JPH11196861A - Microorganism having oil and fat decomposability - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new species of bacterium having oil and fat- decomposability and a decomposing method of oil and fat by using the same. SOLUTION: In this invention, a new species of bacterium belonging to the genus Acinetobacter derived from rice bean, i.e., Acinetobacter sp. YU-1 is provided. This bacterium has an oil and fat-decomposing enzyme (lipase) activity. The lipase is an intracellular enzyme and has a high stability of the enzyme activity. When the bacterium is used in a meat processing factory, a bovine oil and fat is efficiently decomposed and an effect of removing dirts in an underdrain ditch, or the like is confirmed.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a novel microorganism isolated from rice bran, and more particularly to a new bacterial species having fat-and-oil degradability.

[0002]

2. Description of the Related Art In recent years, with increasing interest in environmental protection, techniques for suppressing various environmental pollutants have been developed. Oils and fats discharged from food factories and households are also one of the causes of polluting rivers, lakes, and oceans, and it is desired to control them.

As a technique for disposing of fats and oils from the viewpoint of environmental protection, conventionally, fats and oils are solidified using a solidifying agent,
A technique for performing the same incineration treatment as ordinary solid waste is known. However, this method has a problem that the disposal cost increases, and cannot be applied to oils and fats mixed with water. For this reason, it is difficult to implement it in business establishments such as food factories, and in fact, it is not widely used in homes. Accordingly, there is a need for a technology that biologically or chemically decomposes waste fats and oils and treats them at a low cost so that they can be discharged in a manner that is compatible with the environment.

[0004]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a new kind of bacterium derived from rice bran and having the ability to decompose fats and oils, which is useful for decomposing fats and oils. .

[0005]

Means for Solving the Problems The inventor of the present invention has discovered a new kind of bacterium having the ability to degrade fats and oils during the research on rice bran, and has reached the present invention. Rice bran is conventionally used as a fertilizer, and in particular, is used as a so-called "bokashi" obtained by fermenting rice bran to prevent nitrogen starvation during fertilization. When microorganisms were detected from this blur,
In addition to one type of yeast, one type of bacteria was separated and detected. This bacterium was identified as a bacterium belonging to the genus Acinetobacter based on the taxonomic properties and the nucleotide sequence of the common conserved gene (16S rRNA). Conventionally, only one kind of bacteria belonging to the genus Acinetobacter (A. calcoa
ceticus), but the bacterium identified this time was different from this conventional species. Therefore, the bacteria identified this time were recognized as a new species, and Acinetobacter YU-1 (Acinetoba
cter sp. YU-1). According to the Budapest Treaty on the International Recognition of the Deposit of Microorganisms in Patent Procedures, this bacterium was obtained on January 12, 1998, by the Institute of Biotechnology, Institute of Biotechnology and Industrial Technology, 1-3-1 Higashi, Tsukuba, Ibaraki, Japan. Deposited at the Microbial Depositary Center (Deposit No. FERM B
P-6222).

The Acinetobacter YU-1 bacterium was found to have lipolytic enzyme (lipase) activity. Notably, this bacterium accumulates lipase in the presence of fats and oils (eg, olive oil) around it. This indicates that the rice bran used as the source is Acinetobacter YU-1
This indicates that the environment is suitable for The culture of this bacterium was performed at 37 ° C. using a liquid medium containing 1 to 5% of olive oil.
For 48 hours. Another notable fact is that most of the microbial lipases are secreted extracellularly, whereas the lipase of Acinetobacter YU-1 is an intracellular enzyme and is not secreted extracellularly. . This means that the stability of the enzyme activity is high, which is advantageous for use in wastewater treatment and the like. Actually, when this bacterium was used in a meat processing plant, it was confirmed that the beef oil and fat was efficiently decomposed and the dirt in the drainage ditch could be removed.

[0007]

The present invention will be described in more detail with reference to the following examples. Example 1: Isolation of Acinetobacter YU-1 94 kg of rice bran and 6 kg of firgrass are mixed, lactic acid bacteria and yeast are added using molasses as a medium, fermented to a temperature of about 35 to 50 ° C, and cut back once every three days While doing 7
Bokashi was prepared over a period of days.

