CN117088734A - Preparation method for producing green alga fertilizer synergist for improving soil fertility based on enzyme-fermentation coupling technology - Google Patents
Preparation method for producing green alga fertilizer synergist for improving soil fertility based on enzyme-fermentation coupling technology Download PDFInfo
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- 239000003337 fertilizer Substances 0.000 title claims abstract description 23
- 238000000855 fermentation Methods 0.000 title claims abstract description 21
- 230000035558 fertility Effects 0.000 title claims abstract description 10
- 230000008878 coupling Effects 0.000 title claims abstract description 7
- 238000010168 coupling process Methods 0.000 title claims abstract description 7
- 238000005859 coupling reaction Methods 0.000 title claims abstract description 7
- 238000005516 engineering process Methods 0.000 title claims abstract description 5
- 238000002360 preparation method Methods 0.000 title claims description 7
- 241000195628 Chlorophyta Species 0.000 claims abstract description 19
- 230000004151 fermentation Effects 0.000 claims abstract description 18
- 229920001542 oligosaccharide Polymers 0.000 claims abstract description 10
- 150000002482 oligosaccharides Chemical class 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims abstract description 9
- 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 claims abstract description 9
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000000126 substance Substances 0.000 claims abstract description 9
- 230000000813 microbial effect Effects 0.000 claims abstract description 8
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims abstract description 7
- 230000001105 regulatory effect Effects 0.000 claims abstract description 6
- 238000004519 manufacturing process Methods 0.000 claims abstract description 3
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims description 16
- 229920001282 polysaccharide Polymers 0.000 claims description 12
- 239000005017 polysaccharide Substances 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 102000004190 Enzymes Human genes 0.000 claims description 9
- 108090000790 Enzymes Proteins 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 9
- 102000020244 polysaccharide lyase Human genes 0.000 claims description 8
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- 102000004317 Lyases Human genes 0.000 claims description 6
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- 239000000243 solution Substances 0.000 claims description 6
- 241000278457 Bacillus siamensis Species 0.000 claims description 4
- 241000563903 Bacillus velezensis Species 0.000 claims description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 4
- 229920002521 macromolecule Polymers 0.000 claims description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 239000005955 Ferric phosphate Substances 0.000 claims description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 2
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 229940032958 ferric phosphate Drugs 0.000 claims description 2
- 238000004108 freeze drying Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- WBJZTOZJJYAKHQ-UHFFFAOYSA-K iron(3+) phosphate Chemical compound [Fe+3].[O-]P([O-])([O-])=O WBJZTOZJJYAKHQ-UHFFFAOYSA-K 0.000 claims description 2
- 229910000399 iron(III) phosphate Inorganic materials 0.000 claims description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000006116 polymerization reaction Methods 0.000 claims description 2
- 239000011780 sodium chloride Substances 0.000 claims description 2
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- 238000000108 ultra-filtration Methods 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 241000589563 Alteromonas sp. Species 0.000 claims 1
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- 241001148470 aerobic bacillus Species 0.000 abstract description 3
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- 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
- 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
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- 241000590031 Alteromonas Species 0.000 description 1
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- 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 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
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- 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
- 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
- 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
- 150000001720 carbohydrates Chemical class 0.000 description 1
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- 229930182830 galactose Natural products 0.000 description 1
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Classifications
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- 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
- 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
- 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
- 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.)
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- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
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- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/12—Disaccharides
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- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- 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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y402/00—Carbon-oxygen lyases (4.2)
- C12Y402/02—Carbon-oxygen lyases (4.2) acting on polysaccharides (4.2.2)
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Abstract
The invention provides a method for producing a green alga fertilizer synergist for improving soil fertility based on an enzyme-fermentation coupling technology. The invention takes green algae as a main raw material, adopts an enzyme-ferment coupling liquefaction technology of high-concentration green algae, adopts a degradation mode combining microbial fermentation degradation and biological enzymolysis, rapidly and efficiently releases active substances, and simultaneously can efficiently prepare rhamnose sulfate block oligosaccharides with uniform molecular weight. The rhamnose and organic matters are combined in soil to form a high molecular weight compound, so that soil moisture is maintained, and the aggregate structure is improved; the water, fertilizer, gas, heat and other conditions of the soil are regulated, the soil exchange capacity is improved, and the acid-base balance is achieved. Promote the activity of soil microorganisms, increase the number of aerobic bacteria, actinomycetes and cellulose decomposing bacteria, accelerate the decomposition of organic substances and increase the soil fertility.
Description
Technical Field
The invention relates to a preparation method of a green alga fertilizer synergist, belonging to the technical field of biology.
