Insect body small molecule protein slurry, amino acid fertilizer and production process of insect body small molecule protein slurry and amino acid fertilizer

Abstract

The invention belongs to the field of fertilizer preparation, and particularly relates to polypide micromolecular protein slurry, an amino acid fertilizer and a production process thereof. The crude protein content of the polypide micromolecular protein slurry is greater than or equal to 16.24%, the total amino acid content is greater than or equal to 16.29%, the micromolecular protein slurry is applied to an amino acid fertilizer, and the amino acid fertilizer comprises the following raw materials in parts by mass: 80-100 parts of a fertilizer base material, 1-20 parts of the polypide micromolecular protein slurry, 1-25 parts of a stabilizer and 1-5 parts of a synergist. The amino acid fertilizer is simple in production process, has relatively good stability and can remarkably improve the quality of crops.

Description

Technical Field

[0001] The invention belongs to the field of fertilizer preparation, and in particular relates to an insect body small molecule protein slurry and amino acid fertilizer and a production process thereof. Background Art

[0002] Fertilizer is the cornerstone of agricultural production and is essential for the growth and development of crops. Compound fertilizers are widely used because they contain a variety of nutrients required for plant growth. There are many types of fertilizers on the market, and polypeptide compound fertilizers and ordinary compound fertilizers are two main types. Ordinary compound fertilizers only contain basic nutrients and do not contain polypeptide substances. In addition to basic elements such as nitrogen, phosphorus, and potassium, polypeptide compound fertilizers also contain polypeptide substances. These polypeptide substances are usually extracted from animals and plants and have multiple biological activities such as stimulating plant growth and improving stress resistance. Because polypeptide compound fertilizers contain a certain amount of bioactive polypeptides, they can stimulate plant growth and development, improve stress resistance, promote plant absorption and utilization of nutrients, and improve the quality of agricultural products.

[0003] Patent CN 115094107 A discloses a method for preparing a soybean polypeptide composition and a polypeptide amino acid fertilizer. The preparation steps of the soybean polypeptide composition are: using a composite protease composed of alkaline protease and neutral protease to enzymolyze sterilized soybean meal to obtain a soybean meal hydrolysate; then fermenting the soybean meal hydrolysate for 46-52 hours with Bacillus subtilis to obtain the soybean polypeptide composition. In this application, the protein hydrolysis rate in the soybean polypeptide composition reaches 48.5%. The prepared soybean polypeptide composition is highly stable and is not easy to deteriorate when directly stored. It can be directly used to prepare polypeptide amino acid fertilizers. However, the amount of composite protease used in the enzymolysis in this patent is relatively high, which makes the production cost of the composite protease relatively high. In addition, when the polypeptides and amino acids obtained by microbial fermentation technology are used in fertilizers, problems such as low polypeptide and protein content, instability, slow hydrolysis, and easy occurrence of undecomposed living bodies often occur.

[0004] At present, there are more studies on plant proteins, amino acids and peptides, while there are fewer studies on animal proteins, amino acids and peptides. The white-spotted flower beetle is an insect of the family Scarabaeidae in the order Coleoptera. It is widely distributed in China and neighboring countries and has two growth stages: larvae and adults. In the adult stage, it is an important agricultural pest. However, its larvae do not harm crops and can convert agricultural waste into high-quality insect protein and fat. Given the high protein content of the larvae themselves, it is a meaningful idea to effectively utilize them and convert them into valuable resources.

[0005] Therefore, developing a method for fermenting and preparing small molecule protein slurry using white-spotted flower beetle as raw material and applying it in fertilizer is of great significance for improving resource utilization and crop quality. Summary of the invention

[0006] In view of the deficiencies in the prior art, the present invention provides an insect body small molecule protein slurry and amino acid fertilizer and a production process thereof. The white-spotted flower beetle insect is converted into a small molecule protein slurry by enzymatic hydrolysis and fermentation, and the small molecule protein slurry is made into an amino acid fertilizer. The amino acid fertilizer has a simple production process, good stability, and can significantly improve the quality of crops.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] The first aspect of the present invention provides an insect body small molecule protein slurry, wherein the crude protein content is ≥16.24%; the total content of aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine and arginine is ≥16.29%.

[0009] The second aspect of the present invention provides a production process of insect body small molecule protein slurry, comprising the following preparation steps:

[0010] (1) taking dried and crushed white star flower beetle bodies and banana peels, mixing them evenly, and sterilizing them to obtain composite white star flower beetle powder;

[0011] (2) adding 3-10 times by weight of pure water to the composite white star flower beetle powder of step (1), then adding alkaline protease, stirring evenly, adjusting the pH to 8-10, enzymolyzing at 25-55° C. for 2-8 hours, inactivating the enzyme, and obtaining a white star flower beetle enzymolysis solution;

[0012] (3) The pH of the enzymatic hydrolyzate of the white-spotted flower beetle in step (2) is adjusted to 5-7, and then the mixed bacterial seed solution is inoculated therein, and shake-cultured at 25-40° C. for 3-7 days to obtain a worm body small molecule protein slurry.

