CN111647026B

CN111647026B - Method for preparing water-soluble micromolecular active substance by one-step conversion of shrimp and crab shells and application thereof

Info

Publication number: CN111647026B
Application number: CN202010540023.7A
Authority: CN
Inventors: 胡雪芳; 王士奎
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-06-15
Filing date: 2020-06-15
Publication date: 2021-08-06
Anticipated expiration: 2040-06-15
Also published as: CN111647026A

Abstract

The invention relates to a method for preparing water-soluble micromolecule active substances by one-step conversion of shrimp and crab shells and application thereof, relating to an environment-friendly processing technology, which is characterized in that organic and inorganic matrix materials represented by the shrimp and crab shells are crushed to the granularity of less than 20 meshes, and a polyphosphoric acid-choline chloride eutectic solvent complex system is used for dissolving for 2 to 4 hours under the stirring condition of 80 to 95 ℃; under the condition of heat preservation, 0.1-1% of catalyst aqueous solution is added for catalytic degradation for 1-3 hours to obtain a stable water-soluble active substance solution. The polyphosphoric acid-choline chloride eutectic solvent composite system has good solubility, has good solubility for cellulose, protein, inorganic salt and the like in an organic-inorganic mixed matrix, can dissolve shrimp and crab shells by a one-step method to prepare a stable water-soluble active substance solution rich in amino-oligosaccharin, soluble calcium, soluble small molecular peptide and amino acid, can prepare a product suitable for multiple purposes by taking organic-inorganic matrix waste as a raw material in a low-cost, environment-friendly and efficient mode.

Description

Method for preparing water-soluble micromolecular active substance by one-step conversion of shrimp and crab shells and application thereof

Technical Field

The invention relates to an environment-friendly processing technology, in particular to a method for preparing water-soluble micromolecule active substances by one-step conversion of shrimp and crab shells and application thereof.

Background

A large amount of organic or inorganic wastes, such as marine resource wastes, such as shrimp and crab shells, fishbone, shells, oyster shells, gulfweed, enteromorpha, kelp and the like, are generated in daily production by a plurality of industries; planting waste such as corn stalk, rice stick (core), wheat stalk, wheat bran, rice stalk, rice hull, rice bran, peanut shell, pericarp, fruit shell, and coconut husk; food material processing residue such as mushroom residue, wine residue, vinegar residue, bagasse and Chinese medicinal residue; and microbial fermentation byproducts. These organic and inorganic wastes have reprocessing and recycling value, and organic or inorganic substrates contained therein are extracted or separated and used as fertilizer, feed, animal and plant growth regulator, and even raw material for pharmaceuticals.

For example, the shrimp and crab shells in the marine resource waste are unique organic-inorganic composite matrix rich in amino polysaccharide in the nature, and the matrix is rich in nutrition and contains 20-40% of protein, 20-50% of calcium carbonate and 25-40% of chitin. Shrimp and crab shells are used as a substrate rich in organic matters and inorganic matters, and are mainly used for chitin extraction, chitosan and chitosan oligosaccharide production and used as a feed additive at present.

The shrimp and crab shells are used as feed additives, because the aminopolysaccharide is a nitrogen-containing organic matter which is difficult to digest and absorb by animals, although the nitrogen-containing organic matter in the shrimp and crab shells is very high, more than 80 percent of the nitrogen-containing organic matter is a nitrogen source which is not absorbable by animals, the utilization rate of the nutritional value is very limited, and the small molecular weight and water solubility of the shrimp and crab shells are necessary ways for realizing high value of the shrimp and crab shells as the feed additives. In the prior art for producing chitin, chitosan and chitosan oligosaccharide by using shrimp and crab shells, a large amount of acid and alkali is needed in the processes of deproteinization, decalcification and deacetylation, so that a large amount of organic wastewater is generated, the environmental pollution is serious due to direct discharge, the treatment cost is high, and 80 percent of shrimp and crab shell resources in China are not developed and utilized due to the factors.

In conclusion, a green, environment-friendly and effective method is sought to solve the problems of environmental protection, cost and production efficiency, and the method has a very practical significance for recycling organic and inorganic waste resources including shrimp and crab shells.

