Amino acid water-soluble fertilizer containing chitosan oligosaccharide and salt-tolerant bacillus and preparation method of amino acid water-soluble fertilizer

Abstract

The invention belongs to the technical field of agricultural fertilizers, and particularly relates to an amino acid water-soluble fertilizer containing chitosan oligosaccharide and salt-tolerant bacillus and a preparation method of the amino acid water-soluble fertilizer. According to the method, litopenaeus vannamei heads are used as raw materials, a refined solution is prepared through mixed acid pretreatment, double-enzyme fermentation and nanofiltration membrane desalination, and the refined solution is mixed with a salt-tolerant domesticated bacillus licheniformis / bacillus natto solution, citric acid-malic acid chelated medium elements and a protective agent. According to the method, by optimizing a pretreatment process and a chelating system, synergistic degradation of shrimp head protein and chitin is achieved, the prepared fertilizer has the compound functions of amino acid, chitosan oligosaccharide and a salt-tolerant microbial agent, the problems that a microbial agent in a traditional fertilizer is poor in salt tolerance and prone to precipitation are solved, and the method is suitable for facility agriculture and saline-alkali soil and has a wide application prospect. The crop stress resistance can be enhanced; the growth effect is improved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of agricultural fertilizers, and particularly relates to an amino acid water-soluble fertilizer containing chitooligosaccharide and salt-tolerant bacillus and a preparation method thereof. BACKGROUND

[0002] With the development of modern agriculture towards precision and high efficiency, the demand for amino acid water-soluble fertilizer containing functional components is increasing. The current market products mainly use grain crops such as soybean meal as raw materials, which has two major contradictions: first, the competition between grain raw materials and human food, which increases the production cost (the price of soybean meal has increased by 40% in the past three years); second, the traditional acidolysis process requires high temperature (120-150 DEG C) treatment, resulting in an amino acid loss rate of 15-20%, and the functional components such as chitin cannot be retained.

[0003] The shrimp head waste of white shrimp produced by aquatic processing industry accounts for 30-40% of the weight of raw materials every year, which is rich in protein (50-60%) and chitin (15-20%), but the traditional treatment has multiple technical bottlenecks.

[0004] Synergistic degradation of shrimp head components is difficult: the crystallinity of shrimp head chitin is 40%, the traditional single enzymatic process has a chitin degradation rate of less than 30%, the yield of chitooligosaccharide is less than 5 g / L, and the proportion of macromolecular peptides (>1000 Da) produced by protein degradation is 60%, which has low absorption efficiency for crops.

[0005] The salt tolerance of the microbial agent is insufficient: the survival rate of wild bacillus strains is less than 50% when the salt concentration is greater than 5 g / L, which cannot adapt to the high salt environment (8-12 g / L) of shrimp head fermentation broth, and the secretion amount of biocontrol metabolites is low, and the inhibition rate of pathogenic bacteria is only 50-60%.

[0006] The compatibility of the multi-component system is poor: when there are amino acids (120-150 g / L), medium elements (CaO 25-35 g / L) and microbial agents in the fertilizer, Ca²⁺ and amino acids are easy to form precipitates, metal ions damage the cell membrane, and the number of viable bacteria decreases to less than 30% of the initial value after 3 months of storage.

[0007] The chelation stability is defective: the chelation rate of the traditional normal temperature chelation process is less than 85%, Ca²⁺ precipitates easily after standing, which blocks the drip irrigation system, and the agglomeration of nanoparticles leads to low absorption efficiency of crops.

[0008] The prior art has not achieved the technical breakthrough of "high value of aquatic waste-precise regulation of functional components-stable coexistence of multi-component system", lacks a synergistic degradation process of protein and chitin, has not established a salt-tolerant bacillus domestication system, and the medium element chelation process does not consider the compatibility with microorganisms, resulting in mutual antagonism of functional components. SUMMARY

[0009] The application provides a chitooligosaccharide and salt-tolerant bacillus amino acid water-soluble fertilizer containing chitooligosaccharide and salt-tolerant bacillus, which comprises the following components in mass concentration: Shrimp head fermentation product refined liquid: 70%-85% in volume ratio, containing 120-150 g / L of free amino acids, 10-30 g / L of small molecule peptides less than 1000 Da, and 5-15 g / L of chitooligosaccharide; Bacillus licheniformis / natto bacillus liquid: the number of viable bacteria is greater than or equal to 1x10 10 CFU / mL, and the adding amount is 0.5%-2.0% in volume ratio; Medium element composition: 25-35 g / L of CaO and 5-10 g / L of MgO, which exist in the form of citric acid-malic acid chelation; Protective agent: 1%-3% of trehalose and 0.5%-2% of glycine; Buffer: 0.05-0.1 mol / L of potassium dihydrogen phosphate-potassium hydrogen phosphate, and the pH is adjusted to 6.5-7.0; The rest is water.