[0013] In some embodiments, the mass ratio of the white-spotted flower beetle body to the banana peel is (30-45):1.

[0014] Preferably, the mass ratio of the white-spotted flower beetle body to the banana peel is 40:1.

[0015] In some embodiments, the amount of alkaline protease used in step (2) is 0.5-1.5wt% of the composite white star flower beetle powder, and the enzyme activity of the alkaline protease used is ≥200000 U / g.

[0016] Preferably, the amount of alkaline protease used in step (2) is 1 wt % of the composite white star flower beetle powder.

[0017] In some embodiments, the amount of the mixed bacteria seed solution is 1-5wt% of the enzymatic hydrolyzate of the white star flower beetle; the mixed bacteria seed solution includes a mass ratio of (1-2): 1:1 domesticated Aspergillus oryzae seed solution and domesticated Bacillus plantarum seed solution; the concentrations of the domesticated Aspergillus oryzae seed solution and the domesticated Bacillus plantarum seed solution are both (1-10)×10 8 CFU / mL.

[0018] Preferably, the amount of the mixed bacteria seed liquid is 3wt% of the enzymatic hydrolyzate of the white star flower beetle; the mixed bacteria seed liquid comprises a mass ratio of 1.5:1 domesticated Aspergillus oryzae seed liquid and a domesticated Bacillus plantarum seed liquid; the concentrations of the domesticated Aspergillus oryzae seed liquid and the domesticated Bacillus plantarum seed liquid are both (3-7)×10 8 CFU / mL.

[0019] More preferably, the Aspergillus oryzae is Aspergillus oryzae CICC 40214 and the Bacillus plantarum is Bacillus plantarum CICC 10345.

[0020] In some embodiments, the acclimation step is:

[0021] S1: picking activated Aspergillus oryzae or Bacillus plantarum respectively and inoculating them into a sterilized liquid culture medium with a pH of 5-7 and containing 1-5wt% of composite white star flower beetle powder, and culturing them at 25-40° C. with shaking for 5-10 days to obtain a primary acclimation liquid of Aspergillus oryzae and a primary acclimation liquid of Bacillus plantarum;

[0022] S2: inoculating the primary acclimation liquid of Aspergillus oryzae or the primary acclimation liquid of Bacillus plantarum from step S1 into a sterilized liquid culture medium having a pH of 5-7 and containing 5-10 wt % of the composite white star flower beetle powder, and culturing the liquid culture medium at 25-40° C. with shaking for 5-10 days to obtain the secondary acclimation liquid of Aspergillus oryzae and the secondary acclimation liquid of Bacillus plantarum;

[0023] S3: inoculating the secondary acclimation liquid of Aspergillus oryzae or the secondary acclimation liquid of Bacillus plantarum from step S2 into a sterilized liquid culture medium having a pH of 5-7 and containing 10-20 wt % of the composite white star flower beetle powder, and culturing the liquid culture medium at 25-40° C. with shaking for 5-10 days to obtain the tertiary acclimation liquid of Aspergillus oryzae or the tertiary acclimation liquid of Bacillus plantarum;

[0024] S4: Spread the three-time acclimation liquid of Aspergillus oryzae or the three-time acclimation liquid of Bacillus plantarum in step S3 on a plate culture medium, and culture at 25-40° C. for 1-5 days to obtain acclimated Aspergillus oryzae and acclimated Bacillus plantarum.

[0025] The third aspect of the present invention provides an amino acid fertilizer, which comprises the following raw materials, measured by weight: 80-100 parts of fertilizer base, 1-20 parts of insect body small molecule protein slurry, 1-25 parts of stabilizer, and 1-5 parts of synergist;

[0026] The fertilizer base material is nitrogen, phosphorus, potassium macronutrient fertilizer and / or medium and trace element fertilizer;

[0027] The insect body small molecule protein slurry is the insect body small molecule protein slurry described in the above scheme.

[0028] In some embodiments, the stabilizer comprises cocamide betaine, polyether polyol, polyvinyl pyrrolidone and trehalose.

[0029] Preferably, the stabilizer comprises cocamide betaine, polyether polyol, polyvinyl pyrrolidone and trehalose in a mass ratio of (1-10):(1-10):1:(1-5).

[0030] In some embodiments, the raw materials of the synergist include sodium sulfite solution, dilute sulfuric acid solution, fulvic acid, potassium sulfate, crop straw, nitrification inhibitor, urease inhibitor, river mud, shell powder, medical stone, diammonium phosphate and vitamin B.

[0031] The preparation method of the synergist is referred to patent CN 109627094A.

[0032] A fourth aspect of the present invention provides a production process for an amino acid fertilizer, comprising the following steps:

[0033] Firstly, the stabilizer is evenly mixed with the insect body small molecule protein slurry to obtain a mixture, and then the mixture is evenly mixed with the fertilizer base material and the synergist to obtain the amino acid fertilizer.