Disclosure of Invention

The invention provides a method for preparing water-soluble micromolecule active substances based on organic and inorganic matrix materials represented by shrimp and crab shells, and provides a polyphosphoric acid-choline chloride eutectic solvent composite system which has good solubility, has good solubility for cellulose, protein, inorganic salt and the like in an organic and inorganic mixed matrix, can dissolve organic and inorganic matrixes such as the shrimp and crab shells in a one-step method, and obtains products suitable for multiple purposes, wherein the specific scheme is as follows:

the invention adopts the following technical scheme:

a method for preparing water-soluble micromolecule active substances by one-step transformation of shrimp and crab shells is characterized by comprising the following steps:

drying shrimp and crab shells, crushing the dried shrimp and crab shells to a granularity of less than 20 meshes, and dissolving the shrimp and crab shells for 2 to 4 hours by using a polyphosphoric acid-choline chloride eutectic solvent complex system under the stirring condition of 80 to 95 ℃;

under the condition of heat preservation, adding a catalyst aqueous solution with the mass volume ratio concentration of 0.1-1% for catalytic degradation for 1-3 hours to obtain a stable solution containing the soluble micromolecular active substance.

2. Preferably, the catalyst is hypoiodic or hydroiodic acid, or a mixture thereof.

3. Preferably, the polyphosphoric acid-choline chloride eutectic solvent composite system is formed by mixing choline chloride and polyphosphoric acid according to the mass ratio of 1: 5-10 and stirring for 0.5-1 hour at 85 ℃.

4. Preferably, the mass ratio of the shrimp and crab shells to the polyphosphoric acid-choline chloride eutectic solvent composite system to the catalyst aqueous solution is 0.2-1: 2: 0.7-3;

5. preferably, the shrimp and crab shells are dried to a moisture content of less than 10%.

6. Preferably, the mass fraction of phosphoric acid in the polyphosphoric acid is 105-118%.

7. Preferably, in any of the above embodiments, after the catalytic degradation reaction is completed, the reaction mixture is cooled to room temperature, insoluble substances are removed by filtration, and then centrifugation or filtration is performed to obtain a stable water-soluble active substance solution from the filtrate.

8. The steady-state water-soluble active substance solution prepared by the method has the nitrogen-containing organic water-soluble substance more than 10 percent, the glucosamine total amount more than 5 percent and the nitrogen organic matter content more than 2 percent; the water-soluble phosphatidated fatty acid content is greater than 0.2%; the content of water-soluble calcium is more than 8%. Can be used as mother liquor, raw material or raw material of amino-oligosaccharin, and can also be used as raw material of crustacean stimulin, and can be used as partial substitute of plant hormone for regulating plant growth.

9. The application of any one of the methods in the treatment of organic and inorganic materials is characterized in that shrimp and crab shell materials are replaced by organic and inorganic materials;

the organic and inorganic materials refer to marine wastes, planting wastes or food material processing residues;

the marine waste comprises: fishbone, shell, oyster shell, gulfweed, enteromorpha, kelp, or a mixture of any two or more of them;

the planting waste is: corn stalks, corn cobs, wheat stalks, wheat bran, rice stalks, rice hulls, rice chaff, peanut shells, fruit peels, fruit shells, coconut husks, or a mixture of any two or more of the foregoing;

the food material processing residue: mushroom residue, wine residue, vinegar residue, bagasse, herb residue, or a mixture of any two or more thereof.

10. The liquid product obtained from the aforementioned application.

11. The steady-state water-soluble active substance solution or the liquid product is used as a raw material for preparing amino-oligosaccharin or crustacean stimulin or used as a plant hormone substitute for regulating plant growth.

