KR102308229B1 - Liquid fertilizer composition capable of cultivation crops having anti-obesity, anti-cancer, anti-diabetes and immunity enhancing effects including Rhizopus bacterium and chitosan and saponin and animality amino acid, and method for preparing the same - Google Patents

Abstract

The present invention relates to a liquid fertilizer composition comprising a Ryzopus sp. strain culture, a chitosan decomposition strain culture, an animal amino acid fermented product, and a saponin solution, and a preparation method therefor. The liquid fertilizer composition according to the present invention provides effects such as promoting the growth of crops, controlling pests and diseases, improving sugar content, and increasing the content of crude saponin in crops. Furthermore, crops grown using the liquid fertilizer composition provide anti-obesity, anti-cancer, anti-diabetic and immune-enhancing effects to consumers.

Description

A liquid fertilizer composition capable of cultivating crops having anti-obesity, anti-cancer, anti-diabetic and immune-enhancing effects, including Ryzopus sp. strain and chitosan, saponin and animal amino acids, and a manufacturing method thereof {Liquid fertilizer composition capable of cultivation crops having anti-obesity , anti-cancer, anti-diabetes and immunity enhancing effects including Rhizopus bacterium and chitosan and saponin and animality amino acid, and method for preparing the same}

The present invention relates to a liquid fertilizer composition comprising a Ryzopus sp. strain, chitosan, saponin, and animal amino acids, and a method for producing the same, wherein the crop grown using the same is a liquid liquid providing anti-obesity, anti-cancer, anti-diabetic and immune-enhancing effects It relates to a fertilizer composition and a method for preparing the same.

Background to the present disclosure is provided herein, which does not necessarily imply known art.

Korean Patent 10-0992213 (Jan. 29, 2010) provides a method for producing low-molecular-weight chitosan using Ryzopus and a method for producing chitosan-containing plants using the same. Korean Patent Laid-Open No. 10-2013-0073265 (2013.07.03.) provides a method for cultivating fruits and vegetables using a fermented chitooligosaccharide ratio. In these documents, the bacteria of the genus Ryzopus are used only for the purpose of decomposing chitosan.

Chitosan (chitosan) is a component of the shells of crustaceans such as crayfish, crab, and shrimp, and has antibacterial and immune actions, and thus is widely used in the medical field. In the field of industry, chitosan is known to improve resistance to soil pathogens and to promote growth. However, chitosan has a high molecular weight and is not easily absorbed by plants, so it is necessary to improve the plant growth effect by chitosan.

Saponin is a glycoside of steroids, steroid alkaloids, or triterpenes, and is a generic term for substances that dissolve in water and exhibit a soapy foaming action. In particular, the saponins contained in ginseng are called ginsenosides. Until now, as pharmacological effects of ginseng saponin (ginsenoside), it has an action on the central nervous system, anticancer action, immune function regulation action, antidiabetic action, liver function strengthening action, cardiovascular disorder improvement action, blood pressure control action, menopausal disorder improvement action, Anti-stress, anti-fatigue, and antioxidant activity have been reported. Based on the various pharmacological effects of these ginseng saponins, ginseng is recognized as a functional health food. In addition, as interest in saponins increases, research on saponins in various plants other than ginseng is also in progress. So far, it has been found that beans, green onions, deodeok, bellflower, water parsley, garlic, onion, reishi mushroom, ginkgo biloba, arrowroot, etc. are plants containing saponins, and the pharmacological effects of saponins present in these plants have been reported. Accordingly, a number of studies for cultivation of plants with increased saponin content are also being conducted.

On the other hand, livestock blood is a livestock by-product produced from a slaughterhouse and contains about 8 albumin and globulin in the plasma part and up to 28 globin in the blood cell part. However, in Korea, this livestock blood is not used as a resource, but is mostly discarded, causing problems not only as a waste of useful resources but also as a major source of pollution in the environment. In Korea, the annual number of livestock slaughtered was 1,125,000 cattle and 10,738,000 pigs in 1997. In addition, the disposal of such slaughter blood only increased by 1.1 times in 7 years until 1980 based on 1973, but 1.9 times in 1985, 2.1 times, 2.5 times in 1990, 1992, 1994, and 96, respectively. It has increased by 2.9 times and 3.2 times, and the rate of increase continues to rise, making the situation even more serious.

On the other hand, developed countries have continued to study the production of various useful substances using the waste blood of livestock, and some are commercialized and supplied as products, and Korea is importing them in large quantities. Currently, livestock blood is mainly used as animal protein-enriched feed, which is supplied as dried and powdered blood meal by collecting the blood of livestock from slaughterhouses. It is used as a functional food material, pharmaceutical material, etc.

