KR101832065B1 - Method for Preparing Liquefied Fertilizer - Google Patents

Abstract

The present invention relates to an amino acid liquefied fertilizer obtained by treating tuna processing residues with yeast and fermenting, and a method for preparing the liquefied fertilizer. The amino acid liquefied fertilizer has storability, a function to reduce the repulsion caused by malodor from using the fertilizer, and a microbial pesticide function to reduce diseases by applying an antagonistic function to phytopathogenic microorganisms on major crops.

Description

FIELD OF THE INVENTION [0001] The present invention relates to an amino acid liquid fertilizer and a method for preparing the same,

The present invention relates to an amino acid liquid fertilizer obtained by enzymatic treatment and fermentation of tuna processing residues and a method for preparing the same, and more particularly, to an amino acid liquid fertilizer obtained by fermenting a tuna processing residue with improved plant pathogen resistance and growth promoting function, ≪ / RTI >

As is well known, nutrient management in organic agriculture is based on supplying organic matter such as compost, organic fertilizer, and green grass. However, in order to replenish the nutrients that are released into the surrounding environment or used for growing crops Additional nutrients are regularly required. For example, in the cultivation of organic peppers, green grass crops or organic fertilizers are used as a foundation, and liquid fertilizer (liquid fertilizer: hereinafter referred to as "fertilizer") is produced using various organic resources after the middle of growing season, ). As such, the liquid fertilizer can be regarded as a quick-acting fertilizer.

Meanwhile, the liquid fertilizer manufactured by the present applicant (Hyundai E & C Co., Ltd.) is a product obtained by decomposing wet pulverized tuna byproduct with a protease and controlling inorganic nutrients (K, P) It is a liquid amino acid type fertilizer that receives the list of environmentally friendly organic ingredients for growing crops (Product name: Pungchul-L, Publication No. 11- Organic-2-505) and supplies them to rose and other high-level gardening complexes.

The applicant of the present invention is to develop a functional liquid fertilizer which enhances the shelf life of amino acid liquid fertilizer (product name: Fruit-L) and adds microbial pesticidal function to the existing commercial product, and its significance is as follows.

Lactic acid produced by the present applicant (product name: Pomegranate-L) is produced by proteolytic enzymes, so that the molecular weight of the protein degradation product is high, so that the absorption ability of the crop is partially disturbed and the fish oil of the tuna is not emulsified, There is a disadvantage in use of the spray of the unique odor. Therefore, it is mixed and cultured with a microorganism that secretes a protease and a microorganism having the ability to decompose a fatty acid, and the protein is aged with a more homogeneous amino acid, and the fish oil is emulsified during the fermentation process to reduce the inherent fish oil odor, . In addition, by adding plant pathogenic resistance microorganism, microorganism pesticide function is added. By adding fermentation process and adding processable microorganisms, if high-functioning liquid is supplied, nutritional and pathogenic reduction effect can be realized simultaneously in farming, It is important to reduce the use of pesticides and to make them eco-friendly for pest and disease relief.

On the other hand, the prior art documents related to amino acid fertilizers include the following Patent Documents 1 to 6.

The amino acid fertilizers disclosed in the above Patent Documents 2 to 6 are prior art documents for producing amino acid solution ratio by using mainly livestock blood products, by-products of amino acid seasoning industry, enzymatic hydrolysis solution of hairy or fur, and amino acid added to seaweed extract.

As can be seen from the above patent documents, there is no direct patent literature using tuna by-products. The only patent application of the present inventor is "a method for producing a fertilizer containing tuna by-products as a main component (patent document 1)". The above patent document 1 is a prior art related to a case somewhat different from the present invention in which a tuna by-product and an organic material are mixed and fermented.

On the other hand, as in Patent Document 1, the tuna which is the main ingredient of the amino acid solution of the tuna by-product (product name: Pomegranate-L) and the by-product thereof are mainly used as the tuna fish species processed for edible purposes, Spines, fins, skin, hair (dark), blood (with a lot of flesh), and sawdust (occurring when cutting frozen tuna). These tuna and its by-products contain 53 ~ 55% of protein after drying and 8.0 ~ 12% of ash (Ca, P component) and contain very high content of ingredients compared with general natural organic materials. In addition, tuna by-products contain excellent proteins and contain large amounts of minerals (Ca, Fe, Mg, etc.) as well as vitamins (B1, B2, B6 and E). In addition, 20 kinds of amino acids by enzymatic decomposition promote the decomposition of organic matter by promoting the microbial growth of soil, and ammonia nitrogen is absorbed in roots as amino acid form as well as decomposition and mineralization in soil. Therefore, organic fertilizer using tuna byproducts is very effective in promoting the growth of underground and above ground parts, and preventing cold damage and disease when lack of sunshine.