[0010] On the basis of the above technical scheme, further, the shrimp head fermentation liquid extract is prepared through the following steps: The shrimp head is pretreated with 0.1 mol / L-0.5 mol / L mixed acid for 3-6 hours, filtered, and the filtrate is reserved for later use; wherein the mixed acid is in a weight ratio of citric acid: malic acid 3:2-2:3; The filter residue is added with clean water to form a suspension liquid at 10-30 g / L, and then yeast powder, KH2PO4, MgSO4·7H2O, FeSO4·2H2O and ZnSO4 are added, and the pH is adjusted to 4.5-5.5 to prepare a fermentation culture medium, which is sterilized for later use; Inoculate bacillus licheniformis and bacillus natto for mixed fermentation, convert chitin into chitooligosaccharide, and convert protein into amino acid; The fermentation product is filtered, the filter residue is dried to prepare a solid inoculant, and the supernatant is desalted to a salt content of less than or equal to 5 g / L by using a nanofiltration membrane with a molecular weight cut-off of 200-500 Da to obtain a refined liquid.

[0011] On the basis of the above technical scheme, further, the bacillus licheniformis / natto bacillus is a salt-tolerant domesticated strain, which can survive for more than 80% at a concentration of 5 g / L of NaCl, and the spore rate is greater than or equal to 95%.

[0012] On the basis of the above technical scheme, further, the preparation process of the medium element composition is as follows: The shrimp head is decalcified to obtain a filtrate containing calcium fruit acid, and suspended solids, colloids, bacteria and macromolecular organic substances are removed from the filtrate by using an ultrafiltration membrane. 3-5 g / L citric acid and 2-3 g / L malic acid are used as chelating agents to react with Ca 2+ , Mg 2+ at a molar ratio of 1.5-2:1 at 40-50°C for 30-60 minutes, and the chelation rate is greater than or equal to 95%.

[0013] On the basis of the above technical scheme, further, the protective agent further includes 0.3%-0.4% polyvinylpyrrolidone, which is used to protect the bacteria from direct contact with calcium and magnesium ions.

[0014] On the basis of the above technical scheme, further, in the mixed acid pretreatment, the pretreatment temperature is 40-45°C.

[0015] On the basis of the above technical scheme, further, in the fermentation medium, the addition amount of yeast powder is 5-10 g / L, and the addition amount of KH2PO4 is 1-2 g / L.

[0016] On the basis of the above technical scheme, further, the molecular weight cut-off of the ultrafiltration membrane is 0.1 μm, and the concentration of calcium ions in the desalted filtrate is 30-35 g / L.

[0017] According to the preparation method of the amino acid water-soluble fertilizer containing chitooligosaccharide and salt-tolerant Bacillus according to any of the above technical schemes, the preparation method comprises: Shrimp head fermentation liquid pretreatment: the shrimp head is pretreated by mixed acid, and the filter residue after filtration is used to prepare a fermentation medium, and the fermentation medium is inoculated with Bacillus licheniformis and Bacillus natto for mixed fermentation, and the fermentation liquid is desalted by a nanofiltration membrane to a salt content of less than or equal to 5 g / L to obtain a refined liquid; Medium element combination preparation: a filtrate containing calcium fruit acid is obtained by decalcifying shrimp heads pretreated by mixed acid, and citric acid and malic acid are mixed as chelating agents to react to form a medium element combination; Bacterium preparation: the Bacillus licheniformis / Bacillus natto is salt-tolerant domesticated so that more than 80% of the bacteria survive under a concentration of 5 g / L NaCl and the spore rate is greater than or equal to 95%, and a bacterium liquid with a viable count of greater than or equal to 1×10 0 CFU / mL is prepared; Mixing and blending: the refined liquid, the medium element combination solution, the bacterium agent and the protective agent are mixed below 25°C, and are filtered through a 0.22 μm membrane or screened by an ultrafiltration membrane; Filling in a sterile environment to obtain the amino acid water-soluble fertilizer containing chitooligosaccharide and salt-tolerant Bacillus.

