JPH10146186A - New antibacterial microorganism, antibacterial deodorizing treatment using the microorganism, production of antibacterial substance using the microorganism, antibacterial substance obtained by the method, antibacterial deodorizing processing using the substance, and antibacterial deodorized processed product subjected to the method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new microorganisms having antibacterial properties against Staphylococcus aureus and Klebsiella pneumoniae, safe for human bodies, and useful for antibacterial deodorizing agents, etc. SOLUTION: New microorganisms: Bacillus sp. OYK-03-600 (FERM P-15608) and Bacillus sp. OYK-03-000 (FERM P-15609), which belong to the genus Bacillus and have antibacterial properties against Staphylococcus aureus and Klebsiella pneumoniae. The microorganisms are useful for producing antibacterial deodorizing agents safe (mild) for human bodies, etc. The microorganisms are obtained by screening microorganisms separated from soil by the utilization of the antibacterial properties against the Staphylococcus aureus and the Klebsiella pneumoniae generating malodors due to their proliferation.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel microorganism having antibacterial properties, a method for antibacterial deodorization treatment using the novel microorganism,
The method for producing an antibacterial substance using the novel microorganism, the antibacterial substance obtained by the method, the antibacterial deodorant processing method using the antibacterial substance, and the antibacterial deodorant processed product subjected to the method are described in detail. Bacillus s , a novel microorganism with high antibacterial activity against bacteria that produce bad odors
p. OYK-01-600 (FERMP-15607),
Bacillus sp. OYK-03-600 (FERM P-1
5608), Bacillus sp. OYK-04-000 (FE
RMP-15609) and methods of using the microorganisms, for example, using the microorganisms directly or using an antibacterial substance separated and purified from a culture solution of the microorganisms, to control an odor generated during wastewater treatment, for example, to control odors. Or the antibacterial substance, for example, clothing (shirts, socks, ties, etc.)
The present invention relates to the deodorizing method used in the above.

[0002]

2. Description of the Related Art One of the techniques for treating wastewater is a biological treatment method. Among them, the activated sludge method is particularly well known. This is a method of removing or decomposing pollutants in wastewater using various organisms existing in nature, especially microorganisms.

[0003] There are many microorganisms used for wastewater treatment, and the main ones are as follows. That is, as bacteria, Zooglea , Sphaerotilus
Protozoa include amoebae, ciliates, and flagellates, and algae include cyanobacteria, green algae, and diatoms.

[0004] These can decompose organic substances and some inorganic substances, but at the time of decomposition, they may cause the production of offensive odors (ammonia, hydrogen sulfide, etc.) and offensive odor-producing substances, and are generated from treatment plants. The stench
It is often addressed as a social issue. More specifically, in wastewater treatment based on a biological treatment method, even if organic substances or inorganic substances were efficiently decomposed, the odor generated from the treatment plant remained undisturbed. In particular, activated sludge settling in the sedimentation tank gradually becomes anaerobic, and a large amount of odor is generated with the growth of anaerobic bacteria.When returning this sludge to the aeration tank, a large amount of odor is generated around the wastewater treatment plant. Drifted.

On the other hand, recently, many antibacterial products (eg, clothing articles such as socks and shirts) have been manufactured and sold for the purpose of deodorizing and deodorizing effects. The mechanism will be described in detail as follows. That is, there are many bacteria called indigenous skin bacteria in human skin, which prevents the invasion of malignant bacteria from the outside. However, due to sweating and stuffiness of humans, resident bacteria on the armpits, neck streaks, back, feet, etc. abnormally multiply and produce odors with the growth of the bacteria, but products such as socks have antibacterial properties. If so, abnormal occurrence of bacteria can be suppressed, and at the same time, deodorizing and deodorizing effects are exhibited.

[0006] Antibacterial agents used in antibacterial processing, for example, antibacterial agents for fibers can be roughly classified into piguanide, alcohol, phenol, anilide, iodine, imidal and thiazole, isothiazolone, triazine, nitrile, and the like. Fluorine-based, saccharide-based, tropolone-based, organic metal-based, inorganic metal-based, and the like.

[0007] These are all synthetic chemicals, and although their toxicity to humans is safe for as long as they are within an allowable range, they cannot be said to have no problem in long-term use.
In particular, the persistence of the antibacterial effect may cause various allergies, disturb the balance of indigenous bacteria in the skin, and allow invasion of malignant bacteria from the outside. In other words, currently used antibacterial deodorants have no guarantee that they will maintain their effect while maintaining a balance of normal skin resident bacteria, which may result in antimicrobial allergies, atopic dermatitis, and other malignant bacteria Was causing intrusion.

[Object of the Invention] The present invention has been made in view of the above-mentioned circumstances, and its object is to provide Staphylococcus aurea.
A novel antibacterial substance that shows antibacterial properties to both us and Klebsiella pneumoniae and can prevent bad odors when used in wastewater treatment plants, and that the antibacterial substance separated and purified from itself or the culture solution is safe for the human body In addition to providing microorganisms, in addition to the use of the microorganisms, that is, a method for directly using the microorganisms or an antibacterial deodorizing treatment method using the antibacterial substance, culturing the novel microorganisms, and separating and purifying from the culture solution It is an object of the present invention to provide a method for producing an antibacterial substance, an antibacterial substance obtained by the method, an odor-proofing method using the antibacterial substance in, for example, clothing, and an antibacterial and deodorant-processed product obtained by applying the method.

[0009]

The novel microorganism according to the present invention belongs to the genus Bacillus and is characterized by having antibacterial activity against Staphylococcus aureus and Klebsiella pneumoniae .

The novel microorganism according to claim 2 is the novel microorganism according to claim 1, wherein the microorganism having antibacterial activity against Staphylo coccus aureus and Klebsiella pneumoniae is Ba.
cill us sp. OYK-01-600 (FERM P-15
607).

The novel microorganism according to claim 3 is the novel microorganism according to claim 1, wherein the microorganism having antibacterial activity against Staphylo coccus aureus and Klebsiella pneumoniae is Ba.
cill us sp. OYK-03-600 (FERM P-15
608).

The novel microorganism according to claim 4 is the novel microorganism according to claim 1, wherein the microorganism having antibacterial activity against Staphylo coccus aureus and Klebsiella pneumoniae is Ba.
cill us sp. OYK-04-000 (FERM P-15
609).

According to a fifth aspect of the present invention, there is provided an antibacterial and deodorizing treatment method using the microorganism according to any one of the first to fourth aspects.

According to a sixth aspect of the present invention, there is provided a method for producing an antibacterial substance, wherein the microorganism according to any one of the first to fourth aspects is cultured, and a novel antibacterial substance is separated and purified from the culture solution.

The novel antibacterial substance according to claim 7 is characterized by being obtained by the production method according to claim 6.

An antibacterial and deodorant processing method according to an eighth aspect is characterized in that the novel antibacterial substance according to the seventh aspect is contained in a workpiece.

According to a ninth aspect of the present invention, there is provided an antibacterial and deodorized processed product characterized by being subjected to the method of the eighth aspect.

The antibacterial powder according to the tenth aspect is the first aspect.
A large number of the microorganisms according to any one of Items 1 to 4, each forming a spore.

The tablet according to the eleventh aspect is formed by tableting the antibacterial powder according to the tenth aspect.

A method for producing compost according to a twelfth aspect is characterized in that the microorganism according to any one of the first to fourth aspects is mixed into feces.