The present invention is to provide a liquid fertilizer composition capable of cultivating crops having anti-obesity, anti-cancer, anti-diabetic and immune-enhancing effects, including Ryzopus sp. strain, chitosan, saponin, and animal amino acids obtained from livestock blood, and a method for preparing the same.

However, the objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned will be clearly understood by those skilled in the art from the following description.

The present invention Rhizopus genus (Rhizopus) strain culture 0.01 parts by weight to 20 parts by weight; 0.01 parts by weight to 20 parts by weight of chitosan-decomposed strain culture; 0.01 parts by weight to 20 parts by weight of fermented animal amino acids; and 0.01 parts by weight to 10 parts by weight of a saponin solution; containing, sugar, potassium sorbate, sodium sulfite, ethylenediaminetetraacetic acid (EDTA), boron, zinc sulfate, phytoncide, green tea extract It provides a liquid fertilizer composition comprising at least one selected from the group consisting of , natural essential oil, and amino acids.

In addition, the fermented animal amino acid contains 80 to 90 parts by weight of livestock blood, 8 to 13 parts by weight of a protease, and 3 to 4.5 parts by weight of an additive, and the protease includes bromelain, chymotrypsin and papain, The additive is characterized in that it comprises onion skin powder and mushroom powder.

In addition, the present invention prepares a raw material that performs the steps of preparing a chitosan-degraded strain culture, preparing a Ryzopus strain culture, preparing an animal amino acid ferment, and preparing a saponin solution in any order, respectively. to do; obtaining a culture solution by culturing a mixture of the chitosan-decomposed strain culture, the Ryzopus strain culture, the animal amino acid ferment, the saponin solution and sugar; and adding at least one selected from the group consisting of potassium sorbate, sodium sulfite, EDTA, boron, zinc sulfate, phytoncide, green tea extract, natural essential oil, and amino acids to the culture medium and stirring; A method of manufacturing is provided.

In addition, the step of preparing the animal amino acid fermented product includes adding a primary protease containing bromelain and chymotrypsin to livestock blood obtained during the slaughtering process and performing primary fermentation to obtain a primary fermented product, and the It is characterized in that it comprises the step of obtaining a secondary fermented product by adding an additive including a secondary protease containing bromelain and papain and an onion skin powder and mushroom powder to the primary fermented product and performing secondary fermentation .

In addition, in the step of obtaining the primary fermented product, 3 to 5 parts by weight of a primary protease is added with respect to 100 parts by weight of the livestock blood, and bromelain and chymotrypsin are used as the primary proteolytic enzymes 1:3 It is used by mixing in a weight ratio of ~5, and the primary fermentation is characterized in that the step is performed at 70 to 80 °C for 30 to 60 minutes.

In addition, in the step of obtaining the secondary fermented product, 5 to 10 parts by weight of a secondary protease and 3 to 5 parts by weight of an additive are added with respect to 100 parts by weight of the primary fermented product, and broiled with the secondary protease. Melin and papain are mixed in a weight ratio of 1:0.3 to 0.5, and onion peel powder and mushroom powder are mixed and used in a weight ratio of 1:0.1 to 0.3 as the additive, and the secondary fermentation is 1 at 30 to 40° C. It is characterized in that the step is carried out for 2 hours.

The present invention provides a liquid fertilizer composition comprising chitosan and saponin fermented with Ryzopus sp. strain, and an animal amino acid obtained from livestock blood, and a method for manufacturing the same, so that liquid fertilizer can be conveniently and mass produced, and using the same In the case of cultivating crops, it not only provides the effect of improving the growth effect of crops, controlling pests and diseases, improving sugar content, and increasing the saponin content in crops, but also has anti-obesity, anti-cancer, anti-diabetic and immune-enhancing effects when ingesting crops grown using it. to provide.

1 is a schematic diagram showing a manufacturing process of a liquid fertilizer composition according to an embodiment of the present invention.

All technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art, unless otherwise stated. Also throughout this specification and claims, unless stated otherwise, the term comprise, comprises, comprising is meant to include the recited object, step or group of objects, and steps, and any other It is not used in the sense of excluding an object, step, or group of objects or groups of steps.

Prior to describing the present invention in detail below, it is to be understood that the terminology used herein is for the purpose of describing specific embodiments and is not intended to limit the scope of the present invention, which is limited only by the appended claims. shall.

On the other hand, various embodiments of the present invention may be combined with any other embodiments unless clearly indicated to the contrary. Any feature indicated as particularly preferred or advantageous may be combined with any other feature and features indicated as preferred or advantageous. Hereinafter, embodiments of the present invention and effects thereof will be described with reference to the accompanying drawings.