[0018] The technical scheme provided by the present application has the following beneficial effects: The mixed acid (citric acid: malic acid = 1:1) cooperates with the ultrasonic-assisted enzymatic hydrolysis process, and the degradation rate of shrimp head protein is increased from 65% of the traditional process to 88%, and the yield of chitooligosaccharide is increased from 5g / L to 12g / L (increased by 40%). Each ton of shrimp head can produce 200L of refined liquid, and the small molecule peptides less than 1000Da account for 70%, and the crop absorbability is significantly enhanced. The process synchronously solves the problems of high cost of traditional grain raw materials and waste of aquatic waste, and meets the needs of circular economy.

[0019] Through the 3-5g / L gradient salt acclimation process, the survival rate of Bacillus licheniformis in 5g / L NaCl is 85% (wild strain is only 35%), and the spore rate is greater than or equal to 96%, which can directly adapt to the high salt environment of shrimp head fermentation liquor (salt content is less than or equal to 5g / L).

[0020] In the preferred scheme, the chelation process (molar ratio of citric acid-apple acid 1.8:1) is adopted, the chelation rate is 98.5% (traditional process <85%), the nanoparticle particle size is concentrated in 100-200nm (accounting for 92%), the particle size growth rate is less than 8% after 6 months of storage, and the Ca²⁺ precipitation problem is completely solved. The calcium element absorption rate of crop leaves is increased by 25% compared with the traditional process, and the clogging risk is reduced by 90% when adapting to the drip irrigation system.

[0021] Other features and advantages of the present application will be set forth in the following description, and in part will become apparent to those skilled in the art from the description, or can be learned by practice of the present application. The objects and other advantages of the present application can be realized and achieved by the structure specifically pointed out in the specification, claims and accompanying drawings. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical scheme and advantages of the embodiments of the present application more clear, the following will clearly and completely describe the technical scheme in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all the embodiments. The technical features designed in different embodiments of the present application can be combined with each other as long as they do not conflict with each other. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0023] In the description of the present application, it should be noted that all the terms (including technical terms and scientific terms) used in the present application have the same meaning as that generally understood by ordinary skilled persons in the field to which the present application belongs, and should not be understood as limiting the present application; it should be further understood that the terms used in the present application should be understood as having the same meaning as the terms in the context of the present application and the related field, and should not be understood in an idealized or overly formal sense, unless defined explicitly in the present application.

[0024] The present application provides the following examples and comparative examples: The shrimp used in the following examples and comparative examples is Litopenaeus vannamei.

[0025] Example 1 Formulation (per 1L): Shrimp head fermentation product refined liquid: 800 mL (free amino acid 135 g / L, small molecule peptide 20 g / L, chitooligosaccharide 10 g / L); Salt-tolerant domesticated bacteria liquid (Bacillus licheniformis and Bacillus natto, volume ratio 1:1): 10 mL (viable bacterial count 1×10 0 CFU / mL); Medium element composition: CaO 30 g / L, MgO 8 g / L (citric acid-malic acid chelated state, chelation rate 97%); Protective agent: trehalose 2%, glycine 1.5%, polyvinylpyrrolidone 0.35%; Buffer: 0.08 mol / L potassium dihydrogen phosphate-potassium hydrogen phosphate (molar ratio 1:1, pH 6.8).

[0026] The balance is made up to 1L with water.

[0027] Key parameters for preparation: Shrimp head pretreatment: 0.3 mol / L mixed acid (citric acid: malic acid = 1:1), 42℃ for 4h; Fermentation medium: yeast powder 8 g / L, KH2PO4 1.5 g / L, fermentation temperature 37℃, time 48h; Chelation process: 45℃, citric acid 4g / L + malic acid 2.5g / L, chelation time 45 minutes.

[0028] Performance indicators: Protein degradation rate: 88%, chitooligosaccharide yield: 12 g / L; Bacterial agent salt tolerance: survival rate 85% under 5 g / L NaCl, spore rate 96%; Storage stability: 18% viable bacterial attenuation rate after 6 months of storage at 30℃, no precipitation.

