KR20200037468A - Method for composite liquid fertilizer by microbial mass production having improved durability for microbial with citric acid - Google Patents

Abstract

The present invention relates to a liquid fertilizer production method, and more specifically, to a liquid fertilizer for high-throughput production comprising composite microorganisms, and a method of producing the same, wherein a medium for an initial culture of microorganisms is a batch culture utilizing disaccharides as a carbon source; through a mixture multi-step introduction and mixing step, an initial culture medium establishing step is set forth for enhancing survival durability of effective microorganisms; a medium for high-throughput production of microorganisms is a fed-batch culture utilizing disaccharides as a carbon source; and through a mixture multi-step introduction and stirring step, the survival durability of effective microorganisms can be enhanced using citric acid. The liquid fertilizer of the present invention is a convenient liquid type fertilizer composition that has long-lasting effects for an extended period even after a single application; can activate microorganisms by means of organic matters; and can be used for folial applications, nutrient solution cultivation, irrigation applications, and for flowers. The liquid fertilizer can provide increased absorption and migration of phosphoric acid within plant nutrient channels, can increase a calcium content in fruits during a harvest season; can increase the quality, color, and sugar content of the fruits; and can prevent softening and rotting of the fruits during storage. In the high-throughput production of microorganisms using citric acid according to the present invention, phosphate-solubilizing bacteria can form spores, and thus have high viability, and moreover, the bacteria have excellent phosphate-solubilizing ability as well as excellent antibacterial ability and protein-degrading ability, and thus, when a soil treatment agent comprising the same is used, insoluble phosphates can be solubilized, thereby addressing salt injury of soil, can supply phosphoric acid to crops, and can control plant diseases. By breaking down organic matters, the growth of crops can be promoted through supply of nutrients.

Description

Method for composite liquid fertilizer by microbial mass production having improved durability for microbial with citric acid}

The present invention relates to a liquid microorganism fertilizer for mass production of a composite microorganism composed of lactic acid bacteria and phosphate solubilizing bacteria, and to strengthen the survival durability of microorganisms through citric acid.

The complex fertilizer guarantees at least two or more of the three elements of fertilizer, nitrogen, phosphoric acid, and girly, and is the most commonly used fertilizer as a general term for multi-component fertilizers containing trace elements depending on the species. In the fertilizer process standard, it is classified into first, second, third, fourth, and slow-acting complex fertilizers according to the manufacturing method, main use, and guaranteed ingredients. Particularly, the fourth type fertilizer is a liquid composite fertilizer composition, which is difficult to absorb nutrients from the roots of plants, or increases the taste, color, and marketability of fruit trees and vegetables. It is categorized into cultivation of nutrient solution to supply nutrients, and for irrigation and flowers for fertilizing the soil with fertilizer. As such, liquid fertilizer, that is, liquid fertilizer, can be regarded as a fast-acting fertilizer, and can promote the growth of crops through irrigation or fertilizing on a leaf surface.

In Korean Patent Application No. 10-2000-0044502, acids such as nitric acid or phosphoric acid are added together with water to hydrolyze proteins to a solid organic material such as animal carcasses, slaughter by-products or livestock manure, fish, etc., After adding chymotrypsin, which is a proteolytic enzyme and which hydrolyzes ester bonds, an organic liquid fertilizer is provided, which is characterized by liquefying a workpiece by pressure heating, and in Korean Patent Application No. 10-2001-0006610, It relates to a liquid fertilizer that prevents acidification and neutralizes acidified soil, and a method for manufacturing the same. The method of preparing liquid fertilizer comprises preparing water, fish meal, bone meal, blood meal, rice cake, molasses, crab shells and microorganisms, and mixing water, fish meal, bone meal, blood meal, rice cake, molasses, crab shells and microorganisms at a predetermined ratio. Liquid fertilizer is provided, which includes the steps of fermenting mixed water, fish meal, bone meal, blood meal, fish cake, molasses, crab shells and microorganisms to produce liquid fertilizer, and in Korean Patent Application No. 10-2003-0003832 Liquid fertilizer for adding fermentation bacteria after dissolving germanium powder in natural germanium mineral water is provided,

In Korean Patent Application No. 10-2003-0040883, the liquid composite fertilizer containing nano silver and allicin and its manufacturing method and antibacterial activity are emphasized. In Korean Patent Application No. 10-2007-0131169, the liquid composite fertilizer composition and the same are used. As a plant cultivation method, it is present as a part of the liquid composite fertilizer composition with simple addition of citric acid, not for the purpose of cultivating microbial oil or solubilizing phosphoric acid, and in the case of the above-mentioned existing invention, it is used for various purposes and purposes of liquid fertilizer, animal carcasses. It is used in various ways such as utilization of solid organic substances such as fishmeal & bone meal & blood meal, utilization of germanium powder, utilization of nano-silver and allicin, utilization of liquid fertilizer, etc., but strengthens microbial survival durability through citric acid as in the present invention. And liquid lactic acid fertilizer consisting of lactic acid bacteria and phosphate solubilizing bacteria For cases that can not see find.

