KR100925879B1 - Complex strain for the waste water/sewage treatment and nitrogen treatment process using the same - Google Patents

Abstract

PURPOSE: A method for treating nitrogen and composite microorganism is provided to effectively remove nitrogen in waste water. CONSTITUTION: A composite microorganism (SejungBio-FA100(deposit number: KCTC 11400BP)) for treating organic material of waste water and performing denitrofication comprises: Bacillus licheniformis FE1, Bacillus cereus FE3, Paenibacillus sp. H10-02, Acetobacter pomorum FM2, Acetobacter peroxydans FM3, Bacillus subtilis FM4, Lactobacilus casei FM5, Paenibacillus sp. P33 FM8; Kluyveromyces fragilis FY1, and Debaryomyces hansenii FY2. A composite microorganism formulation for treating organic material of waste water and denitrogenification 0.5-20 weight% of composite microorganism 20-50 weight% of rice bran, 20-50 weight% of food waste residue, 10-30 weight% of rice hull charcoal coarse powder, and 5-10 weight% of humus soil.

Description

COMPLEX STRAIN FOR THE WASTE WATER / SEWAGE TREATMENT AND NITROGEN TREATMENT PROCESS USING THE SAME}

The present invention relates to a composite microorganism for treatment of wastewater and wastewater, and a composition for treatment of wastewater containing the same. The present invention also relates to a method for treating organic matter of wastewater and wastewater using the complex microorganism and a method for treating denitrification under aerobic conditions.

Microbial preparations are direct use of living microorganisms, and their vitality must be maintained until the moment they are consumed, and the living function must be exerted. Microbial preparations are currently required in a wide variety of fields, the use of which is used in a variety of applications, such as deodorants, air purifiers, herbicides, soil pesticides, soil improvers, growth promoting agents, water purification agents, including odor. Bacillus (Bacillus), Pseudomonas (Pseudomonas), photosynthetic various jungyimyeo strains have been developed, such as bacteria, soil microbial formulation as it is yeast, lactic acid bacteria, actinomycetes, fungi, and algae and their kind to protozoa varied to use a zero environmental improvers . Recently, microbial preparations have been immobilized on carriers and sold, with the development of homogenization technology.

At present, foreign countries use microbial preparations for herbicides, soil diseases, harvesting agents, powdery mildews, termites, gray mold, plant growth regulators, and seed treatments. Recently, the development technology of microbial preparations in Korea is showing a lot of improvement. For example, the Korean Life Science Research Institute has developed soil microbial agents and antifungal agents using Bacillus sp. However, compared to foreign countries, the level of development technology is still only about 30% level, and microbial products consumed in Korea tend to be mainly imported from foreign countries.

The discharge of sewage and wastewater, including nitrogen, accelerates the eutrophication of lakes and reservoirs, and promotes the growth of algae and aquatic plants in light rivers, which is not good in aesthetic sense. Nitrogen removal is important for water quality management and sewage treatment plant design. Therefore, various methods for removing substances such as nitrogen have been developed. Initially, biological nitrification for ammonia oxidation and regulation, biological denitrification using methanol as an external carbon source, and chemical precipitation for phosphorus removal were mainly used. Recently, the process of removing nitrogen using microorganisms has been in the spotlight.

Therefore, the present inventors continued to develop a method of effectively removing nitrogen from wastewater by using microorganisms to treat organic matter in wastewater and denitrifying microorganisms. As a result, the present invention treated organic matter in wastewater. In addition to aerobic conditions, denitrification can lead to the development of complex microorganisms with high nitrogen removal efficiency.

The present invention is to provide a complex microorganism for organic matter treatment and denitrification of waste water and waste water.

In addition, the present invention is to provide a composition for organic matter treatment and denitrogenation treatment of waste water.

In addition, the present invention is to provide a method for treating denitrification of sewage and wastewater using the complex microorganism.