[0029] Example 2 Formulation adjustment compared with Example 1: The amount of bacterial solution added was increased to 2.0% (20 mL / L), and the proportions of the remaining ingredients remained unchanged Preparation differences: Two-stage gradient salt acclimation was used in the fermentation stage (3 g / L→5 g / L NaCl) Performance characteristics: Salt-alkali land applicability: 42% increase in crop root activity in 0.5% saline soil Biocontrol effect: 78% inhibition rate on tomato fusarium wilt (10% higher than Example 1) Example 3: Formulation adjustment compared with Example 1: Medium element composition: CaO 35 g / L, MgO 10 g / L Key preparation parameters: Chelation temperature 50°C, molar ratio of chelating agent 1.8:1, reaction time 60 minutes Performance improvement: Chelation rate reached 98.5%, and the nanoparticle particle size distribution was more uniform (92% of 100-200 nm) Crop absorption efficiency: 25% increase in leaf calcium element absorption rate compared with Example 1 Comparative Example 1 Difference from Example 1: Replaced with wild-type bacillus licheniformis (not salt-tolerant).

[0030] Other formulations and processes are the same as Example 1.

[0031] Performance comparison: Salt tolerance: survival rate of only 35% at 5 g / L NaCl, and spore rate of 70%.

[0032] Field effect: 15% reduction in yield of crops in saline-alkali land (20% increase in yield in Example 1).

[0033] Storage problem: 3 months of active bacteria count decayed to 40% of the initial value.

[0034] Comparative Example 2 Difference from Example 1: 0.3 mol / L single citric acid (canceling malic acid complex) was used to treat shrimp heads at 42°C for 4 hours, and the remaining pretreatment steps (filtration, fermentation medium preparation, etc.), as well as the ingredients and preparation parameters of the bacterial agent, medium element composition, and protective agent were the same as Example 1.

[0035] Performance defects: The protein degradation rate decreased from 88% to 65%, the proportion of macromolecular peptides (>1000Da) increased from 30% to 55%, and the content of small molecule peptides absorbed by crops decreased by 40%.

[0036] Due to single acid pretreatment, the decalcification of shrimp heads was not complete, the Ca²⁺ concentration in the filtrate fluctuated by ±15%, and the CaO content after chelation reaction was unstable (fluctuating between 25-32g / L), and the chelation rate decreased from 97% to 85%.

[0037] The initial salt residue in the fermentation broth decreased from 8-12g / L to 10-14g / L (single citric acid had stronger ability to dissolve the salt carried by shrimp heads), resulting in a decrease in the survival rate of salt-tolerant bacteria to 70% (85% in Example 1) during the acclimation stage.

[0038] Comparative Example 3 Differences from Example 1: Removal of polyvinylpyrrolidone in the protective agent Direct mixing of the bacterial agent with the medium elements The contact rate of the bacterial body with calcium and magnesium ions reached 85%, and the viable bacterial count decreased by 50% within 24 hours Storage stability: stratification occurred after 1 month, and the particle size of the chelation system increased to 200% of the initial value.

[0039] The performance comparison of the above examples and comparative examples is shown in the following table: Table 1

[0040] Among them, the protein degradation rate refers to the proportion of protein in shrimp heads that is degraded into free amino acids and small molecule peptides, and the calculation formula is: (initial protein content - residual protein content) / initial protein content x 100%; Directly reflects the utilization efficiency of the pretreatment process (mixed acid pretreatment + double enzyme fermentation) on shrimp head protein; An increase in degradation rate to 88% means that 30kg of absorbable nitrogen can be produced per ton of shrimp heads, which proves the advantages of the process compared to traditional acid hydrolysis process (65% degradation rate).

[0041] Chitooligosaccharide yield refers to the weight (g) of chitooligosaccharides (polymerization degree 2-8) extracted per liter of fermentation broth, which is quantitatively detected by high performance liquid chromatography (HPLC). The yield of 12 g / L is 40% higher than that of the traditional process, which measures the efficiency of conversion of shrimp chitin into functional oligosaccharides; as a plant immune inducer, the content of chitooligosaccharides directly affects the disease resistance effect of the fertilizer (such as the inhibition rate of tomato fusarium wilt); the yield is positively correlated with the proportion of mixed acid (citric acid: malic acid = 1:1) and the activity of the fermentation strain.

[0042] 5g / L NaCl survival rate refers to the percentage of viable bacteria after 24 hours of culture in 5g / L sodium chloride solution, which is used to verify the salt-tolerant domestication effect of the strain: the survival rate of the wild strain is only 35%, and the domesticated strain reaches 85%. Chelation rate refers to the proportion of medium elements (Ca²⁺, Mg²⁺) forming stable chelates with citric acid-malic acid, which is calculated by measuring free metal ions by atomic absorption spectrometry; a chelation rate of more than 97% avoids the formation of precipitates (such as the 85% chelation rate in Comparative Example 2) between Ca²⁺ and amino acids.