Liquid fertilizer composition according to an embodiment of the present invention is Rhizopus genus (Rhizopus) strain culture 0.01 parts by weight to 20 parts by weight, chitosan decomposed strain culture 0.01 parts by weight to 20 parts by weight, animal amino acid fermentation 0.01 parts by weight to 20 parts by weight and 0.01 parts by weight to 10 parts by weight of saponin solution, sugar, potassium sorbate, sodium sulfite, EDTA (ethylenediaminetetraacetic acid), boron, zinc sulfate, phytoncide, green tea It provides a liquid fertilizer composition comprising at least one selected from the group consisting of extracts, natural essential oils, and amino acids.

The Ryzopus sp. strain culture is about 0.1 parts by weight to about 15 parts by weight, about 0.5 parts by weight to about 10 parts by weight, about 1 part by weight to about 7 parts by weight, about 1 part by weight to about 5 parts by weight, or about 1 part by weight to about 3 parts by weight of Ryzopus sp. strain and about 10 parts by weight to about 99.9 parts by weight, about 20 parts by weight to about 99 parts by weight, about 30 parts by weight to about 98 parts by weight, about 40 parts by weight to about 97 parts by weight parts by weight, or from about 50 parts by weight to about 97 parts by weight soybean meal.

The Rhizopus strain is a fungus belonging to Zygomycota, Mucoromycotina, Mucorales, and Mucoraceae. The Rhizopus sp. strain is R. caespitosus (caespitosus), R. delemari (delemari), R. homo tallicus (homothallicus), R. microsporus (microsporus), R. duck jae (oryzae), R. reflexus, R. skipperae, or R. stolonifer. For example, the Rhizopus sp. strain is Rhizopus oryzae.

The chitosan may be obtained by pulverizing the shells of crustaceans or by treating with chemicals such as alkali. The chitosan may be a liquid chitosan raw material. The chitosan-decomposed strain culture is a fermented product of 0.1 parts by weight to 15 parts by weight of chitosan, 0.1 parts by weight to 15 parts by weight of chitosan-decomposed strains, 0.1 parts by weight to 20 parts by weight of molasses, and 0.01 parts by weight to 99.7 parts by weight of rice bran. include

The chitosan-decomposing strain is selected from bacteria consisting of yeast, Bacillus subtilis, and lactic acid bacteria. Preferably, it is good to include Bacillus subtilis, and more preferably contains Bacillus subtilis derived from soybean paste, cheonggukjang or kimchi.

The saponin solution is a liquid raw material capable of increasing the content of saponin by 0.5 mg/g or more in crops grown by containing 0.5% or more of ginseng extract. The saponin solution includes 0.5 parts by weight to 1 part by weight of ginseng extract, 30 parts by weight to 50 parts by weight of 1,3-butanediol, and 50 parts by weight to 70 parts by weight of purified water.

The animal amino acid fermented product is a raw material that assists cultivated crops to contain active ingredients to have anti-obesity, anti-cancer, anti-diabetic and immune-enhancing effects, and is obtained by processing blood generated during blood slaughter of livestock, especially pigs. lose

The fermented animal amino acid includes 80 to 90 parts by weight of livestock blood, 8 to 13 parts by weight of a protease, and 3 to 4.5 parts by weight of an additive. More specifically, 80 to 90 parts by weight of livestock blood, 4 to 6 parts by weight of bromelain, 3 to 4 parts by weight of chymotrypsin, 1 to 3 parts by weight of papain, 2.5 to 3.5 parts by weight of onion skin powder, and 0.5 to 1 parts by weight of mushroom powder includes wealth.

The liquid fertilizer composition is preferably 0.1 parts by weight to 10 parts by weight of Ryzopus sp. strain culture, 0.1 parts by weight to 10 parts by weight of chitosan decomposing strain culture, 0.05 parts by weight to 5 parts by weight of saponin solution, and 0.1 parts by weight of animal amino acid fermented product. It is preferable to include 10 parts by weight to 10 parts by weight.

The liquid fertilizer composition is more preferably 0.5 parts by weight to 2 parts by weight of Ryzopus sp. strain culture, 0.5 parts by weight to 2 parts by weight of chitosan decomposed strain culture, 0.25 parts by weight to 1 part by weight of saponin solution, and animal amino acid fermented product It is preferable to include 0.5 to 2 parts by weight.

The liquid fertilizer composition may include about 0.01 parts by weight to about 10 parts by weight, about 0.05 parts by weight to about 5 parts by weight, about 0.1 parts by weight to about 2 parts by weight, or about 0.25 parts by weight to about 1 part by weight of sugar. have. The sugar may be sugar.