The invention establishes a polyphosphoric acid-choline chloride eutectic solvent composite system, can quickly dissolve organic matters (such as water-insoluble chitin, cellulose, muramic acid, glucan and other organic macromolecules taking beta-1, 4 glycosidic bond, beta-1, 3 glycosidic bond or beta-1, 6 glycosidic bond as main components) in an organic-inorganic mixed matrix represented by shrimp and crab shells at high concentration, and water-insoluble inorganic matters taking calcium carbonate (including calcium sulfate, calcium phosphate and the like) as main components to form stable organic-inorganic mixed dissolved solution; further controlling hydrolysis conditions, and further obtaining the water-soluble small-molecule organic-inorganic compound under the condition of below 100 ℃, so that the full conversion of matrix nutrients is achieved, and the mass of insoluble substances obtained by filtration is less than 10% of the mass of the total initial materials, namely the material conversion rate is more than 90%. When the obtained product is used for developing a plant resistance inducer product, polyphosphoric acid and choline chloride adopted in the product do not need a subsequent separation and recovery step, and the polyphosphoric acid and the choline chloride can be used as plant nutrient components to form a small molecular substance dissolved matter together with materials to form a stable soluble active fertilizer.

When the organic-inorganic matrix is shrimp and crab shells, through detection, the stable water-soluble active substance solution obtained by the method contains more than 10% of nitrogen-containing organic water-soluble substances (wherein, glucosamine phosphate, amino oligosaccharide and soluble nanowire glucosamine are more than 5%, the total content of the glucosamine is more than 2%, the content of amino acid, polypeptide and water-soluble protein and the content of protein nitrogen organic matters is more than 2%), and simultaneously contains more than 0.2% of water-soluble phosphatided fatty acid; the content of water-soluble calcium (ammonia sugar complex calcium, amino acid complex calcium, peptide complex calcium, calcium polyphosphate and the like) is more than 8 percent; meanwhile, the nutrient solution contains soluble mineral nutrients such as zinc, iron, magnesium, potassium, iodine, selenium and the like.

In summary, the technical advantages of the present invention are summarized as follows:

1. the process technology is green, environment-friendly, safe and efficient. Taking shrimp and crab shells as an example, the production efficiency is high: in the traditional technology, three steps of processes (shrimp and crab shells → chitin, chitin → chitosan, chitosan → amino-oligosaccharin) are needed for producing the amino-oligosaccharin from shrimp and crab shells, but the technology of the invention can shorten the process into a one-step conversion process for directly preparing the amino-oligosaccharin from the shrimp and crab shells.

The cost is low: the cost of raw materials for producing 1 ton of amino-oligosaccharide by the traditional process is about 10 ten thousand yuan, the cost of raw materials for producing 1 ton of amino-oligosaccharide by the process is about 4 ten thousand yuan, and the cost can be saved by more than 60%; meanwhile, no waste liquid and waste gas is discharged, the subsequent waste water treatment cost is saved, and the social and economic benefits are very obvious.

The technology is environment-friendly: the green treatment of the shrimp and crab shells is realized, and the adoption of the technology for preparing the amino-oligosaccharin can solve the bottleneck problems that the traditional process has serious environmental pollution, high production cost and the like and restricts the popularization and application of the amino-oligosaccharin in the agricultural planting industry. The components such as chitin, protein, calcium carbonate and the like contained in the raw materials, and phosphate radicals and choline chloride contained in the solvent are all converted into small molecular water-soluble substances which can be absorbed and utilized by plants, so that the technology completely meets the requirements of environment-friendly industrial production, and feasible technical guarantee is provided for the development of the plant resistance inducer industry.

2. The technology of the invention can thoroughly solve the problem of green harmless high-density liquefaction of residues (including microbial residues, straws, seaweed, crustaceans, shells and the like) after biomass processing, and lays an industrial industrialization foundation for rapidly liquefying organic-inorganic mixed matrixes.

3. The method has no discharge of three wastes (no waste gas, waste residue and waste water) and no pollution, and has the advantages of low cost, environmental friendliness and conformity with discharge standards compared with the conventional organic and inorganic waste treatment processes such as fermentation process, enzymolysis process, fixed-length cracking process, carboxylation degradation process and the like, thereby being suitable for large-scale industrialization.

Drawings

FIG. 1 shows the growth promoting effect of the small molecular water-soluble substance obtained in example 1 of the present invention on peanut seedlings.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to be limiting.

Experimental reagent:

polyphosphoric acid (phosphoric acid content 105% -118%): CAS number 8017-16-1 is commercially available

Choline chloride: CAS number 67-48-1, commercially available;

hypoiodic acid: commercial purchase

The methods of measurement and the procedures used in the present application are, unless otherwise specified, indicated by the methods, procedures and reagents commonly used in the art.