According to an aspect of the present invention, the present invention provides Bacillus licheniformis FE1, Bacillus cereus FE3, Paenibacillus sp H10-02 FE4, Acetobacter formo Acetobacter pomorum FM2, Acetobacter peroxydans FM3, Bacillus subtilis FM4, Lactobacilus casei FM5, Paenibacillus sp. P33 FM8, Provides a complex microorganism SejungBio-FA100 (microbial accession number: KCTC 11400BP) for organic matter treatment and denitrification of sewage and wastewater consisting of Kluyveromyces fragilis FY1 and Debaryomyces hansenii FY2 do.

Specifically, the complex microorganism SejungBio-FA100 Bacillus licheniformis FE1 is 9X10 ⁷ cfu / g, Bacillus cereus FE3 is 7.2X10 ⁷ cfu / g, Fanibacillus Esp H10-02 (Paenibacillus sp. H10-02) FE4 is 4.2X10 ⁷ cfu / g, acetonitrile bakteo Formosan column (Acetobacter pomorum) FM2 is 7.2X10 ⁷ cfu / g, acetonitrile bakteo peroxy thiooxidans (Acetobacter peroxydans) FM3 is 7.2X10 ⁷ cfu / g, Bacillus subtilis FM4 at 4.2 × 10 ⁷ cfu / g, Lactobacilus casei FM5 at 6 × 10 ⁷ cfu / g, Paenibacillus sp . P33 FM8 at 4.2 × ^{10 7} cfu / g, Kluyveromyces fragilis FY1 with 5.4 × 10 ⁷ cfu / g and Debaryomyces hansenii FY2 with 5.4 × 10 ⁷ cfu / g.

According to one aspect of the invention, the present invention is a composite microorganism SejungBio-FA100 (Microorganism Accession No .: KCTC 11400BP) culture medium 0.5 to 20% by weight; Rice bran 20-50% by weight; Food waste fermented 20 to 50% by weight; And it provides a complex microbial preparation for organic matter treatment and denitrogenation treatment of sewage and waste water consisting of 10 to 30% by weight of chaff charcoal powder and 5 to 10% by weight of humus.

The complex microorganism SejungBio-FA100 (Microorganism Accession No .: KCTC 11400BP) culture medium is 1 to 5 parts by weight of the SejungBio-FA100, 4 to 15 parts by weight of sucrose, 1 to 5 parts by weight of peptone, 0.05 to 1 parts by weight of lactose and yeast extract 0.5 to 10 parts by weight may be prepared by mixing the remaining water to 100 parts by weight.

According to another aspect of the present invention, the present invention provides a method for treating denitrification of sewage and wastewater, wherein the complex microbial agent is added to an aerobic tank and / or aeration tank.

The complex microorganism according to the present invention has an effect of decomposing organic matter of wastewater and wastewater and denitrification reaction under active aerobic conditions to efficiently remove nitrogen in wastewater.

Conventional biological nitrogen treatment methods are divided into nitrification under aerobic conditions and denitrification under anoxic conditions. In the two reactors, ammonia nitrogen is converted to nitrate nitrogen (nitride) and nitrate nitrogen is nitrogen gas (denitrification). Stepwise removal of nitrogen is achieved. The reason is that denitrification rarely occurs under aerobic conditions. However, this method, as shown in Figures 1a and 1b, the site area and additional costs associated with the use of the two reactors are expensive, there may be a problem of the increase in power costs due to the internal circulation. In addition, a lot of alkalinity (alkalinity) is consumed due to the pH decrease in the nitrification process, there is a disadvantage that the external carbon must be additionally supplied in the denitrification process.

Denitrification usually occurs in two stages. As shown in Scheme 1 below, the first step is a process in which nitric acid is reduced to nitrous acid, and the second step is a process in which nitrous acid is converted to nitrogen gas through two intermediate products.

Scheme 1

Denitrification microorganisms are widely found in the natural environment and vary in metabolism and activity. This denitrification is a widely ubiquitous feature among bacteria that are not related to the taxonomy, which is considered to be the result of gene transfer during evolution. Thus, denitrification is also carried out in some archaeological and fungal mitochondria, in some bacteria nitrate is reduced to only nitrite or N ₂ O, and some species are nitrates as receptors associated with respiration in anaerobic conditions. Some require only nitrate.