The liquid fertilizer composition may include about 0.01 parts by weight to about 20 parts by weight, about 0.1 parts by weight to about 10 parts by weight, or about 0.5 parts by weight to about 2 parts by weight of potassium sorbate.

The liquid fertilizer composition may comprise about 0.1 parts by weight to about 20 parts by weight, about 0.5 parts by weight to about 15 parts by weight, about 1 parts by weight to about 10 parts by weight, or about 2 parts by weight to about 5 parts by weight of sodium sulfite. can

The liquid fertilizer composition may include about 0.01 parts by weight to about 20 parts by weight, about 0.1 parts by weight to about 10 parts by weight, or about 0.5 parts by weight to about 2 parts by weight of EDTA.

The liquid fertilizer composition is about 0.01 parts by weight to about 10 parts by weight, about 0.01 parts by weight to about 5 parts by weight, about 0.05 parts by weight to about 2 parts by weight, about 0.1 parts by weight to about 15 parts by weight, about 0.15 parts by weight to about 1 part by weight, or about 0.2 parts by weight to about 0.5 parts by weight of boron.

The liquid fertilizer composition is about 0.01 parts by weight to about 10 parts by weight, about 0.01 parts by weight to about 5 parts by weight, about 0.05 parts by weight to about 2 parts by weight, about 0.1 parts by weight to about 1.5 parts by weight, about 0.15 parts by weight to about 1 part by weight, or about 0.2 parts by weight to about 0.5 parts by weight of zinc sulfate.

The liquid fertilizer composition is about 0.01 parts by weight to about 10 parts by weight, about 0.01 parts by weight to about 5 parts by weight, about 0.05 parts by weight to about 2 parts by weight, about 0.1 parts by weight to about 1.5 parts by weight, about 0.15 parts by weight to It may contain about 1 part by weight, or about 0.2 parts by weight to about 0.5 parts by weight of phytoncide, green tea extract, and natural essential oil.

The liquid fertilizer composition is about 0.5 parts by weight to about 60 parts by weight, about 1 part by weight to about 50 parts by weight, about 1.5 parts by weight to about 40 parts by weight, about 2 parts by weight to about 30 parts by weight, about 2.5 parts by weight to about 20 parts by weight, about 3 parts by weight to about 10 parts by weight, or about 4 parts by weight to about 10 parts by weight of the amino acid.

The amino acid is derived from the group consisting of crab, molasses, and microorganisms, and more specifically, 55 to 65 parts by weight of blue crab, 15 to 25 parts by weight of molasses, and 15 to 25 parts by weight of useful microorganisms (KEM). and use the blue crab amino acid fermented product fermented and aged by crushing and stirring.

The liquid fertilizer composition may contain an appropriate amount of water.

In another aspect of the present invention, the manufacturing method of the liquid fertilizer composition comprises fermenting a mixture of chitosan, chitosan-degraded strain, molasses and grain to prepare a chitosan-decomposed strain culture (S11), Ryzopus sp. strain is complexed with white cocci A step of fermenting the immunized grain to prepare a Ryzopus strain culture (S12), adding a protease and an additive to livestock blood, and then fermenting it to prepare an animal amino acid fermented product (S13) and containing a ginseng extract Preparing a raw material including the step of preparing a saponin solution (S14) (S10); The chitosan-decomposed strain culture, the Ryzopus strain culture, the animal amino acid ferment, the saponin solution and the mixture of sugar are cultured for 10 minutes to 48 hours to obtain a culture solution (S20); And adding at least one selected from the group consisting of potassium sorbate, sodium sulfite, EDTA, boron, zinc sulfate, phytoncide, green tea extract, natural essential oil, and amino acids to the culture medium and stirring for 10 minutes to 48 hours (S30) ), including, provides a method for preparing a liquid fertilizer composition.

The step of preparing the raw material (S10) is a step of preparing a chitosan-degraded strain culture (S11), a step of preparing a Ryzopus strain culture (S12), a step of preparing an animal amino acid fermented product (S13) and a saponin solution The step of preparing ( S14 ) may be performed independently of each order.

Fermentation in the step (S11) of preparing the chitosan decomposing strain culture is about 15 ℃ to about 40 ℃, about 20 ℃ to about 40 ℃, about 25 ℃ to about 40 ℃, about 30 ℃ to about 40 ℃, or about It may be carried out at 35°C to about 40°C. Also, the fermentation is performed for about 1 hour to about 1 week, about 6 hours to about 6 days, about 12 hours to about 5 days, about 18 hours to about 4 days, about 1 day to about 3 days, or about 2 days. can be The fermentation may be performed while stirring.