Example 1: method 1 for preparing water-soluble micromolecular active substance containing amino-oligosaccharin from shrimp and crab shells

Taking 10kg of polyphosphoric acid with the phosphoric acid content of 105% and 1kg of choline chloride, fully and uniformly mixing, adding the mixture into a 50L reaction kettle, and stirring and heating the mixture for 1 hour at the temperature of 85 ℃ to form a polyphosphoric acid-choline chloride eutectic solvent composite system;

drying shrimp and crab shells until the water content is lower than 10%, crushing, adding 5kg of crushed shrimp and crab shells into the reaction system, and stirring and dissolving for 2 hours at the temperature of 90 ℃ to form a phosphatidization dissolved solution;

adding 10kg of catalyst aqueous solution 1.0% hypoiodic acid aqueous solution under the condition of heat preservation for catalytic degradation for 2 hours, and filtering by a 50-mesh gauze to remove insoluble substances to form a stable water-soluble active substance solution; wherein the weight of insoluble matter obtained by filtration was 0.39 kg.

And (3) product detection:

1. the amino-oligosaccharin content was measured by the procedure described in 4.3 of NY/T2889.1-2016, and the results showed that: the mass fraction of the obtained amino-oligosaccharin is 6.7%;

2. determining the mass fraction of the nitrogen-containing organic water-soluble substance to be 3.5% by adopting a Kjeldahl method;

and 3, determining the mass fraction of the water-soluble calcium to be 9.0 percent by an EDTA method.

Example 2: method for preparing water-soluble micromolecular active substance containing amino-oligosaccharin from shrimp and crab shells 2

Taking 400kg of polyphosphoric acid with the phosphoric acid content of 105% and 10kg of choline chloride, fully and uniformly mixing, adding the mixture into a 2-ton reaction kettle, and stirring and heating the mixture for 0.5 hour at the temperature of 85 ℃ to form a polyphosphoric acid-choline chloride composite system;

drying and crushing shrimp and crab shells, adding 200kg of the crushed shrimp and crab shells into the reaction system, heating to 85 ℃, stirring and dissolving for 4 hours to form a phosphatide dissolved solution;

150kg of 0.5 percent hypoiodic acid aqueous solution of catalyst aqueous solution is added under the condition of heat preservation for catalytic degradation for 3 hours, and after insoluble substances are removed by filtering through a 50-mesh gauze, a stable water-soluble active substance solution is formed. Wherein the weight of insoluble matter obtained by filtration was 9.2 kg.

Measurement of the product by the measurement method described in example 1

The content of amino-oligosaccharin in the product is 6.1 percent; the content of the nitrogen-containing organic water-soluble substance is 4.6 percent, and the content of the water-soluble calcium is 9.8 percent; the fatty acid content was 0.26%.

Example 3: method for preparing water-soluble small-molecule active substance from enteromorpha

Taking 9kg of polyphosphoric acid with phosphoric acid content of 115% and 2kg of choline chloride, fully and uniformly mixing, adding the mixture into a 50L reaction kettle, and stirring and heating the mixture for 1 hour at 85 ℃ to form a polyphosphoric acid-choline chloride composite system;

drying and crushing enteromorpha, adding 4kg of enteromorpha into the reaction system, heating to 85 ℃, stirring and dissolving for 2 hours to form a phosphatide dissolved solution;

adding 5kg of 0.2% hypoiodic acid aqueous solution of catalyst aqueous solution under the condition of heat preservation for catalytic degradation for 2 hours, and filtering by a 50-mesh gauze to form a stable water-soluble active substance solution, wherein the weight of insoluble substances obtained by filtering is 0.15 kg.

And (3) product determination:

the content of soluble total sugar is 9.6% by the anthrone method;

the content of the nitrogen-containing organic water-soluble substance is 1.9 percent by a Kjeldahl method.