In addition, in the case of nitrogen removal by continuous reaction of nitrification and denitrification, appropriate pH adjustment is required for each reaction in order to optimize each reaction.

However, aerobic denitrification may simultaneously occur nitrification and denitrification in an aerobic and / or aeration tank in which organic matter removal takes place. In addition, the aerobic denitrification process does not require a pH adjustment because the pH change is not large, and has the potential to solve some of the disadvantages of the biological denitrification system by the continuous reaction of the existing anoxic-aerobic (see FIGS. 1A and 1B). have.

Various kinds of denitrification bacteria are being separated in aerobic environment, and research on the development of new biological denitrification process using these characteristics is ongoing. In addition, the use of aerobic denitrification bacteria in addition to the existing method to take a biological nitrogen removal method is being actively researched the situation.

In relation to aerobic denitrification, many such as Alcaligenes faecalies , Pseudomonas nautica , Thiosphaera pantotropha , Microvirgula aerodenitrificans , etc. It has been reported that denitrifiers can simultaneously use oxygen and nitrate as electron acceptors to increase their growth rate. Among them, Paracoccus denitrificans and Microvirgula aerodenitrificans have been studied in various aspects.

In addition, in order to economically remove high concentrations of nitrogen compounds in sewage and wastewater, studies on biological denitrification such as independent nutrient denitrification and aerobic denitrification using sulfur have been continuously conducted.

The complex microorganism of the present invention is a complex microorganism capable of decomposing organic matter in wastewater and wastewater, and causing denitrification not only under anaerobic conditions but also under aerobic conditions. Denitrification microorganisms that cause denitrification under aerobic conditions can be said to be an economical and effective method because they can operate a two-step process of nitrogen removal-an anoxic reaction process and an aerobic reaction process in one reactor.

In the present invention, microorganisms were separated by using a fermentation culture made by fermenting plant extracts such as bamboo leaves, buttercups, and wormwood leaves in the vicinity of swamps in the mountainous region of Gyeongsangnam-do, to create a complex microorganism. Specifically, after the fermentation culture was put into a reactor of a leather wastewater treatment plant operated and managed by Sejung Biotech Co., Ltd., Busan, a new type of denitrification treatment occurred under organic matter treatment and aerobic conditions of wastewater. Confirmed. And the strain is isolated and identified from the fermentation culture to prepare a complex microbial and complex microbial formulation.

Specifically, the complex microorganism of the present invention is Bacillus licheniformis FE1, Bacillus cereus FE3, Paenibacillus sp H10-02 FE4, Acetobacter formum ( Acetobacter pomorum ) FM2, Acetobacter peroxydans FM3, Bacillus subtilis FM4, Lactobacilus casei FM5, Paenibacillus sp. P33 FM8, Cluibero Kluyveromyces fragilis FY1 and Debaryomyces hansenii FY2.

Specifically, the complex microorganism according to the present invention, Bacillus licheniformis FE1 is 9X10 ⁷ cfu / g, Bacillus cereus FE3 is 7.2X10 ⁷ cfu / g, Fanibacillus sp H10-02 ( Paenibacillus sp. H10-02) FE4 is 4.2X10 ⁷ cfu / g, Acetobacter pomorum FM2 is 7.2X10 ⁷ cfu / g, Acetobacter peroxydans FM3 is 7.2X10 ⁷ cfu / g g, Bacillus subtilis FM4 is 4.2X10 ⁷ cfu / g, Lactobacilus casei FM5 is 6X10 ⁷ cfu / g, Paenibacillus sp. P33 FM8 is 4.2X10 ⁷ cfu / g, Kluyveromyces fragilis FY1 with 5.4 × 10 ⁷ cfu / g and Debaryomyces hansenii FY2 with 5.4 × 10 ⁷ cfu / g.

The complex microorganism was named SejungBio-FA100, and deposited on October 14, 2008 under the accession number KCTC 11400BP to the Korea Institute of Bioscience and Biotechnology.

The complex microorganism of the present invention can be used as a complex microbial preparation for organic matter treatment and denitrogenation treatment of waste water.

Specifically, the composite microorganism SejungBio-FA100; Rice bran; Food waste fermentation; And chaff charcoal coarse powder.

