CN112919938A - Fertilizer synergist produced by utilizing rape seed meal double-bacterium fermentation, method and application - Google Patents
Fertilizer synergist produced by utilizing rape seed meal double-bacterium fermentation, method and application Download PDFInfo
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- CN112919938A CN112919938A CN202110346051.XA CN202110346051A CN112919938A CN 112919938 A CN112919938 A CN 112919938A CN 202110346051 A CN202110346051 A CN 202110346051A CN 112919938 A CN112919938 A CN 112919938A
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- rapeseed meal
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- 235000019779 Rapeseed Meal Nutrition 0.000 title claims abstract description 68
- 239000004456 rapeseed meal Substances 0.000 title claims abstract description 68
- 238000000855 fermentation Methods 0.000 title claims abstract description 52
- 230000004151 fermentation Effects 0.000 title claims abstract description 52
- 239000003337 fertilizer Substances 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 19
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 claims abstract description 50
- 239000007788 liquid Substances 0.000 claims abstract description 44
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 30
- 238000004519 manufacturing process Methods 0.000 claims abstract description 28
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 claims abstract description 25
- 229960003692 gamma aminobutyric acid Drugs 0.000 claims abstract description 25
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims abstract description 22
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 19
- 239000004220 glutamic acid Substances 0.000 claims abstract description 19
- 239000001963 growth medium Substances 0.000 claims abstract description 19
- 241000186226 Corynebacterium glutamicum Species 0.000 claims abstract description 18
- 239000006185 dispersion Substances 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims abstract description 15
- 229920000053 polysorbate 80 Polymers 0.000 claims abstract description 15
- 239000011780 sodium chloride Substances 0.000 claims abstract description 15
- 239000002699 waste material Substances 0.000 claims abstract description 15
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 14
- 239000000203 mixture Substances 0.000 claims abstract description 12
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims abstract description 11
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims abstract description 11
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims abstract description 11
- 229910004664 Cerium(III) chloride Inorganic materials 0.000 claims abstract description 11
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000005642 Oleic acid Substances 0.000 claims abstract description 11
- 235000019270 ammonium chloride Nutrition 0.000 claims abstract description 11
- 239000002518 antifoaming agent Substances 0.000 claims abstract description 11
- VYLVYHXQOHJDJL-UHFFFAOYSA-K cerium trichloride Chemical compound Cl[Ce](Cl)Cl VYLVYHXQOHJDJL-UHFFFAOYSA-K 0.000 claims abstract description 11
- 235000019864 coconut oil Nutrition 0.000 claims abstract description 11
- 239000003240 coconut oil Substances 0.000 claims abstract description 11
- 239000008367 deionised water Substances 0.000 claims abstract description 11
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 11
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 11
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 11
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 11
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 11
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims abstract description 11
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims abstract description 11
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims abstract description 11
- 229940099596 manganese sulfate Drugs 0.000 claims abstract description 11
- 235000007079 manganese sulphate Nutrition 0.000 claims abstract description 11
- 239000011702 manganese sulphate Substances 0.000 claims abstract description 11
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims abstract description 11
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 11
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 11
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims abstract description 11
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims abstract description 9
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 8
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims abstract description 8
- 238000002156 mixing Methods 0.000 claims description 14
- 238000003756 stirring Methods 0.000 claims description 13
- 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 11
- 239000008103 glucose Substances 0.000 claims description 11
- 239000000843 powder Substances 0.000 claims description 10
- 238000011218 seed culture Methods 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- 238000000227 grinding Methods 0.000 claims description 6
- 239000002245 particle Substances 0.000 claims description 6
