JP2003024051A - New microorganism, oil-and-fat decomposing agent containing the same and method for treating oil-and-fat containing substance with the decomposing agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new microorganism useful for the ready treatment of waste water discharged from a Chinese noodle restaurant, Western style restaurant and Tempura (deeply fried flour coated ingredients) restaurant as well as waste water of a plant using a relatively simple apparatus, an oil-and-fat decomposition agent containing the microorganism, and to provide a method for the treatment of a material containing oil-and-fat using the decomposition agent. SOLUTION: The microorganism is an immotile Gram-negative spherobacillus belonging to the genus Acinetobacter, exhibiting a catalase-positive and oxidase- negative nature and having an oil-and-fat decomposition activity. This invention further provides an oil-and-fat decomposition agent containing the microorganism and a method for the treatment of an oil-and-fat containing material using the agent.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel microorganism, particularly a microorganism having fat and oil degradability, an oil and fat decomposing agent containing the same, and a method for treating an oil and fat-containing substance using the same.

[0002]

2. Description of the Related Art For the treatment of wastewater containing fats and oils, industrially, a treatment facility such as a pressure flotation device, a natural flotation tank (oil pit) or an oil separation tank (grease trap) is used.

However, the installation of conventional oil and fat removing equipment such as a grease trap causes complicated problems such as accumulation of solidified oil and fat, blockage of sewer pipe, generation of bad odor, and exceeding of drainage standard value. . One of the wastewater treatment techniques that solves this problem is a biological treatment method that removes or decomposes pollutants containing fats and oils in wastewater by utilizing various organisms existing in nature, especially microorganisms. Microorganisms for this purpose include Bacillus sp. , Pseudomo
nas sp. SK0401 strain (Microtech Research Institute 1323
1), Acinetobacter sp. SK040
2A strain (Microtechnology Research Institute 13232), S. saprophy
ticus OD-1 (Microtechnological Research Institute 17201) etc.
Protozoa include amoeba, ciliates, and flagellates, and algae include cyanobacteria, green algae, and diatoms.

[0004]

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a novel microorganism useful for easily treating not only industrial wastewater but also wastewater from ramen shops, western food restaurants, tempura shops, etc. with a relatively simple device. Another object of the present invention is to provide an oil and fat decomposing agent containing the same, and a method for treating an oil and fat-containing substance using the decomposing agent.

[0005]