According to a thirteenth aspect of the present invention, there is provided a sheet-like material.
The microorganism according to any one of Items 1 to 4, wherein the microorganism is attached as it is and / or in a sporulated state via a binder.

[0022] A method for removing a malodor according to claim 14 is as follows.
Ponds, water reservoirs, sewage reservoirs, sewers, sewers, etc.
The microorganism according to any one of claims 1 to 4 as it is in a sewage treatment plant, a dairy or beef cattle breeding ground, a poultry farm, a pig farm, a slaughterhouse, an operating room, sanitary sewage, or animal leather being processed. And / or a sporulated state, or an intracellular product and / or an extracellular product of the microorganism are sprayed, mixed or attached.

[0023]

BEST MODE FOR CARRYING OUT THE INVENTION

[0024]

EXAMPLES Examples of the present invention will be described below, but these examples are merely examples for describing the present invention, and do not limit the scope of the present invention.

A. Novel Microbial Screening Three microorganisms having antibacterial properties against bacteria that produce a bad odor by growth were isolated from soil. Soil is generally said to be less toxic and can be said to be safe (friendly) for the three recent humans isolated from such soil.

B. Identification of bacterial strain The mycological properties of the cells isolated and selected here were identified based on the following. That is, Bergey's Mannual of Determinative Bacteriolog
y, Vol. 2 (1986), Williams & Wilkins USA REGordon, WCHaynes, CHPang. (1973) The G
enus Bacillus. Agr. Handbook No.427 United states
department of Agriculture. Identified based on Washington DC. The test was performed by the following method.

1. Morphological properties (the results are shown in Table 1 below)
Be) (1) The size of the cells (2) cell shape (3) is observed with an electron microscope with and without magnification 5,000 to 30,000 times the polymorphic cell. The test bacterium is cultured at 37 ° C. for 1 day on a normal agar plate medium, and is collected from the generated colonies. The normal agar medium used was 5 g of meat extract, 10 g of peptone, 5 g of NaCl,
It consists of 15 g of agar and 1,000 ml of distilled water.

(4) Motility 1) Flagella are stained by the method of Nishizawa and Sugawara and observed with an electron microscope (magnification: 10,000 times). The first liquid was composed of 100 ml of tannic acid, 1.5 ml of ferric chloride solution, 1.5 ml of formalin, 1 ml of 1% NaOH, and 100 ml of distilled water. The second liquid was composed of 2 g of silver nitrate, a small amount of aqueous ammonia, and 100 ml of distilled water. Become. 2) Perform puncture culture on a semifluid ordinary agar high-rise medium, and determine the turbidity of the entire medium. The semi-fluid agar medium used was meat extract 5 g, peptone 10 g, NaCl 5 g, agar 5
g, 1,000 ml of distilled water.

(5) Presence or absence of spores 1) A culture solution shake-cultured in a normal broth medium is used for 8 hours.
After heating at 5 ° C for 15 minutes, pour and culture on a normal agar plate medium,
Judgment is based on the occurrence of colonies. The normal broth medium used was 3 g of meat extract, 5 g of peptone, and 1,000 ml of distilled water.
ml. 2) Wirtz's method (Schaeffer-Fulto)
The spores are stained according to (n modification), and the bacterial cells are judged to be red and the spores are judged to be green. The red color of the cells is from a 0.5% safranin aqueous solution, and the green spores are from a 5% malachite green aqueous solution.

(6) Spore shape (7) Spore shape (8) Spore formation site (9) Spore size Spores are stained by Wirtz's method (a modified method of Schaeffer-Fulton), and magnification is 5, Observe with an electron microscope at 000x. The test bacterium is usually 37 on an agar plate medium.
After culturing at 1 ° C. × 1 day, pick from colonies cooled in a refrigerator at 4 ° C. × 1 day.

(10) Gram staining The modification is performed by the modified method of Hucker. Positives are determined by crystal violet purple and negatives by counterstained safranin red. The test bacterium is cultured at 37 ° C. for 1 day on a normal agar plate medium, and is collected from the generated colonies.

(11) An acid-fast Ziehl-Neiesen staining method is used. Positive is determined by red of fuchsin carbonate and negative is determined by green of counterstained malachite green. The test bacterium is cultured at 37 ° C. for 1 day on a normal agar plate medium, and is collected from the generated colonies.

[0033]

[Table 1] .

[0034] 2. Growth conditions in each medium (results are shown below)
( Shown in [Table 2] and [Table 3]) (1) Normal agar plate culture [Medium composition] (in 1,000 ml of distilled water) Meat extract 5 g, sodium chloride 5 g, peptone 10 g,
15 g of agar (pH 7.0 ± 0.1) [Culture conditions] A medium is fixed on a sterile petri dish, and streaked culture is performed thereon. 37 ° C. × 3 days.

(2) Ordinary agar slant culture [Medium composition] (in 1,000 ml of distilled water) Same as above [Culture conditions] A medium is fixed on a slant in a sterile test tube and streaked on it. 37 ° C. × 3 days.

(3) Ordinary broth liquid culture [Medium composition] (in 1,000 ml of distilled water) 5 g of meat extract, 5 g of sodium chloride, 10 g of peptone,
pH 7.0 ± 0.1 [Culture conditions] The medium is dispensed into a sterile test tube, and the bacteria are suspended therein. Shaking culture at 37 ° C. × 3 days, 170 times / min.

(4) Gelatin stab culture [Medium composition] (in 1,000 ml of distilled water) 5 g of meat extract, 5 g of sodium chloride, 10 g of peptone,
15 g of gelatin (pH 7.0 ± 0.1) [Culture conditions] A medium is fixed in a sterile test tube at a high layer, and puncture culture is performed. 25 ° C. × 7 days [Observation items other than growth] Liquefaction of gelatin by digestion of protein.

(5) Litmus milk liquid culture [Medium composition] (in 1,000 ml of distilled water) 110 g of skim milk powder, appropriate amount of litmus liquid, pH 7.0 ± 0.
1 [Culture conditions] Dispense a medium into a sterile test tube and suspend the bacteria therein. Stationary culture at 25 ° C for 14 days [Observation items other than growth] Coagulation of milk by the production of acid when decomposing lactose Coagulation of milk by the production of lactose When lactose is decomposed Of beef sediment by the production of beef.

[0039]

[Table 2]

[Table 3] .

[0040] 3. Physiological properties (1)
4)) (1) Reduction of nitrate [Medium composition] (in 1,000 ml of distilled water) 1 g of potassium nitrate, 5 g of peptone [Culture conditions] Dispense the medium into a sterile test tube, and place one platinum loop of bacteria therein. Suspended. Culture with shaking at 37 ° C. × 5 days, 170 times / minute [Items to be observed] Detection of nitrite: 1 ml of α-naphthylamine solution and 1 ml of sulfanilic acid solution were mixed well in the medium, and a pink-red color was judged as positive.

(2) Denitrification reaction [Medium composition] (in 1,000 ml of distilled water) 1) 10 g of sodium nitrate, 5 g of meat extract 2) 5 g of meat extract [Culture conditions] Put the above culture mediums 1 and 2 in a sterile test tube. Dispense two tubes at a time and suspend one loopful of bacteria in it. Liquid paraffin is layered on one of the two by 1-2 cm. 37 ℃ ×
Static culture for 1 to 3 days [Observation items] Whether there is anaerobic growth in the presence of nitrate: Judgment is made by observing the presence or absence of nitrate, the turbidity of the four liquid paraffins, and the observation of gas generation.