- 238000009423 ventilation Methods 0.000 claims description 6
- 240000008042 Zea mays Species 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 5
- 235000005822 corn Nutrition 0.000 claims description 5
- 230000012010 growth Effects 0.000 claims description 5
- 239000012137 tryptone Substances 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 239000004202 carbamide Substances 0.000 claims description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L magnesium sulphate Substances [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 4
- 238000007873 sieving Methods 0.000 claims description 4
- 239000002002 slurry Substances 0.000 claims description 4
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 claims description 2
- 239000002609 medium Substances 0.000 abstract description 12
- 239000002994 raw material Substances 0.000 abstract description 5
- 238000003912 environmental pollution Methods 0.000 abstract description 3
- 229960002989 glutamic acid Drugs 0.000 description 17
- 241000318358 Bacillus licheniformis WX-02 Species 0.000 description 8
- 241000759360 Corynebacterium glutamicum S9114 Species 0.000 description 8
- 230000006872 improvement Effects 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- 238000005070 sampling Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 230000001737 promoting effect Effects 0.000 description 5
- 230000001105 regulatory effect Effects 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000011573 trace mineral Substances 0.000 description 3
- 235000013619 trace mineral Nutrition 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000037149 energy metabolism Effects 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 238000010902 jet-milling Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 230000029553 photosynthesis Effects 0.000 description 2
- 238000010672 photosynthesis Methods 0.000 description 2
- PUKLDDOGISCFCP-JSQCKWNTSA-N 21-Deoxycortisone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(=O)C)(O)[C@@]1(C)CC2=O PUKLDDOGISCFCP-JSQCKWNTSA-N 0.000 description 1
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 description 1
- 244000221633 Brassica rapa subsp chinensis Species 0.000 description 1
- 241000186146 Brevibacterium Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- FCYKAQOGGFGCMD-UHFFFAOYSA-N Fulvic acid Natural products O1C2=CC(O)=C(O)C(C(O)=O)=C2C(=O)C2=C1CC(C)(O)OC2 FCYKAQOGGFGCMD-UHFFFAOYSA-N 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000459 effect on growth Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000002509 fulvic acid Substances 0.000 description 1
- 229940095100 fulvic acid Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000011090 industrial biotechnology method and process Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- LPUQAYUQRXPFSQ-DFWYDOINSA-M monosodium L-glutamate Chemical compound [Na+].[O-]C(=O)[C@@H](N)CCC(O)=O LPUQAYUQRXPFSQ-DFWYDOINSA-M 0.000 description 1
- 239000004223 monosodium glutamate Substances 0.000 description 1
- 235000013923 monosodium glutamate Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000009105 vegetative growth Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/10—Fertilisers containing plant vitamins or hormones
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/005—Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Pest Control & Pesticides (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fertilizers (AREA)
Abstract
The invention discloses a fertilizer synergist produced by utilizing double-bacterium fermentation of rapeseed meal, which is characterized in that the fertilizer synergist is a mixture containing poly-gamma-glutamic acid and gamma-aminobutyric acid, which is produced by adopting a culture medium containing the rapeseed meal through fermentation of corynebacterium glutamicum and bacillus by a one-step method; and the components of the culture medium are as follows: 2.0-8.0 percent of rapeseed meal dispersion liquid, 0.2-5.0 percent of yeast production waste liquid, 0.01-6.5 percent of ammonium chloride, 0.1-2.5 percent of monopotassium phosphate, 0.05-1.5 percent of magnesium sulfate heptahydrate, 0.001-1.0 percent of manganese sulfate, 0.05-2.5 percent of ammonium citrate, 0.01-0.05 percent of sodium chloride, 0.001-0.006 percent of ferrous sulfate, 0.001-0.005 percent of cerous chloride, 0.05-0.2 percent of coconut oil oleic acid antifoaming agent, 0.02-0.25 percent of Tween 80 and the balance of deionized water. The invention also discloses a preparation method and application of the fertilizer synergist. The rapeseed meal is used as a raw material of a fermentation medium, and the fertilizer synergist PG containing the mixture of poly-gamma-glutamic acid and gamma-aminobutyric acid is produced by a one-step method, so that the production cost is reduced, the environmental pollution is less, and the method is safe and reliable.
Description
Technical Field
The invention belongs to the technical field of preparation methods of fertilizer synergists, and particularly relates to a fertilizer synergist produced by utilizing double-bacterium fermentation of rapeseed meal, and a preparation method and application thereof.