Also, as citric acid-related prior art, Korean Patent Registration No. 10-1999-0039110 discloses a method for manufacturing oral urinary tract stones using citric acid, and Korean Registered Patent No. 10-2002-0029697 discloses citric acid, zinc, and albumin. Characterized by a method for producing a beverage composition containing a, and Republic of Korea Patent Registration No. 10-2005-0014848 is characterized by a composition for the prevention and treatment of swelling filigree containing citric acid, zinc and arginine as active ingredients In Korea Patent Registration No. 10-2009-0007719, it is disclosed that it is characterized by a method for stabilizing a polluted soil site using a calcium citrate phosphate soil stabilizer. As such, the use of citric acid is widely used as a urinary tract stone, beverage composition, expansion ancestor treatment composition, soil pollution stabilizer, etc., but also consists of lactic acid bacteria and phosphate solubilizing bacteria as well as the present invention. There are no cases where complex microbial mass-producing liquid fertilizers and their manufacturing methods are used.

Republic of Korea Patent Application No. 10-2000-0044502 Republic of Korea Patent Application No. 10-2001-0006610 Republic of Korea Patent Application No. 10-2007-0131169 Republic of Korea Patent Application No. 10-2003-0003832 Republic of Korea Patent Application No. 10-2003-0040883 Republic of Korea Patent Registration No. 10-1999-0039110 Republic of Korea Patent Registration No. 10-2002-0029697 Republic of Korea Patent Registration No. 10-2005-0014848 Republic of Korea Patent Registration No. 10-2009-0007719

The present invention relates to a liquid microorganism fertilizer for mass production of a composite microorganism composed of lactic acid bacteria and phosphate solubilizing bacteria, and to strengthen the survival durability of microorganisms through citric acid.

In order to achieve the above object, the present invention is a method for preparing a liquid fertilizer, the initial culture of microorganisms is applied as a carbon source as a batch culture method, and the mixture is subjected to a multi-stage injection and agitation step to increase the survival durability of effective microorganisms. Create medium establishment stage, mass production medium of microorganism applies disaccharide as carbon source as oil price cultivation method, multi-stage injection and agitation step of mixture to utilize citric acid to enhance survival durability of effective microorganisms and mass of complex microorganisms composed of lactic acid bacteria and phosphate soluble bacteria A liquid fertilizer produced and a method of manufacturing the same are provided.

It is a convenient liquid form fertilizer composition that can be used for both fertilization, nutrient culture, irrigation, and flowers for foliar fertilization, nutrient cultivation, and fertilization for a considerable period of time even after fertilization for a long time. Promotes absorption and migration of phosphoric acid, increases the calcium content of fruits during harvest, increases fruit quality, improves coloration and sugar content, provides fruit softening prevention and anti-corrosion effects during storage, and mass production of microorganisms using citric acid in the present invention. Phosphoric acid soluble bacteria caused by the formation of spores have good viability and excellent phosphate solubility, but also excellent antibacterial and protein decomposition, so when using a soil treating agent containing it, solubilize poorly soluble phosphate to solve the salt disturbance of the soil and phosphate to the crop In addition to supplying nutrients, it is possible to supply nutrients by controlling plant diseases and decomposing organic substances. It can be expected to accelerate the growth of crops.

In the present invention, in the liquid fertilizer manufacturing method,

The initial culture medium for microorganisms is a batch culture method, and disaccharides are applied as a carbon source. Through the multi-stage infusion and agitation steps, an initial culture medium is established to enhance the survival durability of effective microorganisms.

The microbial mass production medium applies disaccharides as a carbon source as a fed-batch culture method, and uses multi-stage infusion and agitation steps to utilize citric acid to enhance the survival durability of effective microorganisms and to produce liquid fertilizers for mass production of complex microorganisms composed of lactic acid bacteria and phosphate solubilizing bacteria. As providing a manufacturing method,

Hereinafter, the configuration and operation of the present invention will be described in detail.

Phosphorus (phosphorus) is an essential nutrient that is very important for plant growth and development. Therefore, in order to increase the yield of major crops in agriculture, the presence or absence of phosphorus available to plants is a very important condition. However, most of the phosphorus present in the soil exists as an insoluble phosphate that cannot be used at all by plants because it exists in a state such as a chemically firmly bound insoluble complex or an organically bound phytate. Therefore, it is necessary to supply inorganic phosphate fertilizer in order to prevent the growth of plants. However, even if an inorganic phosphate fertilizer is supplied, more than 75% of the supplied fertilizer is very rapidly combined with iron, aluminum and calcium ions in alkali and acidic soils to be converted into a form of insoluble phosphate that cannot be used by plants. Accordingly, excessive fertilization is repeated in which excessive inorganic phosphate fertilizer is supplied to the soil to improve the productivity of the crop.