(3) VP test [Medium composition] (in 1,000 ml of distilled water) 5 g of peptone, 5 g of potassium dihydrogen phosphate, 5 g of glucose
g [Culture conditions] A medium is dispensed into a sterile test tube, and one loopful of bacteria is suspended therein. Shaking culture at 37 ° C x 3 days, 170 times / minute [Observation item] acetylmethylca, a glucose degradation product
Investigate the presence or absence of rbinol: 0.6 ml of α-naphthol solution and 0.2 ml of 40% KOH aqueous solution are added to 1 ml of the culture solution, and those which turn dark red are judged as positive.

(4) MR test [Medium composition] (in 1,000 ml of distilled water) 5 g of peptone, 5 g of potassium dihydrogen phosphate, 5 g of glucose
g [Culture conditions] A medium is dispensed into a sterile test tube, and one loopful of bacteria is suspended therein. 37 ° C. × 3 days, shaking culture at 170 times / min [Observation item] Investigate acid formation by glucose decomposition: Methyl red is dropped into 1 ml of culture solution, and red is determined as positive and yellow is determined as negative. Production [Medium composition] (in 1,000 ml of distilled water) 10 g of peptone, 5 g of sodium chloride [Culture conditions] A medium is dispensed into a sterile test tube, and one loopful of bacteria is suspended therein. Shaking culture at 37 ° C. × 3 days, 170 times / minute [Observation items] Detection of indole: The presence or absence of the ability to produce indole from the amino acid tryptophan is examined. 1/5 of culture
Add ~ 1/10 Kovac reagent (see below), shake well, and allow to stand. If the upper layer separated into two layers is reddish, it is determined to be positive; if yellow, it is negative. <Kovac reagent> 5 g of p-dimethylaminobenzaldehyde, amyl alcohol 75 ml, concentrated hydrochloric acid 25 ml.

(6) Production of hydrogen sulfide [Medium composition] (1,000 ml of distilled water, commercially available TSI agar medium) 5 g of meat extract, 1 g of glucose, 5 g of sodium chloride, 0.2 g of ferric citrate, 15 g of peptone, Sodium thiosulfate 0.2 g, lactose 10 g, phenol red 0.0
02 g, sucrose 10 g, agar 15 g [Culture conditions] A medium is fixed on a semi-slope in a sterile test tube, punctured and cultured on a high layer, and applied and cultured on a slope. 37 ° C. × 1 day [Observation items] Detection of hydrogen sulfide: A black spot on the lower slope is judged as positive.

(7) Hydrolysis of starch [Medium composition] (in 1,000 ml of distilled water) 2 g of starch, 5 g of meat extract, 10 g of peptone, 5 g of sodium chloride, 15 g of agar [Culture conditions] The medium was fixed on a sterile petri dish. And streak culture thereon. Room temperature × 5 days [Observation items] Detection of starch: Iodine, iodine potash solution (see below) was dropped on a petri dish in which bacterial growth was observed, and if dark purple disappeared, it was determined to be positive. <Iodine, iodine potash solution> 5 g of potassium iodide, 4 g of iodine / 200 ml.

(8) Liquefaction of casein [Medium composition] (in 1,000 ml of distilled water) 2 g of skim milk, 5 g of agar [Culture conditions] The above media were separately sterilized, mixed and fixed on a sterile petri dish, and placed on top. Streak culture. 37 ° C. × 1 day [Observation item] Casein remaining: If a transparent part was observed around the image area, it was determined to be positive.

(9) Use of citric acid [Medium composition] (1,000 ml of distilled water, commercially available CIT agar medium) 1 g of dipotassium phosphate, 1 g of monoammonium phosphate, 2 g of sodium citrate, 0.2 g of magnesium sulfate, chloride 5 g of sodium, 0.024 g of BTB, 15 g of agar [Culture conditions] A medium is fixed on a sterile petri dish and streaked on it. 37 ° C. × 1 to 3 days [Observation item] Whether to grow using only citric acid as a carbon nutrient: Whether the culture is blue or the growth of bacteria is recognized as positive.

(10) Production of pigment [Medium composition] (in 1,000 ml of distilled water) 20 g of peptone, 10 g of glycerin, K ₂ SO ₄ 10
g, 1.4 g of MgCl _2, 15 g of agar [Culture conditions] A medium is fixed on a sterile petri dish, and streaked culture is performed thereon. 25 ° C. × 5 days [Observation items] Dye formation: When only the periphery of the colony is colored, it is determined that each dye is water-insoluble.

(11) Urease [Medium composition] (in 1,000 ml of distilled water) 20 g of urea, 2 g of peptone, 1 g of glucose, 5 g of sodium chloride, 2 g of potassium dihydrogen phosphate, 6 ml of 0.2% phenol red, 15 g of agar ( However, agar is 121
Sterilize at 15 ° C. × 15 minutes, filter sterilize the others.) [Culture conditions] The medium is fixed on a slope in a sterile test tube, and the medium is applied and cultured. 37 ° C. × 1 day [Observation item] Detection of ammonia: When the culture medium turns red, it is judged to be positive.

(12) Oxytase [Medium composition] (in 1,000 ml of distilled water) 5 g of meat extract, 10 g of peptone, 5 g of sodium chloride,
15 g of agar [Culture conditions] A medium is fixed on a sterile petri dish, and streaked culture is performed thereon. 37 ° C x 1 day [Observation items] Presence or absence of cytochrome: Dimetyl-p
-A 1% aqueous solution of phenylenediamine was dropped, and the color of the drop changed from pink to black.

(13) Catalase [Medium composition] (in 1,000 ml of distilled water) 5 g of meat extract, 10 g of peptone, 5 g of sodium chloride,
15 g of agar [Culture conditions] A medium is fixed on a sterile petri dish, and streaked culture is performed thereon. 37 ° C. × 1 day [Observation items] The presence or absence of catalase: the presence or absence of an enzyme that catalyzes the decomposition of hydrogen peroxide. One drop of a 3% H ₂ O ₂ solution is placed on a slide glass, and one platinum loop of the bacterium is thoroughly mixed therein. It is determined to be positive when a large amount or continuous generation of oxygen bubbles occurs.

[0052]

[Table 4] .

[0053] 4. Physiological properties (2)
5] [Table 6] (1) Growth by pH [Medium composition] (1,000 ml of distilled water, commercially available nutrient broth) 3 g of meat extract, 5 g of peptone, pH adjuster (acid: H ₂ S)
O ₄ 1ml / 100ml, alkali: NaOH 4g
/ 1,000 ml), and the change in pH is 3.6 to 10.9.
[Culture conditions] A medium is dispensed into a sterile test tube, and one platinum loop of bacteria is suspended therein. Shaking culture at 37 ° C x 1 day, 170 times / minute [Observation items] Presence or absence of growth ++: Growing well + ... Growing-+: Growing slightly -... Grows ---......... Does not grow at all.

(2) Growth by Temperature [Medium composition] (1,000 ml of distilled water, commercially available ordinary agar medium) 5 g of meat extract, 5 g of sodium chloride, 10 g of peptone,
15 g of agar (pH 7.0 ± 0.1) [Culture conditions] The medium was fixed on a sterile petri dish, and the streak cultivation temperature was raised thereon to 4,10,20,30,37,40,5.
Change to 0 ° C x 1 day for each [Observation item] Presence / absence of growth +++ ... Grow well + ... Growth-+ ... Grow slightly -... Grow very slightly ---- … They do not grow at all.