In addition, glycine, a component of tuna, promotes the absorption of nitrogen and carrie, while aspartic acid and serine promote absorption of phosphoric acid and caries, and increase taste, sugar and storage by some amino acids The effect is very large. In addition, since it contains a small amount of oil (EPA, DHA, etc.), it is possible to reduce habitat and scattering in stems, leaves and surrounding soil as a repellent of some harmful insects.

Today, such tuna byproducts are decomposed into enzymes (alkalis) and liquid products are separated and commercialized. However, there is a demand in the market for solving the problem of the storage stability of existing products and the environmentally friendly function of the microbial control of the hospital.

Registration No. 10-1187327 (Title of the invention: Method of producing fertilizer containing tuna by-product as a main ingredient) Date of Notification: October 02, 2012 Registration No. 10-1203945 (the name of the invention: compound fertilizer for producing sprout vegetables and young leafy vegetables containing high concentrations of GABA, Tr y-Pro, antioxidant activity and vitamin C using seaweed extract and its preparation method) Date: November 23, 2012) Patent Publication No. 10-2013-0094374 (entitled: Highly efficient batch production system for the production of amino acid fertilizers and feeds using slaughter blood proteins and method for producing the same) Published Date: Aug. 26, 2013 Patent Publication No. 10-2009-0053050 (Title: Method for producing amino acid liquid fertilizer) Published on May 27, 2009 Open No. 10-2002-0076502 (entitled: Fermented Amino Acid Organic Fertilizer, Fermented Amino Acid Liquid Fertilizer and Method for Producing the Same) Published Oct. 11, 2002 Patent Publication No. 10-2001-0081476 (Title of the invention: Method for producing fertilizer using oligopeptide salt and / or amino acid salt mixture.) Published Date: Aug. 29, 2001

It is an object of the present invention to provide an amino acid liquid fertilizer and a method for producing the same, wherein the tuna processing residues are enzymatically treated and fermented with a tuna processing residue, which improves the plant pathogen resistance and growth promoting function of the livestock.

It is an object of the present invention to provide an amino acid liquid fertilizer which is fermented with an organic acid-producing microorganism, which is capable of eliminating a sterilization process and improving storage stability, by enzymatic treatment and fermentation of a tuna processing residue.

An object of the present invention is to provide an amino acid liquid fertilizer and a method for producing the same, wherein the tuna processing residue is easily fermented by reducing the intrinsic odor due to fish oil contained in the tuna processing by-product through microbial fermentation.

It is an object of the present invention to provide an amino acid liquid fertilizer and a method for producing the amino acid liquid fertilizer by enzymatically treating and fermenting tuna processing residues added with a microbial pesticide function using a plant pathogenic microorganism.

The present invention relates to an enzyme treatment of tuna processing residues which can significantly contribute to the growth of plants by containing amino acids in a considerably short period of 4 to 5 days compared with the conventional liquid tanning processing byproducts produced by enzymatic digestion And fermented amino acid liquid fertilizer and a method for producing the same.

The present invention relates to a process for fermenting a liquid fertilizer obtained by enzymatically treating a tuna processing residue by adding 2% each of proteolytic microorganism, phytopathogenic microorganism, plant lactic acid bacterium, 3% molasses and 0.1% .

Herein, the proteolytic microorganism used in the present invention is characterized by being Bacillus sutillus LH1.

As described above, the amino acid liquid fertilizer which has been subjected to the enzymatic treatment and fermentation of the tuna processing residue according to the embodiment of the present invention and its manufacturing method achieve the following many effects.

Firstly, the present invention has a nitrogen component which is enzymatically and microbiologically fermented by amino acid fermentation and is rapidly absorbed by plants, thereby promoting the growth of crops and increasing the yield of the crops. As a result, Can be used. Especially, it is considered to be an excellent liquid fertilizer because it contains a microorganism having an antagonistic function against scalloping glanders.

Secondly, the present invention can increase profit through increased harvesting of agricultural crops, and thus, it is expected that the farmers will have a continuous purchasing effect, and the sales of production companies are expected to increase.

Thirdly, the present invention is environmentally friendly because of the use of highly nutritious and inexpensive tuna processing residues. Value-added ecological virtuous cycle structure, which is an efficient land disposal method for fishery processing residues.

Fourth, the present invention is not only in that the fermentation of high-speed high-speed fermentation by aerobic high-temperature fermentation does not occur, but also has the effect of improving the working environment as well as the conventional liquid fertilizer.