The excessive supply of phosphoric acid as described above causes various problems including environmental pollution. The inorganic phosphate fertilizer supplied to the soil is partially absorbed by the crops, and the rest moves to the surface layer of the soil along with the water, so that the water evaporates and only the salts accumulate, resulting in salty soils where crops cannot grow normally. The soil with excessive accumulation of salt increases the risk of various disease infections by interfering with the nutrient absorption and growth of plants, and causes eutrophication of surface water and contamination of groundwater.

A salt is a chemically combined acid and base, and the calcium sulfate used as fertilizer is a salt in which sulfuric acid (acid) and cali (base) are combined, and the base is an electrically positive cali, lime, magnesium, sodium, iron , Zinc, manganese and other metal elements. Acids include electrically negative nitric acid, sulfuric acid and chlorine ions. Therefore, salt is an essential nutrient for crop growth. However, when these salts are present above the appropriate level in the soil, we say that they are salt accumulation, and the excess salt is absorbed through the root of the plant and causes a fatal injury to the plant.

Salt disturbances occur in three forms: saline soils and sodium soils, and saline-sodic soils. In the first case, the soil contains excessive soluble salts. In severe cases, the soil surface becomes white due to residues, which makes it difficult to absorb moisture and nutrients from plants and has toxicity, so it is essential to filter and drain salts. In the second case, the soil contains excessive exchangeable sodium, which does not directly harm the plant itself, but the soil constituent particles are made of fine soil to narrow the soil gap, which in turn makes water permeation and drainage extremely difficult and Prevents root growth. In the third case, as soil containing soluble salt and exchangeable sodium, the soil structure must be improved because the soil structure deteriorates and the growth of crops becomes impossible. One of the measures to overcome salt disturbances carried out at home and abroad relies on the cultivation control method of growing salt tolerant crops. However, crops with high tolerance are not only limited to a few species, such as cotton and barley, but only to relatively low levels of salt concentration in the soil below the limit in order for seeds to germinate. Most plants are more sensitive to salt concentrations than these.

As a method of treating the soil in which salt disturbance has occurred, a method of fresh water for a long time or landing the upper portion of the plantation soil is known, and recently, a phosphate solubilizing microorganism is inoculated into the soil to promote solubilization of poorly soluble phosphate in the soil. Research is also underway to solve salt disturbances and promote crop growth. Microorganisms with excellent phosphate solubility that can be used for this purpose include Pantoeaagglomerans, Enterobacter aerogenes, Genus Klebsiella, Penicillium oxalicum ), Burkholderia cepacia, and many other microorganisms are known. However, the Pantoea agglomerans and Enterobacter aerogenes strains are difficult to formulate, and even when formulated, the viability of external environment changes is rapidly reduced. In addition, the Burkholderia cepacia strain has recently been known to cause infection in humans and has been known to have problems in agricultural use.

Rezopus oligosporus is a representative nitrogen-fixing microorganism, a bacteria that invades the roots of plants to form humps and symbiotic with the host to fix nitrogen. Originally, the bacteria are bacilli that live freely in the soil, but nitrogen fixation is only possible when they do not live freely in the soil and invade the roots of legumes. Each of these bacteria symbiotic only for a specific host and consumes carbohydrates supplied from the host to fix nitrogen and supply nitrogen compounds to the host plant again.

Aspergillus Oraijae is a representative strain of Koji mold, a mold that has been important for products such as cheonju, miso, soy sauce, persimmon, and pickles. The fungus is white at first, but becomes yellow to yellow-green from the time of conidia, and becomes brown when older. It also has strong starch saccharification and proteolytic power, and is used for saccharification and soybean decomposition. In the present invention, by decomposing the protein, amino acids may be provided to the nitrogen assimilation as described above, or the starch of the crop is decomposed into sugars, thereby improving the sugar and freshness of fruits and vegetables such as watermelon, melon, tomato, and strawberry.

Bacillus subtilis was used as a bacterium of the genus Bacillus. A representative secretion protein of the Bacillus subtilis is an antibiotic, bacitracin, which is a peptide antibiotics that inhibits the cell wall synthesis of pathogenic bacteria or C55-isoprenyl. It exhibits antibacterial activity by blocking dephosphorylation of pyrophosphate (C55-isoprenylpyrophosphate) with C55-isoprenylphosphate.

It has the function of maintaining the efficacy of the antibiotic having the ring. The Lactobacillus permentium is a hetero type lactic acid bacterium that grows by obtaining energy by fermenting lactic acid of sugars, and serves to purify the soil by decomposing carbohydrate organic substances remaining in the soil. As described above, the microorganisms used in the present invention are strains readily available to those skilled in the art.

Accordingly, the phosphate solubilizer applied to the present invention is applied to compensate for the disadvantages of the phosphate solubilization, and various phosphate solubilizers, not specific phosphate solubilizers, can be used as effective microorganisms in oil culture. As a microbial culture method, it is usually divided into batch culture, continuous culture and fed-batch culture according to the method of supplying the substrate.

In the batch culture, the method of fermentation is continued until all the raw material substrates initially supplied are consumed, the concentration of the substrate, the concentration of metabolites, and the concentration of cells are continuously changed over time. Nevertheless, due to the simplicity of the operation, most fermentation industries adopt a batch culture type.