(3) Attitude to oxygen [Medium composition] (1,000 ml of distilled water, commercially available ordinary agar medium) 5 g of meat extract, 5 g of sodium chloride, 10 g of peptone,
15 g of agar (pH 7.0 ± 0.1) [Culture conditions] Culture was performed by fixing the bacteria and the medium in a sterilized test tube in a high layer in a mixed state. 37 ° C x 1 day [Observation items] Growth under aerobic and anaerobic conditions Growing only on the surface ………… Aerobic Growing on the surface and high layers ……… Facultative anaerobic Growing only in the high layers ……… ... obligate anaerobic.

(4) OF test [Medium composition] (in 1,000 ml of distilled water) 2 g of peptone, 5 g of sodium chloride, K ₂ HPO ₄
0.3 g, agar 3 g, 0.2% BTB 15 ml, glucose 10 g (Glucose is sterilized by filtration, and others are sterilized at 121 ° C. × 15 minutes) [Culture conditions] The medium is dispensed into two sterile test tubes, and Fixed to. Puncture culture is performed on each, and one of them is overlaid with liquid paraffin by 1 to 2 cm. 37 ° C x 3 to 4 days [Observation item] Check whether sugar degradation is performed oxidatively or fermentatively. "0" ..... Oxidative sugar degradation (when yellowing occurs only under anaerobic conditions) "F" ..... fermentative sugar decomposition (when both aerobic and anaerobic conditions turn yellow) (5) PPA test [Medium composition] (in 1,000 ml of distilled water) 3 g of yeast extract, 2 g of phenylalanine, 1 g of sodium phosphate, 5 g of sodium chloride 15 ml of agar [Culture conditions] A medium is fixed on a slope in a sterile test tube, and the medium is applied and cultured. 37 ° C. × 1 day [Observation items] Whether or not phenylalanine is deaminated to phenylpyruvic acid: A positive judgment is made when green color changes due to the dropwise addition of a 10% aqueous ferric chloride solution.

(6) Use of propionic acid [Medium composition] (in 1,000 ml of distilled water) 0.2 g of magnesium sulfate, 2 g of sodium propionate,
1 g dipotassium hydrogen phosphate, 1 g monoammonium phosphate, 5 g sodium chloride, 10 g agar, 0.2% BTB solution 1
2 ml [Culture conditions] The medium is fixed on a sterile petri dish, and streaked culture is performed thereon. 37 ° C. × 1 day [Observation items] Whether to grow using only propionic acid as a carbon nutrient: Whether the medium is blue or the growth of bacteria is determined to be positive.

(7) Decomposition of tyrosine [Medium composition] (in 1,000 ml of distilled water) 5 g of L-tyrosine, 3 g of meat extract, 5 g of peptone, 15 g of agar (however, tyrotin and nutrient agar were separately sterilized by wet heat). [Culture conditions] The culture medium is fixed on a sterile petri dish, and streaked culture is performed thereon. 37 ° C. × 7 to 14 days [Observation items] Tyrosine degradation: Whether or not tyrosine present as crystals under the colony is dissolved is judged to be positive.

(8) Egg yolk reaction [Medium composition] (in 1,000 ml of distilled water) 10 g of peptone, 5 g of Na ₂ HPO ₄ , KH ₂ PO ₄
1 g, 2 g of NaCl, 0.1 g of MgSO ₄ , 2 g of glucose, and 15 ml of egg yolk (however, except for the yolk, sterilize by moist heat, add the aseptically sucked egg yolk, chill in the refrigerator all day and night, and prepare broth without yolk) [Culture conditions] Dispense the medium with or without egg yolk into two sterile test tubes and suspend one loopful of bacteria in each of them. At 37 ° C. × 7 days, observations are made on days 1, 3, 5, and 7.
Shaking culture at 170 times / min [Observation item] Presence / absence of white precipitate: Positive when white precipitate is observed in the lower part or surface of test tube with yolk compared to without yolk.

(9) Growth under 2% NaCl [Medium composition] (in 1,000 ml of distilled water) 3 g of meat extract, 5 g of peptone, and 20 g of NaCl [Culture conditions] The medium was dispensed into a sterile test tube and placed in the tube. 1. Suspension of platinum loop bacteria. 37 ° C. × 14 days, shaking culture at 170 times / minute [Observation items] Presence or absence of growth ++: Growing well +: Growing-+: Growing slightly-: Very slight Grows ---......... Does not grow at all.

(10) Growth under 5% NaCl (11) Growth under 7% NaCl (12) Growth under 12% NaCl (13) Growth under 20% NaCl [Medium composition] (distilled water) (In 1,000 ml) Change the above NaCl to 70 g, 120 g, and 200 g. [Culture conditions] Same as 9 [Observation items] Same as 9 above.

(14) Growth in the Presence of Rhizotium [Medium composition] (in 1,000 ml of distilled water) 3 g of meat extract, 5 g of peptone, 0.1 g of lysotium (lysotium was boiled in a 0.01 N HCl solution for 20 minutes) [Culture conditions] A medium is dispensed into a sterile test tube, and one loopful of bacteria is suspended therein. 37 ° C. × 14 days, shaking culture at 170 times / minute [Observation items] Presence or absence of growth ++: Growing well +: Growing-+: Growing slightly-: Very slight Grows ---......... Does not grow at all.

(15) Production of hemolysin [Medium composition] (1,000 ml of distilled water, commercially available sheep blood agar medium) Pancreatic Digest of Casein. 14.5 g, Papaic Dig
est of Soybean Meal. 5.0 g, Sodium Chloride.
0g, Growth Factors 1.5g, Agar 14.0g, Shee
p blood, defibrinated. 5.0% [Culture conditions] The medium is fixed on a sterile petri dish, and streaked on it. 37 ° C. × 1 day [Observation item] Presence or absence of production of hemolysin α-type hemolysis: green zone around colony (degeneration of hemoglobin) β-type hemolysis: clearing around colony (destruction of erythrocyte membrane).

[0064]

[Table 5]

[Table 6] .

[0065] 5. Carbon source acid and gas generation (results below
( Shown in Table 7) (1) D-glucose [Medium composition] (in 1,000 ml of distilled water) 1 g of peptone, 5 g of sodium chloride, 10 g of agar, 0.1 g of agar.
15 ml of 2% BTB, 8 g of D-glucose [Culture conditions] A medium is dispensed into a sterile test tube and fixed on a high layer. Thereafter, it is punctured and cultured. 37 ° C x 14 days [Observation item] Acid generation: Judgment based on yellowing degree of BTB reagent ++ ... well generated + ... ... generated-+ ... weak generated------No generated gas Generation: Judgment based on the presence or absence of a crack in the high-rise part ++: well-formed +: generated-+: weakly generated-: not generated

(2) L-arabinose (3) D-xylose (4) D-mannitol (5) D-mannose (6) D-galactose (7) D-sorbitolose (8) inositol (9) trehalose (10 ) Lactose (11) Maltose (12) Fructose [Medium composition] (in 1,000 ml of distilled water) The sugar (D-glucose) part in (1) above was changed to the sugars in (2) to (12) above. [Culture conditions] Same as above (1) [Observation items] Same as above (1).

[0067]

[Table 7] .