[0008] 0.1 g of this blur is weighed, suspended in 1.0 to 2.0 ml of sterile physiological saline (sterile water is also acceptable), shaken well, and allowed to stand for 4 to 5 minutes. Ten
It was diluted 0-fold. Then, in the clean bench,
50 μl of this diluted solution was added to an agar medium (Nutrient ager, pH 6.
8) and cultured for 24 to 48 hours in a thermostat at 37 ° C. When yeast is isolated, malt extract agar is used as an agar medium, and the culture temperature is
30 ° C.

In a clean bench, the number of colonies formed on the agar medium is counted to calculate the total number of bacteria,
After observing the type of colony, the bacteria were picked from an independent colony and transplanted to an agar medium. After this was cultured at 37 ° C. for 48 hours, one kind of bacteria was isolated. This is stored on a slant agar medium in a clean bench,
The bacterial species was identified. In addition, the frozen storage method (20 ~
50% glycerol, -20 ° C or -80 ° C) or lyophilization method may be used.

Next, in order to identify the preserved bacteria, the taxonomic properties were examined according to a conventional method, and the following results were obtained. 1. 1. Form Gram-negative, facultative anaerobic, non-glucose-fermenting, penicillin-resistant bacillus. Biochemical properties Enzyme production Catalase: + Oxidase:-Urease:-β-galactosidase:-Biochemical reaction VP reaction:-Indole purification:-Hydrogen sulfide generation:-Utilization of citric acid: + Nitrate reduction:-Malonate Availability: + Hydrolytic activity Starch:-Casein:-Gelatin:-Tributyrin: + Olive oil: + Ornithine:-Lysine:-Arginine:-3. GC content of chromosomal DNA 42.4% 4.16S rRNA nucleotide sequence Completely coincided with the nucleotide sequence known for A. calcoaceticus, a known Acinetobacter.

5. Sugar utilization ・ Generates acid from glucose, mannose, arabinose and xylose.

Does not produce acid from maltose, sucrose, fructose, rhamnose, mannitol, adnit, inositol, sorbitol.

6. Growth conditions Temperature range: 20-30 ° C., maximum 52 ° C. (with heat resistance) pH range: 6-8 In addition to the above properties, it was confirmed to have esterase activity and lipase activity.

Based on the above taxonomic properties and characteristics, this isolated bacterium was identified as a new species of the genus Acinetobacter, and Acinetobacter YU-1 (Acinetobacterium) was identified.
bacter sp. YU-1). According to the Budapest Treaty on the International Recognition of the Deposit of Microorganisms in Patent Procedures on January 12, 1998, this fungus was patented by the Institute of Biotechnology and Industrial Technology at the Institute of Industrial Science and Technology, 1-3-1 Higashi, Tsukuba, Ibaraki, Japan. Deposited at Microbial Depositary Center (Deposit No.FE
RM BP-6222).

Note that, from the blur used in this embodiment,
In addition to Acinetobacter YU-1, one type of yeast was detected. The abundances of this yeast and Acinetobacter YU-1 in the oak are as follows.

Acinetobacter YU-1: 7.5 × 10 ³ cells / g Yeast: (4 ± 1) × 10 ⁵ cells / g
g Example 2: Example of culture In order to culture Acinetobacter YU-1 in large quantities, 37
1% of olive oil was added to 1 liter of medium at a temperature of ℃, aeration was performed, and a few minutes later, the inoculum isolated in Example 1 was inoculated. The temperature was maintained at 37 ° C and 10 ⁶ over 48 hours.
The cells were cultured to 10 ⁸ / ml.

Example 3 Preparation of a Composition for Water Treatment 15 kg of the enriched culture of Acinetobacter YU-1 obtained in Example 2 was uniformly mixed with 100 kg of the beaker used in Example 1. This was dried until the water content became 12 to 13% to obtain a water treatment composition.

Example 4: Use example 1 kg of the water treatment composition obtained in Example 3 was placed in 30 liters of hot water at 45 ° C to 50 ° C and allowed to stand for 20 to 30 minutes. Used as body fluid.