Continuous culture is a method of fermenting while maintaining a constant culture medium in a fermenter by constantly discharging the same amount of fermentation broth while supplying fresh medium to the fermenter at a constant rate, and in continuous culture, the concentration of dissolved oxygen. It is characterized by the ability to arbitrarily control the rate of proliferation, since environmental conditions such as concentration and pH of metabolites are always kept constant.

In fed-batch culture, batch culture does not add or remove medium once fermentation begins. Continuous culture feeds the medium continuously and also removes it. The fed-batch culture is a culture method in which the medium is supplied intermittently or continuously, and the substrate concentration in the culture medium can be arbitrarily controlled. That is, the culture medium is not taken out during the culture, and a medium containing a specific substrate is supplied intermittently or continuously to arbitrarily control the concentration of the specific substrate.

Since the substrate is added at a moderate rate and there is no spillage, the substrate can be freely controlled by maintaining a balance between the amount of substrate supplied and consumption by microorganisms.

As a method for cultivating microorganisms used in the present invention, batch culture was applied to the initial culture medium, and milk culture was applied to the milk culture medium. Through this, mass production of microorganisms was possible. When only the batch culture was performed, the used lactic acid bacteria and phosphate solubilizing bacteria were used. It can interfere with the mass production of the composed microbial organisms.

Citric acid, which is contained as a free acid in the seeds or juices of many plants, is a substance that is especially contained in lemon, plum, and citrus fruits. It is a basic organic acid that has been extracted from fruits and used as a soft drink or food additive. In recent years, sugars (eg, starch or starch foil) are used as a substrate for fermentation using microorganisms. In general, citric acid is a crystalline white powder and dissolves well in water. When added to soft drinks, it gives a refreshing taste. In addition, citric acid is a component of the tricarboxylic acid (TCA) circuit in vivo, and plays an important role in metabolism of higher animals. In addition, it is known to exhibit the action and bactericidal effect of promoting calcium absorption in the body. It is mainly used as a food additive in sour juices, soft drinks, and diuretic beverages, and is widely used as pharmaceuticals, analytical reagents, and cosmetic ingredients.

However, citric acid has a strong acidity, and when added to juices, beverages, and foods, there is a disadvantage in that sugar must be added in excess to compensate for the acidity. In addition, the prior art citric acid-related technology is limited to a general citric acid manufacturing method or a method for manufacturing citric acid-containing food, and there is little technology to reduce the strong acidity of citric acid.

On the other hand, citric acid (citric acid) is commonly used as a preservative and acidity increasing agent in foods. It is a polybasic carboxylic acid having a hydroxyl group. It is soluble in water and has a refreshing acidity.

As a component of the tricarboxylic acid (TCA) circuit, it plays an important role in metabolism of higher animals.

The citric acid of the present invention can be included in the form of a free acid or salt, which is usually commercially available. Citric acid is prepared by extracting citric acid, which is present in free form in the seeds or juices of various plants. In addition, citric acid is also prepared by a surface fermentation process or submerged fermentation process using the fungus Aspergillus niger, which can also be used in the compositions of the present invention. . Citric acid used in the present invention contributes to the enhancement of microbial survival durability and the proliferation of mass production of complex microorganisms composed of lactic acid bacteria and phosphate solubilizing bacteria, and contributes to the performance quality of liquid fertilizer, which is the final product. It was confirmed in the present invention that the efficacy applied to both batch culture and mass production medium of oil price culture.

Carbon sources used in microbial initial culture medium and microbial mass production medium are glucose, sucrose, maltose, etc., which are selected and used as the most preferred carbon source for complex microorganisms composed of lactic acid bacteria and phosphate solubilizing bacteria.

Citric acid used in the batch culture method of the initial culture medium of microorganisms is less than 5 mL, it is insufficient in the progress of batch culture, and when used in more than 25 mL may cause trouble in culture due to excessive use.

Citric acid used in the oil culture method of the microbial mass production medium is insufficient in the process of oil culture when used in less than 10mL, and when used in excess of 100mL may cause trouble in culture due to excessive use.

One of the complex microorganisms used in the batch culture method of the initial culture medium of microorganisms, yeast extract, which is a lactic acid bacterium, is less than 0.1 part by weight, does not progress in the batch culture, and when used in more than 5 parts by weight, is cultured due to excessive use. Trouble will occur.

Phosphate solubilizing bacteria, one of the complex microorganisms used in the batch culture method of the initial culture medium for microorganisms, is less than 1 part by weight, insufficient in the progress of batch culture, and when used in more than 10 parts by weight, it is difficult to culture due to excessive use. Will occur.

One of the complex microorganisms used in the fed-batch culture method of the microbial mass production medium, the yeast extract, which is a lactic acid bacterium, is less than 0.1 part by weight, and is insufficient in the process of feeding, and when used in more than 10 parts by weight, it is cultured due to excessive use. Trouble will occur.