Background
Soil fertility is one of the determining factors of crop yield and fertilization economic benefit. In recent years, with the improvement of the living standard of people, the demands for high-yield and high-quality agricultural products and nutritional health-care foods are increasing. It is therefore a great trend in the art to find a new fertilizer species that is environmentally friendly and does not present food safety concerns.
Take Enteromorpha in green algae as an example. It is nutritious, contains protein 9-14%, ash 32-36%, water 6.7-9%, fat 2%, and carbohydrate 40% in balance. Especially, enteromorpha has special seaweed 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 sugar chains, and the monosaccharide composition of the chlorella polysaccharide comprises xylose, galactose, arabinose, rhamnose and glucose, so that the chlorella polysaccharide has the effects of regulating plant growth and enhancing disease resistance, and particularly the rhamnose sulfate block oligosaccharide is combined with organic matters in soil to form a high molecular weight compound, so that soil moisture is maintained, and the aggregate structure is improved; the conditions of water, fertilizer, gas, heat and the like of the soil are regulated, the exchange capacity of the soil is improved, and the acid-base balance is achieved. Promote the activity of soil microorganisms, increase the number of aerobic bacteria, actinomycetes and cellulose decomposing bacteria, accelerate the decomposition of organic substances and increase the soil fertility. The degradation, development and utilization of the chlorella polysaccharide at home and abroad are in an exploration stage.
The conventional degradation methods of the chlorella polysaccharide 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 green alga lyase has very important effect on improving the resource utilization rate of the green alga oligosaccharide, and is also a technical problem to be solved in the field.
Disclosure of Invention
The invention aims at overcoming the defects in the prior art and provides a preparation method of a green algae fertilizer synergist for improving soil fertility.
In order to achieve the aim of the invention, the invention is realized by adopting the following technical scheme:
a method for realizing high-flux degradation of green algae by enzyme-ferment coupling liquefaction is characterized in that mixed strains of Bacillus methylotrophicus LJ and Bacillus siamensis L are used for fermenting to realize the green algae liquefaction, and special green algae 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 microbial fermentation degradation realizes preliminary degradation, the biological enzymolysis damages the cellulose structure cell wall of the green alga, the effective active substances in the cells are dissolved out to cut off the organic active substances, and the macromolecular chain fragments are used for exposing more active groups.
The special lyase is characterized in that the ratio of the chlorella polysaccharide lyase to the rhamnosidase is 1:2. the Chlorella polysaccharide lyase is prepared by fermenting alteromonas species, which is called Catenocum sp.LP214. The special lyase can degrade functional substance rhamnose sulfate block oligosaccharide, the polymerization degree is 2-5, and the molecular weight is 244, 402, 628, 760 and 982Da.
The invention has the advantages and positive effects that:
1. the invention realizes the high-flux degradation of green algae through enzyme-ferment coupling liquefaction. 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 cell, and the macromolecule chain fragments are cut off, so that more active groups are exposed.
2. The green algae fertilizer synergist produced by the invention can be added as a fertilizer synergist in various fertilizers such as high-tower compound fertilizers, macroelement water-soluble fertilizers, intermediate element water-soluble fertilizers, free trace element water-soluble fertilizers and the like, and cannot generate precipitation, so that the soil fertility is increased, and the effect is synergistically multiplied.
3. The green alga fertilizer synergist produced by the invention is rich in functional substances rhamnose sulfate block oligosaccharides, and is combined with organic matters to form a high molecular weight compound in soil, so that soil moisture is maintained, and the granule structure is improved; the water, fertilizer, gas, heat and other conditions of the soil are regulated, the soil exchange capacity is improved, and the acid-base balance is achieved. Promote the activity of soil microorganisms, increase the number of aerobic bacteria, actinomycetes and cellulose decomposing bacteria, accelerate the decomposition of organic substances and increase the soil fertility.
Other features and advantages of the present invention will become more apparent upon reading the detailed description of the invention
Detailed Description
The technical scheme of the invention is further described in detail below with reference to the specific embodiments.
Example 1: preparation of green algae fertilizer synergist
(1) Microbial fermentation: mixing green algae with water, performing microwave treatment, centrifuging the obtained microwave treated matter to obtain first filter residue and first clear liquid, and centrifuging the first clear liquid to obtain second clear liquid and second filter residue. The second supernatant was inoculated with a mixed strain of Alternomonas sp.A321, bacillus methylotrophicus LJ and Bacillus siamensis L, and fermented at 30 ℃.