The first aspect of the present invention is the Aci
Non-motile Gram-negative spheres belonging to the genus netobacter
Spherical bacterium that is positive for catalase and negative for oxidase
It is a rod-shaped bacterium and is characterized by having a fat and oil degradability.
Regarding living things. A second aspect of the present invention is that the microorganism is a colony.
-The shape is circular, all edges are smooth, low convex,
2. A cream having a glossy color.
Of microorganisms. The third aspect of the present invention is the result of physiological property test.
3. The microorganism Acinet according to claim 2, wherein
object. 1. Cultural properties (1) Liquid broth culture: cloudy (2) Litmus milk: unchanged 2. Physiological properties (1) Denitrification reaction- (2) MR test + (3) VP test- (4) Indole generation − (5) Nitrate reduction- (6) Arginine dihydrolase- (7) Urease- (8) Hydrolysis Esculin − Gelatin- Starch- (9) β-galactosidase- (10) β-xylosidase + (11) Hemolytic- (12) Production of hydrogen sulfide − (13) Use of citric acid Koser + Christensen + (14) Use of inorganic nitrogen source NaNO_Three                +_W (NH_Four)_TwoSO_Four          +_W (15) Range of growth 44 ° C + 41 ° C + 37 ° C + (16) Attitude toward oxygen Anaerobic growth −   (17) Acid and gas generation from saccharides by OF medium (acid / gas)       L-arabinose +/- D-xylose +/-       D-glucose +/- D-mannose +/-       D-fructose − / − D-galactose +/−       Maltose-/-Sucrose-/-       Lactose +/- Trehalose-/-       D-sorbitol − / − D-mannitol − / −       Inositol − / − Glycerin − / −       Starch-/- (18) Availability of organic compounds D-glucose + L-arabinose + D-mannose- D-mannitol- N-acetyl-D-glucosamine- Maltose- Potassium gluconate + n-capric acid + Adipic acid + DL-malic acid + Sodium citrate + Phenyl acetate + DL-sodium lactate + Glutaric acid +_W L-phenylalanine- Malonic acid + L-histidine + Azelaic acid + L-aspartic acid + L-leucine + Histamine − L-tyrosine + β-alanine + Ethanol + 2,3-butanediol- trans-aconitic acid- L-arginine- L-ornithine + DL-4-aminobutyric acid- Regarding The fourth aspect of the present invention is an integrated administrative agency for industrial technology.
Laboratory Patent Biological Depository Depositary No. FERM P-
Acinetobacter s indicated by 18378
p. The fine SOD-1 according to claim 3, which is a strain SOD-1.
The organism Acinetobacter. The present invention
5 contains the microorganism according to any one of claims 1 to 4.
And relates to an oil and fat decomposing agent. The sixth aspect of the present invention is
An oil characterized by using the oil-and-fat decomposition agent according to claim 5.
The present invention relates to a method for treating a fat-containing substance. The seventh of the present invention is the above
The fat-and-oil-containing substance according to claim 6, wherein the fat-and-oil-containing substance is waste water.
Regarding the processing method of. The eighth aspect of the present invention is treatment of the wastewater.
The fat-and-oil-containing substance according to claim 7, wherein the temperature is 15 to 45 ° C.
Regarding the processing method of.

The same Acinetobacter as in the present invention
Bacteria belonging to the genus are described in JP-A-6-153922, Acinetobacter sp. SK
Although 0402A (Microtechnological Research Institute 13232) is disclosed, it is different in that it is a motile short bacillus, and that the present invention is a non-motile globular bacillus.

Acineto, which is a representative of the microorganisms of the present invention
bacteria, especially Acinetobacter s
p. Strain SOD-1 and Acinetob
acter sp. Differences from SK0402A are shown in the table below.

[Table 1]

The fats and oils in the present invention are mainly those derived from animals and plants, but are not necessarily limited thereto, and include fats and oils obtained by the synthetic method, their derivatives, and various other hydrocarbons.

The microorganism of the present invention is collected from soil and is highly safe to the human body and the environment.
The microorganism of the present invention can be cultured using, for example, a broth medium. The culture conditions may be set to those suitable for the medium to be used, for example, published by Kodansha in 1993,
The method described in "Microbiology Experimental Method", edited by the Microbial Research Method Roundtable, can be used.

The oil-fat-degrading microorganisms obtained by such culture can be separated by various known methods, and specifically, they can be cultured on an ordinary agar plate medium containing agar. The identification, properties, properties, etc. of the obtained microorganism can be investigated using a generally known identification test method or identification kit. Specifically, "Bergey '
s Manual of Systematic Ba
Cteriology, Holt, J. et al. G. Equal volume, Li
ppincott Williams & Wilkins
(1984) ","Bergey's Manual "
of Determinative Bacterio
logy, Holt, J. et al. G. Equal volume, Lippinco
tt Williams & Wilkins (199
4) ”described above can be used.

In order to provide the microorganism of the present invention as an oil and fat decomposing agent, the microorganism of the present invention may be immobilized on various carriers to prepare a preparation. Specifically, for example, 1993
The formulation methods described in "Baifukan", published by Sawao Murao et al., "Revised version of applied microbiology" and published by Maruzen in 1995, written by Takayuki Uejima, "Industrial enzymes", etc. can be adopted.

