CN116986932A - Preparation method of enteromorpha fertilizer synergist - Google Patents
Preparation method of enteromorpha fertilizer synergist Download PDFInfo
- Publication number
- CN116986932A CN116986932A CN202210444281.4A CN202210444281A CN116986932A CN 116986932 A CN116986932 A CN 116986932A CN 202210444281 A CN202210444281 A CN 202210444281A CN 116986932 A CN116986932 A CN 116986932A
- Authority
- CN
- China
- Prior art keywords
- enteromorpha
- fermentation
- liquid
- polysaccharide
- enzymolysis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000196252 Ulva Species 0.000 title claims abstract description 60
- 239000003337 fertilizer Substances 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000000855 fermentation Methods 0.000 claims abstract description 36
- 230000004151 fermentation Effects 0.000 claims abstract description 35
- 102000020244 polysaccharide lyase Human genes 0.000 claims abstract description 12
- 108091022901 polysaccharide lyase Proteins 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
- 230000015556 catabolic process Effects 0.000 claims description 11
- 238000006731 degradation reaction Methods 0.000 claims description 11
- 230000000813 microbial effect Effects 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 5
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000004108 freeze drying Methods 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims description 2
- 150000008264 rhamnoses Chemical class 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 241000196253 Ulva prolifera Species 0.000 claims 1
- 229920001282 polysaccharide Polymers 0.000 abstract description 12
- 239000005017 polysaccharide Substances 0.000 abstract description 12
- 241000278457 Bacillus siamensis Species 0.000 abstract description 4
- 241000195628 Chlorophyta Species 0.000 abstract description 4
- 102000004190 Enzymes Human genes 0.000 abstract description 4
- 108090000790 Enzymes Proteins 0.000 abstract description 4
- 150000004676 glycans Chemical class 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 abstract description 2
- 210000002421 cell wall Anatomy 0.000 abstract description 2
- 239000001913 cellulose Substances 0.000 abstract description 2
- 229920002678 cellulose Polymers 0.000 abstract description 2
- 244000005700 microbiome Species 0.000 abstract description 2
- 102000004169 proteins and genes Human genes 0.000 abstract description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract 2
- 239000011159 matrix material Substances 0.000 abstract 1
- 235000015097 nutrients Nutrition 0.000 abstract 1
- 239000000843 powder Substances 0.000 abstract 1
- 150000004804 polysaccharides Chemical class 0.000 description 11
- 229920001542 oligosaccharide Polymers 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 241000563903 Bacillus velezensis Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000009264 composting Methods 0.000 description 4
- 230000000593 degrading effect Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 150000002482 oligosaccharides Chemical class 0.000 description 4
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 102000004317 Lyases Human genes 0.000 description 3
- 108090000856 Lyases Proteins 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 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 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 235000010344 sodium nitrate Nutrition 0.000 description 2
- 239000004317 sodium nitrate Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000590031 Alteromonas Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 241000194103 Bacillus pumilus Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 244000247747 Coptis groenlandica Species 0.000 description 1
- 235000002991 Coptis groenlandica Nutrition 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000519590 Pseudoalteromonas Species 0.000 description 1
- 241000223261 Trichoderma viride Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000008233 hard water Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000010496 root system development Effects 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- -1 sulfuric acid oligosaccharide Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 208000016261 weight loss Diseases 0.000 description 1
- 239000013585 weight reducing agent Substances 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
-
- 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
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/50—Treatments combining two or more different biological or biochemical treatments, e.g. anaerobic and aerobic treatment or vermicomposting and aerobic treatment
-
- 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
- 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
- C05G3/80—Soil conditioners
-
- 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
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Soil Sciences (AREA)
- Virology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Fertilizers (AREA)
Abstract
The invention provides a preparation process of an enteromorpha fertilizer synergist, which takes enteromorpha powder as a raw material and utilizesAlteromonas cdwelliana A321And (3) preparing enteromorpha polysaccharide lyase. By passing throughBacillus methylotrophicus LJ、Bacillus siamensis L13The enteromorpha is treated by large-scale fermentation of the same microorganism, so that the effective components of the enteromorpha are dissolved out, and the enteromorpha polysaccharide is adopted for pyrolysisThe enzyme efficiently degrades characteristic polysaccharide, protein and cellulose in the cell matrix and the cell wall of the enteromorpha, fully releases nutrient elements in the enteromorpha, improves the fermentation speed of the enteromorpha, prepares the green algae-derived biostimulant, and is used as a raw material for producing novel green environment-friendly fertilizer suitable for modern agriculture.