[0068] 6. Identification results OYK-01-600, OYK-03-600, and O
Three strains of YK-04-000 were classified according to the following literature.

[0069] Bergey's Mannual of Determinative
Bacteriology, Vol.2 (1986), Williams & Wilkins US
A REGordon, WCHaynes, CHPang. (1973) The G
enus Bacillus. Agr. Handbook No.427 United states
department of Agriculture. Washington DC

[Table 8]

[Table 9]

[Table 10] .

As a result, all of the above three strains showed that Ba
cillus , then [Table 9] [Table 10]
It is considered to be a related bacterium of B. subtilis , but it grows under anaerobic conditions, the production of acid from sugar is weak,
And did not grow in 7% NaCl, the identification was not completed, and it was judged to be a new species, and it was named Bacillus sp .

In addition, Bacillus sp. OYK-01-600
Was registered with the Patent Microorganisms Depositary Center of the Institute of Biotechnology and Industrial Technology, Ministry of International Trade and Industry on April 30, 1996, at FERM.
Deposited as P-15607 and OYK-03-60
0 is FERM P-15608, OYK-04-00
0 has also been deposited as FERM P-15609. Hereinafter, OYK-01-600, OYK-03-
Each of 600 and OYK-04-000 is also simply referred to as “OYK bacteria”.

The above mutant strain has antibacterial properties against bacteria that generate a bad odor with growth. Further, the microorganism can be cultured, and an antibacterial substance can be separated and purified from the culture solution. Separated and purified antibacterial substance as it is, or dissolved or dispersed in a suitable solvent such as water,
As it is or appropriately diluted, the mixture alone or in combination with other substances, spraying, coating, impregnation, immersion,
Antibacterial, odor-preventing articles, or articles for which these properties are to be provided in advance, using at least one of attachment, adhesion, adhesion, mixing, addition, oral administration, and injection. Products, bedding, clothing, medical products,
Sanitary equipment and supplies, sanitary products, footwear, various air conditioners and air conditioners (including air purifiers)
Filters, building materials including interior and exterior, furniture, animals, animal breeding grounds, animal breeding equipment, animal breeding water and feed, tableware, chemicals, sewage or sewage treatment plants or equipment, microcapsules, passenger transportation vehicles or ships or aircraft It is applied directly or indirectly to interior materials including seats, morgues and funeral supplies.

Further details will be described in the following examples. C. Confirmation of antibacterial active substance possessed by microorganisms (results are shown in [Table 11] below) Bacillus sp. OYK-01-600 (FERM P-1
5607), Bacillus sp. OYK-03-600 (FE
RM P-15608), Bacillus sp. OYK-04-
000 (FERM P-15609) secreted an antibacterial active substance by the following experiment.

1. Preparation of microbial suspension (1) Microorganisms OYK-01-600, OYK-03-600, and O
Three strains of YK-04-000.

(2) Preculture [Medium composition] (in 1,000 ml of distilled water) 10 g of peptone, 5 g of meat extract, 5 g of sodium chloride,
Agar 10 g [Culture conditions] A medium is fixed on a sterile petri dish, and the stored bacteria are streaked thereon. 37 ° C. × 1 day.

(3) Main culture [Medium composition] (in 1,000 ml of distilled water) 10 g of peptone, 5 g of meat extract, 5 g of sodium chloride [Culture conditions] The medium was dispensed into a sterilized test tube, and one platinum loop was placed therein. Resuspend. Incubate at 37 ° C x 1 day with shaking 170 times / min.

[0077] 2. Preparation of suspension of antibacterial test bacteria (1) Antibacterial test bacteria Klebsiella pneumoniae ATCC 4352 Staphylococcus aureus ATCC 6538P.

(2) Preculture [Medium composition] (in 1,000 ml of distilled water) Calf brain leachate 200 g, bovine heart leachate 250 g, proteose peptone 10 g, glucose 2 g, sodium chloride 5 g, disodium phosphate 2.5 g [Agar] 15 g [Culture conditions] A medium is fixed in a sterile petri dish, and the stored bacteria are streaked thereon. 37 ° C. × 1 day.

(3) Main culture [Medium composition] (in 1,000 ml of distilled water) 10 g of peptone, 5 g of meat extract, and 5 g of sodium chloride [Culture conditions] The medium was dispensed into a sterile test tube, and put into the tube. Incubate with shaking.

Klebsiella pneumoniae … 37 ° C. × 20
Time, 170 times / minute Staphylococcus aureus … 37 ° C. × 10 hours, 170
Times / minute.

(4) Adjustment of the number of bacteria The number of bacteria was 5 to 30 × 10 ⁵ cells /
Adjust to make ml.

[0082]

[0083] 3. Antimicrobial test (1) Preparation of test medium 1) 10 g of peptone,
A medium (in 1,000 ml of distilled water) consisting of 5 g of meat extract, 5 g of sodium chloride, and 10 g of agar was added in an amount of 0.1% each.
Pour 2 ml and fix on sterile Petri dish. 2) Peptone 10g, meat extract 5g, sodium chloride 5
medium containing 10 g of agar and 10 g of agar (1,000 m of distilled water).
is fixed on a sterile petri dish, and the bacterial solution of the two test bacteria of the above (4) is inoculated on the entire surface thereof with a 0.2 ml spreader.

[0084] (2) Preparation of bacteria for comparison of microorganisms and antimicrobial OYK-01-600, OYK-03-600, Baci llus subtilis , including natto for comparison with three strains of OYK-04-000 Three strains of each of A, B, and C were prepared.

(3) Inoculation of Microorganisms and Antibacterial Comparative Bacteria OYK- (3) of the above “1. Preparation of Microbial Suspension”
01-600, OYK-03-600, OYK-04-000 and the above (2)
A total of 6 strains, 3 strains of each of A, B and C, are inoculated on the test medium prepared in the above (1) by the following methods 1) and 2). 1) Place a penicillin cup at the center of the Petri dish and pour 0.05 ml of each bacterial solution into it. 2) 0.02 ml of each bacterial solution is dropped at the center of the Petri dish. The inoculated one is cultured at 37 ° C. for 1 day.

(4) Judgment method ++: No growth of test bacteria was observed below and around the colonies of the test bacteria, and a growth inhibition zone (halo) of 2 mm or more was observed. No growth of test bacteria, no growth inhibition zone (halo) of 0 to 2 mm is observed + -... No growth inhibition zone (halo) is observed, but no growth of test bacteria is observed in the test colony -------------------------------------------------------------------------------------------------------------Test

[0087]

[Table 11] .

As shown in [Table 11], in the growth phase of the microorganism, Klebsiella pneumoniae and Staphylococcus au
reus was found to secrete a substance showing antibacterial activity against both.

D. Odor prevention test of wastewater treatment plant by microorganisms (results are shown in the following [Table 12]) Bacillus sp. OYK-01-600 (FERM P-1
5607), Bacillus sp. OYK-03-600 (FE
RM P-15608), Bacillus sp. OYK-04-
The following experiment confirmed that putting 000 (FERM P-15609) into the wastewater treatment plant was effective in deodorizing around the treatment plant.

1. Outline of wastewater treatment plants used for testing (1) Location and type of industry Mie Prefecture, laundry industry (2) Wastewater facilities and treatment capacity See Figure 1.

[0091] 2. Deodorization test method (1) Preparation of suspension of microorganisms In this test, Bacillus sp. OYK-01-600 (FER
MP-15607) was selected, and a suspension of microorganisms was prepared in the same manner as in “1. Preparation of suspension of microorganisms” described above.

