JP2011074047A - Process of humus extract - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a process for a humus extract without toxicity to human, which can be used as a microbicide having excellent safety and stably obtained with a high rate of eradication. <P>SOLUTION: The process for the humus extract comprises (1) light-reacting a humic soil while keeping its moisture content at 50-80% and (2) mixing the resulting humic soil with water and a magnesium calcium acetate hydrate, and fermenting and maturing it. Preferably, in process (2), a fat-free milk and a pumice are mixed with the humus. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method for producing a humus extract, and more particularly, to a method for producing a humus extract exhibiting a higher effect as a growth promoter or fungicide.

Humus is a substance contained in humus soil that has been produced by repeated fermentation and ripening of fallen leaves and organic and inorganic substances buried in the ground for approximately 8,000 years. Humus contains humic acid, fulvic acid, minerals, amino acids, vitamins, and other useful bacteria. Humus has many characteristics:
(1) pH buffering action is large,
(2) chelate reaction,
(3) High cation exchange capacity,
(4) forming a soil aggregate structure;
(5) has a physiologically active function,
(6) suppress pathogenic bacteria,
(7) It has the effect of preventing the growth of plants and animals.
(8) Deodorizing action
(9) There is a cleansing action of sewage,
(10) There is a sludge reduction effect in the sewage treatment process,
(11) Has an antioxidant effect.

  Depending on the intended use of the humus, various forms and compositions of the humus are applied so that at least one of the above characteristics can be exhibited to the maximum. For example, sewage purification accelerator containing various forms of humus for sewage treatment, humus powder as sludge reduction agent, humus pellet / humus powder as deodorizing agent, humus extract as soil conditioner, sterilization / control for pathogenic bacteria, virus As a lotion having a virucidal agent humus extract, moisturizing, sterilizing and skin conditioning effects, humus extract and the like can be mentioned.

  Humus is used in the form of pellets, powders and extracts. However, since humus resources produced in nature are limited, it is desirable to use them as efficiently as possible by increasing the effects of humus and reducing the amount used. It is. Furthermore, in addition to the conventional fields, it is also desired to contribute to society in new fields and applications.

  Sterilization, disinfection, and cleaning are important to control pathogens, especially infectious diseases and influenza viruses. Conventional sterilization and disinfection methods include physical methods such as superheating, filtration, and ultraviolet rays, and chemical methods include disinfectants such as glutaraldehyde, sodium hypochlorite, ethanol, and chlorhexidine. The disinfection effect changes depending on which method is adopted.

  Depending on the method, there is a limitation in use. For example, physical methods such as boiling and hot water can be applied to steel and magnetic accessories and tableware, but cannot be used for living bodies and the environment.

  Chemical disinfectants are generally concerned with toxicity to the human body, damage to the skin, and residual toxicity. For example, glutaraldehyde exhibits a high level of disinfection effect, but is highly toxic to the human body. Well-known sodium hypochlorite and ethanol show a moderate level of disinfection, but cannot be used on mucous membranes or damaged skin. In addition, chemical disinfectants also cause metal corrosion problems.

Thus, there is a need for a safe disinfectant that has no residual toxicity to the human body compared to a disinfectant that has restrictions on use. As one of the measures, Patent Documents 1 to 4 propose techniques for using a humus extract as an antibacterial agent. In particular, Patent Document 2 is obtained by performing extraction by adding water to humus soil by rough drying and precision drying, and filtering the obtained extract using a 0.1 to 0.3 μm filter. An aqueous solution containing a humus extract material consisting of a filtrate is described. Patent Document 4 discloses a humus produced by keeping a humus soil at a moisture content of 50 to 80% and causing a bright reaction, and a humus extract produced by extracting the humus into water. Yes.
JP-A-2004-51590 (culture solution production method, culture solution, and lotion using the same) JP 2000-136140 (humus soil extractable substance-containing aqueous liquid) JP 2003-267821 (Cosmetic raw material) JP 2006-273734 (humus, humus extract and moisturizing liquid, and production method and use thereof)

  However, with the conventional antibacterial agent using the humus extract, viable bacteria are detected against E. coli, Salmonella and Staphylococcus aureus even after 2 to 24 hours have elapsed from the start of use, and antibacterial properties are satisfied. Not obtained.

  In the present situation where the sterilization rate is not always satisfactory even when the conventional humus extract is used as a bactericidal agent, the object of the present invention is that it is not toxic to the human body, can be used as a bactericidal agent with excellent safety, and has high sterilization efficiency. An object of the present invention is to provide a method for producing a bactericide that can be stably obtained immediately after the start of use.