Description
Technical Field
The invention relates to a preparation method of special enteromorpha polysaccharide lyase and a preparation method of an enteromorpha fertilizer synergist, belonging to the technical field of biology.
Background
The enteromorpha is rich in nutrition, contains 9-14% of protein, 32-36% of ash, 6.7-9% of water, 2% of fat and the balance of carbohydrate, and is about 40%. Especially, enteromorpha has special algal polysaccharide, highly unsaturated fatty acid, alginic acid, natural growth regulating substances and the like of marine organisms, is a plant with wide distribution and huge reserves in the ocean, and is one of plants with development value in the ocean. Compared with terrestrial plants, the enteromorpha contains rich specific polysaccharide chains, and the monosaccharide composition of the enteromorpha polysaccharide comprises xylose, galactose, arabinose, rhamnose and glucose, has the effects of regulating plant growth and enhancing disease resistance, and accounts for 10-40% of dry basis. At present, the degradation, development and utilization of green algae moss at home and abroad are still in an exploration stage.
The conventional enteromorpha polysaccharide degradation methods comprise an acidolysis method, a microwave-assisted acidolysis method and the like, but the oligosaccharide obtained by the acidolysis method has wider molecular weight distribution and the sugar chain groups and structures are affected to a certain extent; the oligosaccharide obtained by the microwave-assisted acidolysis method has lower yield and high production cost. Compared with physical and chemical methods, the biological enzyme method has the characteristics of strong specificity, mild enzymolysis conditions, simple process, controllable hydrolysis and the like, and becomes the optimal means for degrading the polysaccharide. Enteromorpha polysaccharide is used as a polyanionic heterogeneous polysaccharide, the sugar chain structure is special, and the existing tool enzyme can not hydrolyze the polysaccharide effectively. Therefore, developing a special enteromorpha lyase plays an important role in improving the recycling utilization rate of enteromorpha oligosaccharides, and is also a technical problem to be solved in the field.
Through searching, zhao Ming and the like, enteromorpha composting conditions are researched by using microbial agents, and the composting temperature of pure enteromorpha is less than 50 ℃. The composting conditions of the enteromorpha are researched by using natural bacteria such as coptis light, the highest temperature of the pure enteromorpha composting is only 54 ℃, and 7 d is required to reach the highest temperature. Zhu Jiang in the patent, a mixture of enteromorpha and straw is fermented by a compound microbial agent consisting of bacillus subtilis, pseudoalteromonas, bacillus pumilus, bacillus licheniformis and trichoderma viride. Shan Junwei and the like develop a method for fermenting enteromorpha by utilizing a microbial agent composed of bacteria, fungi and actinomycetes. There are no reports and patents related to the bacterial fermentation of green algae related to this patent.
Disclosure of Invention
Aiming at the defects in the technology, the invention provides a preparation method of the enteromorpha fertilizer synergist, which utilizes a pure biological method to ferment and degrade enteromorpha and release functional substances, and has the advantages of high efficiency, thoroughly, no pollution and mild action condition.