(2) Synchronous enzymolysis: when fermenting for 60 hours, adding water into the system for dilution, wherein the mass ratio of the green algae degradation mass to the water is preferably 5g:2L, adding special chlorella polysaccharide lyase in an amount of 0.5%, adjusting the pH of the system to 6, maintaining the temperature of the enzymolysis system at 30 ℃, fermenting and synchronously carrying out enzymolysis for 5h, and centrifugally separating while the solution is hot to obtain chlorella enzymolysis solution.
(3) Preparing a green algae fertilizer synergist: after the enzymolysis is finished, heating to 100 ℃, preserving heat for 1-2min to inactivate enzymes, cooling, centrifuging for 10min at 8000r/min, and taking supernatant enzymolysis liquid; removing macromolecular substances from the supernatant enzymolysis liquid by using an ultrafiltration membrane with the molecular weight cutoff of 3000Da to obtain rhamnose sulfate-rich block oligosaccharide, and obtaining the chlorella fertilizer synergist after freeze drying.
Example 2: preparation of Chlorella polysaccharide lyase
Inoculating the strain Catenocum sp.LP214 into an enzyme-producing culture medium for fermentation, wherein the fermentation condition is 32 ℃ for 36 hours; wherein the enzyme-producing culture medium comprises 10g of chlorella polysaccharide, 3g of yeast extract, 4g of magnesium sulfate, 0.5g of calcium chloride, 20g of sodium chloride, 1.5g of ammonium sulfate, 0.2g of ferric phosphate, 1000mL of tap water and pH7.5; centrifuging the fermentation broth at 10000r/min for 5min to obtain supernatant, maintaining the supernatant at 40deg.C for 30-45min, and cooling to room temperature to obtain Chlorella polysaccharide lyase.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; while the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to one skilled in the art that modifications may be made to the embodiments described, or equivalents may be substituted for elements thereof; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (5)
1. A preparation method of a green algae fertilizer synergist for improving soil fertility is characterized by utilizing enzymolysis and fermentation coupling technology; the method comprises the following specific steps:
(1) Microbial fermentation: mixing green algae with water, performing microwave treatment, centrifuging the obtained microwave treated matter to obtain first filter residue and first clear liquid, and centrifuging the first clear liquid to obtain second clear liquid and second filter residue. Inoculating mixed strains of Alternomonas sp.A321, bacillus methylotrophicus LJ and Bacillus siamensis L into the second clear liquid, and fermenting at 25-35 ℃.
(2) Synchronous enzymolysis: in the fermentation process, water is added into the fermentation broth for dilution, and the mass ratio of the green algae degradation product to the water is preferably (5-10) g: (0.1-2) L, then adding 0.5% of special lyase, regulating pH of the system to 5-7.5, maintaining the temperature of the enzymolysis system to 25-45 ℃, fermenting and synchronously carrying out enzymolysis for 1-15h, and centrifuging while the solution is hot to obtain the green algae enzymolysis solution.
(3) Preparing a green algae fertilizer synergist: after the enzymolysis is finished, heating to 100 ℃, preserving heat for 1-2min to inactivate enzymes, cooling, centrifuging for 10min at 8000r/min, and taking supernatant enzymolysis liquid; removing macromolecular substances from the supernatant enzymolysis liquid by using an ultrafiltration membrane with the molecular weight cutoff of 3000Da to obtain rhamnose sulfate-rich block oligosaccharide, and obtaining the chlorella fertilizer synergist after freeze drying.
2. The mixed strain for microbial fermentation according to claim 1, wherein the ratio of Alteromonas sp.a321, bacillus methylotrophicus LJ and Bacillus siamensis L is 3:2:1.
3. the special lyase of claim 1 is a complex enzyme, and is characterized in that the ratio of the chlorella polysaccharide lyase to the rhamnosidase is 1:2.
4. the process for preparing a polysaccharide lyase for green algae of claim 3, wherein:
inoculating the strain Catenocum sp.LP214 into an enzyme-producing culture medium for fermentation, wherein the fermentation condition is 32 ℃ for 36 hours; wherein the enzyme-producing culture medium comprises 10g of chlorella polysaccharide, 3g of yeast extract, 4g of magnesium sulfate, 0.5g of calcium chloride, 20g of sodium chloride, 1.5g of ammonium sulfate, 0.2g of ferric phosphate, 1000mL of tap water and pH7.5; centrifuging the fermentation broth at 10000r/min for 5min to obtain supernatant, maintaining the supernatant at 40deg.C for 30-45min, and cooling to room temperature to obtain Chlorella polysaccharide lyase.
5. The synergist according to claim 1, characterized by containing functional substances of rhamnose sulfate block oligosaccharides with a degree of polymerization of 2-5 and molecular weights of 244, 402, 628, 760, 982Da.
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