  Bacteria contained in the humus extract (hereinafter referred to as humus extract A) obtained by the production method described in Patent Document 4 presented by the present inventor was sterilized, and the effect on the antibacterial action was examined. . The results are shown in Table 1.

1) The number of viable control bacteria immediately after inoculation with the bacterial solution was measured and set as the start time.
2) Sterilization of the humus extract was performed at 121 ° C. for 15 minutes.

  From Table 1, it can be seen that the bacteria that survive in the humus extract A have an effect on the antibacterial action. It was confirmed by other experiments that the bacterium was mainly Bacillus. Since this bacterium is a viable bacteria peculiar to a lot of humus components, the antibacterial action of the humus extract is thought to be due to the humus component, which is an environmental component where bacteria are the main cause, but the bacteria survive. .

  In the experience of the present inventors, the occupancy rate of fusarium (gram-negative bacteria) is high and the humic component is reduced where the pathogenic bacteria grow. In the absence of pathogenic bacteria, there are many Gram-positive bacteria such as actinomycetes and Bacillus bacteria, and there are many amino acids, organic acids, vitamins, and other physiologically active substances, and the humic component tends to increase. Although all of these tendencies do not match, if there are many humus components that indicate the relationship between pathogenic bacteria and humus soil, pathogens are suppressed.

  On the other hand, it was also found that pathogenic bacteria are suppressed by Bacillus bacteria present in an environment rich in humic components. We tried to improve the antibacterial properties of the humus extract by repeating the fermentation and aging to meet these conditions. As a result, it has been found that the following problems can be solved according to the following invention. That is, the present invention comprises the following steps: (1) light reaction with humus soil kept at a moisture content of 50 to 80%, (2) the obtained humus is mixed with water and magnesium calcium acetate hydrate, and fermented And a method for producing a humus extract comprising aging.

  In the present specification, the light reaction means that the excavated humus soil is not dried under sunlight and the moisture content is maintained at 50 to 80% and fermented and matured. The reaction of humus in the soil is called dark reaction.

  In step (2), it is preferable that skim milk is further mixed with the humus.

  In the step (2), it is preferable to further mix pumice with the humus.

  In the present invention, a humus extract having a higher antibacterial property than before can be produced by adding specific additives to humus obtained by light reaction of humus and fermenting and aging.

  Conventionally, the disinfectant spraying method has not been used much because the human body inhales the disinfectant. However, when the humus extract obtained by the production method of the present invention is used as a disinfectant in the spray method, there is no toxicity to the human body, and the space can be easily sterilized.

The method for producing a humus extract of the present invention includes the following steps:
(1) The humus soil is kept at a moisture content of 50 to 80% to cause a light reaction,
(2) The obtained humus is mixed with water and magnesium calcium acetate hydrate, fermented and matured. Details thereof will be described below.

  Since step (1) is the same as the method for producing humus described in Patent Document 4 (Japanese Patent Laid-Open No. 2006-273734), the contents of Patent Document 4 are incorporated herein by reference. Below, the outline | summary of a process (1) is shown.

  The humus soil used in step (1) is a deciduous, fallen leaf, decomposed or biosynthetic material derived from organic matter such as trees that has been buried in the basement for, for example, about 8000 years or more. . The age of the humus can be obtained, for example, by measuring the years with coexisting pollen.

  In step (1), a light reaction is performed on the excavated humus soil. That is, it is usually left under sunlight for 1 to 12 months, preferably 3 to 12 months, particularly preferably 6 to 12 months.

  In step (1), the moisture content of the humus soil on the ground is 50 to 80%, preferably 50 to 70%, more preferably 50 to 65%, and it is necessary to maintain a moisture content suitable for fermentation. . The moisture content of the underground humus is preferably maintained at 50-80%.

  In order to make the whole humus soil lightly react uniformly, it is sometimes preferable to cut back.

  The humus soil is neutral at a pH of about 7 when excavated, but when the light reaction on the ground is advanced, the pH becomes about 3 after one month. Even after one year, there is no significant change in pH, and the pH is 2.6 to 3.3.

  In the step (2), humic components are extracted into water from the humus obtained in the step. The humus soil used in step (1) and the humus obtained in step (1) have the effects of promoting plant growth and bioactive functions, and are similar to indoleacetic acid (auxin) known as a plant hormone. . Although the chemical structure of humus is not constant, a typical chemical structure model by Schulten et al. (1993) is shown below.

  Although this molecule is a polymer, its structural formula includes a structure similar to auxin of the following structural formula.