[0034] The present invention utilizes an enzymatic method combined with a fermentation method to convert the white-star flower beetle into a small-molecule protein slurry. The applicant has found that strictly controlling the process parameters during the enzymatic hydrolysis and fermentation process is beneficial to increasing the content of amino acids and crude protein in the insect body small-molecule protein slurry. First, the present invention selects a certain amount of alkaline protease to enzymatically hydrolyze the broken white-star flower beetle body and banana peel, and regulates the enzymatic hydrolysis process by controlling the enzyme concentration, enzymatic hydrolysis time, enzymatic hydrolysis temperature, environmental pH and substrate concentration, so as to degrade the macromolecules that are difficult to degrade in the white-star flower beetle body and banana peel into small molecules, which is beneficial to the subsequent absorption of small molecules by Aspergillus oryzae and Bacillus plantarum, and converts them into amino acids and polypeptides, thereby improving the utilization rate of raw materials. Secondly, the invention domesticates Aspergillus oryzae and Bacillus plantarum first, and increases the composite white star flower beetle powder during the domestication period to improve the adaptability and selectivity of Aspergillus oryzae and Bacillus plantarum to the composite white star flower beetle powder. The banana peel in the composite white star flower beetle powder is rich in carbohydrates, cellulose, minerals, etc., and the mass ratio of the banana peel to the white star flower beetle powder is controlled, so that the banana peel can provide carbon sources and nutrient elements for Aspergillus oryzae and Bacillus plantarum and promote their biological utilization of the white star flower beetle powder, while avoiding competitive inhibition of microorganisms due to excessive nutrition, reducing the conversion rate of the white star flower beetle powder, and further reducing the content of crude protein and amino acids in the insect body small molecule protein slurry.

[0035] In addition, the general fermented polypeptide or protein liquid has poor stability and is not easy to store. The present invention uses cocamide betaine, polyether polyol, polyvinyl pyrrolidone and trehalose together in a certain mass ratio to develop a stabilizer, and uses it together with the insect body small molecule protein slurry. The effective substances in the stabilizer can form a stable complex or chelate structure with the polypeptides, amino acids, etc. in the insect body small molecule protein slurry, significantly improving the stability of the insect body small molecule protein slurry, thereby extending the storage time of the amino acid fertilizer.

[0036] Finally, the present invention compounds the fertilizer base material, the insect body small molecule protein slurry, the stabilizer and the synergist to obtain an amino acid fertilizer with high stability and capable of significantly improving the quality of agricultural products, which is beneficial to the development of agriculture.

[0037] Compared with the prior art, the present invention has the following beneficial effects:

[0038] 1. The present invention uses white star flower beetle body and banana peel as raw materials, and ferments them by using enzymatic hydrolysis combined with microbial fermentation to obtain a small molecule protein slurry with high protein and amino acid content. The production process of the small molecule protein slurry is mild, environmentally friendly and pollution-free, and has a high cost performance.

[0039] 2. The present invention domesticates the microorganisms Aspergillus oryzae and Bacillus plantarum, improves the adaptability and selectivity of Aspergillus oryzae and Bacillus plantarum to the composite white star flower beetle powder, has a good bioavailability of the white star flower beetle powder, and further increases the content of crude protein and amino acids in the insect body small molecule protein slurry.

[0040] 3. The present invention applies the insect body small molecule protein slurry obtained by fermentation to fertilizer to obtain an amino acid fertilizer, which has good stability and has a good improvement effect on the quality of crops. DETAILED DESCRIPTION

[0041] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0042] In order to better illustrate the specific implementation mode of the present invention, further examples are given below.

[0043] The white-spotted flower beetle insect body used in the present invention is the white-spotted flower beetle larvae; the tryptone, PDA slant culture medium and LB plate culture medium used are all purchased from Guangdong Huankai Microbiological Technology Co., Ltd.; the enzyme activity of the trypsin used is 250000 U / g, and the enzyme activity of the alkaline protease used is 200000 U / g, both of which are purchased from Shanghai Yuanye Biotechnology Co., Ltd.; the Aspergillus oryzae used is Aspergillus oryzae CICC 40214, and the plant lactobacillus plantarum is plant lactobacillus plantarum CICC 10345; the cocamide betaine used is cocamide propyl betaine, which is purchased from Suzhou Minghua Sugar Alcohol Co., Ltd.; the polyether polyol used is the general polyether H4110 purchased from Hongbaoli Group Co., Ltd.; and the polyvinyl pyrrolidone used is polyvinyl pyrrolidone K30, which is purchased from Shanghai Maokang Biotechnology Co., Ltd.