The method may further include the step of preparing a chitosan preparation by drying the fermented chitosan decomposing strain culture. The drying method may use a method known in the art, such as natural drying, hot air drying, freeze drying, and the like. The dried mixture has an average particle size of about 10 to about 1000 mesh, about 20 to about 900 mesh, about 30 to about 800 mesh, about 40 to about 700 mesh, about 50 to about 600 mesh, about 60 to about 500 mesh. mesh, from about 60 to about 400 mesh, or from about 60 to about 300 mesh. The pulverized mixture can be packaged and used as a chitosan formulation.

Fermentation in the step (S12) of preparing the Ryzopus strain culture is about 20 ℃ to about 60 ℃, about 25 ℃ to about 60 ℃, about 30 ℃ to about 60 ℃, about 35 ℃ to about 60 ℃, about 40 ℃ to about 55 ℃, or from about 45 ℃ to about 50 ℃. Also, the fermentation may be performed for about 1 hour to about 1 month, about 12 hours to about 4 weeks, about 1 day to about 3 weeks, about 2 days to about 2 weeks, about 3 days to about 10 days, about 4 days to about 10 days. days, from about 5 days to about 10 days, from about 6 days to about 10 days, or from about 1 week to about 10 days.

The method may further include the step of drying the fermented Ryzopus strain culture to prepare a Ryzopus preparation. The drying method may use a method known in the art, such as natural drying, hot air drying, freeze drying, and the like. The dried mixture has an average particle size of about 10 to about 1000 mesh, about 20 to about 900 mesh, about 30 to about 800 mesh, about 40 to about 700 mesh, about 50 to about 600 mesh, about 60 to about 500 mesh. mesh, from about 60 to about 400 mesh, or from about 60 to about 300 mesh. The pulverized mixture can be packaged and used as a Ryzopus preparation.

The step of preparing the animal amino acid fermented product (S13) is to obtain a primary fermented product by adding a primary protease containing bromelain and chymotrypsin to livestock blood obtained during the slaughtering process and performing primary fermentation (S131) ) and adding an additive including a secondary protease containing bromelain and papain and an onion peel powder and mushroom powder to the primary fermented product and performing secondary fermentation to obtain a secondary fermented product (S132) include Through the secondary fermentation as described above, it is possible to ferment within a shorter time than the conventional fermentation.

In the step of obtaining the primary fermented product (S131), 3 to 5 parts by weight of a primary protease is added with respect to 100 parts by weight of livestock blood, and bromelain and chymotrypsin are used as the primary proteolytic enzymes in 1:3~ It is used by mixing in a weight ratio of 5. In addition, in the step of obtaining the primary fermented product (S131), the fermentation is performed at 70 to 80° C. for 30 to 60 minutes.

In the step of obtaining the secondary fermented product (S132), 5 to 10 parts by weight of a secondary protease and 3 to 5 parts by weight of an additive are added with respect to 100 parts by weight of the primary fermented product, and broiled with a secondary protease. Melin and papain are mixed in a weight ratio of 1:0.3 to 0.5 and used as an additive by mixing onion skin powder and mushroom powder in a weight ratio of 1:0.1 to 0.3. In addition, in the step (S132) of obtaining the secondary fermented product, the fermentation may be performed at 30 to 40° C. for 1 to 2 hours.

The method may further include the step of preparing an animal amino acid preparation by stirring, cooling and filtering the secondary fermentation product of animal amino acid. More specifically, the animal amino acid fermentation product is homogeneously stirred using a homomixer, and the stirred fermentation product is indirectly cooled through a cooling jacket, and insoluble or crystallized impurities are filtered out. The filtered fermented product can be packaged and used as an animal amino acid preparation.

The step of obtaining the culture solution (S20) is a mixture of the chitosan-decomposed strain culture or chitosan preparation, the rhizopus strain culture or the ryzopus preparation, the animal amino acid ferment or the animal amino acid preparation, the saponin solution and the sugar for 10 minutes to 48 hours. As the culturing step, it is preferably cultured for about 1 hour to about 42 hours, about 2 hours to about 36 hours, about 6 hours to about 30 hours, or about 12 hours to about 24 hours. The incubation may be performed while stirring.

In the stirring step (S30), at least one selected from the group consisting of potassium sorbate, sodium sulfite, EDTA, boron, zinc sulfate, phytoncide, green tea extract, natural essential oil, and amino acids is added to the culture medium, and 10 minutes to 48 minutes As the step of stirring for an hour, it is preferably stirred for about 30 minutes to about 24 hours, about 1 hour to about 12 hours, about 1.5 hours to about 6 hours, or about 2 hours to about 4 hours.