Fifthly, the present invention is based on the fact that since the fermentation production period of the liquid fertilizer (product name: PURPOSE-BL) is considerably short, the manufacturing productivity is increased, and the production unit cost can be lowered in the production plant, And high-quality industrialization of related industries.

Sixth, the present invention is expected to enable the production of high-quality horticultural crops through high-level flowers and the activation of urban plant agriculture in the future along with the participation of the well-being period in the diet requiring eco-friendly agricultural products by organic farming.

FIG. 1 is a diagram sequentially showing an amino acid liquid fertilizer obtained by enzymatic treatment and fermentation of a tuna processing residue according to a preferred embodiment of the present invention and a method for producing the same.
Figure 2a shows pathogenic microorganisms distributed in the Center for Agricultural Genetic Resources, specifically, KACC 40457 ( Sclerotinia sclerotiorum ), KACC 40124 ( Rhizoctonia solani ), KACC 40241 ( Fusarium sp ., ), KACC 40180 ( hytophthora capsici ), which is a pathogenic microorganism.
Figure 2b sprocket pathogenic microorganisms in the microorganism pre-sale received resource center, specifically, KCTC 6084 (Fusarium oxysporum f. Sp . Cucumerinum), KCTC 6104 (Glomerella cingulata ), respectively.
FIG. 2C is a photograph showing the submerged foliar fungus separated from the subspecies field.
FIG. 3 is a graph showing the distribution of antagonistic microorganisms prevalent in the microbial resource center, specifically, KCTC 3554 ( Paenibacillus polymyxa ), KCTC 1643 ( Pseudomonas putida ), KCTC 9204 ( Streptomyces sp . ).
Figure 4 shows the results of the paper disc method of various crab microbes on plant pathogens, specifically 1 in Fusarium oxysporum f. sp . cucumerinum , 2 shows the results of Fusarium sp., 3 , and 3 shows the results of Rhizoctonia solani , and 4 is Phytophthora capsici , and 5 shows Glomerella cingulata , 6 shows the results of Sclerotinia sclerotiorum , and 7 shows the results of the Fungus of field-dropwort. A shown in the figure is Pseudomonas putida , B is Bacillus subtilis , C is Streptomyces sp., D is Bacillus subtilis , E is Paenibacillus polymyxa , F is Bacillus sp. LH1.
Figure 5 shows the time course of Bacillus sp. This graph shows the growth and pH change of LH1.
Fig. 6 is a graph showing the distribution of the Bacillus sp. This is a photograph showing the result of the dual culture method of LH1.
FIG. 7 is a photograph showing purely isolated bacteria isolated from pure cultures cultured on a medium, specifically, 1, 2, 1, 2, and 1 from the left.
8 is a graph showing the growth and pH change of Lactobacillus curvatus with time.
FIG. 9 is a photograph showing the strains of cultivated sorted isolated microorganisms, specifically, on the left, Bacillus subtilis LH1, and on the right is a photograph showing strains of Lactobacillus curvatus .
Fig. 10 is a test report for analyzing an active ingredient and a harmful component of a liquid fertilizer (product name: Pomegranate-BL) obtained by enzymatically treating a tuna processing residue produced by the method of the present invention.
11 is a test report of an effective microorganism according to the present invention.
12 is a test report of a pathogenic microorganism according to the present invention.
Fig. 13 is an ineffective and comparative test report according to the present invention.

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. In the following detailed description, one exemplary embodiment of the present invention will be described in order to accomplish the above-mentioned technical problems. And other embodiments which may be presented by the present invention are replaced by descriptions in the constitution of the present invention.

In the present invention, a method of converting an amino acid solution obtained by enzymatically digesting a tuna by-product into a protein fermentation in which the amino acid is improved in an absorption-type manner, a resistance to plant pathogens and a unique odor, We suggest a way to do this.

Specifically, the present invention relates to a method for producing an amino acid liquid fertilizer obtained by enzymatically digesting a tuna by-product, which comprises fermenting a protein-degrading microorganism, a plant pathogenic antibacterial agent and molasses into a fermentation process, In addition to the performance of the product, it further enhances the growth of crops by increasing the bioavailability and enhances the resistance to plant diseases, as well as the enzymatic treatment of tuna processing residues with ease of application by reducing the intrinsic odor due to microbiological emulsification of oil And fermented amino acid liquid fertilizer and a method for producing the same.

The production process to be implemented in the present invention is characterized in that after the enzymatic reaction process, secondary fermentation with amino acid and fermenting microorganism and sterilization treatment prior to storage are performed by the organic acid produced by fermentation with an organic acid-producing microorganism, The storability is automatically generated as well as improved. Therefore, in the present invention, the conventional sterilization step is omitted, and microorganism pesticide functions are also shown by adding microorganisms resistant to the pathogenic microorganisms in the fermentation process.