Background
Research finds that poly-gamma-glutamic acid (gamma-PGA) and gamma-aminobutyric acid (GABA) both have promotion effect on growth of the poly-gamma-glutamic acid (gamma-PGA) and the gamma-aminobutyric acid (GABA); wherein, the poly-gamma-glutamic acid (gamma-PGA) has the functions of promoting the development of root systems, enhancing the efficiency, increasing the yield and improving the quality, and obviously improves the utilization rate of the fertilizer. The fertilizer consumption is reduced by more than 10% when the same yield is obtained by planting and cultivating the pakchoi, the corn and the tobacco leaves, or the crop yield is improved by more than 10% on the premise of the same fertilizer consumption; gamma-aminobutyric acid (GABA) has dual identities of a metabolic substance and a signal substance in a plant body, has the functions of promoting photosynthesis and energy metabolism, promoting vegetative growth and reproductive growth of plants, improving the resistance of the plants to different stresses, promoting the absorption of trace elements in the plants, regulating the endogenous hormone level of the signal substance in the plants and the like, can stimulate the growth and stress resistance of crops, and remarkably promote the absorption of the trace elements in the crops.
And the poly-gamma-glutamic acid (gamma-PGA) and gamma-aminobutyric acid (GABA) are mixed to form a fertilizer synergist PG for promoting the growth of crops and improving the yield (the synergist is formed by mixing the poly-gamma-glutamic acid gamma-PGA and the gamma-aminobutyric acid GABA in a ratio of 1:0.1-10, and is named as the fertilizer synergist PG by initials). The fertilizer synergist PG is very soluble in water, can synergistically promote photosynthesis and energy metabolism, promote growth and stress resistance, and obviously improve the utilization rate of the fertilizer.
The preparation method of the conventional fertilizer synergist PG comprises the following steps: respectively producing poly-gamma-glutamic acid and gamma-aminobutyric acid by adopting a microbial fermentation method, and then mixing, stirring and packaging to obtain a finished product. On one hand, the traditional method needs to respectively ferment and then mix, and is complicated in process; on the other hand, the conventional fermentation method has a relatively expensive culture medium, and requires expensive raw materials such as L-glutamic acid, tryptone, glucose, refined milk powder and the like, so that the production cost is always high.
Disclosure of Invention
Aiming at one or more of the defects or the improvement requirements in the prior art, the invention provides a fertilizer synergist produced by utilizing the double-bacteria fermentation of rapeseed meal, a method and application thereof.
In order to achieve the above objects, according to one aspect of the present invention, there is provided a fertilizer synergist produced by using two-strain fermentation of rapeseed meal, wherein the fertilizer synergist is a mixture containing poly-gamma-glutamic acid and gamma-aminobutyric acid, which is produced by using a culture medium containing rapeseed meal through fermentation of corynebacterium glutamicum and bacillus in a one-step process.
As a further improvement of the invention, the rapeseed meal-containing medium comprises the following components by weight of dry matter: 2.0-8.0 percent of rapeseed meal dispersion liquid, 0.2-5.0 percent of yeast production waste liquid, 0.01-6.5 percent of ammonium chloride, 0.1-2.5 percent of monopotassium phosphate, 0.05-1.5 percent of magnesium sulfate heptahydrate, 0.001-1.0 percent of manganese sulfate, 0.05-2.5 percent of ammonium citrate, 0.01-0.05 percent of sodium chloride, 0.001-0.006 percent of ferrous sulfate, 0.001-0.005 percent of cerous chloride, 0.05-0.2 percent of coconut oil oleic acid antifoaming agent, 0.02-0.25 percent of Tween 80 and the balance of deionized water.
As a further improvement of the invention, the ratio of the mixed liquid of the corynebacterium glutamicum seed liquid and the bacillus seed liquid inoculated into the culture medium is 1: 8-10.
The rapeseed meal which is the main raw material of the invention is a high-protein feed with great utilization value, generally contains 35 to 40 percent of protein, 0.61 percent of calcium, 0.95 percent of phosphorus, trace elements such as iron, copper, manganese, selenium and the like and a plurality of vitamins, and the digestion energy is 10.46 to 12.55 kj/kg.