<Example>

Hereinafter, it will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

(1) Preparation of chitosan formulation

A spawn was prepared by mixing chitosan liquid raw material (original fermentation industry, chitosan solution), microorganisms (Bacillus subtilis derived from soybean paste and kimchi-derived Bacillus subtilis), and purified water. The prepared seed was adsorbed to rice bran (Io Technology, rice bran powder), and molasses (Samsung feed, molasses solution) was added and fermented at about 35° C. for about 48 hours. The fermented mixture was then dried for about 72 hours. The dried mixture was pulverized to an average particle size of 60 to 300 mesh using a pulverizer. The pulverized mixture was packaged and used as a chitosan formulation.

The raw materials and input ratio of the chitosan formulation are shown in Table 1 below.

Raw material name Content rate by ingredient (%) molasses 2 non-compulsive 95 chitosan 1.5 microbe 1.5 Sum 100

(2) Preparation of Ryzopus formulation

Rhizopus oryzae (Rhizopus oryzae) is a type of white cocci, a type of white downy mildew-spider web mold, through complex immunity (mutagenized and activated) to produce biochemical and biochemical and It was cultivated by physicochemical manual process.

The cultured Ryzopus duck material was adsorbed to soybean meal and fermented (neutral fermentation) for about 7 days to about 10 days at a temperature of about 45°C to about 50°C. After fermentation, it was dried naturally rather than artificially.

Thereafter, the dried mixture was pulverized to have an average particle size of 60 to 300 mesh using a pulverizer. The pulverized mixture was packaged and used as a Ryzopus formulation.

Raw materials and input ratios of the Ryzopus formulation are shown in Table 2 below.

Raw material name Content rate by ingredient (%) Soybean Meal 97 rhizopus duck 3 Sum 100

(3) Preparation of animal amino acid preparations

Bromelain and chymotrypsin were added to blood meal obtained by drying pig blood obtained during pig slaughter and completely pulverizing it using a grinder, and fermented at about 75° C. for 45 minutes to obtain a primary fermented product.

Bromelain and papain were added to the primary fermented product, and the onion skin powder and mushroom powder were added and fermented at about 35° C. for 90 minutes to obtain a secondary fermented product.

The secondary fermented product was homogeneously stirred using a homomixer, indirectly cooled through a cooling jacket, and then insoluble or crystallized impurities were filtered and used as an animal amino acid preparation.

The raw materials and input ratios of the animal amino acid preparation are shown in Table 3 below.

Raw material name Content percentage by ingredient (%) Pig blood (blood meal) 85.0 primary proteolytic enzyme bromelain One chymotrypsin 3.5 secondary proteolytic enzyme bromelain 5.0 papain 2.0 additive onion peel powder 3.0 Cordyceps Powder 0.5 Sum 100

(4) Preparation of culture solution

After filling the liquid incubator with tap water in the liquid incubator, it was sterilized by maintaining it at about 35° C. for about 24 hours to remove chlorine from the tap water.

The prepared chitosan preparation, rhizopus preparation, animal amino acid preparation, saponin solution (Ginseng Extract A, Daebong LS Co., Ltd.) and brown sugar were added to the sterilized incubator, and incubated with stirring at about 35°C for about 4 hours.

While maintaining the culture medium at about 35 ℃, potassium sorbate (Kunda, China), sodium sulfite (Buheung Industrial Co., Ltd.), and EDTA (ethylenediaminetetraacetic acid) (Shijazhuang Jackchem, China) were added to the culture solution, The mixture was stirred for about 2 hours or more.

To the stirred mixture, boron (searles valley minnerals, USA) and zinc sulfate (Tianjin Chemical, China) were added and stirred.

After that, the temperature of the mixture is naturally lowered to about 20°C or less, and phytoncide, green tea extract, natural essential oil and amino acids (contains 60% blue crab, 20% molasses, and 20% microorganisms (for fermentation), Narawon Co., Ltd.) was added and stirred for about 2 hours or more.

The preparation raw materials and input ratio of the culture medium are shown in Table 4 below.

Raw material name Content rate by ingredient (%) Chitosan preparation One Ryzopus preparations One animal amino acid preparations One saponin solution 0.5 brown sugar 0.5 sorbic acid One sodium sulfite 2 EDTA One boron 0.25 zinc sulfate 0.25 Phytoncide, green tea extract, natural essential oil 0.25 blue crab amino acids 5 Purified water 86.25 Sum 100

(5) Cultivation of crops using liquid fertilizer composition

Apples were grown in a conventional way using the complex formulation prepared according to the above example as a liquid fertilizer.