The invention solves the technical problems by the following technical proposal:
a method for realizing high-flux degradation of enteromorpha through enzyme-fermentation coupling liquefaction is characterized in that the enteromorpha is realized through mixed strain fermentation of Bacillus methylotrophicus LJ and Bacillus siamensis L, and a special enteromorpha lyase is used for further degrading fermentation liquor to release functional substances. The degradation mode of combining microbial fermentation degradation and biological enzymolysis is used for the first time, the primary degradation is realized by the microbial fermentation degradation, the cellulose structure cell wall of the enteromorpha is damaged by the biological enzymolysis, the organic active substances are dissolved out from the cells, and the macromolecule chain fragments are cut off, so that more active groups are exposed.
The microbial strains comprise Bacillus methylotrophicus LJ and Bacillus siamensis L, the bacillus methylotrophicus (Bacillus methylotrophicus) LJ and the bacillus Sims (Bacillus siamensis) L13 are respectively CGMCC No. 7570 and CGMCC No.7285, and can be directly purchased from the common microorganism center of China Committee for culture Collection of microorganisms, and the optimal compatibility ratio is 2:1.
The special enteromorpha polysaccharide lyase is characterized by comprisingAlteromonas cdwelliana A321The alteromonas strain is prepared by fermentation and is screened and preserved by a laboratory of the university of ocean food science and engineering. The molecular weight of the chlorella polysaccharide is 244 to Da, the extraction rate of active substances is high, the variety is many, the molecular weight is small, and the chlorella polysaccharide is easy to be absorbed by crops.
The preparation method of the enteromorpha lyase is characterized by simple production process, high yield of enteromorpha polysaccharide lyase, and the highest enzyme activity in fermentation broth of 72.8U/mL, which is remarkably superior to Li Yin.
The enteromorpha fertilizer synergist is rich in enteromorpha sulfuric acid oligosaccharide and uronic acid, and has the advantages of complete water solubility, hard water resistance, acid and alkali resistance, phosphorus and calcium activation, and good chelating ability for trace elements such as Cu, fe, zn, mn. The fertilizer can be used as a fertilizer synergist in various fertilizers such as high-tower compound fertilizer, macroelement water-soluble fertilizer, intermediate element water-soluble fertilizer, free trace element water-soluble fertilizer and the like, does not generate precipitation, has synergistic multiplication effect, can also be used as a fertilizer singly, and has obvious effects in repairing soil, stimulating photosynthesis of crops, improving seedling yield and promoting root system development. The product has wide application range, and is a novel fertilizer synergist suitable for sustainable development of weight reduction and synergy.
Detailed description of the preferred embodiments
The present invention will be further described with reference to specific examples, which are not intended to limit the scope of the invention.
Example 1: preparation of enteromorpha fertilizer synergist
(1) Microbial fermentation: mixing enteromorpha with water according to (2-4) g: uniformly mixing (20-40) mL, degrading for 20 min by using a microwave reactor under the condition of 180 ℃ and microwave power of 1000W, and centrifugally separating the extract when the reaction end system is cooled to 60 ℃, wherein the centrifugal speed is 10000 rpm, and the time is 20 min. And centrifuging the obtained extract again to obtain the final extract. Inoculating into the second clear liquidBacillus methylotrophicus LJAndBacillus siamensis L13the ratio of the mixed strain is 2:1, fermenting 120 h at 25-30 ℃ to obtain enteromorpha fermentation liquor.
(2) Degradation of enteromorpha polysaccharide lyase: conveying the enteromorpha fermentation liquor obtained in the step (1) to a reaction kettle, and adding water for dilution in the conveying process, wherein the mass ratio of enteromorpha degradation to water is preferably (5-10) g: (0.1-2) L, regulating the pH of an enzymolysis system to 5.5-6.5, then adding 0.5% of special enteromorpha polysaccharide lyase, maintaining the temperature of the enzymolysis system to 33-35 ℃, and carrying out enzymolysis on 2-3 h to obtain the enteromorpha fertilizer synergist.