Example 4: method for preparing water-soluble micromolecular active substance by citric acid fermentation waste thalli

Taking 8kg of polyphosphoric acid with the phosphoric acid content of 118% and 2kg of choline chloride, fully and uniformly mixing, adding the mixture into a 50L reaction kettle, and stirring and heating the mixture for 1 hour at the temperature of 85 ℃ to form a polyphosphoric acid-choline chloride composite system;

drying and crushing the waste thalli, adding 5kg of the waste thalli into the reaction system, heating to 90 ℃, stirring and dissolving for 2 hours to form a phosphatidization dissolved solution;

adding 4kg of 0.2% hypoiodic acid aqueous solution of catalyst aqueous solution under the condition of heat preservation for catalytic degradation for 2 hours, and filtering through a 50-mesh gauze to remove insoluble substances to form a stable water-soluble active substance solution, wherein the weight of the insoluble substances obtained by filtering is 0.31 kg.

Product assay

The content of soluble total sugar is 6.8% by anthrone method, the content of nitrogen-containing organic water soluble substance is 4.4% by Kjeldahl method, and the content of fatty acid is 0.4% by gas chromatography.

Example 5: method for preparing water-soluble micromolecular active substance from bagasse

10kg of polyphosphoric acid with the phosphoric acid content of 110% and 1.5kg of choline chloride are taken, fully and uniformly mixed and then added into a 50L reaction kettle, the mixture is stirred and heated for 1 hour at the temperature of 85 ℃ to form a polyphosphoric acid-choline chloride composite system, bagasse is dried and crushed, 4.5kg of bagasse is taken and added into the reaction system, the temperature is raised to 95 ℃, the mixture is stirred and dissolved for 3 hours to form a polyphosphonized dissolved solution, 5kg of catalyst aqueous solution of 1% hypoiodic acid aqueous solution is added under the condition of heat preservation for catalytic degradation for 3 hours, a gauze is used for filtering and removing insoluble substances to form a stable water-soluble active substance solution, and the weight of the insoluble substances obtained by filtering is 0.40 kg.

The content of soluble total sugar is 8.1% by anthrone method, and the content of nitrogen-containing organic water soluble substance is 1.5% by Kjeldahl method. Application case 1: growth promoting effect of small molecule soluble substance containing amino-oligosaccharin resistance inducer on tomato seedling (foliage spray method)

The container is a flowerpot with the caliber of 10cm and the depth of 10cm, and each pot is filled with the equal amount of sterilized nutrient soil. Soaking the seeds in clear water at 55 ℃ for 5h, then putting the seeds in a culture dish for accelerating germination until the seeds are exposed to the white, planting the seeds in a flowerpot, and putting the flowerpot in an illumination incubator for culturing with the photoperiod of 12h each at day and night. The light intensity in daytime is 100 mu mol m-2 s-1. The temperature was 25 ℃ (day)/20 ℃ (night).

Clear water was set as a Control (CK), and 5g of the steady-state water-soluble active substance solution prepared in example 1 was diluted to four concentration gradients of 500, 1000, 2000, and 3000 times, respectively.

Spraying foliage to the seedling when the seedling grows to have three leaves and one heart, spraying for 5 days at intervals, setting each concentration for 3 times, repeating, recording the phenotype change of the seedling during the period, and measuring the biomass and biochemical indexes of the seedling on the 45 th day.

The result shows that compared with a control group, the obtained water-soluble micromolecule active substance product is diluted by 2000 times, the height of the treated seedlings, the stem thickness and the leaf number reach peak values, the peak values are respectively 3.50cm, 1.12mm and 5.33 higher than the control group, and the growth of tomato seedlings is remarkably promoted.

Application case 2: influence of small molecule soluble substance containing amino-oligosaccharin inducer prepared from shrimp and crab shell on POD enzyme activity and MDA content in tomato leaf

POD is an important active oxygen scavenger in plant cells and has close relation with plant disease resistance, and the increase of POD activity is an important expression for enhancing the plant disease resistance and is positively related to the plant disease resistance. Malondialdehyde content (MDA) affects mitochondrial respiratory chain complexes and mitochondrial intrinsic critical enzyme activity in vitro. MDA is used as the final product of membrane lipid peroxidation, and the content of MDA can be used as an important index for measuring the degree of membrane lipid peroxidation and is directly related to the damage degree of cell membranes. The decrease of MDA content indicates the increase of antioxidant capacity of cells, and is in negative correlation with POD activity. The invention selects two indexes of POD activity and MDA content as the detection standard of plant disease resistance, and verifies the effect of resistance inducer on improving plant disease resistance and bioactivity from the perspective of plant physiology and biochemistry.

