CN114901615A

CN114901615A - Method for enzymatic production of high-activity bio-organic fertilizer

Info

Publication number: CN114901615A
Application number: CN202080091270.1A
Authority: CN
Inventors: 周亚仙; 彭可村; 陈丽珍
Original assignee: Guangxi Shenguan Collagen Biological Group Co ltd
Current assignee: Guangxi Shenguan Collagen Biological Group Co ltd
Priority date: 2019-12-31
Filing date: 2020-12-14
Publication date: 2022-08-12
Also published as: CN111018578A; WO2021135911A1; SG11202107331VA

Abstract

The invention discloses a method for enzymatically producing a high-activity bio-organic fertilizer, which comprises the following steps: (1) pretreating waste protein; (2) activating a fermentation strain; (3) and (5) fermenting. The method has the advantages of high activity of the fermented strains, good fertilizer efficiency of the obtained organic fertilizer, low production cost and environment-friendly production process.

Description

Method for enzymatic production of high-activity bio-organic fertilizer

Technical Field

The invention relates to the technical field of biological fermentation, in particular to a method for producing a high-activity biological organic fertilizer by enzyme.

Background

The biological organic fertilizer is a fertilizer which is compounded by microorganisms with specific functions and organic materials mainly prepared from animal and plant residues (such as livestock and poultry manure, crop straws and the like) through harmless treatment and decomposition. It is characterized in that functional microorganisms are added (secondarily added) into high-quality organic fertilizer (after decomposition). The main functions of the biological organic fertilizer comprise increasing the crop yield and improving the crop quality; improving soil fertility and improving a soil micro-ecosystem; reduce the occurrence of plant diseases and insect pests. The execution standard of the bio-organic fertilizer is NY884-2012, and the core index of the bio-organic fertilizer is that the effective viable count is more than or equal to 0.20 hundred million cfu/g. The prior technical proposal adopts a groove type stacking fermentation method.

The prior art scheme mainly has the following problems:

1. the fermentation substrate is limited. The microbial inoculum and the process are mainly designed aiming at livestock and poultry manure, crop straws, agricultural organic wastes and the like, and cannot effectively treat macromolecular proteins with complex conformations such as collagen, keratin and the like.

2. Functional microorganisms have a slow action. Many functional microorganisms added twice are dormant bodies such as spores or spores, and since heterotrophic bacteria are the main, activation is difficult after application to soil, resulting in a rather slow action.

3. The cost is high. Generally, a microbial agent is directly added to produce a biological organic fertilizer, so that the cost and time burden of purchasing the microbial agent are heavy, and about 150 ten thousand yuan is required for producing ten thousand tons of biological organic fertilizer every year.

Disclosure of Invention

In order to solve the problems existing in the prior art, the invention provides a method for enzymatically producing a high-activity bio-organic fertilizer, which comprises the following steps:

(1) pretreatment of waste protein: adding protease into the waste protein to obtain a fermentation material;

(2) activating a fermentation strain: respectively activating bacillus subtilis, lactobacillus and trichoderma, and then combining, wherein the number of effective viable bacteria is more than or equal to 0.40 hundred million cfu/g after combination;

(3) fermentation: mixing the fermented material with the activated strain, fermenting, and adding the strain for the second time when the temperature is below 40 ℃.

In one embodiment of the present invention, the waste protein material is a material containing high molecular proteins with complex conformation such as collagen and keratin, such as ground cow leather, cow hair, cow hoof nail, waste meat residue, and keratin. Can be used for cutting and pulverizing waste protein.

As a specific embodiment of the invention, in the step (1), before adding the protease, a fermentation substrate is also added; the fermentation substrate is one or more of the following: edible fungus chaff, rice bran, peanut bran, soybean meal, sugarcane straw powder, mushroom residue and the like, preferably one or more of the edible fungus chaff, the rice bran, the peanut bran and the soybean meal;

preferably, the mass ratio of the waste protein material to the fermentation substrate is 1 (1-3), preferably 1 (1.2-2.5), more preferably 1 (1.5-2), and even more preferably 1: 2.

As a particular embodiment of the invention, the protease is an alkaline protease.

As a specific embodiment of the present invention, in the step (1), the protease is added in an amount of 0.08% to 0.15%, preferably 0.10% to 0.12%, and more preferably 0.10% of the amount of the waste protein; preferably, the protease is added, and simultaneously, dry intestine-coating powder can be added, wherein the addition amount of the dry intestine-coating powder is 3-6% of the amount of the waste protein, and 5% of the waste protein is preferred.