When the liquid fertilizer is produced by the same method as the present invention, it is converted into an amino acid form having a higher absorption rate of low molecular weight and is endowed with the introduction of plant pathogenic antimicrobial. When fermentation is performed by feeding nutrient source and molasses, It is possible to prevent deterioration of amino acid and to reduce input energy by omitting the sterilization process due to lowering of the deterioration property as well as to enhance the self-storage property of the organic acid fermentation broth generated in the fermentation process. Further, It is a method capable of producing a further improved amino acid solution in which the amino acid is reacted through the fermentation process with the sugar to reduce the intrinsic odor (i.e., odor).

In the meantime, in the present invention, when preparing a liquid fertilizer obtained by enzymatically treating a tuna processing residue, the pathogenic microorganism having an antagonistic property is introduced in the fermentation stage in order to prevent diseases and pests of agricultural crops. It searches the homepage of the Resource Center and the Agricultural Genetic Resources Information Center to search for microorganisms causing major microbiological pathogenesis of large-scale agricultural crops such as leek, onion, pepper and parsley, which are typically commercialized. They are sold as follows and used as pathogenic microorganisms Respectively.

There are seven types of plant pathogens used in the present invention, and they are specifically shown below. Each photograph of the plant pathogenic bacteria that has been put into sale is shown in FIGS. 2A to 3C.

① Fusarium oxysporum f. sp . Cucumerinum : A causative organism of leek and onion was sold at the Microbiological Resource Center.

② Fusarium sp . : It was sold at the Agricultural Genetic Resources Information Center as a cause of wilt disease of leek and onion.

③ Rhizoctonia solani : Leek and onion are the causative agents of the disease and have been sold at the Agricultural Genetic Resources Information Center.

④ Phytophthora capsici : It was sold at the Agricultural Genetic Resources Information Center as a cause of P. aeruginosa .

⑤ Glomerella cingulata : Anthracnose caused by red pepper was distributed at the microbial resource center.

⑥ Sclerotinia sclerotiorum: Sclerotinia sclerotiorum was licensed from the Agricultural Genetic Resources Information Center.

⑦ Jaguksan mildew: It was collected directly from the diseased buttercups of Mt. As mildew fungi cause diseases of the buttercup jagulsan classification of 5.8S rRNA of fungi Ascomycetes separated from jagulsan buttercup field, Sclerotinia causing gyunhaekbyeong sclerotiorum , and showed the nucleotide sequence of 5.8S rRNA.

Sclerotinia The gene bank and gene sequence of 5.8S-RNA of sclerotiorum strain XJSHZ-3 are as follows.

Sclerotinia sclerotiorum strain XJSHZ-3 internal transcribed spacer 1, partial sequence; 5.8S ribosomal RNA gene, complete sequence; and internal transcribed spacer 2, partial sequence

GenBank: JN012606.1

There are six kinds of antagonistic microorganisms used in the present invention, and will be specifically described below. The photograph of the pre-cultured antagonistic microorganism is shown in FIG.

① Pseudomonas putida (A): It is distributed at the microbial resource center. It produces catechol 2,3-oxygenase as a product and produces enzyme of cellulase and protease.

② Bacillus subtilis (B): extracted from Chungkookjang.

③ Streptomyces sp . (C): It is distributed at the microbial resource center. It produces lincomycin as a product and produces cellulase and protease enzymes.

④ Bacillus subtilis KCCM 41462 (D): It was sold at the Korea Microorganism Conservation Center.

⑤ Paenibacillus polymyxa (E): It is distributed in the microbial resource center. It is a product of production of enzyme of cellulase and protease.

⑥ Bacillus SP. LH1 (F) is a self-contained strain isolated from rice straw.

In the present invention, antagonistic microorganisms against pathogenic microorganisms were tested and selected in the following manner.

1. Experimental Method

end. Medium and culture conditions

Growth of plant pathogens Potato Dextrose Agar (Potato dextrose borth 24 g / L, agar 15 g / L; 25 ℃) was used as the growth of the antagonistic microorganism is Bacillus genus Luria Bertani (Tryptone 10 g / L , Yeast extract 5 g / Pseudomonas putida was prepared by adding Benzoate Medium ((NH4) 2HPO4 3 g / L, KH2PO4 1.2 g / L, NaCl 5 g / L, MgSO47H2O 0.2 g / L, Yeast extract 0.5 g / L, NaCl 10 g / L; L, Na-benzoate 3 g / L, AGAR 20 g / L; 25 ° C), Strepomyces sp. Nutrient Agar (Beef extract 3 g / L, peptone 5 g / L, agar 15 g / L; 30 ° C) was used as the panibacillus polyxa . Liquid cultures of antagonistic microorganisms were cultured in a shaking incubator using Nutrient broth (rpm: 130)