The waste liquid produced in the yeast production is the waste liquid discharged after being separated by a separator in the process of molasses fermentation in the yeast manufacturing industry, the year-round production of one production line can produce more than 5 million tons of waste liquid, and the method belongs to the problem of environmental protection treatment for a long time. The waste liquid can be obtained freely, is rich in active yeast and fulvic acid, and has a solid organic matter content of more than 50%.
According to another aspect of the present invention, there is provided a method for producing a fertilizer synergist by using double-bacterial fermentation of rapeseed meal, comprising the steps of:
adding a culture medium containing rapeseed meal into a fermentation tank, mixing and inoculating corynebacterium glutamicum seed liquid and bacillus seed liquid into the fermentation tank according to the ratio of 1: 8-10, stirring at the rotating speed of 200-700 r/min, the ventilation volume of 3L/(L-min), and the pH of 6.5-7.0, culturing at the temperature of 30-45 ℃, feeding 30g/L glucose liquid after 24 hours, and performing fermentation culture for 36-72 hours to obtain a finished product.
As a further improvement of the invention, the rapeseed meal-containing medium comprises the following components by weight of dry matter: 2.0-8.0 percent of rapeseed meal dispersion liquid, 0.2-5.0 percent of yeast production waste liquid, 0.01-6.5 percent of ammonium chloride, 0.1-2.5 percent of monopotassium phosphate, 0.05-1.5 percent of magnesium sulfate heptahydrate, 0.001-1.0 percent of manganese sulfate, 0.05-2.5 percent of ammonium citrate, 0.01-0.05 percent of sodium chloride, 0.001-0.006 percent of ferrous sulfate, 0.001-0.005 percent of cerous chloride, 0.05-0.2 percent of coconut oil oleic acid antifoaming agent, 0.02-0.25 percent of Tween 80 and the balance of deionized water.
As a further improvement of the invention, the preparation method of the rapeseed meal dispersion liquid comprises the following steps: crushing rapeseed meal, collecting rapeseed meal particles, sieving to obtain rapeseed meal powder, stirring and mixing the rapeseed meal powder and water according to a mass ratio of 1:5, grinding the mixture into slurry by a grinder, adding Tween 80, performing ultrasonic dispersion for 5-10 min at 200-300W, and collecting rapeseed meal dispersion liquid.
As a further improvement of the invention, the ratio of the corynebacterium glutamicum seed culture medium is as follows: glucose 25g/L, corn steep liquor 30g/L, K2HPO41.5g/L，MgSO40.8g/L, 5.5g/L urea, and pH7.0-7.3.
As a further improvement of the invention, the ratio of the bacillus seed culture medium is as follows: 12g/L of tryptone, 5g/L of yeast extract, 40g/L of sodium chloride, 25g/L of agar and 7.1-7.6 of pH.
According to another aspect of the invention, the invention provides an application of the fertilizer synergist PG produced by utilizing the double-bacteria fermentation of rapeseed meal as a fertilizer synergist in crop growth.