(2) Injection of bacteria into drainage facility 2.5 liters of the microorganism suspension prepared in the above (1) was introduced into the raw water tank of the wastewater treatment facility in FIG.

(3) Judgment of effect 1) Judge: 3 employees of the same factory 2) Judgment criteria: 6-step odor intensity display method Odor intensity 0 = odorless 1 = odor that can be finally detected (detection threshold density) 2 = what Weak odor (cognitive threshold concentration) that can be detected. Number of bacteria per ml of input bacteria

[Table 12] .

As shown in [Table 12], it was confirmed that the intensity of the odor decreased as the microorganisms grew. It is considered that the above results were obtained because this microorganism did not generate ammonia, hydrogen sulfide, and the like at the time of growth, and performed sugar decomposition without generating gas. When other microorganisms grow,
No confirmation was made as to whether the generated gas would be consumed nutritionally.

In addition, Bacillu sp. OYK-03-600
(FERM P-15608) and Bacillus sp. OYK
-04-000 (FERM P-15609) was confirmed to provide the same good results as above.

E. Separation and purification of secretions produced by microorganisms Bacillus sp. OYK-01-600 (FERM P-1
5607), Bacillu sp. OYK-03-600 (FE
RM P-15608), Bacillus sp. OYK-04-
000 (FERM P-15609) is available from Klebsiella p.
Substances showing antibacterial activity against neumoniae and Staphylococcus aureus were separated and purified as follows.

1. Preparation of microorganism suspension (1) Microorganisms OYK-01-600, OYK-03-600, OYK
Baci containing Bacillus natto for comparison
Three strains of A, B and C strains of llus subtilis performed the same operation.

(2) Pre-culture [Medium composition] (in 1,000 ml of distilled water) 10 g of peptone, 5 g of meat extract, 5 g of sodium chloride,
Agar 10 g [Culture conditions] A medium is fixed on a sterile petri dish, and the stored bacteria are streaked thereon. 37 ° C. × 1 day.

(3) Main culture [Medium composition] (in 1,000 ml of distilled water) 10 g of peptone, 5 g of meat extract, 5 g of sodium chloride [Culture conditions] A medium was dispensed into a sterilized Sakaguchi flask, and one loop of platinum was added. Resuspend. 37 ° C x 2 days, 170 times /
Incubate with shaking for a minute.

[0100] 2. Separation of bacterial cells and secretions The obtained culture solution was centrifuged at 4 ° C and 12,000 RPM for 1 hour.
Centrifuge for 4 minutes in a cryogenic centrifuge. Only the supernatant is taken out, and the solution is subjected to suction filtration with a 0.45 μm membrane filter.

3. Concentration of secretions The obtained filtrate was transferred to an eggplant type flask, and after the filtrate was completely frozen on the wall of the flask, it was dried with a freeze vacuum dryer for 48 hours to obtain a purified powder.

F. Antibacterial activity test of substances produced by microorganisms (results are shown in [Table 13] below) Bacillus sp. OYK-01-600 (FERM P-1
5607), Bacillu sp. OYK-03-600 (FE
RM P-15608), Bacillus sp. OYK-04-
000 (FERM P-15609), the antibacterial activity of the separated and purified product which is considered to have antibacterial activity was confirmed by the following method.

1. Sample: Secreted substance Bacillus sp. OYK-01-600 (FERM) obtained in “3 .
P-15607) Bacillus sp. OYK-03-600 (FERM
P-15608) Bacillus sp. OYK-04-000 (FERM
P-15609) Bacillus sp. A Bacillus sp. B Six secretions of Bacillus sp. C and the following eight samples used as antibacterial reference agents were used. Chloramphenicol (Antibiotics manufactured by Wako Pure Chemical Industries, Ltd., Mw: 323.13 CAS: 56-75-7, which has an inhibitory effect on protein synthesis, Gram-positive bacteria, Gram-negative bacteria, rickettsia,
Effective against viruses, antibacterial, usually bacteriostatic. 0.2 ml of a 10 mg / 100 ml aqueous solution was used for the antibacterial test). Nicanon RB (a quaternary ammonium-based synthetic antibacterial agent manufactured by Nikka Kagaku Co., Ltd., which has excellent antibacterial properties against Staphylococcus aureus and has antifungal properties. 0.2 ml of a 1 ml / 100 ml aqueous solution). A total of 8 samples are performed.

[0104] 2. Antibacterial test bacteria Two kinds of Klebsiella pneumoniae ATCC 4352 Staphylococcus aureus ATCC 6538P obtained in "2. Preparation of suspension of antibacterial test bacteria" described above.

[0105] 3. Antibacterial test method (1) Method for determination based on turbidity of culture solution 1) [Medium composition] (1,000 ml of distilled water, commercially available nutrient broth) 5 g of meat extract, 10 g of peptone,
5 g of sodium chloride 2) [Culture conditions] Dispense 5 ml of the medium into a sterile test tube, and add
2 g and 0.1 ml of the test bacterial suspension are added and poured. 37
C. × 1 day, 170 times / minute shaking culture 3) [Observation items] Model 100-20, manufactured by Hitachi, Ltd.
The transmittance of the culture solution is measured with a spectrophotometer. Measurement wavelength 4
75 nm, the display unit% (by Klebsiella pneumonia e, about 5 to 30 × 10 ⁸ number of bacteria, showing transmittance of 52%).

(2) Method for determination by measuring the number of bacteria 1) [Medium composition] (commercially available agar medium in 1,000 ml of distilled water) 5 g of meat extract, 10 g of peptone, 5 g of sodium chloride,
15 agar agar 2) [Culture conditions] 15 ml of the medium is dispensed into a sterile petri dish, and 0.02 g of the sample is pulverized and dissolved therein, and then fixed. A test bacterial suspension (0.2 ml) is uniformly applied to the surface and cultured. 37 ℃ ×
1 day 3) [Observation items] The number of colonies grown on the petri dish is counted. Display unit: exponent part (for example, “B” is displayed for A × 10 ^B ).

(3) Method of Judgment by Observation of Growth Inhibition Zone (Halo) 1) [Medium composition] (1,000 ml of distilled water, commercially available ordinary agar medium) 5 g of meat extract, 10 g of peptone, 5 g of sodium chloride,
15 g of agar 2) [Culture conditions] 15 ml of the medium was dispensed and fixed in a sterile petri dish, 0.2 ml of the test bacterial suspension was uniformly applied to the surface, and after the surface was half-dried, the sample was solid in the center of the petri dish. And static culture. 37 ° C x 1 day 3) [Observation item] The maximum diameter of the generated halo is measured. Display unit: m
m.

(4) Penicillin cup method 1) [Medium composition] (commercially available nutrient broth in 1,000 ml of distilled water) Lower medium: meat extract 5 g, peptone 10 g, sodium chloride 5 g, agar 15 g Upper layer medium: meat extract 5 g, peptone 10 g, sodium chloride 5 g, agar 7 g 2) [Culture conditions] 15 ml of the lower medium is dispensed and fixed in a sterile petri dish. On top of this, 0.1 ml of the test bacterial suspension is mixed with 4 ml of the upper layer medium and layered. After the surface is semi-dried, a stainless steel cylinder (inner diameter 6 mm, outer diameter 8 mm, height 10)
mm) is dropped vertically from a height of 10-13 mm.
A sample prepared by dissolving 0.02 g of the sample in 0.2 ml of sterilized water is placed in the cylinder, and the culture is allowed to stand. 37 ° C x 1 day 3) [Observation items] Observe the growth of test bacteria inside and outside the cup. Display unit---The growth of test bacteria is seen in the entire cup. Proliferation is observed +: No growth of test bacteria is observed in the cup A: When a halo having a maximum diameter of Amm (for example) is observed outside the cup.