I. Antagonistic test

In order to increase the activity of plant pathogens, antagonistic microorganisms, and direct - harvested Mycobacterium tuberculosis pathogens, they were subcultured in pre - solid medium. Antagonistic microorganisms were cultured in liquid medium and subjected to paper disc method. The biomass of the antagonistic microorganism was measured by measuring the OD value of the culture solution at 600 nm. When the OD value was 0.4, 32 .mu.l was injected, and the OD of each antagonistic microorganism was adjusted to the OD value. On the other hand, the microorganism culture of the control experiment was not added and the same conditions were used.

A part of the cultured plant mycelium was inoculated into the center of the PDA medium, and six paper disks (Diameter: 0.5 cm) were placed in six directions 2.5 cm away from the center of the medium. The effect of antagonistic microorganisms on plant pathogens was investigated on the 7th day after culturing at 25 ℃ for 3 days by wetting 6 liquid cultured antagonistic microorganisms on paper discs.

2. Experimental results

end. Antagonistic strain

Table 1 shows the results of investigating the inhibition of mycelial growth by the paper disc method in order to find strains showing pathway resistance in plant pathogenic bacteria. Table 1 shows the results of inhibition of antagonistic strains against plant pathogenic bacteria.

1) Fusarium oxysporum f. sp . cucumerinum : causative bacteria of leek and onion

2) Fusarium sp . : Wiltful disease caused by leek and onion

3) Rhizoctonia solani : the quality of leek and onion

4) Phytophthora capsici : Plague causative bacteria of pepper

5) Glomerella cingulata : Anthracnose causing bacteria of pepper

6) Sclerotinia sclerotiorum : Sclerotinia causing bacteria

7) Jagul Mt. Fungus : Fungus isolates from mallow field

* 1) A, Pseudomonas putida

* 2) B, Bacillus subtilis

* 3) C, Streptomyces sp.

* 4) D, Bacillus subtilis

* 5) E, Paenibacillus polymyxa

* 6) F, Bacillus sp. LH1

As shown in Table 1, Fusarium oxysporum f. sp . cucumerinum had include A, B, C, D, E, F, respectively 4, 6, 9, 6, 10, born this way receive drag of 13mm (see Fig. 1 of 4), Fusarium sp . , A, C, E, and F were 2, 5, 4, and 7 mm, respectively (see FIG. Rhizoctonia solani, the A, B, C, D, E, F This was 4, the road drag of 11, 10, 5, 4, 13mm appeared respectively (see 3 in Fig. 4), Phytophthora C, E, and F were found in the glomerella cingulata (Fig. 4), and the A, B, C, D, E, and F of the capsici showed 15, 15, 15, 15, 16, (See 5 in Fig. 4), and Sclerotinia < RTI ID = 0.0 > Sclerotiorum showed B, C, F of 9, 1, and 10 mm, respectively (see 6 in FIG. 4). Finally, in the submicron fungus, B, C, D, E, and F were 10, 2, 7, 11, and 12, respectively.

As shown in Table 1 and FIG. 4, Bacillus sp. LH1 showed a high level of inhibition of growth on plant pathogens 1, 2, 3, 4, 5, 6 and 7, with a high level of 7 ~ 16mm. Therefore, in the present invention, Bacillus sp. LH1 was selected as an antagonistic strain.

The microorganism deposit number of " Bacillus sp. LH1" is as follows.

In the present invention, the selected antagonistic strain, Bacillus sp. For the production of LH1 cells, a growth curve was prepared in the fermentation culture medium (Luria Bertani (Tryptone 10 g / L, Yeast extract 5 g / L, NaCl 10 g / L; And the end stage of maximum proliferation was calculated and collected as inoculum cells. That is, as shown in the growth curve graph of FIG. 5, the maximum proliferation state was observed at about 4 hours, and the cells were collected at this time.

I. Identification of selected microorganisms

In the present invention, when preparing a liquid fertilizer obtained by enzymatically treating a tuna processing residue, the antagonistic microorganism to the plant pathogenic bacteria introduced in the fermentation step is preferably Bacillus sp. LH1, and the Bacillus sp. The 16S rRNA classification of LH1 identified Bacillus subtilis as " Bacillus subtilis LH1".

Bacillus sp. The gene bank and gene sequence for 5.8S rRNA analysis of BAB-3807 are as follows.