Generally, compared with the prior art, the above technical solution conceived by the present invention has the following beneficial effects:
the invention uses rapeseed meal as raw material of fermentation medium, and by fermentation of corynebacterium glutamicum and bacillus, the corynebacterium glutamicum can ferment to produce poly-gamma-glutamic acid, and the bacillus can continue to ferment on the fermentation medium to produce gamma-aminobutyric acid; the mixture ratio of poly-gamma-glutamic acid gamma-PGA and gamma-aminobutyric acid GABA in different proportions can be obtained by adjusting the proportion according to production requirements, namely the fertilizer synergist PG (the proportion is 1: 0.1-10). The invention produces the poly-gamma-glutamic acid and gamma-aminobutyric acid mixture fertilizer synergist PG by a one-step method, reduces the production cost, has little pollution to the environment, and is safe and reliable.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The invention provides a preparation method of a one-step fertilizer synergist PG, which has the advantages of high quality, low price, simple process, low production cost and less environmental pollution, and aims to overcome the defects of the preparation method of the fertilizer synergist PG in the prior art. The invention relates to a method for producing a fertilizer synergist by utilizing double-bacterium fermentation of rapeseed meal, which comprises the following steps:
(1) preparing rapeseed meal dispersion liquid;
selecting fresh rapeseed meal, performing jet milling, collecting rapeseed meal particles, grinding the rapeseed meal particles, sieving with a 200-mesh sieve to obtain rapeseed meal powder, stirring and mixing the rapeseed meal powder and water according to a mass ratio of 1:5, grinding the mixture into slurry by a grinder, adding Tween 80, performing ultrasonic dispersion at 200-300W for 5-10 min, and collecting a rapeseed meal dispersion liquid;
(2) respectively preparing corynebacterium glutamicum seed liquid and bacillus seed liquid;
wherein the ratio of the corynebacterium glutamicum seed culture medium is as follows: glucose 25g/L, corn steep liquor 30g/L, K2HPO41.5g/L，MgSO40.8g/L, 5.5g/L urea, and pH7.0-7.3;
the ratio of the bacillus seed culture medium is as follows: 12g/L of tryptone, 5g/L of yeast extract, 40g/L of sodium chloride, 25g/L of agar and 7.1-7.6 of pHs;
the inoculation amounts of corynebacterium glutamicum and bacillus are 3-6% (volume ratio);
(3) the fermenter was charged with a medium which (on a dry basis) had the following composition: 2.0-8.0 percent of rapeseed meal dispersion liquid, 0.2-5.0 percent of yeast production waste liquid, 0.01-6.5 percent of ammonium chloride, 0.1-2.5 percent of monopotassium phosphate, 0.05-1.5 percent of magnesium sulfate heptahydrate, 0.001-1.0 percent of manganese sulfate, 0.05-2.5 percent of ammonium citrate, 0.01-0.05 percent of sodium chloride, 0.001-0.006 percent of ferrous sulfate, 0.001-0.005 percent of cerous chloride, 0.05-0.2 percent of coconut oil oleic acid antifoaming agent, 0.02-0.25 percent of Tween 80 and the balance of deionized water.
(4) Mixing corynebacterium glutamicum seed liquid and bacillus seed liquid in a ratio of 1: 8-10, and inoculating into a fermentation tank; stirring at a rotation speed of 200-700 r/min and a ventilation volume of 3L/(L-min); the pH value is 6.5-7.0, the culture is carried out at the temperature of 30-45 ℃, 30g/L glucose solution is fed after 24 hours, the finished product is obtained after fermentation culture is carried out for 36-72 hours, and in the fermentation process, sampling is carried out at intervals of a plurality of hours to measure and regulate various parameters.
To better illustrate the product and the method of preparation of the invention, the following specific examples are provided:
example 1
Selecting fresh rapeseed meal, performing jet milling, collecting rapeseed meal particles, grinding the rapeseed meal particles, sieving with a 200-mesh sieve to obtain rapeseed meal powder, stirring and mixing the rapeseed meal powder and water according to the mass ratio of 1:5, grinding the rapeseed meal powder into slurry by a grinder, adding Tween 80, performing ultrasonic dispersion for 8min at 200-300W, and collecting rapeseed meal dispersion liquid;
the medium (dry matter basis) was added to a 10L fermentor and was composed of: 6.0% of rapeseed meal dispersion, 3.0% of yeast production waste liquid, 2.5% of ammonium chloride, 1.5% of potassium dihydrogen phosphate, 1.1% of magnesium sulfate heptahydrate, 0.5% of manganese sulfate, 1.5% of ammonium citrate, 0.02% of sodium chloride, 0.003% of ferrous sulfate, 0.003% of cerous chloride, 0.1% of coconut oil oleic acid defoaming agent, 0.15% of tween 80 and the balance of deionized water;
preparing corynebacterium glutamicum seed liquid and bacillus seed liquid; in the specific embodiment of the present invention, the selection of Corynebacterium glutamicum S9114 and Bacillus licheniformis WX-02 from Bacillus may be performed, but other species of Corynebacterium glutamicum and Bacillus may be used as long as the technical effects of the present invention are achieved.