[0044] Preparation Example 1

[0045] Domestication of Aspergillus oryzae:

[0046] In a sterile environment, the preserved Aspergillus oryzae mother strain was inoculated into a 0.5 cm×0.5 cm bacterial block on a PDA slant medium, and cultured at 37°C until the hyphae were full, thereby obtaining activated Aspergillus oryzae;

[0047] S1: Pick the activated Aspergillus oryzae and inoculate it into a 500mL inorganic salt liquid culture medium which is sterilized, has a pH of 6, contains 3wt% of composite white star flower beetle powder crushed to 40 mesh, and culture it at 37°C and 150r / min shaking for 7 days to obtain a primary acclimation liquid of Aspergillus oryzae;

[0048] S2: inoculate 1 mL of the primary acclimation liquid of Aspergillus oryzae obtained in step S1 into 500 mL of a sterilized inorganic salt liquid culture medium having a pH of 6 and containing 8 wt % of composite white star flower beetle powder, and culture the medium at 37° C. with shaking for 7 days to obtain a secondary acclimation liquid of Aspergillus oryzae;

[0049] S3: inoculate 1 mL of the secondary acclimation liquid of Aspergillus oryzae obtained in step S2 into 500 mL of a sterilized inorganic salt liquid culture medium having a pH of 6 and containing 15 wt % of composite white star flower beetle powder, and culture the medium at 37° C. with shaking for 7 days to obtain a tertiary acclimation liquid of Aspergillus oryzae;

[0050] S4: Take 100 μL of the three-time acclimation solution of Aspergillus oryzae in step S3 and spread it on a PDA plate culture medium, and culture it at 37° C. for 3 days to obtain acclimated Aspergillus oryzae.

[0051] Preparation Example 2

[0052] Acclimation of Lactobacillus plantarum:

[0053] Under a sterile environment, 100 μL of bacterial solution was drawn from a glycerol tube storing the plant lactobacillus strain, and evenly spread on an LB plate medium by a plate streak method, and inverted and cultured at 37° C. until colonies grew out, thereby obtaining activated plant lactobacillus;

[0054] S1: picking the activated plant lactobacillus and inoculating it into a sterilized inorganic salt liquid culture medium with a pH of 6.5 and containing 3 wt % of composite white star flower beetle powder crushed to 40 mesh, and culturing it at 37° C. with shaking for 7 days to obtain a primary acclimation solution of plant lactobacillus;

[0055] S2: inoculate 1 mL of the plant lactobacillus primary acclimation solution of step S1 into a sterilized inorganic salt liquid culture medium having a pH of 6.5 and containing 8 wt % of composite white star flower beetle powder, and culture at 37° C. with shaking for 7 days to obtain a plant lactobacillus secondary acclimation solution;

[0056] S3: 1 mL of the secondary acclimation solution of Bacillus plantarum obtained in step S2 is inoculated into a sterilized inorganic salt liquid culture medium having a pH of 6.5 and containing 15 wt % of composite white star flower beetle powder, and cultured at 37° C. with shaking for 7 days to obtain a tertiary acclimation solution of Bacillus plantarum;

[0057] S4: Take 100 μL of the three-time acclimation solution of plant lactobacillus in step S3, spread it on the LB plate culture medium, and culture it at 37° C. for 1 day to obtain the acclimated plant lactobacillus.

[0058] Preparation Example 3

[0059] Preparation of acclimated Aspergillus oryzae seed solution:

[0060] In a sterile environment, the domesticated Aspergillus oryzae in Preparation Example 1 was inoculated into 500 mL of potato culture medium, shaken and cultured at 37°C for 3 days, and then centrifuged at 5000 r / min to obtain the bacterial cells, which were diluted with sterile water to 6 × 10 8 CFU / mL, and the domesticated Aspergillus oryzae seed liquid was obtained.

[0061] Preparation Example 4

[0062] Preparation of the acclimated Lactobacillus plantarum seed solution:

[0063] In a sterile environment, the plant lactobacillus acclimated in Preparation Example 2 was inoculated into 500 mL of liquid culture medium, shaken and cultured at 37 ° C for 1 day, and then centrifuged at 5000 r / min to obtain the bacteria, which were diluted with sterile water to 6 × 10 8 CFU / mL, and the acclimated Lactobacillus plantarum seed solution was obtained.

[0064] Preparation Example 5

[0065] Preparation of Aspergillus oryzae seed solution:

[0066] In a sterile environment, the activated Aspergillus oryzae in Preparation Example 1 was inoculated into 500 mL of potato culture medium, and after shaking culture at 37°C for 3 days, the culture was centrifuged at 5000 r / min to obtain the bacterial cells, which were diluted with sterile water to 6×10 8 CFU / mL, and Aspergillus oryzae seed liquid was obtained.

[0067] Preparation Example 6

[0068] Preparation of Lactobacillus plantarum seed solution:

[0069] In a sterile environment, the activated plant lactobacillus in Preparation Example 2 was inoculated into 500 mL of liquid culture medium, shaken and cultured at 37°C for 1 day, and then centrifuged at 5000 r / min to obtain bacterial cells, which were diluted with sterile water to 6×10 8 CFU / mL, and the Lactobacillus plantarum seed solution was obtained.