Phosphate solubilizing bacteria, one of the complex microorganisms used in the fed-batch culture method of microbial mass production medium, is less than 1 part by weight, and is insufficient in the process of oil-value culture. When used in more than 15 parts by weight, it is difficult to culture due to excessive use. Will occur.

Calcium phosphate used in the batch culture method of the initial culture medium of microorganisms is less than 1 part by weight, it is insufficient in the progress of batch culture, and when used in more than 15 parts by weight, trouble occurs in culture due to excessive use.

Calcium phosphate used in the oil price cultivation method of the microbial mass production medium is less than 1 part by weight, insufficient in the process of oil cultivation, and when used in more than 20 parts by weight, it causes trouble in culture due to excessive use.

The temperature range of the aging tank used in the oil price culture method of the microbial mass production medium is less than 20 ℃, and the aging is insufficient.

In the case of the citric acid input time used in the oil price cultivation method of the microbial mass production medium, the maturation is insufficient in less than 3 hours, and in the case of more than 9 hours, the culture is over-cultured and culture trouble occurs.

Applying the batch culture method through the multi-stage mixture and agitation step of the mixture to the initial culture medium of the microorganism, and applying the fed-batch culture method through the multi-stage mixture and the agitation step of the mixture to the microbial mass production medium strictly follow each step and sequence. In the end, it was proved through the results of the present invention that it is possible to invent the liquid microorganism fertilizer for mass production of a complex microorganism composed of lactic acid bacteria and phosphate solubilizing bacteria and strengthen the survival durability of the effective microorganism of the present invention. Specific examples and comparative examples are as follows.

Example 1

In the liquid fertilizer manufacturing method, the initial culture medium for microorganisms is a batch culture method, and disaccharide glucose is applied as a carbon source. In a 30 ° C stirrer, 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, and calcium chloride Stirring the mixture to add 1.0 parts by weight (1); A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed;

A glucose input step (3) in which 60 parts by weight of glucose is added to the mixture stirred after the mixture stirring step (2) is completed and maintained at room temperature for 1 day at a rate of 100 rpm; An effective microorganism (1) input step (4) for introducing 1.0 part by weight of yeast extract, which is an effective microorganism, to the mixture after the glucose input step (3); An effective microorganism (2) injecting step (5) of adding 1.0 part by weight of phosphate solubilizing bacteria, which is effective microorganism (2), to the mixture after the effective microorganism (1) input step (4); And a calcium phosphate input step (6) in which 5.0 parts by weight of calcium phosphate is added to the mixture after the effective microorganism (2) input step (5). To the mixture after completing the calcium phosphate input step (6), 10 mL of citric acid (80%) is added to make an initial culture medium establishment step for enhancing the survival durability of effective microorganisms.

Microbial mass production medium applies disaccharide glucose as a carbon source as a fed-batch culture method, and agitator at 30 ° C is stirred with a mixture of 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, and 1.0 parts by weight of calcium chloride. Step (1); A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed; A glucose input step (3) in which 60 parts by weight of glucose is added to the mixture stirred after the mixture stirring step (2) is completed and maintained at room temperature for 1 day at a rate of 100 rpm; An effective microorganism (1) injecting step (4) in which 3.0 parts by weight of yeast extract, which is an effective microorganism, is added to the mixture after the glucose injecting step (3); An effective microorganism (2) injecting step (5) in which 3.0 parts by weight of phosphate solubilizing bacteria, effective microorganism (2), is added to the mixture after the effective microorganism (1) input step (4); And

A calcium phosphate input step (6) in which 7.0 parts by weight of calcium phosphate is added to the mixture after the effective microorganism (2) input step (5);

A citric acid input step (7) for adding 50 mL of citric acid (80%) to the mixture after completing the calcium phosphate input step (6) to increase the survival endurance and maximize liquid fertilization performance of effective microorganisms; And liquid fertilizer was prepared by maintaining citric acid for 2 days while continuously and intermittently adding and stirring citric acid every 6 hours in a aging tank at 30 ℃, and lettuce liquid fertilizer prepared at a nearby crop plantation in Chungju City. Spraying on the magnetic field and examining the physiologically active state of the plant. As a result of liquid fertilizer performance, Table 1. As shown in Table 2, the storage period of the liquid fertilizer to determine whether germs are generated after storing at room temperature in relation to the microbial survival durability applied to the liquid fertilizer is shown in Table 2.Also, it is possible to comprehensively judge the microbial survival durability and performance applied to the liquid fertilizer. As a judgment on maintaining freshness after lettuce extraction, the lettuce distribution period was shown in Table 3. The results were all excellent.