As a specific embodiment of the invention, in the step (1), before adding the protease, the waste protein is added with alkali and boiled; preferably, the mass concentration of the alkali is 0.03-0.08%, preferably 0.05-0.08%, and more preferably 0.05%; further preferably, the boiling time is 25 to 40 minutes, preferably 30 to 35 minutes, and more preferably 30 minutes. Preferably, the waste protein material is subjected to a high temperature drum for primary dissolution prior to addition of the protease, followed by alkali boiling.

In one embodiment of the invention, the carbon-nitrogen mass ratio of the fermentation material is 20-30: 1, preferably 25-28: 1, and more preferably 25: 1.

As a specific embodiment of the invention, the moisture can be controlled by matching the materials or fermentation substrates to better adjust the carbon nitrogen ratio required by the fermentation of the strain, and preferably, the moisture content of the fermentation material is 58-68%, preferably 60-65%.

As a specific embodiment of the invention, in the step (2), bacillus subtilis is proliferated by NB culture solution, lactobacillus is proliferated by MRS culture solution, and then the lactobacillus is respectively added into the composition of the bran and the bean pulp for propagation activation; adding Trichoderma into the mixture of testa Tritici and soybean meal for activating.

In the step of activating the fermentation strains, bacillus subtilis and lactobacillus are respectively proliferated by using culture solutions and then respectively added into the composition of the bran and the bean pulp for propagation activation, so that the strains are activated by adopting a liquid-solid two-phase fermentation method, and the effect of getting twice the result with half the effort can be achieved.

As a specific embodiment of the invention, the water content of the composition of the bran and the soybean meal is 50-70%, and the mass ratio of the bran to the soybean meal is (8-10): 1.

As a specific embodiment of the invention, in the secondary bacterium adding in the step (3), one or more of bacillus subtilis, bacillus mucilaginosus, bacillus amyloliquefaciens, schizophyllum freundii, photosynthetic bacteria and the like can be added; preferably, the bacillus subtilis, the bacillus mucilaginosus, the bacillus amyloliquefaciens and the schizomycete fradiae can be added after being activated by NB culture solution respectively, and the photosynthetic bacteria can be directly sprayed; more preferably, each of the above strains is added after being activated.

During the second bacteria adding, part of the activated beneficial microorganisms are added again, so that the biological activity of the fertilizer is quick and durable.

The NB culture solution and the MRS culture solution used in the invention are culture solutions commonly used in the field, and can be obtained commercially.

Another object of the invention is to provide a high-activity bio-organic fertilizer produced by the method.

In the technical scheme of the invention, the enzyme and the microorganism are adopted to act together, namely, the enzyme-promoted biotechnology is adopted, so that the macromolecular proteins with complex conformations, such as collagen, keratin and the like, are effectively degraded.

According to the technical scheme, the protein waste raw material is subjected to enzymolysis pretreatment, and the activated microbial strains are added, so that the fermentation time is shortened, and the fermentation efficiency is improved.

The invention has at least one of the following beneficial effects:

1. the activity is high. The effective viable count can reach 0.40 hundred million cfu/g, which is far higher than 0.20 hundred million cfu/g of the basic requirement of NY884-2012 standard.

2. The fertilizer efficiency is good. The nutrient body of the effective live bacteria and the dormant body are mixed, after the nutrient body is applied to soil, the nutrient body can immediately play a role, and the dormant body can play a role in a subsequent long-term effect.

3. The cost is low. The enzymatic biotechnology is adopted, the fermentation time is saved by about 10 days, and meanwhile, the cost of the commercial microbial inoculum can be saved by more than half by adopting the activated strains.

4. The production is environment-friendly. Lactic acid bacteria, photosynthetic bacteria and the like are added in the fermentation process, so that ammonia nitrogen and hydrogen sulfide can be effectively degraded, and the method is environment-friendly.

Drawings

FIG. 1 is a flow chart of the method for producing high-activity bio-organic fertilizer by the enzymatic biotechnology of the invention.

Detailed Description

The present invention is further illustrated by the following examples, it is to be understood that the preparation methods described in the examples are illustrative only and not limiting, and that simple modifications of the preparation methods of the present invention based on the concepts of the present invention are intended to be within the scope of the invention as claimed.