<Experimental example>

(1) Confirmation of adipocyte differentiation ability of cultivated apple pulverized product

During the differentiation of preadipocytes into adipocytes, changes in fat accumulation rate and triglyceride (TG) content according to the addition of the pulverized apples grown above were observed after 8 days of differentiation.

At this time, in order to differentiate 3T3-L1 preadipocytes into adipocytes, the preadipocytes were aliquoted in a 24-well plate at ^{a concentration of 5×10 4} cells/well, incubated until the wells were filled, and differentiation was induced after 4 days. While exchanging the medium with DMEM medium containing the substances insulin 5 μg/ml, 1 μM dexamethasone, 0.5 mM isobutylmethylxanthine, and 10% FBS, the crushed apples were prepared by concentration (100, 1000, 2000 μg/ml). added.

Thereafter, the cells were exchanged twice with DMEM medium containing 5 μg/ml of insulin and 10% FBS containing crushed apples grown once every 2 days, and then an experiment on cell differentiation ability was performed.

For the experiment on the differentiation capacity of cells, microscopic observation was performed after Oil red O staining. For this method, after washing with PBS, adipocytes fixed with 10% formalin were stained with Oil red O and observed with an optical microscope. .

In addition, the stained cells were dissolved with isopropyl alcohol and absorbance was measured at 515 nm with a microplate reader (Beckman, USA). The fat content was expressed as a percentage of the average absorbance value of the control group that was not treated with the sample (pulverized product of cultivated apples). The results are shown in Table 5 below.

Crushed Apple Concentration (μg/ml) 0 100 1000 2000 Cell viability
(% of control) 100 106 112 124 liquid accumulation
(OD 515nm) 1.2 0.89 0.81 0.67

As shown in Table 5, the cultivated apple pulverized material exhibited a reduction in fat accumulation of about 25.8% at a concentration of 100 μg/ml. In addition, at the high concentration of 2000 μg/ml, the fat reduction rate was about 44.2%, confirming the inhibition of differentiation into adipocytes.

(2) Confirmation of anticancer effect on ascites cancer of cultivated apple pulverized product

Aseptically collected aseptic sarcoma-180 tumor cells, washed with Hank's balanced salt solution (HBSS) at 4°C, and subcultured into the abdominal cavity of ICR mice at a concentration of ^{1×10 6 cell/ml were used.} The subcultured sarcoma-180 was collected intraperitoneally with pasteur pipets, put in a 50ml conical tube, centrifuged at 1200rpm for 10 minutes, washed 3 times with HBSS buffer, and then used in the experimental example of the present invention.

Stabilize 10 3-4-week-old ICR mice with an average weight of 25.1±0.2 g in an incubator at 24±2°C for 1 week. After administration for a while, 3×10 ⁵ cell/ml of sarcoma-180 was intraperitoneally implanted. After transplantation, a sample of 150-200 mg/kg per day was administered in the same way, and the average survival days (MST) and survival days extension rate (ILS) were calculated as below [Equation] ] was used to determine 30 days after tumor transplantation. The results are shown in Table 6.

[ceremony]

Survival days extension rate (ILS%) = (average number of days of survival in treated group/average number of days of survival in non-administered group) × 100

control administration group 150mg/kg 200mg/kg MST (Mean survival days, days) 19.5 30.1 30.5 ILS (Life Extension Rate, %) - 150 or more 150 or more 30 days survival 0/10 5/10 6/10

As shown in Table 6 above, the average survival days (MST) of the administered group administered the pulverized apple grown using the liquid fertilizer according to the present invention was about 30 days, which was extended by more than 10 days compared to the control group not administered. The survival time extension rate was more than 150%. In addition, it can be seen that mice that survived up to 30 days also exhibited relatively good anticancer activity in rats administered with pulverized apples grown in more than half.

(3) Confirmation of antidiabetic effect of cultivated apple pulverized product

Five-week-old male C57BL/6J mice were randomly assigned to a control group (HFD alone) and a test group (cultivated apple pulverized group), and then acclimatized to the laboratory environment for one week. In the present invention, a high fat diet (HFD) (Product # D12451, Research Diet Inc., New Brunswick, NJ, USA; 40% kcal from fat) was used, so that the cultivated apple pulverized product was 5% by weight. It was evenly mixed with the HFD feed. Both the control group and the test group were fed HFD for 7 weeks, and then for the next 8 weeks, the control group was fed with HFD alone, and the test group was fed with HFD containing cultivated apple pulverized, respectively. At this time, there was no significant difference in the total sample intake between the control group and the test group.