Example 2

In the liquid fertilizer manufacturing method, the initial culture medium for microorganisms is a batch culture method, and a disaccharide sucrose is applied as a carbon source, to a stirrer at 40 ° C, 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, Stirring the mixture to add 1.0 part by weight of calcium chloride (1); A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed; A sucrose input step (3) in which 60 parts by weight of sucrose is added to the mixture stirring material after the mixture stirring step (2) is completed, and maintained at room temperature for 1 day at a rate of 100 rpm; An effective microorganism (1) input step (4) of adding 1.0 part by weight of yeast extract, which is an effective microorganism, to the mixture after the sucrose input step (3); An effective microorganism (2) injecting step (5) of adding 1.0 part by weight of phosphate solubilizing bacteria, which is effective microorganism (2), to the mixture after the effective microorganism (1) input step (4); And

Calcium phosphate input step (6) for adding 5.0 parts by weight of calcium phosphate to the mixture after the effective microorganism (2) input step (5);

To the mixture after completing the calcium phosphate input step (6), 10 mL of citric acid (80%) is added to make an initial culture medium establishment step for enhancing the survival durability of effective microorganisms.

The microbial mass production medium is a fed-batch culture method, in which a disaccharide sucrose is applied as a carbon source, and a mixture of 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, and 1.0 parts by weight of calcium chloride is added to a stirrer at 40 ° C. Stirring step (1); A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed; A sucrose input step (3) in which 60 parts by weight of sucrose is added to the mixture stirring material after the mixture stirring step (2) is completed, and maintained at room temperature for 1 day at a rate of 100 rpm; An effective microorganism (1) injecting step (4) in which 3.0 parts by weight of yeast extract, which is an effective microorganism, is added to the mixture after the sucrose addition step (3); An effective microorganism (2) injecting step (5) in which 3.0 parts by weight of phosphate solubilizing bacteria, effective microorganism (2), is added to the mixture after the effective microorganism (1) input step (4); And a calcium phosphate input step (6) in which 7.0 parts by weight of calcium phosphate is added to the mixture after the effective microorganism (2) input step (5). A citric acid input step (7) for adding 50 mL of citric acid (80%) to the mixture after completing the calcium phosphate input step (6) to increase the survival endurance and maximize liquid fertilization performance of effective microorganisms; And liquid fertilizer was prepared by maintaining citric acid for 2 days while continuously and intermittently adding and stirring citric acid every 6 hours in a aging tank at 30 ℃, and lettuce liquid fertilizer prepared at a nearby crop plantation in Chungju City. After spraying on the magnetic field and examining the physiological activity of plants, the results of liquid fertilizer performance are shown in Table 1. As shown in Table 2, the storage period of the liquid fertilizer to determine whether germs are generated after storing at room temperature in relation to the microbial survival durability applied to the liquid fertilizer is shown in Table 2.Also, it is possible to comprehensively judge the microbial survival durability and performance applied to the liquid fertilizer. As a judgment on maintaining freshness after lettuce extraction, the lettuce distribution period was shown in Table 3. The results were all excellent.

Example 3

In the liquid fertilizer manufacturing method, the initial culture medium for microorganisms is a batch culture method, and a disaccharide maltose is applied as a carbon source. In a stirrer at 50 ° C, 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, Stirring step (1) of adding 1.0 part by weight of calcium chloride; A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed; A maltose input step (3) of adding 60 parts by weight of maltose to the mixture agitated product after the mixture stirring step (2) and maintaining the mixture at a rate of 100 rpm for 1 day at room temperature; An effective microorganism (1) input step (4) for introducing 1.0 part by weight of yeast extract, which is an effective microorganism, to the mixture after the maltose input step (3); An effective microorganism (2) injecting step (5) of adding 1.0 part by weight of phosphate solubilizing bacteria, which is effective microorganism (2), to the mixture after the effective microorganism (1) input step (4); And a calcium phosphate input step (6) in which 5.0 parts by weight of calcium phosphate is added to the mixture after the effective microorganism (2) input step (5). To the mixture after completing the calcium phosphate input step (6), 10 mL of citric acid (80%) is added to make an initial culture medium establishment step for enhancing the survival durability of effective microorganisms.

Microbial mass production medium is a fed-batch culture method, in which disaccharide maltose is applied as a carbon source, and a mixture of 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, and 1.0 parts by weight of calcium chloride is added to a stirrer at 50 ° C. Stirring step (1); A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed; A maltose input step (3) of adding 60 parts by weight of maltose to the mixture agitated product after the mixture stirring step (2) and maintaining the mixture at a rate of 100 rpm for 1 day at room temperature; An effective microorganism (1) injecting step (4) in which 3.0 parts by weight of yeast extract, which is an effective microorganism, is added to the mixture after the maltose injecting step (3); An effective microorganism (2) injecting step (5) in which 3.0 parts by weight of phosphate solubilizing bacteria, effective microorganism (2), is added to the mixture after the effective microorganism (1) input step (4); And a calcium phosphate input step (6) in which 7.0 parts by weight of calcium phosphate is added to the mixture after the effective microorganism (2) input step (5). A citric acid input step (7) for adding 50 mL of citric acid (80%) to the mixture after completing the calcium phosphate input step (6) to increase the survival durability of active microorganisms and maximize liquid fertilization performance; And liquid fertilizer was prepared by maintaining citric acid for 2 days while continuously and intermittently adding and stirring citric acid every 6 hours in a aging tank at 30 ℃, and lettuce liquid fertilizer prepared at a nearby crop plantation in Chungju City. After spraying on the magnetic field and examining the physiological activity of plants, the results of liquid fertilizer performance are shown in Table 1. As shown in Table 2, the storage period of liquid fertilizer to determine whether germs are generated after storage at room temperature in relation to the microbial survival durability applied to liquid fertilizer is shown in Table 2. Also, it is possible to comprehensively judge the microbial survival durability and performance applied to liquid fertilizer. As a judgment on maintaining freshness after lettuce extraction, the lettuce distribution period was shown in Table 3. The results were all excellent.