[0070] The preparation steps of the inorganic salt liquid culture medium used in Preparation Examples 1 and 2 are as follows: dissolve 2 g of (NH4)2SO4, 2 g of Na2HPO4·12H2O, 0.5 g of MgSO4·7H2O, 0.5 g of KH2PO4, 0.5 mL of trace element solution and 0.5 mL of vitamin solution in 500 mL of deionized water, adjust the pH to 7, sterilize at 120°C for 20 min, and cool naturally to room temperature to obtain an inorganic salt liquid culture medium;

[0071] Trace element solution formula: CaCl2 25 mg, MnCl4·4H2O 5 mg, deionized water 1 L;

[0072] Vitamin liquid formula: riboflavin 200 mg, para-aminobenzoic acid 200 mg, thiamine hydrochloride 400 mg, niacin 400 mg, pyridoxine hydrochloride 400 mg, calcium pantothenate 400 mg, folic acid 2 mg, biotin 2 mg, inositol 2000 mg, deionized water 1L.

[0073] The steps for preparing the potato culture medium used in Preparation Examples 3 and 5 are as follows: peel the potatoes, cut them into pieces of about 1 cm 3 100 g of potato pieces were put into 500 mL of deionized water and boiled for 30 min, filtered with double-layer gauze, 10 g of glucose was added to the filtrate, and deionized water was added to 500 mL, the pH was adjusted to 7 with 1 mol / L sodium hydroxide solution, sterilized at 120 ° C for 20 min, and naturally cooled to room temperature to obtain potato culture medium;

[0074] The preparation steps of the liquid culture medium used in Preparation Examples 4 and 6 are as follows: 5 g of yeast powder, 10 g of tryptone and 10 g of NaCl are dissolved in 1 L of deionized water, sterilized at 121° C. for 20 min, and naturally cooled to room temperature to obtain a liquid culture medium.

[0075] Example 1

[0076] A production process of insect body small molecule protein slurry, comprising the following preparation steps:

[0077] (1) 400 g of dried and crushed white star flower beetle bodies and 10 g of banana peels were mixed evenly, and sterilized at 120° C. for 20 min to obtain a composite white star flower beetle powder;

[0078] (2) adding 2050 g of pure water to the composite white star flower beetle powder of step (1), and then adding 4.1 g of alkaline protease, stirring evenly, adjusting the pH to 9, performing enzymolysis at 45° C. for 5 h, and deactivating the enzyme at 100° C. for 15 min to obtain a white star flower beetle enzymolysis solution;

[0079] (3) The pH of the enzymatic hydrolyzate of the white-spotted flower beetle in step (2) is adjusted to 6, and then 3 wt % of the mixed bacterial seed solution is inoculated therein, and shake-cultured at 37° C. for 3 days to obtain a small molecule protein slurry of the insect body.

[0080] The mixed bacterial seed solution used was composed of the Aspergillus oryzae seed solution domesticated in Preparation Example 3 and the Bacillus plantarum seed solution domesticated in Preparation Example 4 in a mass ratio of 1.5:1.

[0081] Example 2

[0082] A production process of insect body small molecule protein slurry, comprising the following preparation steps:

[0083] (1) 350 g of dried and crushed white star flower beetle bodies and 10 g of banana peels were mixed evenly, and sterilized at 120° C. for 20 min to obtain a composite white star flower beetle powder;

[0084] (2) adding 1080 g of pure water to the composite white star flower beetle powder of step (1), and then adding 1.8 g of alkaline protease, stirring evenly, adjusting the pH to 8, performing enzymolysis at 25° C. for 8 h, and deactivating the enzyme at 100° C. for 15 min to obtain a white star flower beetle enzymolysis solution;

[0085] (3) The pH of the enzymatic hydrolyzate of the white-spotted flower beetle in step (2) is adjusted to 5, and then 1 wt % of the mixed bacterial seed solution is inoculated therein, and shake-cultured at 25° C. for 7 days to obtain a worm body small molecule protein slurry.

[0086] The mixed bacterial seed solution used is composed of the Aspergillus oryzae seed solution domesticated in Preparation Example 3 and the Bacillus plantarum seed solution domesticated in Preparation Example 4 in a mass ratio of 1:1.

[0087] Example 3

[0088] A production process of insect body small molecule protein slurry, comprising the following preparation steps:

[0089] (1) 450 g of dried and 60-mesh crushed white star flower beetle bodies and 10 g of banana peels were mixed evenly, and sterilized at 120° C. for 20 min to obtain a composite white star flower beetle powder;

[0090] (2) adding 4600 g of pure water to the composite white star flower beetle powder of step (1), and then adding 7.63 g of alkaline protease, stirring evenly, adjusting the pH to 10, performing enzymolysis at 55° C. for 2 h, and deactivating the enzyme at 100° C. for 15 min to obtain a white star flower beetle enzymolysis solution;

[0091] (3) The pH of the enzymatic hydrolyzate of the white-spotted flower beetle in step (2) is adjusted to 7, and then 5 wt % of the mixed bacterial seed solution is inoculated therein, and shake-cultured at 40° C. for 3 days to obtain a worm body small molecule protein slurry.