The complex microorganism SejungBio-FA100 can be used directly in the form of a seed or in the form of a culture solution. The preferred amount is 0.5 to 20% by weight based on the total weight of the complex microbial agent. In addition, the culture medium of the complex microorganism SejungBio-FA100 can be obtained through a general strain culture method. This is a method well known in the art.

In one embodiment of the present invention, the culture solution was obtained by the following method, but is not necessarily limited thereto. 1 to 5 parts by weight of the composite microorganism SejungBio-FA100 seedlings, 4 to 15 parts by weight of sucrose, 1 to 5 parts by weight of peptone, 0.05 to 1 parts by weight of lactose, and 0.5 to 1 parts of yeast extract, which were refrigerated (4 to 5 ° C.) 10 parts by weight of the remaining water is mixed to 100 parts by weight (pH 6.8) and incubated for 6 to 48 hours with a strong air supply at a temperature of 20 to 35 ℃. The reason for the strong air supply at this time is to cultivate aerobic microorganisms. Then, the air supply was stopped and further cultured under stirring for 2 to 10 days, and the obtained culture was used as SejungBio-FA100 culture. At this time, the temperature is preferably incubated at 30 ° C. for 24 hours and incubated with stirring for 4 days.

In addition to this, other known culture media may be added to prepare a microbial culture. For example, PYK medium and Succinate medium may be used as a culture medium, but are not necessarily limited thereto. The medium composition (g / L), PYK medium (pH 7) is 5 g peptone; 3 g yeast extract; 2 g KNO ₃ and succinic acid medium (pH 7), 4 g succinic acid; 1 g (NH ₄ ) 2 SO ₄ ; 3 g KH ₂ PO ₄ ; 0.1 g K ₂ HPO _4; 0.2 g MgSO ₄ .7H ₂ O; 2g KNO ₃ .

Rice bran is a mixture of various husks that occurs during the process from brown rice to white rice. Rice bran is known to be rich in rice bran fat, protein, essential fatty acids, essential amino acids, vitamins, dietary fiber and trace elements. Therefore, by using this as an additive, when the complex microbial preparation for wastewater treatment is solid fermentation, it provides nutrition and serves as a moisture control agent. In addition, rice bran has the advantage that it contains an effective microorganism that decomposes various organic matters and has a bad smell reduction effect.

In the present invention, the rice bran was purchased and used in the market, specific components of the rice bran is water 13.5%, fat 18.4%, sugar 38.3%, fiber 7.8%, ash 8.9%, vitamin B 12.5%, other 0.6%.

The food waste fermentation product may be any food waste food fermentation known in the art. In addition, by using the present composite microorganism as a seed, it is also possible to use a fermentation at 45 ℃ for about 5 to 10 days with food waste in a solid fermenter. Specifically, the food residue may include 100 to 500 parts by weight and 300 to 1000 parts by weight of sawdust. Through the fermentation process of food waste (45 ℃, stirred fermentation for 4 to 5 days), microorganisms capable of degrading and extinguishing solid organic substances including protein compounds present in the waste water and extinguishing can be proliferated and activated.

The chaff charcoal powder is a powder made by grinding the chaff charcoal charcoal 100% with a mixer grinder, it is added to further suppress the decay fermentation and to enhance the useful fermentation. The term 'crude powder' refers to coarse, ie, rough, powder. Therefore, the rice husk charcoal powder of the present invention refers to crushed chaff charcoal.

The humus soil, more specifically, is a humus-containing soil of the following composition: 60 to 70% by weight of cocoite, 10 to 20% by weight of peat moss, 7 to 10% by weight of pearlite, 5 to 8% by weight of vermiculite and 3 to 5 of zeolite. Wt%, deciduous broadleaf soils 1-2 wt%. Such humus can be used commercially available or manufactured directly.