Comparative Example 1

In the liquid fertilizer manufacturing method, the initial culture medium for microorganisms is a batch culture method, and disaccharide glucose is applied as a carbon source. In a 50 ° C stirrer, 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, and calcium chloride Stirring the mixture to add 1.0 parts by weight (1); A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed;

A glucose input step (3) in which 60 parts by weight of glucose is added to the mixture stirred after the mixture stirring step (2) is completed and maintained at room temperature for 1 day at a rate of 100 rpm; An effective microorganism input step (4) of adding 1.0 part by weight of yeast extract, which is an effective microorganism, to the mixture after the glucose input step (3); And a calcium phosphate input step (5) for adding 5.0 parts by weight of calcium phosphate to the mixture after the effective microbial input step (4); A single microbial liquid fertilizer was prepared by adding 10 mL of citric acid (80%) to the mixture after completing the calcium phosphate input step (5) to make an initial culture medium for effective microorganisms, and lettuce liquid fertilizer prepared at a nearby crop plantation in Chungju City After spraying on the magnetic field and examining the physiological activity of plants, the results of liquid fertilizer performance are shown in Table 1. As shown in Table 2, the storage period of the liquid fertilizer to determine whether germs are generated after storing at room temperature in relation to the microbial survival durability applied to the liquid fertilizer is shown in Table 2.Also, it is possible to comprehensively judge the microbial survival durability and performance applied to the liquid fertilizer. After the lettuce was released, the lettuce distribution period was shown in Table 3.

Comparative Example 2

In the liquid fertilizer manufacturing method, the initial culture medium for microorganisms is a batch culture method, and disaccharide glucose is applied as a carbon source. In a 50 ° C stirrer, 2.0 parts by weight of ammonium sulfate, 1.5 parts by weight of sodium chloride, 0.5 parts by weight of magnesium sulfate hydrate, and calcium chloride Stirring the mixture to add 1.0 parts by weight (1); A mixture stirring step (2) in which 1.5 mL of manganese sulfate first hydrate and 1.3 mL of ferric sulfate hexahydrate are added to the mixture stirred after the mixture stirring step (1) is completed;

A glucose input step (3) in which 60 parts by weight of glucose is added to the mixture stirred after the mixture stirring step (2) is completed and maintained at room temperature for 1 day at a rate of 100 rpm; An effective microorganism input step (4) of adding 1.0 part by weight of phosphate solubilizing bacteria, which is effective microorganism (2), to the mixture after the glucose input step (3); And a calcium phosphate input step (5) for adding 5.0 parts by weight of calcium phosphate to the mixture after the effective microbial input step (4); A single microbial liquid fertilizer was prepared by adding 10 mL of citric acid (80%) to the mixture after completing the calcium phosphate input step (5) to make an initial culture medium for effective microorganisms, and lettuce liquid fertilizer prepared at a nearby crop plantation in Chungju City After spraying on the magnetic field and examining the physiological activity of plants, the results of liquid fertilizer performance are shown in Table 1. As shown in Table 2, the storage period of the liquid fertilizer to determine whether germs are generated after storing at room temperature in relation to the microbial survival durability applied to the liquid fertilizer is shown in Table 2.Also, it is possible to comprehensively judge the microbial survival durability and performance applied to the liquid fertilizer. After the lettuce was released, the lettuce distribution period was shown in Table 3.

Liquid fertilizer performance-related effects division Leaf length (cm) Blade width (cm) Chlorophyll (mg / cm ^ 2) Example 1 35 9 4.90 Example 2 32 9 4.53 Example 3 33 10 4.68 Comparative Example 1 22 5 2.37 Comparative Example 2 25 6 2.59

Storage period of liquid fertilizer division 6 months 9 months 12 months 24 months Example 1 none none none none Example 2 none none none none Example 3 none none none Occurs slightly Comparative Example 1 none Occur Occur Occur Comparative Example 2 none Occur Occur Occur

Lettuce distribution period (whether maintaining freshness after shipment of lettuce) division 1 week 2 weeks 3 weeks 4 weeks Example 1 Good Good Good Good Example 2 Good Good Good Good Example 3 Good Good Good Slightly insufficient Comparative Example 1 Good Bad Bad Bad Comparative Example 2 Good Good Bad Bad

Claims (3)