[0092] The mixed bacterial seed solution used was composed of the Aspergillus oryzae seed solution domesticated in Preparation Example 3 and the Bacillus plantarum seed solution domesticated in Preparation Example 4 in a mass ratio of 2:1.

[0093] Example 4

[0094] A production process of insect body small molecule protein slurry, the specific implementation method is the same as that of Example 1, except that the amount of alkaline protease used is 8.2g.

[0095] Example 5

[0096] A production process for insect body small molecule protein slurry, the specific implementation method is the same as that of Example 1, except that the mixed bacteria seed liquid used is composed of the Aspergillus oryzae seed liquid of Preparation Example 5 and the Bacillus plantarum seed liquid of Preparation Example 6 in a mass ratio of 1.5:1.

[0097] Example 6

[0098] A production process of insect body small molecule protein slurry, the specific implementation method is the same as that of Example 1, the difference is that the mixed fungus seed liquid used is 7wt%.

[0099] Comparative Example 1

[0100] A production process of insect body small molecule protein slurry, comprising the following preparation steps:

[0101] (1) 400 g of dried and crushed white star flower beetle bodies to 40 mesh are sterilized at 120° C. for 20 min to obtain white star flower beetle powder;

[0102] (2) adding 2000 g of pure water to the white star flower beetle powder of step (1), and then adding 4 g of alkaline protease, stirring evenly, adjusting the pH to 9, performing enzymolysis at 45° C. for 5 h, and inactivating the enzyme at 100° C. for 15 min to obtain a white star flower beetle enzymolysis solution;

[0103] (3) The pH of the enzymatic hydrolyzate of the white-spotted flower beetle in step (2) is adjusted to 6, and then 3 wt % of the mixed bacterial seed solution is inoculated therein, and shake-cultured at 37° C. for 3 days to obtain a small molecule protein slurry of the insect body.

[0104] The mixed bacterial seed solution used was composed of the Aspergillus oryzae seed solution domesticated in Preparation Example 3 and the Bacillus plantarum seed solution domesticated in Preparation Example 4 in a mass ratio of 1.5:1.

[0105] Comparative Example 2

[0106] A production process of insect body small molecule protein slurry, the specific implementation method is the same as that of Example 1, the difference is that trypsin with equal enzyme activity is used instead of alkaline protease.

[0107] Example 7

[0108] A production process of amino acid fertilizer comprises the following steps:

[0109] First, 10 g of stabilizer and 10 g of insect body small molecule protein slurry are uniformly mixed to obtain a mixture, and then the mixture is uniformly mixed with 800 g of fertilizer base material and 10 g of synergist to obtain the amino acid fertilizer.

[0110] The insect body small molecule protein slurry used is obtained from Example 1;

[0111] The fertilizer base used was 18-10-20 compound fertilizer;

[0112] The stabilizer used consists of cocamide betaine, polyether polyol, polyvinyl pyrrolidone and trehalose in a mass ratio of 1:1:1:1.

[0113] Example 8

[0114] A production process of amino acid fertilizer comprises the following steps:

[0115] First, 20 g of stabilizer and 25 g of insect body small molecule protein slurry are uniformly mixed to obtain a mixture, and then the mixture is uniformly mixed with 1000 g of fertilizer base material and 50 g of synergist to obtain the amino acid fertilizer.

[0116] The insect body small molecule protein slurry used is obtained from Example 1;

[0117] The fertilizer base used was 18-10-20 compound fertilizer;

[0118] The stabilizer used is composed of cocamide betaine, polyether polyol, polyvinyl pyrrolidone and trehalose in a mass ratio of 10:10:1:5.

[0119] Example 9

[0120] A production process of amino acid fertilizer comprises the following steps:

[0121] First, 15 g of stabilizer and 20 g of insect body small molecule protein slurry are uniformly mixed to obtain a mixture, and then the mixture is uniformly mixed with 900 g of fertilizer base material and 30 g of synergist to obtain the amino acid fertilizer.

[0122] The insect body small molecule protein slurry used is obtained from Example 1;

[0123] The fertilizer base used was 18-10-20 compound fertilizer;

[0124] The stabilizer used is composed of cocamide betaine, polyether polyol, polyvinyl pyrrolidone and trehalose in a mass ratio of 5:5:1:3;

[0125] The preparation method of the synergist used in Examples 7-9 refers to the preparation of Example 1 in patent CN 109627094 A; in terms of weight parts, the raw materials of the synergist used include: 100 parts of 2wt% sodium sulfite solution, 30 parts of 1wt% dilute sulfuric acid solution, 5 parts of fulvic acid, 8 parts of potassium sulfate, 30 parts of wheat crop straw, 3 parts of nitrogen pyridine, 2 parts of sulfur phosphoramide, 10 parts of shell powder, 14 parts of medical stone, 6 parts of ammonium dihydrogen phosphate, and 5 parts of vitamin B.