Wherein the ratio of the corynebacterium glutamicum S9114 seed culture medium (g/L) is as follows: glucose 25, corn steep liquor 30, K2HPO41.5，MgSO40.8, 5.5 of urea, 7.0-7.3 of pHs, and the proportion of a bacillus licheniformis WX-02 seed culture medium (g/L) is as follows: tryptone 12, yeast extract 5, NaCl 40, agar 25, pH7.1-7.6.
The corynebacterium glutamicum S9114 is a strain which is widely applied in the monosodium glutamate industry in China at present and is a mutant strain of Brevibacterium tianjingensis T613. The collection is available in key laboratories of the department of industrial biotechnology education of southern Yangtze university and in key laboratories of the microbial technology country of Shandong university. The common market is known to be sold, and the common commercial route can be obtained. The Bacillus licheniformis WX-02 is screened and preserved by the national emphasis laboratory of agricultural microbiology of Huazhong university of agriculture, and the preservation number is CCTCC M208065.
Mixing the seed solution of corynebacterium glutamicum S9114 and the seed solution of bacillus licheniformis WX-02 in a ratio of 1:9, and inoculating into a 10L fermentation tank; stirring at 380r/min, ventilating at 3L/(L-min) and pH 6.5-7.0, culturing at 37 deg.C, adding 30g/L glucose solution after 24 hr, sampling every 6 hr during fermentation to determine and regulate various parameters, and fermenting for 48 hr to obtain the final product.
Through detection, in the finished fertilizer synergist PG, the content of poly-gamma-glutamic acid is 55.88g/L, and the content of gamma-aminobutyric acid is 9.32 g/L.
Example 2
The present embodiment is different from embodiment 1 in that,
the medium (dry matter basis) was added to a 10L fermentor and was composed of: 2.0% of rapeseed meal dispersion, 0.2% of yeast production waste liquid, 0.01% of ammonium chloride, 0.5% of monopotassium phosphate, 0.5% of magnesium sulfate heptahydrate, 0.01% of manganese sulfate, 0.1% of ammonium citrate, 0.01% of sodium chloride, 0.004% of ferrous sulfate, 0.002% of cerous chloride, 0.05% of coconut oil oleic acid defoaming agent, 0.1% of tween 80 and the balance of deionized water;
mixing the seed solution of corynebacterium glutamicum S9114 and the seed solution of bacillus licheniformis WX-02 in a ratio of 1:8, and inoculating into a 10L fermentation tank; stirring at the rotation speed of 200r/min, ventilation volume of 3L/(L-min), pH of 6.5-7.0, culturing at the temperature of 30 ℃, feeding 30g/L glucose solution after 24 hours, sampling at intervals of 6 hours during the fermentation process, measuring and regulating various parameters, and performing fermentation culture for 36 hours to obtain the finished product.
Through detection, in the finished fertilizer synergist PG, the content of poly-gamma-glutamic acid is 50.65g/L, and the content of gamma-aminobutyric acid is 10.34 g/L.
Example 3
The present embodiment is different from embodiment 1 in that,
the medium (dry matter basis) was added to a 10L fermentor and was composed of: 8.0% of rapeseed meal dispersion, 5.0% of yeast production waste liquid, 4.0% of ammonium chloride, 2.5% of potassium dihydrogen phosphate, 1.5% of magnesium sulfate heptahydrate, 1.0% of manganese sulfate, 0.05% of ammonium citrate, 0.05% of sodium chloride, 0.001% of ferrous sulfate, 0.001% of cerous chloride, 0.2% of coconut oil oleic acid defoaming agent, 0.25% of tween 80 and the balance of deionized water;
mixing a corynebacterium glutamicum S9114 seed solution and a bacillus licheniformis WX-02 seed solution according to the proportion of 1:10, and inoculating into a 10L fermentation tank; stirring at the rotating speed of 700r/min, ventilation volume of 3L/(L-min), pH of 6.5-7.0, culturing at the temperature of 45 ℃, feeding 30g/L glucose solution after 24 hours, sampling at intervals of 6 hours during the fermentation process, measuring and regulating various parameters, and performing fermentation culture for 72 hours to obtain the finished product.