In the liquid fertilizer manufacturing method,
The initial culture medium for microorganisms is a batch culture method, and disaccharides are applied as a carbon source. In a stirrer at 30 ° C to 50 ° C, 0.1 to 5.0 parts by weight of ammonium sulfate, 0.1 to 3.0 parts by weight of sodium chloride, 0.1 to 10.0 parts by weight of magnesium sulfate hydrate, and calcium chloride Stirring the mixture to add 0.1 to 3.0 parts by weight (1); A mixture stirring step (2) in which 0.1 to 5.0 mL of manganese sulfate first hydrate and 0.1 to 5.0 mL of ferric sulfate heptahydrate are added to the mixture stirred after the mixture stirring step (1) is completed;
A disaccharide input step (3) in which 50 to 80 parts by weight of disaccharide is added to the mixture stirred after the mixture stirring step (2) is completed and maintained at room temperature for 1 to 3 days at a rate of 50 to 150 rpm; An effective microorganism (1) input step (4) of adding 0.1 to 5 parts by weight of the yeast extract, which is an effective microorganism, to the mixture after completing the disaccharide input step (3); An effective microorganism (2) injecting step (5) in which 1 to 10 parts by weight of phosphate solubilizing bacteria, which is effective microorganism (2), is added to the mixture after the effective microorganism (1) input step (4); And a calcium phosphate input step (6) in which 1 to 15 parts by weight of calcium phosphate is added to the mixture after the effective microorganism (2) input step (5); 5-5 mL of citric acid (80%) is added to the mixture after completing the calcium phosphate input step (6) to make an initial culture medium establishment step for enhancing the survival durability of effective microorganisms.
Microbial mass production medium applies disaccharides as a carbon source as a fed-batch culture method, in a stirrer at 30 ° C to 50 ° C, 0.1 to 5.0 parts by weight of ammonium sulfate, 0.1 to 3.0 parts by weight of sodium chloride, 0.1 to 10.0 parts by weight of magnesium sulfate hydrate, and calcium chloride Stirring the mixture to add 0.1 to 3.0 parts by weight (1); A mixture stirring step (2) in which 0.1 to 5.0 mL of manganese sulfate first hydrate and 0.1 to 5.0 mL of ferric sulfate heptahydrate are added to the mixture stirred after the mixture stirring step (1) is completed;
A disaccharide input step (3) in which 50 to 150 parts by weight of disaccharide is added to the mixture stirred after the mixture stirring step (2) is completed and maintained at room temperature for 1 to 3 days at a rate of 50 to 150 rpm; An effective microorganism (1) input step (4) of adding 0.1 to 10 parts by weight of yeast extract, which is an effective microorganism, to the mixture after the disaccharide input step (3); An effective microorganism (2) injecting step (5) in which 1 to 15 parts by weight of phosphate solubilizing bacteria, effective microorganism (2), is added to the mixture after the effective microorganism (1) input step (4); And a calcium phosphate input step (6) in which 1 to 20 parts by weight of calcium phosphate is added to the mixture after the effective microorganism (2) input step (5); A citric acid input step (7) for increasing the durability of effective microorganisms and maximizing liquid fertilization performance by adding 10 to 100 mL of citric acid (80%) to the mixture after the calcium phosphate input step (6); And microbial survival through citric acid, characterized in that citric acid is prepared in a aging tank at 20-40 ° C and continuously and intermittently added and stirred at intervals of 3 to 9 hours, and aged for 1 to 3 days while maintaining the oil price culture method. Liquid fertilizer for mass production of complex microorganisms composed of lactic acid bacteria and phosphoric acid solubilizing agents with enhanced durability and a method for manufacturing the same


The method according to claim 1,
Disaccharides, selected from the group consisting of sucrose called sugar, sucrose, sucrose, maltose, maltose, etc., lactose, lactulose, trehalose, cellobiose, etc. It is characterized by being used as citric acid, citric acid, citric acid, calcium citrate, potassium citrate, iron citrate, sodium citrate, magnesium citrate, sodium citrate, anhydrous citric acid, citrate, etc. Or characterized in that it is used in the form of a mixture of two or more types, and as an organic acid, citric acid, malic acid, maleic acid, succinic acid, oxalic acid, cinnamic acid ( Cinamic acid, acetic acid, sulfonic acid, formic acid, sulfinic acid, butyric acid ylic acid, lactic acid, fumaric acid, propionic acid, ketoglutaric acid, glutaric acid, tartaric acid, gluconic acid ), Selected from the group consisting of ascorbic acid, palmitic acid, and sulfamic acid, microorganisms through citric acid, characterized by being used alone or in a mixture of two or more types Liquid fertilizer for mass production of complex microorganisms consisting of lactic acid bacteria and phosphate solubilizing bacteria, with enhanced durability and method for manufacturing the same
The method according to claim 1, wherein the phosphate solubilizing agent is Rhizopus oligos porous, Aspergillus oryzae, Bacillus sp, Cellulomonas sp, Lactobacillus permentium (Lactobacillus fermentum) is a composite microbial mass-producing liquid fertilizer composed of lactic acid bacteria and phosphate solubilizing bacteria and strengthening the durability of microorganisms through citric acid.