[0126] Testing of insect body small molecule protein slurry

[0127] 1. The insect body small molecule protein slurry prepared in Examples 1-6 and Comparative Examples 1 and 2 was sent to Guangdong Microbiological Analysis and Testing Center for crude protein content testing. The crude protein content test method was based on GB / T6432-2018 (7.2). The specific test results of the crude protein content test are shown in Table 1.

[0128] Table 1

[0129]

[0130] As shown in Table 1, the crude protein content in the insect body small molecule protein slurry prepared in Examples 1-3 is relatively high. Compared with Example 1, the amount of alkaline protease used in Example 4 is changed, and the amount of mixed bacteria seed liquid added in Example 6 is also changed, which will cause enzymes and microorganisms to competitively inhibit the utilization of the composite white-star flower beetle powder, resulting in a decrease in crude protein content; Aspergillus oryzae and Bacillus plantarum in the mixed bacteria seed liquid used in Example 5 are not domesticated, so that their ability to utilize the composite white-star flower beetle powder is reduced, thereby reducing the crude protein content; the raw materials of Comparative Example 1 are added with banana peels, resulting in a lack of nutrients, limiting the activity of microorganisms, and the crude protein content is significantly reduced; in Comparative Example 2, trypsin is used to enzymolyze the composite white-star flower beetle powder, and its enzymolysis ability for the composite white-star flower beetle powder is weaker than that of alkaline protease, so that the crude protein content is reduced.

[0131] Based on the crude protein content data, the insect small molecule protein slurry prepared in Example 1 was sent to the Guangdong Microbiological Analysis and Testing Center for amino acid content testing. The amino acid content test method was based on GB / T18246-2019. The specific test results of the amino acid content test are shown in Table 2.

[0132] Table 2

[0133]

[0134]

[0135] As can be seen from Table 2, the content of each amino acid in the insect body small molecule protein slurry prepared in Example 1 is the highest, so the insect body small molecule protein slurry obtained in this example is applied to amino acid fertilizer.

[0136] 2. Stability of insect body small molecule protein slurry

[0137] Equal volumes of the insect body small molecule protein slurry prepared in Example 1 and the mixture obtained in the preparation process of the amino acid fertilizer in Examples 7-9 were placed in sealed containers at 25° C. for static storage, and the generation of odor and the presence of barrel swelling were observed every day.

[0138] After observation, the mixtures prepared in Examples 7-9 had no peculiar smell, no obvious odor after storage for more than 30 days, and no barrel swelling; while the insect body small molecule protein slurry in Example 1 produced a foul odor and severe barrel swelling on the 4th day, and as the storage time increased, the odor deepened and there was severe barrel swelling. This shows that the prepared insect body small molecule protein slurry has poor stability and a short storage time, but the mixture obtained by mixing the insect body small molecule protein slurry with the stabilizer developed by the present invention has good stability and storage resistance, and can be well applied to amino acid fertilizers to reduce the production cost of fertilizers.

[0139] 3. Crop Experiment

[0140] (1) Experimental group: The amino acid fertilizer obtained in Example 9 was used for Hami melon planting experiment:

[0141] The amount of fertilizer applied during the planting period: 1500 kg / mu of decomposed organic fertilizer, 100 kg / mu of amino acid fertilizer obtained in Example 9, and 80 kg / mu of calcium magnesium phosphate fertilizer;

[0142] Variety: Fort Red Hami melon; 3 vines and 1 melon were left on each plant, and three parallel groups were set up. 30 plants were selected from each treatment, and the average value was calculated as the single melon weight. 1,500 plants were planted per mu.

[0143] (2) Control group:

[0144] Fertilizer application during planting: 1500 kg / mu of decomposed organic fertilizer, 100 kg / mu of 18-10-20 compound fertilizer, 80 kg / mu of calcium magnesium phosphate fertilizer;

[0145] Other conditions and operations were the same as those of the experimental group.

[0146] The sugar content of Hami melon was tested using a digital saccharimeter. The specific test index results are shown in Table 3.

[0147] Table 3

[0148] serial number Single melon weight (kg) Sugar content (°Bx) Experimental Group 2.48 12.5 Control group 2.16 10.8

[0149] As shown in Table 3, the amino acid fertilizer prepared in the present invention can well improve the fruit quality of Paotaihong Hami melon, and the single melon weight and sugar content are significantly improved.

[0150] The above is only a preferred embodiment of the present invention and does not constitute any form of limitation to the present application. Although the present application is disclosed as above in the preferred embodiment, it is not intended to limit the present application. Any technician familiar with the profession, without departing from the scope of the technical solution of the present application, using the technical content disclosed above to make slight changes or modifications are equivalent to equivalent implementation cases. However, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution.