The flow chart of the following example is shown in FIG. 1.

Example 1

Taking the production of the high-activity biological organic fertilizer by processing 20 tons of waste protein per day as an example:

1. pretreatment of waste protein: cutting 20 tons of waste protein (broken cowhide, cattle hair, and cattle nail) into pieces of 1-2 cm, placing in a plastic tank, adding 0.05% NaOH solution, boiling for 30 min, mixing with 30 tons of edible fungus bran and 10 tons of rice bran, and adding 20 kg of alkaline protease.

2. Adjusting the moisture content: and (3) adjusting the moisture of the pretreated material to 60% by using edible fungus bran to obtain a fermented material, wherein the carbon-nitrogen ratio in the fermented material is 25: 1.

3. Activating a fermentation strain: respectively taking 10 g of bacillus subtilis and lactobacillus strains, respectively proliferating by 5000 ml of NB culture solution and MRS culture solution, and then respectively adding into 50 kg of bran with water content of 60% and soybean meal (9:1) for propagation to obtain 100 kg of activated strains; adding Trichoderma 1 kg into 20 kg bran with water content of 60% and soybean meal (9:1) for activation; the activated strains are combined, and the effective viable count is more than or equal to 0.40 hundred million cfu/g.

4. Pile building and fermentation: mixing the fermented material and the activated strain, uniformly mixing, stacking, turning over and throwing at proper time until the temperature does not exceed 70 ℃, slowly reducing the temperature after 7 days, adding the bacillus subtilis, the jelly bacillus, the amylolytic bacillus and the schizomycete for the second time when the temperature is reduced to about 40 ℃, adding the bacillus subtilis, the jelly bacillus, the amylolytic bacillus and the schizomycete after the NB culture solution is activated, and directly spraying the photosynthetic bacteria.

5. And (5) processing after decomposition. Drying at low temperature (below 40 deg.C) until the water content is less than or equal to 30%, pulverizing, inspecting, and packaging. And finally 50 tons of high-activity bio-organic fertilizer can be obtained.

Example 2

Take the production of high-activity biological organic fertilizer by daily processing 25 tons of waste protein as an example:

1. pretreatment of waste protein: 25 tons of waste protein (broken cowhide, cattle hair, and cattle nail) are cut, pulverized to 2cm, placed in a plastic tank, boiled with 0.06% NaOH solution for 35 min, mixed with 30 tons of edible fungus bran and 7.5 tons of bran, and added with 30 kg of alkaline protease.

2. Adjusting the water content: adjusting the moisture of the pretreated material to 65% by using bran to obtain a fermented material, wherein the carbon-nitrogen ratio in the fermented material is 23: 1.

3. Activating a fermentation strain: taking 12 g of bacillus subtilis and lactobacillus strains respectively, proliferating by 5000 ml of NB culture solution and MRS culture solution respectively, and then adding into 50 kg of bran with the water content of 65% and bean pulp (8:1) respectively for propagation to obtain 100 kg of activated strains; adding Trichoderma 1.2 kg into 20 kg testa Tritici containing water 65% and soybean meal (8:1) for activation; the activated strains are combined, and the effective viable count is more than or equal to 0.42 hundred million cfu/g.

5. And (5) processing after decomposition. Drying at low temperature (below 40 deg.C) until the water content is less than or equal to 30%, pulverizing, inspecting, and packaging. And finally 55 tons of high-activity bio-organic fertilizer can be obtained.

Example 3

Taking the production of the high-activity biological organic fertilizer by processing 15 tons of waste protein per day as an example:

1. pretreatment of waste protein: cutting 15 tons of waste protein (broken cowhide, cattle hair, and cattle hoof) into pieces of 1cm, placing in a plastic tank, adding 0.03% NaOH solution, boiling for 30 min, mixing with 10 tons of bran coat and 8 tons of mushroom residue, and adding 12 kg of alkaline protease.

2. Adjusting the moisture content: and (3) adjusting the moisture of the pretreated material to 58% by using edible fungus bran to obtain a fermented material, wherein the carbon-nitrogen ratio in the fermented material is 28: 1.