Example 2: preparation of enteromorpha polysaccharide lyase
(1) Activating: will beAlteromonas cdwelliana A321Inoculating the strain to an activation culture medium, adjusting the initial pH to 7.0, and fermenting at 28-35 ℃ for 6-10 h to obtain fermentation seed liquid.
The composition of the activation culture medium is as follows: glucose 2%, enteromorpha polysaccharide 1%, peptone 0.5%, sodium nitrate 0.5%, sodium chloride 1%, mgSO 4 0.1 %，K 2 HPO 4 0.2 %。
(2) Preparing fermentation liquid: preparing fermentation liquor, adjusting the initial pH to 7.0, and sterilizing at 121 ℃ for 30 min.
The composition of the fermentation liquor is as follows: 2% of enteromorpha, 0.5% of peptone, 0.5% of sodium nitrate, 1% of sodium chloride and MgSO (MgSO) 4 0.1 %，K 2 HPO 4 0.2 %。
(3) Inoculating: inoculating the fermentation seed liquid into the fermentation liquid, wherein the inoculation amount is 6-10%.
(4) Fermentation: fermenting 36 h under the condition of controlling the temperature to be 28-35 ℃ to obtain fermentation liquor.
(5) Concentrating: concentrating the fermentation liquor to a solid content value of 30-35% to obtain the enzymolysis liquor.
(6) And (3) drying: and (3) spray drying or freeze drying the enzymolysis liquid to obtain the solid chlorella polysaccharide lyase.
Example 3: product inspection
Weighing 10 g enteromorpha fermentation end product, and dissolving in 90 mL distilled water. Adding 400 mL absolute ethyl alcohol, standing for 30 min, centrifuging at 3000 rpm for 10 min, collecting supernatant, and concentrating by rotary evaporation to 10 mL to obtain characteristic oligosaccharide concentrate for degrading green algae. GC and ESI-MS analysis are carried out on the concentrated solution to obtain a specific product, namely sulfated rhamnose, the molecular weight of the specific product is 244 Da, and the specific product can be used as a characteristic index for identifying enteromorpha fermentation of the strain.
Claims (4)
1. The preparation method of the enteromorpha fertilizer synergist is characterized by comprising the following steps of:
microbial fermentation: mixing Enteromorpha prolifera with water, performing microwave treatment, centrifuging the obtained microwave treated matter to obtain first residue and first clear liquid, centrifuging the first clear liquid to obtain second clear liquid and second residueThe method comprises the steps of carrying out a first treatment on the surface of the Inoculating into the second clear liquidBacillus methylotrophicus LJAndBacillus siamensis L13fermenting 120 h at 25-30 ℃ to obtain enteromorpha fermentation liquor,
degradation of enteromorpha polysaccharide lyase: conveying the enteromorpha fermentation liquor obtained in the step (1) to a reaction kettle, and adding water for dilution in the conveying process, wherein the mass ratio of enteromorpha degradation to water is preferably (5-10) g: (0.1-2) L, regulating the pH of an enzymolysis system to 5.5-6.5, then adding 0.5% of special enteromorpha polysaccharide lyase, maintaining the temperature of the enzymolysis system to 33-35 ℃, and carrying out enzymolysis on 2-3 h to obtain the enteromorpha fertilizer synergist.
2. A mixed strain for microbial fermentation according to claim 1, whereinBacillus methylotrophicus LJAndBacillus siamensis L13the proportion is 2:1.
3. the method for preparing enteromorpha polysaccharide lyase according to claim 1, which is characterized in that:
(1) Activating: will beAlteromonas cdwelliana A321Inoculating the strain to an activation culture medium, adjusting the initial pH to 7.0, and fermenting at 28-35 ℃ for 6-10 h to obtain fermentation seed liquid;
(2) Preparing fermentation liquid: preparing fermentation liquor, adjusting the initial pH value to 7.0, and sterilizing for 30 min at 121 ℃;
(3) Inoculating: inoculating the fermentation seed liquid to the fermentation liquid, wherein the inoculation amount is 6-10%;
(4) Fermentation: fermenting 36 h at the temperature of 28-35 ℃ to obtain fermentation liquor;
(5) Concentrating: concentrating the fermentation liquor to a solid content value of 30-35% to obtain an enzymolysis liquor;
(6) And (3) drying: and (3) spray drying or freeze drying the enzymolysis liquid to obtain the solid chlorella polysaccharide lyase.