The following method can be used to prepare a liquid preparation. (1) The microorganism of the present invention is cultured in a general nutrient medium such as a broth medium for about 12 to 36 hours, and if necessary, a pH adjuster or the like is added thereto to prepare a preparation. (2) The bacterial cells are recovered from the culture of (1) by centrifugation or the like, suspended in a medium such as physiological saline so as to have an appropriate concentration, and if necessary, a pH adjuster or the like is added thereto. To prepare a formulation. (3) The culture of (1) above is appropriately concentrated by freeze-drying or the like, and if necessary, a pH adjuster or the like is added to prepare a preparation. (4) The bacterial cells are collected from the culture of (1) above by centrifugation, suspended in a medium such as a fresh broth medium, and a pH adjusting agent or the like is added to the medium as necessary to prepare a preparation. (5) The above (4) is further appropriately concentrated by freeze-drying or the like to give a preparation.

The following method can be adopted for preparing a powder formulation. (A) The microorganism of the present invention is cultivated in a general nutrient medium such as a broth medium for about 12 to 36 hours, and if necessary, a pH adjusting agent or the like is added thereto, and the product is dried by freeze-drying or the like to give a preparation. (B) The bacterial cells are recovered from the culture of (a) by centrifugation or the like, suspended in a medium such as physiological saline or a solution containing skimmyrum and sodium glutamate, etc., at an appropriate concentration, and if necessary, Then, a pH adjusting agent is added thereto, and the product is dried by freeze-drying or the like to prepare a preparation. (C) Cells are collected from the culture of (a) by centrifugation, suspended in a medium such as a fresh broth medium, and if necessary, a pH adjuster or the like is added thereto and dried by freeze-drying or the like. And make a formulation. (D) A formulation is prepared by appropriately adding fine powder such as fiber scraps and sawdust to the above-mentioned products (a) to (c). (E) For example, the microorganism of the present invention can be granulated by the method described in “Industrial Enzyme” by Takayuki Uejima, published by Maruzen in 1995. Specifically, (a) to (d)
The microbial cell suspension prepared in the same manner as described above is used as prim granules, malum granules, film-coated malum granules, cellulose fiber-containing T granules, and the like.

In addition to the above methods, the microorganisms of the present invention can be immobilized using various immobilizing materials by known techniques such as a carrier binding method, a cross-linking method, an encapsulation method, and a composite method. In addition, the microorganism of the present invention can be tableted by a known tableting technique.

In using the microorganism of the present invention,
It is preferable to employ an aeration means for blowing air in order to maintain a high activity of the microorganisms, that is, an aeration means. Further, in the treatment apparatus using the microorganism of the present invention, it is preferable to attach a trap for preventing the microorganism of the present invention or the oil / fat decomposition agent of the present invention from flowing out and separating the oil / fat from the trap. As the trap, one having a membrane filter having a predetermined pore size (which does not allow microorganisms to permeate but allows only water and low-molecular compounds to pass) is preferable. Further, in the case of wastewater containing a large amount of animal fats and oils that solidify at low temperatures, a higher effect can be obtained by controlling the wastewater temperature in the trap with a heater or the like.

Acinetobacter s of the present invention
p. Oil decomposition rate of strain SOD-1 (% / 24
The following experiment was conducted to investigate the relationship between the time) and the amount of fat and oil present. First, the following medium: diammonium phosphate 0.2% by weight dipotassium phosphate 0.2% by weight monosodium phosphate 0.1% by weight magnesium sulfate heptahydrate 0.02% by weight yeast extract 0.01% by weight % Water Residual pH 7.0 A component containing 6000 ppm of salad oil in 5 ml was put into a test tube having a diameter of 18 mm, and reciprocal shaking culture (170 Stokes / min) was performed at 28 ° C. for 24 hours. O at this time
D660 is about 0.50, and the number of bacteria is 3 × 1 on average.
0 ⁸ was cells / milliliter. Thus pre-cultured Acinetobacter sp. strai
The nSOD-1 culture solution was added to 1% by volume of 100 ml of the above-mentioned medium containing the amount of fats and oils in the table below, and kept at 28 ° C.
0r. p. culture for 24 hours, after autoclaving, J
Each decomposition rate was determined based on the IS official method (JIS K0102). The decomposition rate was calculated based on a control experiment in which no bacterium was inoculated. The results are shown in Table 2 and FIG.