3. Activating a fermentation strain: respectively taking 10 g of bacillus subtilis and lactobacillus strains, respectively proliferating by 5000 ml of NB culture solution and MRS culture solution, and then respectively adding into 50 kg of bran with the water content of 50% and bean pulp (10:1) for propagation to obtain 100 kg of activated strains; adding Trichoderma 1 kg into 20 kg bran with water content of 50% and soybean meal (10:1) for activation; the activated strains are combined, and the effective viable count is more than or equal to 0.40 hundred million cfu/g.

5. And (5) processing after decomposition. Drying at low temperature (below 40 deg.C) until the water content is less than or equal to 30%, pulverizing, inspecting, and packaging. And finally 28 tons of high-activity bio-organic fertilizer can be obtained.

Example 4

Take the production of high-activity biological organic fertilizer by processing 30 tons of waste protein per day as an example:

1. pretreatment of waste protein: cutting 30 ton waste protein (broken cowhide, cattle hair, and cattle nail) into 2cm pieces, placing in a plastic tank, adding 0.08% NaOH solution, boiling for 40 min, mixing with 40 ton edible fungus bran and 20 ton rice bran, and adding 45 kg alkaline protease.

2. Adjusting the moisture content: and (3) adjusting the moisture of the pretreated material to 68% by using edible fungus bran to obtain a fermented material, wherein the carbon-nitrogen ratio in the fermented material is 20: 1.

3. Activating a fermentation strain: respectively taking 15 g of bacillus subtilis and lactobacillus strains, respectively proliferating by 5000 ml of NB culture solution and MRS culture solution, and then respectively adding into 50 kg of bran with water content of 60% and soybean meal (9:1) for propagation to obtain 100 kg of activated strains; adding Trichoderma 1.5 kg into 20 kg testa Tritici containing water 60% and soybean meal (9:1) for activation; the activated strains are combined, and the effective viable count is more than or equal to 0.45 hundred million cfu/g.

4. Pile building and fermentation: mixing the fermented material and the activated strain, uniformly mixing, stacking, turning and throwing at proper time until the temperature is not more than 70 ℃, slowly reducing the temperature after 7 days, adding the bacillus subtilis, the bacillus mucilaginosus, the bacillus amyloliquefaciens and the schizomycete for the second time after the temperature is reduced to about 40 ℃, and directly spraying the photosynthetic bacteria.

5. And (5) processing after decomposition. Drying at low temperature (below 40 deg.C) until the water content is less than or equal to 30%, pulverizing, inspecting, and packaging. 80 tons of high-activity bio-organic fertilizer can be finally obtained.

Example 5

The method is the same as the method of the example 1 by taking the example of producing the high-activity bio-organic fertilizer by processing 20 tons of waste protein per day, and is different from the method in the step (1) in that the following operation is carried out: cutting 20 tons of waste protein (cow hair, cow nail and waste meat residue), pulverizing to 1-2 cm, placing in a high-temperature rotary drum for primary dissolution, adding 0.05% NaOH alkali solution, boiling for 30 min, mixing with 30 tons of edible fungus bran and 10 tons of rice bran, adding 20 kg of alkaline protease and 1 ton of dried enteric-coated powder, and adjusting pH to not higher than 8.5.

Experimental example 1

The fertilizer application amount is 500 kg/mu by using farmyard organic fertilizer to test on radishes. The radish yield is 5000 kg/mu.

The organic fertilizer of the embodiment 1 of the application is used for testing on radish, and the fertilizer application amount is 350 kg/mu. The radish yield reaches more than 5000 kg/mu, and the quality and the weight are superior to those of farmyard organic fertilizers.

Experimental example 2

The fertilizer application amount is 400 kg/mu by performing experiments on Chinese cabbages by using farmyard organic fertilizers. The yield of the Chinese cabbage is 2500 kilograms per mu.

The organic fertilizer of the embodiment 2 is used for tests on Chinese cabbages, and the fertilizer application amount is 300 kg/mu. The yield of the Chinese cabbage reaches 3500 kg/mu, and the quality and the weight of the Chinese cabbage are superior to those of farmyard organic fertilizers.

Experimental example 3

The fertilizer application amount is 500 kg/mu by performing experiments on the Chinese chives by using farmyard organic fertilizers. The yield of the Chinese chives is 3500 kg/mu.

The organic fertilizer of the embodiment 3 of the application is used for testing on the Chinese chives, and the applied fertilizer amount is 400 kg/mu. The yield of the Chinese chives reaches 4500 kilograms per mu, and the quality and the weight of the Chinese chives are superior to those of farmyard organic fertilizers.