Bacillus sp. BAB-3807 16S ribosomal RNA gene, partial sequence

GenBank: KJ588869.1

In the present invention, it is considered that the antagonistic to the sclerotinia of the subspecies of the subspecies is very important in view of the demand for the main application of the liquid fertilizer and the marketability for the hydroponic cultivation in the future. As shown in FIG. 6, Antagonistic microorganism Bacillus subtilis . The inhibitory effect of LH1 on the antagonistic activity of LH1 was inhibited by the inhibition of growth of pathogenic microorganisms.

On the other hand, in the present invention, in order to improve the shelf life of the fermented amino acid solution, it is intended to supplement the strains producing organic acids and to selectively add plant viable bacteria having high viability.

That is, lactic acid bacteria were isolated from ripe kimchi which is known to be rich in lactic acid bacteria and commercially available lactic acid bacteria beverages. Two kinds of kimchi were isolated from lactic acid bacteria using three kinds of kimchi, .

The medium was diluted 1,000 times by serial dilution with 0.85% physiological saline. The diluted samples were spread at 20 each at 30 째 C, streaked with characteristic colonies, Respectively. The pure isolates were five kinds (1, 2, 2) in mulch kimchi, 2 (1 and 2) in cabbage kimchi, and 1 (1) in lactic acid bacteria beverage.

Antimicrobial susceptibility test of 5 kinds of pure isolates was carried out in the same manner as the antagonistic microorganism screening method. The experimental results showed that none of 5 species showed inhibitory effect on the growth of plant microbes.

As a result of 16S rRNA classification of lactic acid bacteria 2 isolated in mugwort , Lactobacillus curvatus was identified, and gene bank and gene sequence for 16S rRNA analysis of Lactobacillus curvatus were as follows.

Lactobacillus curvatus gene for 16S rRNA, partial sequence, strain: Ni998

GENbank: AB600200.1

Growth curves of Lactobacillus curvatus selected by germination were examined and the degree of viability was determined. As shown in FIG. 8, the maximum growth was observed at about 14 hours. The pH was decreased to 6.0 during the 12 hour incubation period. If it is in a rich state, it can produce a higher amount of organic acid and exhibit a lower pH state.

In the present invention, a freeze-dried microorganism sample was prepared in order to facilitate the transfer of strains of the selected microorganism, Bacillus subtilis LH1, and lactic acid bacteria Lactobacillus curvatus . Bacillus subtilis LH1 and Lactobacillus curvatus were cultured in the same medium and culture conditions as in the selection of antagonistic microorganisms and selection of lactic acid bacteria. The microorganism strains cultured before the preparation of the lyophilized sample are shown in Fig. Microbial lyophilization was commissioned by the Microbiological Resource Center and 10 lyophilized samples were prepared.

In addition, in the present invention, the selection of a strain that performs amino acid fermentation with protein decomposition added to the amino acid fermentation by microorganisms after enzymatic degradation of the tuna by-product is performed using a protein of Bacillus subtilis LH1, a pathogenic microorganism, Lipid degradation ability was used as a proteolytic microorganism. In order to select Bacillus bacterium, which is a soil microorganism preparation using microorganism as a microbial fertilizer, the microorganism was selected as a soil microbial agent.

In the present invention, in order to realize a method of producing a liquid fertilizer in which a tuna processing residue is subjected to enzymatic treatment, it is preferable to use an apparatus for supplying oxygen to inoculate microorganisms, A device was used to prevent problems and foaming during fermentation.

That is, since the operation of the fermenter is operated at 45 to 55 ° C, if the micro-bubbles are supplied by the physical method, the oxygen transfer rate in the fermenter is increased. Thus, in the present invention, fine air bubbles through a high-pressure disperser were supplied to the fermenter. At this time, the air pressure of the high-pressure disperser is 3 L / min · ea and the air pressure is 1.5 kg / cm 2.

In order to increase the dissolved oxygen in the fermenter and to prevent foaming, the oxygen concentration is supplied to 90% or more through a conventional pure oxygen generator. The amount of air supplied is reduced so that the fermentation Water loss and management were facilitated.

Thus, sufficient dissolved oxygen (preferably, 7 mg / L or more) was maintained, and the problem of the disappearance of the culture liquid and contamination due to the generation of massive bubbles as in the conventional microbial fermenter operation test was solved.

As described above, the method for preparing the liquid fertilizer by treating the tuna processing residues according to the preferred embodiment of the present invention comprises inoculating proteolytic microorganisms, pathogenic microorganisms, organic acid-producing microorganisms, and phytopathogenic microorganisms, And 10% of whole milk powder for the initial induction culture. The conventional sterilization process was omitted in order to save microorganisms. In addition, during the process of preparing the liquefied soybeans, it was maintained at over 2 mg / L of dissolved oxygen, and the final pH was reduced to about 4.0, thereby reducing the strong fish meal (ie, smell) by 60%.