Through detection, in the finished fertilizer synergist PG, the content of poly-gamma-glutamic acid is 56.36g/L, and the content of gamma-aminobutyric acid is 12.16 g/L.
Example 4
The present embodiment is different from embodiment 1 in that,
the medium (dry matter basis) was added to a 10L fermentor and was composed of: 4.0% of rapeseed meal dispersion, 2.0% of yeast production waste liquid, 6.5% of ammonium chloride, 0.1% of monopotassium phosphate, 0.05% of magnesium sulfate heptahydrate, 0.001% of manganese sulfate, 2.5% of ammonium citrate, 0.03% of sodium chloride, 0.006% of ferrous sulfate, 0.005% of cerous chloride, 0.15% of coconut oil oleic acid defoaming agent, 0.02% of tween 80 and the balance of deionized water;
mixing the seed solution of corynebacterium glutamicum S9114 and the seed solution of bacillus licheniformis WX-02 in a ratio of 1:9, and inoculating into a 10L fermentation tank; stirring at 580r/min, ventilating 3L/(L-min), pH 6.5-7.0, culturing at 40 deg.C, adding 30g/L glucose solution after 24 hr, sampling every 6 hr during fermentation, measuring and regulating parameters, and fermenting for 60 hr to obtain the final product.
Through detection, in the finished fertilizer synergist PG, the content of poly-gamma-glutamic acid is 56.12g/L, and the content of gamma-aminobutyric acid is 9.05 g/L.
Example 5
The present embodiment is different from embodiment 1 in that,
the medium (dry matter basis) was added to a 10L fermentor and was composed of: 5.0% of rapeseed meal dispersion, 1.0% of yeast production waste liquid, 1.0% of ammonium chloride, 1.1% of potassium dihydrogen phosphate, 0.1% of magnesium sulfate heptahydrate, 0.1% of manganese sulfate, 0.8% of ammonium citrate, 0.04% of sodium chloride, 0.002% of ferrous sulfate, 0.004% of cerous chloride, 0.08% of coconut oil oleic acid defoaming agent, 0.2% of tween 80 and the balance of deionized water;
mixing the seed solution of corynebacterium glutamicum S9114 and the seed solution of bacillus licheniformis WX-02 in a ratio of 1:9, and inoculating into a 10L fermentation tank; stirring at the rotating speed of 350r/min, ventilation volume of 3L/(L-min), pH of 6.5-7.0, culturing at the temperature of 35 ℃, feeding 30g/L glucose solution after 24 hours, sampling at intervals of 6 hours during the fermentation process, measuring and regulating various parameters, and performing fermentation culture for 55 hours to obtain the finished product.
Through detection, in the finished fertilizer synergist PG, the content of poly-gamma-glutamic acid is 55.96g/L, and the content of gamma-aminobutyric acid is 8.13 g/L.
According to the invention, rapeseed meal is used as a raw material of a fermentation medium, and the fertilizer synergist PG containing a mixture of poly-gamma-glutamic acid and gamma-aminobutyric acid can be produced by a one-step method through fermentation of corynebacterium glutamicum and bacillus, so that the production cost is reduced, the environmental pollution is less, and the method is safe and reliable.
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (9)
1. A fertilizer synergist produced by utilizing double-bacterium fermentation of rapeseed meal is characterized in that the fertilizer synergist is a mixture containing poly-gamma-glutamic acid and gamma-aminobutyric acid, which is produced by adopting a culture medium containing the rapeseed meal through fermentation of corynebacterium glutamicum and bacillus and a one-step method.