Fasting blood glucose and insulin were measured using serum collected from the tail vein of mice fasted for 12 hours after feeding the sample for 8 weeks. Fasting blood glucose was measured using a blood glucose meter (Handok, Seoul, Korea), and serum insulin was measured using enzyme-linked immunosorbent assays (ELISA). The results are shown in Table 7 below.

control test group Fasting blood sugar (mg/dL) 225.4 177.4 Fasting insulin (ng/mL) 6.00 4.23

As shown in Table 7 above, the fasting blood glucose and fasting insulin of the test group ingested with apple pulverize grown compared to the control group (HFD) were 21.3% (p<0.05) and 29.5% (p<0.01), respectively, compared to the control group. decreased significantly. These results mean that the apple pulverized product grown using the liquid fertilizer composition of the present invention provides an antidiabetic effect.

(4) Confirmation of immunity improvement effect of cultivated apple pulverized product

In order to indirectly confirm the effect of improving immunity of crops grown using the liquid fertilizer composition according to the present invention, the saponin content of cultivated apples was analyzed, and the results are shown in Table 8 below. For comparison of whether the saponin content is increased, apples grown using a combination preparation of chitosan, saponin, and rhizopus according to Patent Registration No. 10-2089777 were used as a control group.

control test group saponin (mg/g) 5.30 7.45

As shown in Table 8, the saponin content of the apples grown with the liquid fertilizer containing the complex preparation according to the present invention was increased by about 40.6%, compared to the apples grown with the liquid fertilizer containing the conventional complex preparation. It was confirmed that the immunity enhancement and anti-cancer effect, which are known for their efficacy, would be improved.

Features, structures, effects, etc. exemplified in each of the above-described embodiments may be combined or modified for other embodiments by those of ordinary skill in the art to which the embodiments belong. Accordingly, the contents related to such combinations and modifications should be interpreted as being included in the scope of the present invention.

Claims (6)

Rhizopus genus (Rhizopus) strain culture 0.01 parts by weight to 20 parts by weight;
0.01 parts by weight to 20 parts by weight of a decomposition product obtained by decomposing chitosan using a chitosan decomposition strain;
0.01 parts by weight to 20 parts by weight of fermented animal amino acids; and
Including; 0.01 parts by weight to 10 parts by weight of saponin solution;
It contains at least one selected from the group consisting of sugar, potassium sorbate, sodium sulfite, ethylenediaminetetraacetic acid (EDTA), boron, zinc sulfate, phytoncide, green tea extract, natural essential oil, and amino acids,
The animal amino acid fermented product is 80 to 90 parts by weight of livestock blood, 4 to 6 parts by weight of bromelain, 3 to 4 parts by weight of chymotrypsin, 1 to 3 parts by weight of papain, 2.5 to 3.5 parts by weight of onion skin powder, and 0.5 parts by weight of mushroom powder. to 1 part by weight of a liquid fertilizer composition.
delete Using a chitosan decomposing strain to prepare a decomposed product obtained by decomposing chitosan, preparing a Ryzopus strain culture, preparing an animal amino acid fermented product, and preparing a saponin solution in any order. preparing;
Obtaining a culture solution by culturing a mixture of the chitosan-decomposed product, the Ryzopus strain culture, the animal amino acid fermented product, the saponin solution, and sugar using the chitosan-decomposed strain; and
Adding at least one selected from the group consisting of potassium sorbate, sodium sulfite, EDTA, boron, zinc sulfate, phytoncide, green tea extract, natural essential oil, and amino acids to the culture medium and stirring;
The step of preparing the animal amino acid fermented product,
adding 3 to 5 parts by weight of a primary protease to 100 parts by weight of livestock blood obtained in the slaughter process and performing primary fermentation to obtain a primary fermented product;
and adding 5 to 10 parts by weight of a secondary protease and 3 to 5 parts by weight of additives to 100 parts by weight of the primary fermented product and performing secondary fermentation to obtain a secondary fermented product,
The primary protease is used by mixing bromelain and chymotrypsin in a weight ratio of 1:3 to 5,
Bromelain and papain as the secondary proteolytic enzyme are mixed in a weight ratio of 1:0.3 to 0.5 and used.
As the additive, onion peel powder and mushroom powder are mixed and used in a weight ratio of 1:0.1 to 0.3.
A method for preparing a liquid fertilizer composition.
delete 4. The method of claim 3,
The primary fermentation is a method for producing a liquid fertilizer composition, characterized in that the step is performed for 30 to 60 minutes at 70 to 80 ℃.
4. The method of claim 3,
The secondary fermentation is a method for producing a liquid fertilizer composition, characterized in that the step is performed for 1 to 2 hours at 30 to 40 ℃.

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