Experimental example 4

The farmyard organic fertilizer is used for testing on the water spinach, and the fertilizer application amount is 500 kg/mu. The yield of the water spinach is 3500 kg/mu.

The organic fertilizer of the embodiment 4 of the application is used for testing on water spinach, and the applied fertilizer amount is 400 kg/mu. The yield of the water spinach reaches 4500 kg/mu, and the quality and the weight of the water spinach are superior to those of farmyard organic fertilizers.

Experimental example 5

The fertilizer application amount is 500 kg/mu by performing experiments on white gourds by using farmyard organic fertilizers. The yield of wax gourd is 5000 kg/mu.

The organic fertilizer of the embodiment 5 of the application is used for testing on wax gourd, and the applied fertilizer amount is 400 kg/mu. The yield of the white gourd is more than 5000 kg/mu, and the quality and the weight of the white gourd are superior to those of farmyard organic fertilizers.

The above test results are summarized in table 1.

TABLE 1 comparative test results of organic fertilizer and farmyard manure of this application

As can be seen from the results in Table 1, compared with farmyard manure, the application amount of the organic fertilizer is saved by 20-30% per mu, and the yield per mu is increased by 10-40%. For agricultural production, the cost is saved, the yield is improved, and considerable economic benefits are achieved.

The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are all within the protection scope of the present invention.

It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.

In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Claims

A process for the enzymatic production of a highly active bio-organic fertilizer, characterized in that it comprises the following steps:

(1) pretreatment of waste protein: adding protease into the waste protein to obtain a fermentation material;

(2) activating a fermentation strain: respectively activating bacillus subtilis, lactobacillus and trichoderma, and then combining, wherein the number of effective viable bacteria is more than or equal to 0.40 hundred million cfu/g after combination;

(3) fermentation: mixing the fermented material with the activated strain, fermenting, and adding the strain for the second time when the temperature is below 40 ℃.
The method according to claim 1, wherein in step (1), a fermentation substrate is further added before the protease is added; the fermentation substrate is one or more of the following: edible fungus chaff, rice bran, peanut bran, soybean meal, sugarcane straw powder and mushroom residue; preferably one or more of edible fungus bran, rice bran, peanut bran and soybean meal;

preferably, the mass ratio of the waste protein material to the fermentation substrate is 1 (1-3); more preferably 1 (1.2-2.5).
The method of claim 1, wherein the protease is an alkaline protease.
The method according to any one of claims 1 to 3, wherein the protease is added in the step (1) in an amount of 0.08 to 0.15%, preferably 0.10 to 0.12% by weight based on the amount of the waste protein.
A method according to any one of claims 1 to 3, wherein in step (1), the waste protein is boiled with alkali before the protease is added; preferably, the mass concentration of the alkali is 0.03-0.08%, more preferably 0.05-0.08%; more preferably, the boiling time is 25 to 40 minutes, and still more preferably 30 to 35 minutes.
The method according to any one of claims 1 to 3, wherein the fermentation material has a carbon-nitrogen mass ratio of 20 to 30:1, preferably 25 to 28:1, more preferably 25: 1; further preferably, the moisture content of the fermentation material is 58% to 68%, more preferably 60% to 65%.
The method according to any one of claims 1 to 3, wherein in the step (2), Bacillus subtilis is proliferated by NB culture solution, lactobacillus is proliferated by MRS culture solution, and then the proliferated lactobacillus is added into the composition of bran and bean pulp for propagation activation; adding Trichoderma into the mixture of testa Tritici and soybean meal for activating.
The method according to claim 7, characterized in that the water content of the composition of the bran and the bean pulp is 50-70%, and the mass ratio of the bran to the bean pulp is (8-10): 1.
The method according to any one of claims 1 to 3, wherein in the secondary adding of the step (3), one or more of Bacillus subtilis, Bacillus mucilaginosus, Bacillus amyloliquefaciens, Schizophyllum freundii and photosynthetic bacteria are added; preferably, the bacillus subtilis, the bacillus mucilaginosus, the bacillus amyloliquefaciens and the schizomycete fradiae are added after being activated by NB culture solution respectively, and the photosynthetic bacteria are directly sprayed.
The high-activity bio-organic fertilizer produced by the method of any one of claims 1-9.