Experimental materials: selecting the tomato seedlings in the application case I, and determining the contents of Peroxidase (POD) and Malondialdehyde (MDA) in the leaves of the tomato seedlings by a spectrophotometry.

The results show that:

the POD enzyme activity reaches 6547.21 Ug/fresh weight under the condition of dilution by 2000 times, and is 5685.44 Ug/fresh weight higher than that of a control group; the Malondialdehyde (MDA) content in the leaves is just the opposite trend, with 2000 times the liquid MDA content being the lowest, 43.12nmol/g fresh weight, 19.87nmol/g lower than the control.

The steady-state water-soluble active substance solution rich in the amino-oligosaccharin inducer can induce tomato seedlings to release more POD enzyme, and the MDA content is effectively reduced.

Application case 3: method for preparing micromolecular soluble substance containing amino-oligosaccharin resistance inducer by using shrimp and crab shells to promote growth of peanut seedlings

Reagent to be tested: the product obtained in example 1 was diluted to four concentration gradients of 500, 1000, 2000 and 3000 times by 3.5g, respectively, and the diluted product was used as a Control (CK) with clear water.

The peanut seeds with consistent sizes and full seeds are selected to accelerate germination for 2d under the humid condition of 30 ℃, then the peanut seeds with better germination potential are selected to be sowed in flowerpots, 3 seeds are sowed in each pot, each pot is respectively watered with 500, 1000, 2000 and 3000 times of the stable-state water-soluble active substance solution prepared in the embodiment 1 in the seedling emergence period, the stable-state water-soluble active substance solution is placed in a light incubator to be cultured for 25 days, and the growth condition of the peanut seedlings is observed, as shown in figure 1, the result shows that compared with the clear water contrast, the small molecular soluble substance product (amino-oligosaccharin resistance inducer) 1000-fold and 2000-fold solution prepared from shrimp and crab shells can obviously promote the growth of the peanut seedlings.

The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.

Claims

1. A method for converting shrimp and crab shells into water-soluble products in one step is characterized by comprising the following steps:

adding a catalyst aqueous solution with the mass volume ratio concentration of 0.1-1% for catalytic degradation for 1-3 hours under the condition of heat preservation; after the catalytic degradation reaction is finished, cooling to room temperature, filtering to remove insoluble substances, and then centrifuging or filtering to obtain a filtrate to obtain a water-soluble product;

wherein the catalyst is hypoiodic or hydroiodic acid, or a mixture thereof;

the weight ratio of the shrimp and crab shells to the polyphosphoric acid-choline chloride eutectic solvent composite system to the catalyst water solution is 0.2-1: 2: 0.7-3.

2. The method as claimed in claim 1, wherein the polyphosphoric acid-choline chloride eutectic solvent composite system

The choline chloride-polyphosphoric acid composite material is formed by mixing choline chloride and polyphosphoric acid according to the mass ratio of 1: 5-10 and stirring for 0.5-1 hour at the temperature of 85 ℃.

3. The method as claimed in claim 1, wherein the shrimp and crab shells are dried to the water content of less than 10%, and the mass fraction of phosphoric acid in the polyphosphoric acid is 105-118%.

4. A water-soluble product prepared by the process of any one of claims 1 to 3, wherein:

wherein the content of nitrogen-containing organic water-soluble substances is more than 10 percent, the total content of glucosamine is more than 5 percent, and the content of nitrogen organic matters is more than 2 percent;

the water-soluble phosphatidated fatty acid content is greater than 0.2%;

the content of water-soluble calcium is more than 8%.

5. Use of the method of any one of claims 1 to 3 for the treatment of organic inorganic material, wherein shrimp and crab shell material is replaced with organic inorganic material;

the planting waste is: corn stover, corn cobs, wheat straw, rice straw, fruit peels, fruit shells, or a mixture of any two or more thereof;

6. Use of the water-soluble product of claim 4 for the preparation of amino-oligosaccharins or for the preparation of plant hormone substitutes for the regulation of plant growth.