[0017]

[Table 2]

Next, the relationship between the treatment temperature and the decomposition rate was investigated.
Salad oil addition amount in Table 2 0.3 g / 100 ml
Table 3 and FIG. 2 show the results of investigating the decomposition rate in the medium by changing the liquid temperature variously.

[0019]

[Table 3]

Next, the relationship between the pH at the treatment temperature of 20 ° C. and the decomposition rate (%) was investigated. Table 4 and FIG. 3 show the results of investigating the decomposition rate in various media in which the amount of salad oil added in Table 2 was 0.3 g / 100 ml at various pH values.

[0021]

[Table 4]

Furthermore, the relationship between the number of inoculum and the decomposition rate (%) at the treatment temperature of 20 ° C. was also investigated. In the salad oil addition amount 0.3 g / 100 ml medium in Table 2 above,
Table 5 and FIG. 4 show the results of examining the number of inoculum per 100 ml medium (expressed in log) and the decomposition rate corresponding thereto.

[0023]

[Table 5]

[0024]

The present invention will be described below with reference to examples.
The present invention is not limited thereby.

Example 1 At an okonomiyaki store in Komatsu City, Ishikawa Prefecture, about the actual wastewater containing oil for one week from April 1, 2001 to April 8, 2001, Acinetact of the present invention
er sp. Water temperature, pH, normal hexane extract (n-hex value), ammoniacal nitrogen, with and without addition of strain SOD-1 preparation
Five items of dissolved oxygen (DO) were measured. The above-mentioned microbial preparation is added twice a day, and the addition amount is 15 g.
(6 × 10 ⁹ cells) each. The first time is 9: 3 AM
0 and the second time was PM 4:30. The sampling time is AM 9:00 for the first time and PM 4:00 for the second time.
The actual drainage was sampled every day.

The experimental schedule is as follows.

[Table 6] * Indicates Acinetobacter sp. strai
n SOD-1.

Experimental Method As shown in FIGS. 5 and 6, the oil-and-fat-containing wastewater generated from the kitchen is stored in a grease trap tank. From here, one is used as a control tank and the other is uniformly used as a microorganism tank by using a submersible pump. I sent water. 8 control tanks and microorganism tanks
A size of 80 × 430 × 700 (h) mm with an effective water depth of 500 mm was used, and an air flow rate was 80 liters / minute. As for the drainage from these tanks, the amount of the input liquid from the creep strap tank is drained each time.

When the microorganism of the present invention was added, it was confirmed that the microorganism adhered to the wall surface of the tank. The amount of n-hexane extract in the control tank greatly fluctuates day by day because the oil content in the wastewater changes day by day, but the amount of n-hexane extract in the microorganism tank is almost the same.ヾ The value is stable and low, demonstrating the effectiveness of this microorganism. n-
A significant difference was found by t-test comparing the hexane value between the microbial treatment group and the control group. The water temperature of the control tank was the same as that of the microorganism tank for 2 days, but after the 3rd day, it was within 1 ° C, but a slight temperature difference was observed, suggesting the activity of the microorganisms. Since this experiment used actual wastewater, a large amount of detergent flowed into the wastewater from the 2nd to 3rd days, so both the control tank and the microorganism tank were significantly foamed by aeration, and the detergent was present on the 3rd day. A condition has occurred in which microorganisms have been killed or incapacitated. Aeration should ensure the amount of dissolved oxygen in the system, and keep the oil on the surface of each tank by aeration so that the oil does not flow into the first tank to the second tank or the second tank to the third tank as much as possible. There is also a meaning to keep. In addition, since microorganisms can grow in the treatment tank, if the amount of oil that flows in is constant, the number of bacteria will naturally be higher after the number of days has passed than when the experiment started, so the degree of oil decomposition will be small. improves.