Claims

1. A small molecule protein slurry of insect body, characterized in that: Its crude protein content is ≥16.24%; the total content of aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine and arginine is ≥16.29%.

2. A production process of the insect body small molecule protein slurry according to claim 1, characterized in that: The method comprises the following preparation steps: (1) taking dried and crushed white star flower beetle bodies and banana peels, mixing them evenly, and sterilizing them to obtain composite white star flower beetle powder; (2) adding 3-10 times by weight of pure water to the composite white star flower beetle powder of step (1), then adding alkaline protease, stirring evenly, adjusting the pH to 8-10, enzymolyzing at 25-55° C. for 2-8 hours, inactivating the enzyme, and obtaining a white star flower beetle enzymolysis solution; (3) The pH of the enzymatic hydrolyzate of the white-spotted flower beetle in step (2) is adjusted to 5-7, and then the mixed bacterial seed solution is inoculated therein, and shake-cultured at 25-40° C. for 3-7 days to obtain a worm body small molecule protein slurry.

3. The production process of the insect body small molecule protein slurry according to claim 2, characterized in that: The mass ratio of the white-spotted flower beetle body to the banana peel is (30-45):

1.

4. The production process of insect body small molecule protein slurry according to claim 2, characterized in that: The amount of alkaline protease used in the step (2) is 0.5-1.5wt% of the composite white star flower beetle powder, and the enzyme activity of the alkaline protease used is ≥200000U / g.

5. The production process of insect body small molecule protein slurry according to claim 2, characterized in that: The dosage of the mixed bacteria seed liquid is 1-5wt% of the enzymatic hydrolyzate of the white star flower beetle; the mixed bacteria seed liquid includes the tamed Aspergillus oryzae seed liquid and the tamed plant lactobacillus seed liquid in a mass ratio of (1-2):1; the concentrations of the tamed Aspergillus oryzae seed liquid and the tamed plant lactobacillus seed liquid are both (1-10)×10 8 CFU / mL.

6. The production process of insect body small molecule protein slurry according to claim 5, characterized in that: The steps of domestication are: S1: picking activated Aspergillus oryzae or Bacillus plantarum respectively and inoculating them into a sterilized liquid culture medium with a pH of 5-7 and containing 1-5wt% of composite white star flower beetle powder, and culturing them at 25-40° C. with shaking for 5-10 days to obtain a primary acclimation liquid of Aspergillus oryzae or a primary acclimation liquid of Bacillus plantarum; S2: inoculating the primary acclimation liquid of Aspergillus oryzae or the primary acclimation liquid of Bacillus plantarum from step S1 into a sterilized liquid culture medium having a pH of 5-7 and containing 5-10 wt % of the composite white star flower beetle powder, and culturing the liquid culture medium at 25-40° C. with shaking for 5-10 days to obtain the secondary acclimation liquid of Aspergillus oryzae or the secondary acclimation liquid of Bacillus plantarum; S3: The Aspergillus oryzae secondary acclimation liquid and the plant lactobacillus plantarum secondary acclimation liquid of step S2 are inoculated into a sterilized liquid culture medium having a pH of 5-7 and containing 10-20wt% of the composite white star flower beetle powder, and cultured at 25-40° C. with shaking for 5-10 days to obtain the Aspergillus oryzae tertiary acclimation liquid or the plant lactobacillus plantarum tertiary acclimation liquid; S4: Spread the three-time acclimation liquid of Aspergillus oryzae or the three-time acclimation liquid of Bacillus plantarum in step S3 on a plate culture medium, and culture at 25-40° C. for 1-5 days to obtain acclimated Aspergillus oryzae and acclimated Bacillus plantarum.

7. An amino acid fertilizer, characterized in that The raw materials include the following according to the weight ratio: 80-100 parts of fertilizer base material, 1-20 parts of insect body small molecule protein slurry, 1-25 parts of stabilizer, and 1-5 parts of synergist; The fertilizer base material is nitrogen, phosphorus, potassium macronutrient fertilizer and / or medium and trace element fertilizer; The insect body small molecule protein slurry is the insect body small molecule protein slurry described in claim 1 or the insect body small molecule protein slurry prepared by the production process described in any one of claims 2-6.

8. The amino acid fertilizer according to claim 7, characterized in that The stabilizer comprises cocamide betaine, polyether polyol, polyvinyl pyrrolidone and trehalose.

9. The amino acid fertilizer according to claim 7, characterized in that The raw materials of the synergist include sodium sulfite solution, dilute sulfuric acid solution, fulvic acid, potassium sulfate, crop straw, nitrification inhibitor, urease inhibitor, river mud, shell powder, medical stone, diammonium phosphate and vitamin B.

10. A process for producing the amino acid fertilizer according to any one of claims 7 to 9, characterized in that: The following steps are involved: Firstly, the stabilizer is evenly mixed with the insect body small molecule protein slurry to obtain a mixture, and then the mixture is evenly mixed with the fertilizer base material and the synergist to obtain the amino acid fertilizer.

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