2. The fertilizer synergist produced by utilizing the double-bacterial fermentation of rapeseed meal according to claim 1, characterized in that the culture medium containing rapeseed meal comprises the following components by weight of dry matter: 2.0-8.0 percent of rapeseed meal dispersion liquid, 0.2-5.0 percent of yeast production waste liquid, 0.01-6.5 percent of ammonium chloride, 0.1-2.5 percent of monopotassium phosphate, 0.05-1.5 percent of magnesium sulfate heptahydrate, 0.001-1.0 percent of manganese sulfate, 0.05-2.5 percent of ammonium citrate, 0.01-0.05 percent of sodium chloride, 0.001-0.006 percent of ferrous sulfate, 0.001-0.005 percent of cerous chloride, 0.05-0.2 percent of coconut oil oleic acid antifoaming agent, 0.02-0.25 percent of Tween 80 and the balance of deionized water.
3. The fertilizer synergist produced by utilizing the double-bacterial fermentation of rapeseed meal according to claim 1 or 2, characterized in that the ratio of the mixed liquid of the corynebacterium glutamicum seed liquid and the bacillus seed liquid inoculated into the culture medium is 1: 8-10.
4. A method for producing a fertilizer synergist by utilizing double-bacterium fermentation of rapeseed meal is characterized by comprising the following steps:
adding a culture medium containing rapeseed meal into a fermentation tank, mixing and inoculating corynebacterium glutamicum seed liquid and bacillus seed liquid into the fermentation tank according to the ratio of 1: 8-10, stirring at the rotating speed of 200-700 r/min, the ventilation volume of 3L/(L-min), and the pH of 6.5-7.0, culturing at the temperature of 30-45 ℃, feeding 30g/L glucose liquid after 24 hours, and performing fermentation culture for 36-72 hours to obtain a finished product.
5. The method for producing fertilizer synergist by utilizing double-bacterial fermentation of rapeseed meal according to claim 4, wherein the rapeseed meal-containing culture medium comprises the following components: 2.0-8.0 percent of rapeseed meal dispersion liquid, 0.2-5.0 percent of yeast production waste liquid, 0.01-6.5 percent of ammonium chloride, 0.1-2.5 percent of monopotassium phosphate, 0.05-1.5 percent of magnesium sulfate heptahydrate, 0.001-1.0 percent of manganese sulfate, 0.05-2.5 percent of ammonium citrate, 0.01-0.05 percent of sodium chloride, 0.001-0.006 percent of ferrous sulfate, 0.001-0.005 percent of cerous chloride, 0.05-0.2 percent of coconut oil oleic acid antifoaming agent, 0.02-0.25 percent of Tween 80 and the balance of deionized water.
6. The method for producing the fertilizer synergist by utilizing the double-bacterial fermentation of the rapeseed meal according to claim 5, wherein the preparation method of the rapeseed meal dispersion liquid comprises the following steps: crushing rapeseed meal, collecting rapeseed meal particles, sieving to obtain rapeseed meal powder, stirring and mixing the rapeseed meal powder and water according to a mass ratio of 1:5, grinding the mixture into slurry by a grinder, adding Tween 80, performing ultrasonic dispersion for 5-10 min at 200-300W, and collecting rapeseed meal dispersion liquid.
7. The method for producing fertilizer synergist by utilizing rape seed meal double-bacterium fermentation according to any one of claims 4-6, wherein the ratio of the corynebacterium glutamicum seed culture medium is as follows: glucose 25g/L, corn steep liquor 30g/L, K2HPO41.5g/L，MgSO40.8g/L, 5.5g/L urea, and pH7.0-7.3.
8. The method for producing fertilizer synergist by utilizing rape seed meal double-bacterial fermentation according to claim 7, wherein the ratio of the bacillus seed culture medium is as follows: 12g/L of tryptone, 5g/L of yeast extract, 40g/L of sodium chloride, 25g/L of agar and 7.1-7.6 of pH.
9. Use of the fertilizer synergist produced by the double-bacterial fermentation of rapeseed meal according to claims 1-3 as fertilizer synergist PG in crop growth.
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