As shown in FIG. 1, 300 g of tuna by-product was wet-milled and 300 L of hot water at 40 ° C was placed in a fermenter (i.e., a deep-incubator type, capacity 800 L) and the temperature was set to 40 ° C. ), 0.2% of commercial protease is added, and then physicochemical liquefaction and amino acidization of solid protein are sufficiently generated. At this time, the determination of completion of the enzyme decomposition was performed by sampling the sample in units of 2 hours and observing with a microscope (x 200) until a large solid was not seen. Enzyme decomposition was completed at about 12 ~ 15hrs elapsed time.

Then, 2% of proteolytic and antagonistic microorganisms ( Bacillus sutillus LH1) and plant lactic acid bacteria were added, and 3% of molasses and 0.1% of skim milk powder were added as nutrients. And stirred at a speed of 30 rpm while supplying 8 L / min. As a result, odor of fresh fish and smell of dissolved oxygen were maintained at 5 ~ 8mg / L. Therefore, it can be judged that sufficient oxygen is supplied and the organic fermentation process proceeds smoothly at high temperature and high concentration.

At this time, the growth of the microorganism was observed by a microscope, and it was found to be about 10 ⁶ CFU / ㎖ or more, and it was operated until the final pH was about 4.0, and the target reached about 40 hrs later. The pH was lowered to such an extent that the microbial growth could be suppressed by the organic acid sufficiently produced by the microorganism, and the sterilization step was omitted and filtration was performed with a filter press (200 / hr) to prepare the liquid fertilizer of the present invention.

Table 2 summarizes the fermentation conditions as described above. Table 2 shows operating factors of a pilot plant fermenter for functional liquefaction of porridge processing by-products.

Fermenting factor Operating condition Goal of operation and fermentation Judgment Remarks Dissolved oxygen (DO) 7 ~ 8L / min
Pure oxygen supply DO 2mg / L or more
maintain Good 5 ~ 8mg / L maintained and reduced in half Temperature (℃) 40 40 to 50 Good Nutrient Molasses 3 ~ 5% Microbial growth
10 ⁶ CFU / ml or more Good Initial fermented milk also increased by 0.1% and nutrients by 10% pH Organic acid-producing bacteria
input 4.0 Good Improved storage smell Exhaled amino acid
Fermenting bacteria input Odor reduction
(Fresh smell) Good

The present invention also provides a method for producing a liquid fertilizer obtained by enzymatically treating a tuna processing residue as described above, wherein the fermentation step (protein-digesting microorganism, phytopathogenic microorganism, plant lactic acid bacteria 2% each, molasses 3% %). The liquid fertilizer produced by omitting the conventional sterilization process was subjected to an official test in accordance with the list of environmentally friendly farm products list. Specific test methods and results are as follows.

1. Principal component inspection

We commissioned Jeil analysis center, an authorized analytical agency, to test the validity items and harmful components designated as eco - friendly farm products according to the eco - list disclosure.

The nitrogen content of the active ingredient was 1.54%, which was slightly higher than the nitrogen content (1.45%) of the conventional enzyme-digested amino acid solution (trade name: PF-L). Phosphoric acid contained 0.32% and 0.25% of glutinous rice. All of the harmful components are suitable within the standard values (both the efficacy-BL efficacy and harmfulness indexes), and are shown in Tables 3 to 7 below, in which all of arsenic, cadmium, mercury, lead, chromium, copper, . The above Tables 5 and 6 are prepared based on the test results shown in FIG. 10, preferably on the basis of the active ingredient and the harmful component pumice test results of the pomegranate-BL (liquid fertilizer obtained by enzymatically treating the tuna processing residue produced by the method of the present invention) Respectively.

The active ingredient of the amino acid solution (trade name: Pomegranate-L) Test date Test institute Nitrogen total amount All phosphoric acid Whole volume salt moisture Organic matter July 15, 2014. First Analysis Center 1.45% 0.30% 0.33% 0.40% 88.27% 10.48%

Amino Acid Liquid (Product name: Pomegranate-L) Hazardous ingredient Test Year Test institute Analysis item Results of analysis (unit: mg / kg)


July 15, 2014.