[0029]

[Table 7]

[0030]

Industrial Applicability According to the present invention, it is possible to provide a novel microorganism having the ability to decompose oils and fats, an oil and fat decomposing agent containing the same, and a method for treating an oil and fat-containing substance using the same.

[Brief description of drawings]

FIG. 1 is a graph showing the relationship between the oil and fat decomposition rate of the microorganism of the present invention and the oil and fat content of the system.

FIG. 2 is a graph showing the relationship between the treatment temperature (culture temperature) and the decomposition rate in the decomposition of fats and oils by the microorganism of the present invention.

FIG. 3 is a pH of the system in fat and oil decomposition by the microorganism of the present invention
It is a graph which shows the relationship of a decomposition rate.

FIG. 4 is a graph showing the relationship between the number of inoculum (log) in the medium and the decomposition rate in the oil and fat decomposition by the microorganism of the present invention.

FIG. 5 is a plan view showing a device arrangement according to an embodiment of the present invention.

FIG. 6 is a cross-sectional view showing a device arrangement according to an embodiment of the present invention.

Claims (8)

[Claims] 1. A member of the genus Acinetobacter,
A non-motile gram-negative coccobacillus that is catalase-positive and oxidase-negative, and has a fat and oil degradability. 2. The microorganism according to claim 1, wherein the microorganism has a colony shape of a circle, smooth edges, low convexity, and a glossy cream color. 3. A contract in which the physiological property test results are as follows:
The microorganism Acinetobacter according to claim 2. 1. Cultural properties (1) Liquid broth culture: cloudy (2) Litmus milk: unchanged 2. Physiological properties (1) Denitrification reaction- (2) MR test + (3) VP test- (4) Indole generation − (5) Nitrate reduction- (6) Arginine dihydrolase- (7) Urease- (8) Hydrolysis Esculin − Gelatin- Starch- (9) β-galactosidase- (10) β-xylosidase + (11) Hemolytic- (12) Production of hydrogen sulfide − (13) Use of citric acid Koser + Christensen + (14) Use of inorganic nitrogen source NaNO_Three                +_W (NH_Four)_TwoSO_Four          +_W (15) Range of growth 44 ° C + 41 ° C + 37 ° C + (16) Attitude toward oxygen Anaerobic growth −   (17) Acid and gas generation from saccharides by OF medium (acid / gas)       L-arabinose +/- D-xylose +/-       D-glucose +/- D-mannose +/-       D-fructose − / − D-galactose +/−       Maltose-/-Sucrose-/-       Lactose +/- Trehalose-/-       D-sorbitol − / − D-mannitol − / −       Inositol − / − Glycerin − / −       Starch-/- (18) Availability of organic compounds D-glucose + L-arabinose + D-mannose- D-mannitol- N-acetyl-D-glucosamine- Maltose- Potassium gluconate + n-capric acid + Adipic acid + DL-malic acid + Sodium citrate + Phenyl acetate + DL-sodium lactate + Glutaric acid +_W L-phenylalanine- Malonic acid + L-histidine + Azelaic acid + L-aspartic acid + L-leucine + Histamine − L-tyrosine + β-alanine + Ethanol + 2,3-butanediol- trans-aconitic acid- L-arginine- L-ornithine + DL-4-aminobutyric acid- 4. The Acinetobacter sp. Indicated by the Deposit No. FERM P-18378 of the National Institute of Advanced Industrial Science and Technology, National Institute of Advanced Industrial Science and Technology. stra
The microorganism Acin according to claim 3, which is in SOD-1.
etobacter. 5. An oil and fat decomposing agent comprising the microorganism according to any one of claims 1 to 4. 6. A method for treating an oil / fat-containing substance, which comprises using the oil / fat decomposer according to claim 5. 7. The oil-and-fat-containing substance is waste water.
A method for treating an oil-and-fat-containing substance as described above. 8. The method for treating an oil / fat-containing substance according to claim 7, wherein the treatment temperature of the waste water is 15 to 45 ° C.