After cleaning the floor of the working place of the meat processing plant, the above-mentioned bacterial cell fluid was evenly applied. When this operation was continued for one week, the adhesion of oil on the floor of the work place and the outlet of the drainage ditch began to be removed, and the dirt on the lower surface and the side surface of the drainage grew up and began to be removed.

In the same manner as above, 15 liters of the bacterial cell liquid was prepared and sprayed repeatedly over two weeks. As a result, the lower surface of the drainage gutter was removed, and the dirt on the side surface became foamy. In addition, before the grease trap and at the outlet of the drain, the lump of tallow accumulated in the mesh net gradually became smaller. This lump was initially about 1 cm square,
It was broken down into a foam and reduced to about 2-3 mm square.

Next, 15 liters of the bacterial cell fluid was prepared in the same manner, and sprayed repeatedly for three weeks every day.
As a result, a lot of small lump of beef oil and fat floated on the grease trap almost completely after one week, and only a part remained in the form of foam. As a result, the water in the grease trap became clear.

Further, a 15-liter cell suspension was prepared in the same manner and sprayed every day for 4 weeks. By the 4th week, beef oil and fat was completely decomposed, and the grease trap and drains were also cleaned. became.

Example 5: Use Example The effect of the present invention was tested in a factory where meat was sliced and processed meat was packed. Animal waste accounts for most of the SS in the effluent of this plant. Wastewater from the factory is temporarily stored in an underground tank with a capacity of about 80 tons, subjected to sedimentation treatment, and then discharged from the factory. Further, after passing through a simple screen (net) outside the factory, the water is directly passed through an agricultural waterway. Before this test, animal fat was thick and floating on the surface of the tank, and the odor was quite severe.

In the same working process as in Example 4, a bacterial cell solution was prepared by dissolving only 20 kg in 800 liters of hot water and filtering, and the solution was directly poured from a manhole into an underground storage tank.

The cell solution was introduced in the morning of the first day of the test, and when the inside of the tank was observed on the evening of that day, the floating oil was dissolved and the color was dark, but the water was dark. It was separated and the odor was almost gone.

When the method of introducing the bacterial cell fluid was changed so that it was introduced from the drain in the factory after the operation of the factory was completed,
One week later, oily dirt that had adhered to the inside of the pipe from the factory to the storage tank was washed away and turned dry. As a result of continued continuation for 8 months, remarkable reduction of odor was confirmed, and effects such as reduction of the number of vacuums due to elimination of clogging of the filtration layer were also confirmed.

[0027]

As described in detail above, the present invention provides a novel microorganism having the ability to decompose fats and oils. By using this novel microorganism, it is possible to decompose oils and fats contained in wastewater from food factories to reduce environmental problems. Remarkable effects such as prevention of contamination can be obtained.

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[Procedure amendment]

[Submission date] December 21, 1998

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Claims

[Correction method] Change

[Correction contents]

[Claims]

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0010

[Correction method] Change

[Correction contents]

Next, in order to identify the preserved bacteria, the taxonomic properties were examined according to a conventional method, and the following results were obtained. 1. 1. Form Gram-negative, facultative anaerobic, non-glucose-fermenting, penicillin-resistant bacillus. Biochemical properties Enzyme production Catalase: + Oxidase:-Urease:-β-galactosidase:-Biochemical reaction VP reaction:-Indole formation :-Hydrogen sulfide formation:-Utilization of citric acid: + Nitrate reduction:-Malonate Availability: + Hydrolytic activity Starch:-Casein:-Gelatin:-Tributyrin: + Olive oil: + Ornithine:-Lysine:-Arginine:-3. GC content of chromosomal DNA 42.4% 4.16S rRNA nucleotide sequence Completely coincided with the nucleotide sequence known for A. calcoaceticus, a known Acinetobacter.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI C12R 1:01)

Claims (2)

[Claims] 1. A novel microorganism derived from rice bran, having lipolytic activity. 2. Acinetobact YU-1 (Acinetobact)
er sp. YU-1) (deposit number FERM BP-6222).