；Miscellaneous bacteria rate is 4.6~5.2%；Arsenic and its change
Conjunction object content is 6~9 mg/kg, and mercury and mercuric compounds content is 0.7~0.8 mg/kg, lead and its compounds content 9~11
mg/kg；Shelf-life is 20~24 months；
The bacillusmusilaginosiengineering (Bacillus mucilaginosus) deposit number be ACCC10013；
The bacillus subtilis (Bacillus subtilis) deposit number be ACCC01175；
The bacillus licheniformis (Bacillus licheniformis) deposit number be ACCC11091.
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| CN201610061792.2A CN105441370B (en) | 2016-01-29 | 2016-01-29 | A kind of microbial bacterial agent and its production method |
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| CN106701628B (en) * | 2017-01-09 | 2020-10-09 | 杨丁元 | Rural domestic garbage fermentation microbial inoculum and use method and application thereof |
| CN106978374A (en) * | 2017-05-10 | 2017-07-25 | 上海创博土壤修复工程有限公司 | A kind of microbe soil conditioner and preparation method thereof |
| CN107593770A (en) * | 2017-08-25 | 2018-01-19 | 河南中麦生物科技有限公司 | A kind of microbial inoculum and its application for being used to set withered retrogradation greatly |
| CN108342445B (en) * | 2018-04-09 | 2021-09-21 | 中国科学院合肥物质科学研究院 | Method for producing beta-carotene by Blakeslea trispora fermentation and beta-carotene |
| CN114717006A (en) * | 2022-03-11 | 2022-07-08 | 四川爱隆植物营养科技有限公司 | Microbial soil loosening agent |
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| CN102229900A (en) * | 2011-05-20 | 2011-11-02 | 周剑平 | Compound probiotics preparation for biological organic fertilizers and preparation method thereof |
| CN102816726A (en) * | 2012-09-10 | 2012-12-12 | 临沂中磷生物科技有限公司 | Novel compound microbial agent and applications thereof |
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| CN102229900A (en) * | 2011-05-20 | 2011-11-02 | 周剑平 | Compound probiotics preparation for biological organic fertilizers and preparation method thereof |
| CN102816726A (en) * | 2012-09-10 | 2012-12-12 | 临沂中磷生物科技有限公司 | Novel compound microbial agent and applications thereof |
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| 微生物肥料菌种应用与效果分析;陈慧君;《中国优秀硕士学位论文数据库 农业科技辑》;20140215;全文，特别是摘要 |
| 微生物肥料菌种的应用及效果分析;李晓娇;《农业与技术》;20160108;34 |
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