4. The enteromorpha synergist of claim 1, characterized by containing a functional characteristic substance sulfated rhamnose and having a molecular weight of 244 Da.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210444281.4A CN116986932A (en) | 2022-04-26 | 2022-04-26 | Preparation method of enteromorpha fertilizer synergist |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210444281.4A CN116986932A (en) | 2022-04-26 | 2022-04-26 | Preparation method of enteromorpha fertilizer synergist |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116986932A true CN116986932A (en) | 2023-11-03 |
Family
ID=88527118
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210444281.4A Pending CN116986932A (en) | 2022-04-26 | 2022-04-26 | Preparation method of enteromorpha fertilizer synergist |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116986932A (en) |
-
2022
- 2022-04-26 CN CN202210444281.4A patent/CN116986932A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105647832B (en) | One plant of high temperature resistant garden waste decomposer FHM1 and its application | |
| CN110218667B (en) | Bacterial strain SH-1 for producing alginate lyase and application thereof | |
| CN105754881B (en) | A kind of Irpex lacteus of degradable lignin and its application | |
| CN105431534A (en) | beta-1,3-glucanase, polynucleotide, recombinant vector, transformant, production method for beta-1,3-glucanase, enzyme preparation, and production method for paramylon having reduced molecular weight | |
| CN111073839B (en) | Siam bacillus, microbial inoculum and application thereof | |
| CN101289680B (en) | Process for producing 2,3-butanediol using american artichoke as raw material by fermentation | |
| CN113880612A (en) | Method for preparing seaweed biological organic liquid fertilizer, seaweed biological organic liquid fertilizer and application | |
| CN109721421A (en) | A kind of preparation and application containing algal polysaccharides and fulvic acid composite synergistic phosphate fertilizer | |
| CN113321563A (en) | Seaweed organic water-soluble fertilizer | |
| CN106915987A (en) | A kind of preparation method of the special alga fertilizer of grape | |
| CN114561327B (en) | Cellulose degradation composite microbial inoculant, and preparation method and application thereof | |
| CN105670966B (en) | One plant of high temperature resistant garden waste decomposer ST4 and its application | |
| CN110668896A (en) | Method for preparing organic fertilizer by using activated sludge of corn starch plant | |
| CN111302855A (en) | Oligosaccharide organic peptide fertilizer and preparation method thereof | |
| CN106348887A (en) | Method for preparing amino acid liquid water-soluble fertilizer through compound stepped fermentation of seaweed | |
| CN111154661B (en) | Complex microbial inoculant and application thereof | |
| CN110396483B (en) | High-temperature straw degradation bacterium B-8, and microbial inoculum and application thereof | |
| CN111484368A (en) | Solid fermentation production method of microbial fertilizer and solid composite microbial fertilizer | |
| CN114989984B (en) | Method for preparing seaweed extract by enzymolysis | |
| CN113564215B (en) | Preparation method of biosurfactant with carbon dioxide and/or lignocellulose as substrate | |
| CN116986932A (en) | Preparation method of enteromorpha fertilizer synergist | |
| CN105567608B (en) | One plant of high temperature resistant garden waste decomposer ST1 and its application | |
| CN113430126A (en) | Aureobasidium pullulans and method for preparing melanin polysaccharide by using same | |
| CN111205146A (en) | Compound protein microbial fertilizer and preparation method thereof | |
| CN110551640A (en) | Preparation method of composite microbial inoculum capable of efficiently degrading corn straws |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication |