JPH09103290A - Noble microorganism capable of deodorization in sewage treatment, its culture medium composition and cultivation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a noble microorganism developing the deodorizing effect in sewage treatment, comprising Bacillus subtilis (KCTC 8662P), utilizable under aerobic conditions, well growable in an aqueous enriched medium containing carbon source and nitrogen source. SOLUTION: This noble microorganism comprises Bacillus subtilis (KCTC 8662P), utilizable under aerobic conditions, well growable in an aqueous enriched culture medium containing carbon source and nitrogen source and developing the deodorizing effect in sewage treatment. This microorganism decomposes sewage to remove foul smell and is useful in sewage treatment. In addition, it can be utilized in fresh water ponds, fish farming, reservoirs, poultry houses, livestock barns, incineration plants, refrigerators, clothe-changing rooms, cars, boats for deodorization. This novel microorganism has been found near the fresh water fish farming ponds or at the pace of dumping ground. Because the places were filled with malodor at the beginning, but 15 days later, it was observed that the malodor dissipated and swage reduced and a strain of a microorganism having excellent deodorizing effect has been isolated.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[Field of Industrial Application] The present invention relates to a microorganism having a deodorizing ability for treating sewage, a medium composition and a culturing method thereof. More specifically, it decomposes waste water debris to remove a bad odor, and The present invention relates to a sewage treatment and a deodorizing method for decomposing oil and fat components.

[0002]

2. Description of the Related Art As already developed sewage treatment preparations,
Although many microorganisms such as yeast, vibrio bacterium, and activated sludge are known, the conventional sewage treatment bacteria have a problem in that they cannot decompose debris accumulated in the wastewater pipe and cannot deodorize. The organic sludge generated at the biological wastewater treatment plant is also treated by the methods such as concentration, stabilization, improvement, dehydration and incineration, and the cost required for this treatment is high. .

[0003]

Problems relating to environmental protection have recently been investigated in various fields of industry, but sewage debris and offensive odors are also a type of pollution, and are problematic in various industrial fields. It has become. In particular, in the service industry such as a cafeteria, the number of cases in which a bad odor is generated and causing a big problem is actually increasing year by year. In addition, the sewage of the city is an organic substance such as soluble, colloidal, or suspended solid, and drainage pipes are gradually narrowed by spilled oil and fats, and eventually closed. In such a case, the problem of sewage treatment has become a major problem to be solved.

An object of the present invention is to provide a microbial preparation containing a novel microorganism having sewage treatment and deodorizing ability which can solve the above-mentioned problems.

Another object of the present invention is to provide a medium composition for culturing the above-mentioned microorganism and a culturing method using the same, and a sewage treatment method and a deodorizing method using the above-mentioned microorganism preparation. Is also intended to be provided.

[0006]

[Means for Solving the Problems] A microbial preparation of the present invention for achieving the above-mentioned object was obtained from the Bacillus subtilis bacterium (KCTC 86) at the Korea Institute of Science and Technology on April 26, 1995.
62P).

The medium composition for cultivating the above-mentioned bacteria is such that the starch is 8 to 15 g, and the bifix tract is added to 1 liter of distilled water.
0.5-4g, peptone 7-14g and sodium chloride 2-
Composed of 7g.

[0008] As a culture method, the above composition is inoculated with the above microorganisms and then cultured at a temperature range of 25 to 32 ° C and a pH value of 6.5 to 8.5.

Hereinafter, the structure of the present invention will be described more specifically with reference to the accompanying drawings.

The present invention is a Bacillus sulcus which has sewage treatment and deodorizing ability that conventional Bacillus subtilis does not have.
It was deposited as KCTC 8662P at the Korea Institute of Science and Technology on April 26, 1995, regarding Subtilis.

As a result of efforts to find a sewage treatment bacterium and a strain having a deodorizing ability, the present inventors have discarded sludge at the bottom of a fish pond in the vicinity of a freshwater fish farm.
The odor was bad at first, but after about 15 days, the odor disappeared and the amount of dregs was reduced, and the strain of the present invention having a remarkable deodorizing effect was isolated in the soil. The above-mentioned isolates belong to Bacillus subtilis morphologically, biologically and biochemically. However, in terms of their usefulness in reducing waste debris and deodorizing ability, they are usually Bacillus subtilis. Since it was not found in the strain, it was judged that the above-mentioned isolated strain is a new strain of Bacillus subtilis.

The morphological, biological and biochemical properties of the strain according to the invention are as follows.

Morphological characteristics The above-mentioned bacterium is a rod-shaped bacterium and has a size of 1.0 to 1.5 x 3.5 to 4.0.
μm, surrounded by the cell membrane and having a nucleus. Moreover, they are characterized by having spores, which form spores when the culture conditions holding the diaphragm are insufficient, and the cells themselves have about 10 flagella (see Fig. 1).

Biological characteristics The above-mentioned bacteria are motile, and even when they were inoculated on a broth agar plate in a state of being suspended in physiological saline and cultured at 37 ° C for 24 hours, the growth was good.

The peripheral edge of the medium-shaped colony is not circular but rough, and the colony is opaque and has a white or light yellowish tint depending on the type of medium.

Regarding the growth state in each medium, normal agar medium, cotton sheep blood medium, heart inpuzon medium, plain heart inpuzon medium, trypticosoi agar medium showed good growth, but saline yolk agar medium, Vitibitibor medium and Zover. It did not grow on 2216 medium.

As shown in FIGS. 2 and 3, the temperature-dependent growth was well grown at 25 ° C., 30 ° C. and 35 ° C., and the pH was 6.5.
The growth was good at ˜8.5 (preferably pH 6.5 to 7.0), but the growth was poor at pH 6.0 or less. It was aerobic for oxygen utilization and not hemolytic.

Biochemical properties The above-mentioned bacteria are acid-fast bacteria which are Gram-positive and stain red with Dier-Nilsson stain. In addition, nitrate was reduced and oxalic acid was used, and it was also positive for oxidase, catalase and VP reaction. However, neither indol nor hydrogen sulfide was produced, nor did it react negatively with urease. Statin, esculin and cellulose were hydrolyzed but arginine could not be hydrolyzed.

Utilization of Carbohydrate In order to assay the carbohydrate utilization ability of the above-mentioned bacteria, a certain amount of carbohydrate was added to Penolredo liquid medium as a basal medium, and the above-mentioned bacteria were inoculated.

The above-mentioned fungus decomposed D-glucose, D-fructose and trehalulose to produce an acid.
L-arabinose, D-xylose, lactose, D-
Sollittle, D-mannitol and inositol could not be decomposed.

Stability test As a result of oral administration of only the bacterial cells of the above-mentioned bacteria to mice,
The half-maximal lethal concentration was almost infinite, and there was no reaction in the mouse epidermal application test.

According to the measurement results of the morphological, biological and biochemical characteristics of the above-mentioned bacteria, the ordinary Bacillus subtilis strain was found to be a cell size, a fermentative decomposition of glucose and a hydrolysis of arginine. Although it was the same in general aspects, growth temperature (5-55 ℃), growth pH value (5.5-8.5)
There was a difference in deodorization and sludge decomposition ability from the existing Bacillus subtilis. Therefore, the above-mentioned bacterium is judged to be a new strain of Bacillus subtilis.

A preferable medium composition for culturing the above-mentioned bacterium in a large amount is starch 8 to starch per 1 liter of distilled water.
15g, biffix tract 0.5-4g, peptone 7-14
g and 2 to 7 g sodium chloride are preferred. If the above starch is 8 g or less with respect to the whole medium composition, the growth is poor,
If it was more than 15g, the whole starch could not be used until the algebraic growth phase due to excessive nutrient supply. In addition, if the above-mentioned bifix tract is 0.5 g or less, the bacterial growth is not smooth, and if it is 4 g or more, it is overdosed beyond the optimal growth range of the bacterial cells, and if peptone is 7 g or less, the growth of the above bacteria is poor. If the amount is 14 g or more, the maximum growth time will be reached sooner, but the nature of the cells will tend to decline. If the sodium chloride content is 2 g or less, the growth will be poor. It goes beyond the range.

After inoculating the above-mentioned medium composition with the above-mentioned bacteria, it can be cultured in a temperature range of 25 to 35 ° C. and a pH value of 6.5 to 8.5 to mass-proliferate.

Since the culture thus grown contains a large amount of cells, it can be added as it is to a sewage treatment plant or a septic tank. Further, the culture can be centrifuged to obtain pure cells, and after washing with sterile physiological saline, a mixture with an excipient such as starch can be used as a microbial preparation.

The effects of the present invention can be more specifically recognized through the following examples, but the following examples do not limit the scope of the present invention.

[0027]

[Examples] Example 1 Medium composition of a large-scale culture of Bacillus subtilis (KCTC 8662P) and an effect of reducing total organic matter concentration. The medium composition is 12 g of starch, 2 g of bifix tract, 10 g of peptone.
After dissolving 4.5 g of sodium chloride in 1 liter of distilled water, the pH value was adjusted to 7.0. The medium thus constructed was placed in a 500 ml flask, inoculated with Bacillus subtilis (KCTC8662P), and cultured by shaking at 35 ° C. for 24 hours.

During this culture, oxygen may be supplied and introduced considering that Bacillus subtilis (KCTC8662P), which is the core of the present invention, is an aerobic bacterium.
As a method of introduction, it is practical to supply by pump or to supply solid oxygen.

Here, the obtained culture solution was centrifuged at 3,000 rpm for 30 minutes by a low-temperature centrifuge to collect 5 g of bacterial cells, which was then lyophilized at -75 ° C. to produce it. The powder was mixed with 200 g of starch and used as a microbial preparation for the total organic matter reduction test.

100 g of the microbial preparation was added to a settling tank containing 5 tons of starch waste liquid discharged from the starch manufacturing plant, and the concentration of total organic matter was measured according to the elapsed days. In the control area, the total organic matter concentration in the sedimentation tank containing no microbial preparation was also measured, and the results are shown in FIG. As can be seen from Fig. 4, the total organic concentration in the sedimentation tank containing the above-mentioned microbial preparation decreased sharply to 78 mg / l 3 days after the addition, but it was 145 mg / l in the non-addition area, which was more than double the addition area. It was 5
After a day, the spiked area decreased sharply to 30 mg / l, while the spiked area was 110 mg / l. After 10 days, the added area was 23 mg / l, but in the non-added area it was 92 mg / l, and after 15 days,
The results were found to be 14 mg / l in the added region and 81 mg / l in the non-added region. After 20 days, the final measurement shows that the added area is 11 mg.
In contrast to / l, the non-added area was 78 mg / l.

From the above results, it was found that the strain according to the present invention has an excellent effect of decomposing and reducing many organic substances even in a small amount.

Example 2 Deodorizing effect of Bacillus subtilis (KCTC 8662P). Powder production method: The temperature of the high-speed centrifuge was adjusted to 10 ° C. and the rotation speed was 3,000 revolutions / minute in order to prevent the components of the cells from destroying the culture that was cultured from the medium of Example 1 above. After centrifuging once for 30 minutes, the cells collected in 0.85 sterilized physiological saline are centrifuged and washed three times for 30 minutes at 3,000 rpm. The washed cells are placed in a 5 ml-dose freeze-drying test tube, rotated 30 minutes per minute, pre-frozen at -50 ° C, and then frozen at -75 ° C for 40 minutes.

The lyophilized cells are pulverized with a sterilized grinder to give a powder.

Liquid manufacturing method: Bacteria 2.0 × 10 ⁶ ce was prepared from the above powder.
It is suspended in 0.85% sterilized physiological saline so that the concentration becomes ll / ml, and stored in a fixed container.

Deodorizing effect: An experiment was carried out by administering 2 liters of the liquid preparation of the above strain and 5 g of the powder preparation together to a water tank having a dose of 10 tons in the underground space. One month after the administration, the malodor concentration was measured. When the sewage was taken up by the pump before the administration, the malodor concentration was 40,000 reverse odor (threshold THRESHOLD), but one month after the administration, the odor concentration when the sewage was taken dropped to 2,000 reverse odor. When not picked up,
The malodor concentration before administration of the above formulation was 250 reverse odor, but only one reverse odor was produced one month after the administration.

Example 3 Bacillus subtilis (KCTC 8662) on cotton
P) adsorption method. In order to produce a practical product by utilizing the deodorizing effect of the above-mentioned bacteria, cotton was used as a microbial cell adsorption substance. Mold cotton into a cylinder with a diameter of 1 cm and a length of 3 cm.
Sterilize at 15 ℃ for 15 minutes to remove bacteria on the raw cotton. Cylinder-shaped cotton from which germs have been removed is dried at 50 ° C in an aseptic state and then stored in a sterile container.

Bacillus produced from Example 2 above
Cylindrical cotton is soaked in a liquid substance of subtilis bacteria (KCTC 8662P), and soaked for about 1 hour so that the bacteria can fully penetrate into the inside of the cotton. Then, the cylindrical cotton infiltrated with the bacterial cells was dried in an aseptic dryer at 30 ° C., put in an open plastic container, and placed in a place with a bad odor. After that, it was recognized that the bad odor significantly decreased.

As described above, the microbial Bacillus of the present invention
Subtilis (KCTC 8662P) grows well in any medium as long as it is an aqueous nutrient supplemented medium. The range of culture temperature is 25-40 ℃, but the optimum growth temperature is 30
~ 35 ° C. Further, the range of the culture pH value was within the range of 6.5 to 8.5, and it was found that when the optimum growth pH was around 7.0, it was proliferated well by culturing for 10 to 24 hours.

[0039]

EFFECTS OF THE INVENTION Bacillus subtilis (KCTC)
8662P) is well suited for a variety of forms, while exerting an outstanding function in cleaning water, air and soil. For example,
In the sewage treatment, sewage residue is decomposed to remove offensive odors, and oil and fat components attached to the sewer pipe are decomposed. With such an action, the same effect can be obtained in the septic tank.

If the above-mentioned bacteria are added to the fresh water pond, the fish farm, the water storage tank, the cooling tower water storage tank, and the air conditioner water storage tank, the generation of microalgae and miscellaneous bacteria is suppressed and the water quality is maintained. In addition, if the liquid formulation dispersion liquid of the above bacteria is directly sprayed on a place where bad odors frequently occur, such as a chicken house or a livestock house, the bad odor can be suppressed, and odors such as garbage incinerators, refrigerators, dressing rooms, cars, and ships will be generated. It is possible to deodorize by sprinkling the above-mentioned bacteria on the place or by putting it in a container and leaving it open.

[Brief description of the drawings]

FIG. 1 Bacillus subtilis according to the present invention.
Subtilis) (KCTC 8662P) micrograph (X1
000).

FIG. 2 is a Bacillus subtilis bacterium (KCT) according to the present invention.
It is a graph showing the growth degree according to temperature of C8662P).

FIG. 3 is a Bacillus subtilis bacterium (KCT) according to the present invention.
It is a graph showing the growth degree by each pH value of C8662P).

FIG. 4 is a Bacillus subtilis bacterium (KCT) according to the present invention.
It is a graph showing the organic substance resolution of C 8662P).

Claims (5)

[Claims] 1. A Bacillus subtilis bacterium (KCT) which is available under aerobic conditions and is characterized by good growth in an aqueous nutrient medium containing a carbon source and a nitrogen source.
C 8662P), a novel microorganism having deodorizing ability for sewage treatment. 2. The novel microorganism according to claim 1, which has a deodorizing ability for treating sewage, characterized in that the growth temperature of the bacterium is 5 to 55 ° C. and the growth pH is 5.5 to 8.5. . 3. Starch 8-15 per liter of distilled water
g, bifix tract 0.5-4g, peptone 7-14g
And Bacillus subtilis (KCTC 8662P) characterized by being composed of 2 to 7 g of sodium chloride
Medium composition for use. 4. Bacillus subtilis (KCTC 86)
62P) to 1 liter of distilled water, 8 to 15 g of starch,
After inoculating a medium composed of 0.5 to 4 g of bifix tract, 7 to 14 g of peptone and 2 to 7 g of sodium chloride,
A method for culturing a microorganism, which comprises culturing at a temperature range of 25 to 35 ° C. and 6.5 to 8.5 pH. 5. Bacillus subtilis (KCTC 86)
62P) to 1 liter of distilled water, 8 to 15 g of starch,
After inoculating a medium composed of 0.5 to 4 g of bifix tract, 7 to 14 g of peptone and 2 to 7 g of sodium chloride,
When culturing in the temperature range of 25-35 ° C and 6.5-8.5 pH,
A method for culturing a microorganism, which comprises introducing oxygen by a pump-type supply system or solid oxygen introduction means.