[0109]

[Table 13] As shown in [Table 13], it was confirmed that the substance secreted by the microorganism had an antibacterial effect.

G. Antibacterial activity test of OYK cell product As an OYK bacterium, Bacillu sp. OYK-01-600 was used, but Bacillu sp. OYK-03-600 (FERM
P-15608) and Bacillus sp. OYK-04-0
Similar good results were obtained for 00 (FERM P-15609).

[0111] 1. Separation and purification of OYK cell product Extracellular product (1) 1 ml of the seed cell cryopreserved at -82 ° C was thawed. (2) A sterilized BGG liquid medium (a medium comprising soy protein extract, glucose and sodium glutamate,
4 ml was taken, and the bacterium obtained in the above (1) was added thereto and seed-cultured at 37 ° C. for 24 hours. (3) BGG liquid medium 100 sterilized in Sakaguchi flask
Then, the bacteria of the above (2) were added thereto, followed by main culture at 37 ° C. for 72 hours. (OD = 10, pH = 7.9) (4) After centrifugation, the supernatant was collected. (12,000
(0 rpm × 10 min, 4 ° C.) (5) The above (4) was filtered with a membrane filter. (0.2 μm filter) (6) The filtrate in the above (5) was freeze-dried. (7) The product freeze-dried in (6) above was dissolved in 3 to 4 ml of physiological saline. (8) In the antibacterial test described later, 50 μl of the solution of the above (7) was dropped.

Intracellular Product (1) 1 ml of a seed strain cryopreserved at -82 ° C was thawed. (2) Take 4 ml of the sterilized BGG liquid medium into a test tube, put the bacteria obtained in the above (1) into it,
Seed culture was performed for hours. (3) BGG liquid medium 100 sterilized in Sakaguchi flask
Then, the bacteria of the above (2) were added thereto, followed by main culture at 37 ° C. for 72 hours. (OD = 10, pH = 7.9) (4) After that, the mixture was centrifuged to remove a precipitate. (12,000
(0 rpm × 10 min, 4 ° C.) (5) 5 ml of physiological saline was added to the precipitate to disperse the precipitate. (6) 20 ml of n with respect to the dispersion of the precipitate
Add butanol and stir for 1 hour and wait until separated into aqueous and alcohol layers. (7) The alcohol layer of (6) above was taken out, and n-butanol was evaporated by evaporation. (8) This was transferred to a desiccator and further dried. (9) 1 ml of physiological saline was added to the dried product to disperse the purified product. (10) This dispersion was filtered with a membrane filter. (0.2μm filter) (11) In the antibacterial test described in the next section, the filtrate of (10) was
One liter was dropped.

[0113] 2. Antimicrobial test of OYK bacterial cell product Test sample bacterial product (1) OYK extracellular product obtained from (2) Intracellular product of OYK bacteria obtained from the target test bacteria (1) Staphylococcus aureus
(ATCC 25923, Staphylococcus aureus) (2) Klebsiella pneumoniae
(ATCC13883, Klebsiella pneumoniae) (3) MRSA1002 (isolate, methicillin-resistant Staphylococcus aureus) (4) Escherichia coli O-157
(S180, E. coli O-157) (5) Legionella pneumophill
a (OCI 88171, Legionella) (6) Pseudomonas aeruginosa
(IFO3452, Pseudomonas aeruginosa (producing red pigment)) (7) Pseudomonas aeruginosa
(ATCC 27853, Pseudomonas aeruginosa (producing blue pigment)) (8) Fuzarium Proliferatorum
Nirenberg S12 (IFO 6349, Fusarium (Akamold)) (9) Aspergillus niger van
Tiehem S1 (IFO6341, koji mold) (10) Penisillium cittrinum
Thom S5 (IFO6352, blue mold) (11) Trichophyton mentagrop
hytes (IFO5466, Trichophyton) (12) Rhizopus stolonifer Li
nd S7 (FERMS-7, spider mold).

Test Method (1) The storage test bacteria were seed-cultured under the optimum conditions for each of the test bacteria. (2) A fixed amount of the test bacteria solution of a certain number of bacteria was applied on an optimal solid medium of each of the test bacteria, and allowed to stand until it became semi-dry. (3) 50 μL of the two kinds of test sample products (see above) were dropped on the medium of (2). (4) The test bacteria were cultured under optimal culture conditions. (5) If there is antibacterial activity, a growth inhibition zone (halo) is observed at the dropping point. The diameter (minor axis, major axis) of the inhibition zone was measured (unit: cm). The results are shown in [Table 14] below.

[0115]

[Table 14] .

H. Use and application of OYK bacteria In addition, as OYK bacteria, Bacillu sp. OYK-01-60
0 was used, but Bacill u sp. OYK-03-600 (F
ERM P-15608) and Bacillus sp. OYK-
Similar good results were obtained for 04-000 (FERM P-15609).

(Powdering OYK Bacteria) Hereinafter, the step of powdering OYK bacteria will be described. (1) 1 ml of the inoculum frozen and stored at -82 ° C was thawed. (2) Take 4 ml of the sterilized BGG liquid medium into a test tube, put the bacteria thawed in the above (1) into it,
For 24 hours. (3) BGG liquid medium 100 sterilized in Sakaguchi flask
Then, the bacteria of the above (2) were added thereto, followed by main culture at 37 ° C. for 24 hours. At this time, the OD value was about 10, and the number of bacteria was 1 × 10 ¹⁰ cells / ml. (4) The liquid of the above (3) was freeze-dried to obtain 1 g of bacterial powder (including a medium component). 1 × 10 ¹⁰ bacteria /
g.

[0118] In the obtained bacterial powder, all OYK bacteria were sporulated. Thereby, the following operation and effect can be obtained.
That is, since it is sporulated, it can be stored at room temperature if moisture proof is taken into consideration, and there is no decrease in the number of bacteria or deterioration of the bacteria. Tableting can be performed with a tableting machine having a compression strength of about 10 tons. There is no decrease in the bacterial count by tableting. It is possible to add chemicals necessary for tableting, such as fragrances, coagulants, and slow solvents.

(Utilization as a tablet) The bacterial powder obtained in the above example was mixed with an additive (eg, glucose, sodium glutamate, extract of soy protein,
After adding a medium component), the mixture was tableted with a tableting machine to obtain tablets. When this tablet was put into the garbage, the odor generated from the garbage could be prevented as a result. This is because the OYK bacterium grows at a rate that exceeds the growth of putrefactive bacteria that emit water and the spores start to act as bacteria by dissolving the spores by adding water to the garbage. it is conceivable that. Also, OYK
Since the fungus also has antibacterial activity on Escherichia coli O-157, the use of the above-mentioned tablet around the kitchen can be applied to prevention of food poisoning. (Manufacturing Example of Compost and Effect of Manufactured Compost) Further, the OYK bacteria of the present invention exhibit an effective action also in the process of producing compost. In particular, during the production process, the compound has an effect of raising the initial fermentation temperature. An example is shown below.