[0043] 6-month viable bacteria decay rate refers to the reduction in viable bacteria after 6 months of storage at 30°C compared to the initial value, which is detected by plate counting method. It is used to evaluate the synergistic effect of the protective agent system (trehalose + glycine + PVP): the decay rate is 45% without PVP (Comparative Example 3), which reflects the compatibility of the "microbe-nutrient" system: a decay rate of less than 22% ensures the stability of the product's shelf life.

[0044] Field saline soil effect refers to the field measured data such as crop yield increase and root activity improvement after applying the fertilizer in soil containing 0.3-0.5% salt. A 20% increase in yield in saline soil compared to a 15% decrease in yield with traditional fertilizer (Comparative Example 1) proves the practicality of the salt-tolerant system; a 42% increase in root activity is due to the root growth promoting effect of chitooligosaccharides and the biological stimulation effect of the bacterial agent; the biocontrol effect (fusarium wilt inhibition rate 78%) reflects the synergistic mechanism of "chitooligosaccharide-induced immunity + Bacillus biocontrol" Comparative Example 1 refers to the fact that the amino acid water-soluble fertilizer containing chitooligosaccharides and salt-tolerant Bacillus bacteria shows obvious phase separation after 1 month of storage.

[0045] In the above examples and comparative examples: The survival rate of the comparative example 1 is only 35% with 5 g / L NaCl, and the viable bacteria attenuate severely during storage. The wild strain lacks the expression regulation mechanism of salt tolerance genes, and cannot activate Na⁺ / H⁺ antiporter in high salt environment, resulting in ion imbalance in cells; the spore cortex of the bacillus not subjected to domestication is thin, and the content of osmotic protectants (such as pyridinedicarboxylic acid) is low, and the stress resistance decreases significantly.

[0046] In the comparative example 2, the protein degradation rate is reduced by 23% by using single citric acid pretreatment, and the yield of chitooligosaccharides is reduced by 33%. The acid buffer range of single citric acid is relatively narrow (pH 4.0-5.0), and it cannot stably maintain the optimal activity pH (4.5-5.5) of protease and chitinase, resulting in blocked synergistic degradation of shrimp head protein and chitin. In addition, the complexing ability of citric acid to Ca²⁺ is weaker than that of malic acid, resulting in incomplete decalcification of shrimp heads, large fluctuations in Ca²⁺ concentration in the filtrate, and further affecting the stability of subsequent medium element chelation; at the same time, the dissolution ability of single acid to the salt in shrimp heads is too strong, which makes the initial salt content of the fermentation broth increase, and inhibits the growth of bacterial cells.

[0047] In the comparative example 3, the viable bacteria attenuate by 50% within 24 hours in the group without protective agent isolation, and stratification occurs after 1 month of storage. The absence of polyvinylpyrrolidone (PVP) makes the bacterial cells lose steric hindrance protection, and the calcium and magnesium ions directly combine with the phospholipid molecules on the cell membrane, destroying the membrane integrity; PVP, as a hydrophilic polymer, can form a hydration protection layer on the surface of the bacterial cells, isolating the destructive effect of metal ions, and its absence leads to irreversible aggregation of chelated nanoparticles and bacterial cells, causing stratification of the system.

[0048] Finally, it should be noted that: the above examples are only used to illustrate the technical solutions of the present application, and not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. An amino acid water-soluble fertilizer containing chitosan oligosaccharides and halotolerant Bacillus, characterized in that: The following components are included in the following mass concentrations: Shrimp head fermentation product refined liquid: volume ratio is 70%-85%, containing 120-150g / L of free amino acids, 10-30g / L of small molecular peptides less than 1000Da, and 5-15g / L of chitosan oligosaccharides; Bacillus licheniformis and Bacillus natto bacterial solution: viable cell count ≥ 1×10 10 CFU / mL, added at a volume ratio of 0.5%-2.0%; Middle element composition: CaO 25-35g / L, MgO 5-10g / L, present in the form of citric acid-malic acid chelate; Protective agent: trehalose 1%-3%, glycine 0.5%-2%; Buffer: 0.05-0.1 mol / L potassium dihydrogen phosphate - dipotassium hydrogen phosphate, adjust pH to 6.5-7.0; The remainder is water.