  In order to amplify the auxin-like effect of the humus obtained in step (1), it is desired to add acetic acid. However, if acetic acid is added directly to the humus, the acetic acid is decomposed by a biological reaction. In the present invention, in the step (2), the humic component extracted in water is chelated with magnesium calcium acetate hydrate (hereinafter referred to as CMA) to prevent the decomposition reaction of acetic acid. Addition of CMA amplifies the bactericidal action of pathogenic bacteria as shown in the Examples. It also promotes growth and promotes wound healing effects.

  The amount of CMA added is usually 50 to 1000 ppm, preferably 100 to 200 ppm, based on the total amount of humus and water.

  In step (2), it is preferable to add skim milk as a nutrient source to the humus. Glucose grows conventionally in glucose, which is known as a nutrient source, whereas skin milk has little growth of bacteria. Moreover, skin milk grows Bacillus layer bacteria that survive in humus. Skimmed milk grows Bacillus and amplifies the antibacterial action of this bacterium. The skim milk is not particularly limited and may be commercially available.

  The amount of skim milk added is usually 50 to 200 ppm, preferably 50 to 100 ppm, based on the total amount of humus and water.

  In step (2), it is preferable to mix pumice with the humus in order to allow the microorganisms to grow smoothly. In particular, pumice containing as many kinds of minerals as possible is preferable. Table 2 shows typical components of pumice.

  The addition amount of the pumice is usually 0.1 to 1.0% by weight, preferably 0.1 to 0.2% by weight, based on the total amount of humus and water.

  At the time of fermentation and aging in step (2), in addition to the above components, for example, about 100 ppm of cysteine which is a kind of amino acid may be added and mixed.

  In the step (2), the moisture content of the humus is usually 15 to 50%, preferably 15 to 40%, particularly preferably 20 to 30%.

  The fermentation and aging temperature in step (2) may be room temperature. The period of fermentation and aging is usually 2 to 12 months, preferably 3 to 4 months.

  During the ripening and fermentation in step (2), it is preferable to continue aeration and stirring in the tank for the first 2 to 4 months, and to perform static sedimentation for the remaining 1 to 2 months.

  During the step (2), the humic component dissolves and elutes in water, and fermentation and biosynthesis proceed in water, increasing physiologically active substances such as amino acids and vitamins. In particular, a mixed solution that exhibits a yellowish brown color at the initial stage of extraction shows red coloration seen in arginine, cysteine, histidine, tyrosine, etc. after 6 months or more, and all amino acids increase. The initial pH of the mixture of humus and water is 3 to 3.5, but it becomes about pH = 2 to 3 by fermentation for 6 to 12 months.

  The humus supernatant or filtrate obtained in step (2) is used as the humus extract. For the filtration, a filter paper or a cassette filter may be used.

  The purpose of disinfectants is to block infection routes and eliminate pathogens. To prevent infection, sterilization, disinfection and / or cleaning are precautions. The action time, action concentration, and action temperature are important for effective disinfection. Commercial chemical disinfectants must be cleaned after disinfection to remove residual toxicity.

  The humus extract obtained by the production method of the present invention has been confirmed to be safe in many safety tests such as an acute oral toxicity test in mice, a skin irritation test in mice, an ocular mucosal irritation test, and a medaka toxicity test. If this material is used as a disinfectant, there is no residual toxicity and no cleaning is required. Therefore, since it is not necessary to clean it by using it on the human body or metal, the disinfectant adheres to the object for a long time, and the antibacterial action increases. Furthermore, since it has an antioxidant action, metal corrosion is avoided.

  The humus extract obtained by the production method of the present invention is sterilized by spraying, coating and dipping on equipment and the human body as a disinfectant, adding to the water of a humidifier, spray sterilization, working on an air circulator, air sterilization It can be used for addition sterilization to impregnated sterilization lotion on mask, addition deodorization sterilization to bathtub, impregnation and sterilization to linens.

  The use concentration of the humus extract obtained by the production method of the present invention may be a humus extract undiluted solution (supernatant solution or filtrate), and further exhibits antibacterial effects even when diluted up to 20 times. Therefore, according to the said use, what is necessary is just to use normally 1-10 times dilution liquid of a humus extraction undiluted solution, Preferably 5-10 times dilution liquid may be used.

Hereinafter, the present invention will be described in more detail with reference to Examples and Comparative Examples. However, the present invention is not limited to the following examples.
[Example 1]
(Process 1)
The humus soil that had been buried underground for about 8,000 years at the site of Enzyme's Nagasaki Plant was excavated and light reaction was performed for about 12 months at room temperature under sunlight. Table 3 shows the physical properties of humus in which the content of enzymes and coenzymes was increased by this light reaction.