First Analysis Center arsenic Non-detection cadmium Non-detection Mercury Non-detection lead Non-detection chrome 6.64 Copper 13.22 nickel 3.91 zinc 7.9

The active ingredient of the amino acid solution (trade name: SW-BL) Test date Test institute Nitrogen total amount All phosphoric acid Whole volume salt moisture Organic matter October 21, 2015. First Analysis Center 1.54% 0.32% 0.25% - - -

Hazardous components of amino acid solution (trade name: fertilizer-BL) Test Year Test institute Analysis item Results of analysis (unit: mg / kg)


October 21, 2015.


First Division Center arsenic Non-detection cadmium Non-detection Mercury Non-detection lead Non-detection chrome 18.58 Copper Non-detection nickel 3.27 zinc Non-detection

Criteria for harmful ingredients of eco-friendly farm materials Item unit standard arsenic mg kg ^-1 20 or less cadmium mg kg ^-1 2 or less Mercury mg kg ^-1 1 or less lead mg kg ^-1 Less than 50 chrome mg kg ^-1 90 or less Copper mg kg ^-1 120 or less nickel mg kg ^-1 20 or less zinc mg kg ^-1 400 or less

As shown in Tables 3 to 7, the disclosure of the list of environmentally friendly farming materials does not have a guideline for guaranteeing the effective ingredient or the content, and since the hazard component is within the standard value, it is possible to register the environmentally friendly farming materials in the above two items. Satisfaction.

2. Microbiological tests

Effective microbial test and pathogenic microorganism test contained in the functional liquid fertilizer product (trade name: FUKUSHI-BL) according to the present invention were submitted to Mokpo National University Institute of Microbiology and Ecology and received a test report. Identification of effective microorganisms and identification of viable microorganisms were carried out by sequencing analysis of 16S rRNA gene sequences. Bacillus subtilis subsp . subtilis (99% similar to the data obtained from Donga University), and the population was in agreement with the effective microorganism of the prototype. The population of the sample was 2.4 × 10 ⁸ CFU / ㎖, which was quantified as 10 ⁸ CFU / ㎖ (Refer to the test report of effective microorganism in Fig. 11) and satisfied the quantitative index of the number of effective microorganisms.

The pathogenic microorganism test results contained in the functional liquid fertilizer product of the present invention (trade name: FUKUSHI-BL) were examined by pathogenic Escherichia coli not detected, pathogenic Salmonella not detected, S. aureus not detected, Listeria monocytogenes not detected, and Bacillus cereus not detected, And met the quantitative index of non-detection of pathogenic microorganisms (see the test results of pathogenic microorganisms in FIG. 12).

3. Ineffective and comparative inspection

The non-fermentation test was conducted at Mokwon University's microbiological ecological resources research center on the growth and yield of lettuce for the functional livestock product of the present invention (trade name: Pungjak-BL), and compared with 453-4 Pyeongchon-dong, Seo-gu, Daejeon House from September 08, 2015 to October 21, 2015.

The results showed that the growth pattern of lettuce was slightly improved, the yield of lettuce was improved by 7 ~ 10%, and there was no difference between lettuce and lettuce. Therefore, it is considered that the list of environmentally friendly agricultural materials is possible, and it is considered that the microbial pesticide function should be further tested (refer to the test report of the ineffectiveness and the ineffectiveness in FIG. 13).

Claims (5)

The present invention relates to an amino acid liquid fertilizer obtained by enzymatic treatment and fermentation of tuna processing residues,
Wherein the liquid fertilizer is produced by adding a proteolytic microorganism, a plant pathogenic antibacterial agent, a plant lactic acid bacterium, molasses, and whole milk powder to ferment the tuna processing residue, and producing the fermented amino acid liquid fertilizer Way.
The method according to claim 1,
When preparing the liquid fertilizer, 2% by weight of each of proteolytic microorganism, phytopathogenic microorganism, and plant lactic acid bacteria;
3% by weight of molasses,
And adding 0.1 wt% of powdered milk to ferment the tuna processing residue. The method for producing amino acid liquid fertilizer according to claim 1,
3. The method according to claim 1 or 2,
Wherein the proteolytic microorganism is Bacillus sutillus LH1. The method for producing amino acid liquid fertilizer by enzymatic treatment and fermentation of tuna processing residues.
3. The method according to claim 1 or 2,
Wherein the fermentation process is carried out by introducing 7 to 8 L / min of pure oxygen. The method for producing amino acid liquid fertilizer according to claim 1,
In the amino acid liquid fertilizer which has been subjected to the enzymatic treatment and fermentation of the tuna processing residue,
Which is a liquid fertilizer obtained by fermenting a tuna by-product, Bacillus sutillus LH1 (Accession No: KCTC13030BP), a proteolytic microorganism, a plant pathogenic bacterium, a plant lactic acid bacterium, molasses, whole milk powder, Liquid fertilizer. "

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