2. The amino acid water-soluble fertilizer containing chitosan oligosaccharides and halodurable Bacillus according to claim 1, characterized in that: The shrimp head fermentation broth extract is prepared by the following steps: Pre-treat the shrimp heads with 0.1 mol / L to 0.5 mol / L mixed acid for 3 to 6 hours, filter, and keep the filtrate for later use; wherein the mixed acid is citric acid: malic acid in a weight ratio of 3:2 to 2:3; Add clean water to the filter residue to make a suspension at 10-30g / L, then add yeast powder, KH2PO4, MgSO4・7H2O, FeSO4・2H2O, and ZnSO4, adjust the pH to 4.5-5.5, and prepare the fermentation medium. Sterilize and set aside. Inoculate Bacillus licheniformis and Bacillus natto for mixed fermentation to convert chitin into chito-oligosaccharides and protein into amino acids; The fermentation product is filtered, and the filter residue is dried to prepare a solid bacterial agent. The supernatant is desalted to a salt content of ≤5g / L using a nanofiltration membrane with a molecular weight cutoff of 200-500Da to obtain a refined liquid.

3. The amino acid water-soluble fertilizer containing chitosan oligosaccharides and halodurable Bacillus according to claim 1, characterized in that: The Bacillus licheniformis and Bacillus natto are salt-tolerant acclimated strains, which can survive more than 80% at a NaCl concentration of 5 g / L, and have a spore rate of ≥95%.

4. The amino acid water-soluble fertilizer containing chitosan oligosaccharides and halotolerant Bacillus according to claim 1, characterized in that: The preparation process of the intermediate element composition is as follows: The filtrate containing calcium hydroxybenzoate is obtained by decalcifying shrimp heads and then filtered through an ultrafiltration membrane to remove suspended solids, colloids, bacteria, and macromolecular organic matter. 3-5g / L citric acid and 2-3g / L malic acid are used as chelating agents, and Ca 2+ Mg 2+ React at 40-50℃ for 30-60 minutes at a molar ratio of 1.5-2:1, and the chelation rate should be ≥95%.

5. The amino acid water-soluble fertilizer containing chitosan oligosaccharides and halodurable Bacillus according to claim 1, characterized in that: The protective agent also includes 0.3% - 0.4% polyvinyl pyrrolidone, which is used to protect the bacteria from direct contact with calcium and magnesium ions.

6. The amino acid water-soluble fertilizer containing chitosan oligosaccharides and halodurable Bacillus according to claim 2, characterized in that: In the mixed acid pretreatment, the pretreatment temperature is 40°C-45°C.

7. The amino acid water-soluble fertilizer containing chitosan oligosaccharides and halodurable Bacillus according to claim 2, characterized in that: In the fermentation medium, the added amount of yeast powder is 5-10 g / L, and the added amount of KH2PO4 is 1-2 g / L.

8. The amino acid water-soluble fertilizer containing chitosan oligosaccharides and halodurable Bacillus according to claim 4, characterized in that: The molecular weight cut-off of the ultrafiltration membrane is 0.1 μm, and the calcium ion concentration in the filtrate after desalination is 30-35 g / L.

9. The method for preparing the amino acid water-soluble fertilizer containing chito-oligosaccharide and halodurable Bacillus according to any one of claims 1 to 8, characterized in that: The preparation method comprises: Pretreatment of shrimp head fermentation liquid: The shrimp heads are pretreated with mixed acid, and the filter residue is prepared into a fermentation medium, which is inoculated with Bacillus licheniformis and Bacillus natto for mixed fermentation. The fermentation liquid is desalted to a salt content of ≤5g / L through a nanofiltration membrane to obtain a refined liquid; Preparation of the intermediate element composition: Pre-treating shrimp heads with mixed acid to decalcify to obtain a filtrate containing calcium hydroxybenzoate, and reacting with a mixture of citric acid and malic acid as a chelating agent to form the intermediate element composition; Preparation of inoculant: Bacillus licheniformis and Bacillus natto were acclimated to salt tolerance, so that they could survive more than 80% at a concentration of 5g / L NaCl and the spore rate was ≥95%, and the number of viable bacteria was ≥1×10¹ 0 CFU / mL of bacterial solution; Mixing and preparation: Mix the refined liquid, medium element composition solution, bacterial agent and protective agent at a temperature below 25°C, and filter through a 0.22μm membrane or ultrafiltration membrane; By filling under a sterile environment, the amino acid water-soluble fertilizer containing chito-oligosaccharide and halotolerant Bacillus is obtained.

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