(Process 2)
The humus obtained in step 1 and water were mixed at a weight ratio of 30:70 (the moisture content of the humus was 50%). 100 ppm of commercially available skin milk as a nutrient, 100 ppm of CMA as a bioactive agent, and 0.1% by weight of pumice as a support for microbial growth were added to the mixture, and fermentation was matured at room temperature for 3 to 12 months. Aeration and stirring in the tank were continued for 2 to 11 months from aging, and static sedimentation was performed for the remaining 1 month. The supernatant was filtered through a filter paper or a cassette filter to obtain a humus extract. Table 4 shows the physical properties of the humus extract.

[Comparative Example 1]
For comparison (positive control), a humus extract was prepared according to the production method described in JP-A-2006-273734 using humus soil obtained in the site of Enzyme Nagasaki Factory. Specifically, a humus extract was prepared in the same procedure as in Example 1 except that skin milk, CMA, and pumice were not mixed in Step (2) of Example 1. The components of this humus extract are shown in Table 5. Although the contents of both organic and inorganic substances in Table 5 are small, the humus extract can be said to be an activated liquid containing humic components, minerals, microorganisms, enzymes, and coenzymes.

  There is almost no difference in components between the humus extract of Example 1 (Table 4) and the humus extract of Comparative Example 1 (Table 5). However, in Example 1, since a difference was observed in the bacterial flora during the fermentation, it was confirmed in the following antibacterial test whether or not there was a difference in the antibacterial action of both.

(Antimicrobial test)
After inoculating Escherichia coli, Pseudomonas aeruginosa, Salmonella or Staphylococcus aureus into the humus extract of Example 1 and Comparative Example 1, it was stored at 25 ° C., and after 60 seconds, 6 hours, and 24 hours, respectively. The number of viable bacteria in the test solution was measured. The results are shown in Table 6. As a comparison (negative control), a test in which the test solution was changed to purified water was added.

1) Purified water, physiological saline: negative control 2) Starting time: The number of viable bacteria of the control immediately after inoculation with the bacterial solution was measured and set as the starting time.

  From Table 6, it can be seen that both Example 1 and Comparative Example 1 exhibit high antibacterial activity. The humus extract of Example 1 showed almost the same numerical value in terms of the sterilization rate of Escherichia coli as compared with the humus extract of Comparative Example 1, but showed a high value in the sterilization rate of other bacteria.

(Virus inactivation test)
Prior to this test, a preliminary test was conducted to examine a method for measuring the viral infectivity titer. Since sufficient inactivation could be predicted with the stock solution of the humus extract of Example 1, it was decided to carry out this test by preparing a 5-fold solution of the humus extract using water for injection. Furthermore, the infectivity titer was measured by diluting the working fluid 100 times with a cell maintenance medium. As a comparison (negative control), a test in which the test solution was changed to purified water was added.

  A virus suspension of influenza virus (obtained from Japan Food Analysis Center) was mixed with a 5-fold diluted solution of the humus extract of Example 1 to obtain a working solution. The solution was allowed to act at room temperature, and after 60 seconds, 6 hours, and 24 hours, the virus infectivity of the working solution was measured. Table 7 shows the measurement results.

_{TCID 50: median tissue culture infectious dose} , 50% tissue culture infectious dose <2.5: logTCID Not detected _50: when logarithm 1) initiation of TCID ₅₀ per working solution 1 ml: the control TCID ₅₀ immediately after the start of action Measured and started.
2) Purified water: negative control

  In Table 7, in the 5-fold dilution of the humus extract of Example 1, the infectivity removal rate of influenza A virus was 99.9% after 60 seconds and 99.999% or more after 6 hours, with a high rate Inactivated the virus.

  As described above, the difference between the humus extract of Example 1 and the humus extract of Comparative Example 1 appears to be minute, and the antibacterial property of the humus extract of Example 1 is higher than that of the humus extract of Comparative Example 1. Excellent, and also exhibits a very good virus inactivation action in virus inactivation tests. Since the pathogen countermeasures for infectious diseases must be blocked by sterilization (sterilization to 1 / 1,000,000) and disinfection, the production method of the present invention has greatly improved the antibacterial properties of the humus extract. It's a harvest. The manufacturing method of the problem which it tried to interpret in order to obtain the favorable antibacterial action of Table 6 and 7 can be said to be a new invention succeeded from a hypothesis to practical use.

Claims (3)

The following steps:
(1) The humus soil is kept at a moisture content of 50 to 80% to cause a light reaction,
(2) A method for producing a humus extract comprising mixing the obtained humus with water and magnesium calcium acetate hydrate, fermenting and aging.   The method for producing humus according to claim 1, wherein skim milk is further mixed with the humus in step (2).   The method for producing humus according to claim 1, wherein in the step (2), pumice is further mixed with the humus.