The result of measuring the number of days for 700 g of cow dung to rise to a temperature of 50 ° C. is 8 days, whereas 700 g of cow dung was added to OYK bacterial solution (OYK-01-) at a concentration of 1 × 10 ⁸ cells / ml.
600) It takes only 3 days to add 50 ml,
Time savings were seen. In addition, the time for which the temperature is maintained at 40 ° C. or higher is 6 days when no addition is performed,
With the addition of OYK bacteria, the warming time was extended to 17 days. It is said that in the compost production process, a large number of bacteria and molds repeatedly grow and die in each temperature range to reduce the molecular weight of high-molecular organic matter. 70 ° C in this cycle
Survive as spores at the highest temperature of
The cycle is shortened by actively growing around 0 ° C. Furthermore, since no odor is generated during the growth, which is a characteristic of OYK bacteria, the emission of ammonia odor during the production process was prevented.

[0121] The spores of the OYK fungus are present in the fully aged cow dung compost produced in this manner.
Since Fusarium, which is the cause of the wilt, has antibacterial activity, it can prevent the wilt of the plant after fertilization.

(Sheet Formation of OYK Bacteria and Use Thereof) An attempt was made to easily utilize the effects of the OYK bacteria of the present invention in a medical practice. The details will be described below. Bacteria (OYK-01
-600) A 100 g / m ² nonwoven fabric was printed with a bacterial solution having a concentration of 1 × 10 ⁷ cells / ml and a dispersion solution containing 10% of PVA (polyvinyl alcohol) as a binder, using a screen printing machine having a density of 1500 mesh. After evaporating the water, the non-woven fabric was observed with a microscope.
It was confirmed that the YK bacteria were sporulated and immobilized in a state of being entangled with the fibers of the nonwoven fabric.

This non-woven fabric was cut into a suitable size, and the non-woven fabric was spread on sheets to observe the back of a patient suffering from bedsores caused by Pseudomonas aeruginosa. It has been confirmed that the bedsore has been improved and that the bedsores are also improving.

Although a nonwoven fabric is used in the above embodiment, the present invention is not limited to this, and a woven or knitted fabric made of synthetic resin fibers or natural fibers, a synthetic resin sheet, a synthetic resin foam sheet, or the like may be used.

(Utilization of Pond Water as Purification) Since a large amount of fertilizer is put into the turf in a pond in a golf course, the fertilizer flows into the pond and is in a eutrophic state. In such ponds, sometimes reddish phytoplankton grows abnormally, which further reduces the dissolved enzymes in the pond. As a result, the growth of anaerobic bacteria is promoted, causing the generation of offensive odor. Pond in such a state (400t water volume scale)
OYK bacterial solution (OYK-01-600, bacterial concentration 1 × 10
⁽⁸ pieces / ml) When 10 liters were introduced, the redness of the surface disappeared in 2 weeks, and the transparency increased. Although this mechanism has not been elucidated yet, it can be inferred that nutrients in the pond water have decreased due to the growth of OYK bacteria, and phytoplankton has been killed.

(Utilization as a deodorant) Water purification effect in a eutrophic pond, or deodorant effect of a wastewater (wastewater) treatment plant described in the above section "D. Deodorization test of wastewater treatment plant by microorganisms" As described above, odorous odors in dairy and beef cattle breeding farms, poultry farms, and pig farms can be prevented by dispersing OYK bacteria as it can prevent the emission of ammonia odor, as shown in the compost production examples. Deodorant is possible. In addition, in slaughterhouses, operating rooms, sanitary filth, and animal leather during processing, as described in the above [Table 12] and the next paragraph, OYK bacteria are required to degrade proteins, lipids, carbohydrates and the like. Proliferates faster than bacteria that generate bad odors and can prevent odors. The OYK bacterium to be sprayed, mixed or attached may be a solution of the bacterium as it is or a sporulated powder, and the same effect can be obtained even if it is an intracellular product or an extracellular product.

[0127]

As described above, the microorganism of the present invention is excellent in antibacterial and deodorant effects. Therefore, for example, by using the wastewater treatment plant, wastewater can be treated without emitting bad odor.

The novel antibacterial substance of the present invention is isolated and purified from soil-derived microorganisms, which are considered to be less toxic, and is relatively safe for the human body.
Enables antibacterial processing in fields that cannot be used conventionally.
Further, the safety of products that have been conventionally subjected to antibacterial processing, such as products in the field of clothing, for example, can be improved by using the antibacterial substance.

[Brief description of the drawings]

FIG. 1 is an explanatory diagram showing drainage facilities and treatment capacity in a wastewater treatment plant subjected to an odor control test.

──────────────────────────────────────────────────の Continued on front page (51) Int.Cl. ⁶ Identification symbol FI (C12N 1/20 C12R 1:07) (C12P 1/04 C12R 1:07) (54) [Title of Invention] Has antibacterial properties A novel microorganism, an antibacterial / deodorant treatment method using the novel microorganism, a method for producing an antibacterial substance using the novel microorganism, an antibacterial substance obtained by the method, an antibacterial / deodorant processing method using the antibacterial substance, and Antibacterial and deodorized processed products

Claims (14)

[Claims] 1. A bacterium belonging to the genus Bacillus and comprising Staphylococcus aureu.
s and novel microorganism characterized by having antimicrobial properties to Klebsiella pneumoniae. 2. Staphylococcus aureus and Kleb described above.
microorganism having antimicrobial properties to siella pneumoniae is, Bacill
us sp. OYK-01-600 (FERM P-1560
The novel microorganism according to claim 1, wherein the microorganism is (7). 3. The above Staphylococcus aureus and Kleb
microorganism having antimicrobial properties to siella pneumoniae is, Bacill
us sp. OYK-03-600 (FERM P-1560
The novel microorganism according to claim 1, wherein the microorganism is (8). 4. The above Staphylococcus aureus and Kleb
microorganism having antimicrobial properties to siella pneumoniae is, Bacill
us sp. OYK-04-000 (FERM P-1560
The novel microorganism according to claim 1, wherein the microorganism is (9). 5. A method for antibacterial and deodorant treatment, which is carried out using the microorganism according to any one of claims 1 to 4. 6. A method for producing an antibacterial substance, comprising culturing the microorganism according to any one of claims 1 to 4, and isolating and purifying a novel antibacterial substance from the culture solution. 7. A novel antibacterial substance obtained by the production method according to claim 6. 8. A method for antibacterial and deodorant processing, comprising adding the novel antibacterial substance according to claim 7 to a workpiece. 9. An antibacterial and deodorant processed product obtained by performing the method according to claim 8. 10. An antibacterial powder comprising a large number of microorganisms according to any one of claims 1 to 4, each of which forms a spore. 11. A tablet formed by tableting the antibacterial powder according to claim 10. 12. A method for producing compost, comprising mixing the microorganism according to claim 1 in feces. 13. A sheet comprising the microorganism according to any one of claims 1 to 4 attached as it is and / or in a sporulated state via a binder. 14. Ponds, water reservoirs, sewage reservoirs, sewers, sewage reservoirs, sewage treatment plants, dairy cattle and beef cattle breeding grounds, poultry farms, pig farms, etc. The microorganism according to any one of claims 1 to 4 as it is and / or in a sporulated state, or a cell of the microorganism, in a place, a slaughterhouse, an operating room, a sanitary waste or an animal leather being processed. A method of spraying, mixing or attaching a product and / or an extracellular product to remove a bad smell.