The complex microbial preparation, the complex microorganism SejungBio-FA100 (Microbial Accession Number: KCTC 11400BP) or its culture solution 0.5 to 20% by weight; Rice bran 20-50% by weight; Food waste fermented 20 to 50% by weight; It consists of 10 to 30% by weight of chaff charcoal powder and 5 to 10% by weight of humus, but is not necessarily limited thereto. The composition ratio corresponds to the numerical range in which the activity of the complex microorganism SejungBio-FA100 can optimally occur.

The composition containing the microorganism SejungBio-FA100, rice bran, food waste fermented product, rice husk charcoal powder and humus soil was solid-fermented at 40 to 45 ° C. for 3 to 5 days while drying the water content to 40-50% or less. Complex microbial preparations for organic matter treatment and denitrogenation of wastewater can be prepared.

The complex microbial agent, Hyphomicrobium X ( Alphogeneb faecalis ), Nitrosomonas (nitrosomonas), denitrifying bacteria DCB-T6, T23, T25 (Denitrifying bacteria DCB-T6, T23, T25 ), At least one strain selected from the group consisting of Thiosphaera pantotropha , and Pseudomonas stutzer , each 0.5 to 1 part by weight based on 100 parts by weight of the complex microbial agent. It may include. The strains mentioned here are well known and can be distributed and used by strain preservation centers in each country (Korea KCCM, USA ATCC, Japan JCM or IFO, Germany DSM, Netherlands CBS, UK NCIBM, etc.).

According to another aspect of the present invention, the present invention can treat the nitrogen of the waste water by adding the complex microbial agent to the aerobic tank and / or aeration tank.

The complex microbial agent can be added to an aerobic and / or aeration tank for treating wastewater and wastewater to treat nitrogen of the wastewater. In general, when a reaction tank becomes aerobic through aeration, it is separately called "aerobic tank or aerobic aeration tank". The aerobic tank and / or aeration tank includes both aerobic aeration tanks in a general wastewater and wastewater treatment reactor and aeration tanks that may be operated in a quasi-anaerobic state. Since the complex microorganism of the present invention can perform denitrification even under aerobic conditions, it can be added to an aerobic tank and / or aeration tank to treat nitrogen in wastewater and wastewater.

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example 1 Isolation and Identification of Microorganisms

1-1. Pure separation of microorganisms

In order to purely separate the microorganisms which play a major role in wastewater treatment from fermentation cultures made by fermenting plant extracts of soil, bamboo leaves, buttercups and wormwood leaves in the swamps of the mountainous region of Gyeongsangnam-do, first, the fermentation culture samples were vortexed. After sufficient mixing, 1 ml is taken and added to sterile 0.1% NaCl solution, and the solution is mixed well with Vortex, and then placed on 3 times on 0.8% Nutrient broth agar medium. It was divided and spread. The plated agar plates were incubated in an incubator at 30 ° C., and when colonies were formed on the agar plates, colonies of different shapes, colors, and sizes of colonies were marked and collected to determine whether the same bacteria were observed under a microscope. Repeated subculture from the indicated colonies to fresh agar plates was used to isolate the finally isolated pure microorganisms.

1-2. Identification of Selected Microorganisms

Selected useful strains were identified via 16S-rDNA sequencing. First, DNA extraction was performed using a DNA extraction kit (Bioneer Accuprep, Genomic DNA extraction kit, Korea), and extracted DNA was placed in 0.5 ml tube with 10x PCR buffer, 2.5 mM dNTP, each primer 10 pmol, DNA template 1 ug, 5U. Taq polymerase (G-tag) was added and the rest was added to distilled water to make a total volume of 20 μl. PCR amplification was performed for 5 minutes of incubation at 95 ° C, denaturation at 95 ° C for 30 seconds, annealing at 55 ° C for 30 seconds, extension at 72 ° C for 30 seconds, and at 72 ° C. The final extension was run for 10 minutes. At this time, the primers used for the PCR reaction were 27F (5'-AGAGTTTGATCCTGGCTCAG-3 ') and 1429R (5'-GGTTACCTTGTTACGACTT-3), and GeneAmp PCR system 2400 (Perkin Elmer applied Biosystems, USA) was used. The amplified PCR product was electrophoresed on a 1% agarose gel to confirm PCR amplification, and then the PCR product was purified (Generalbiosystem, USA).

The purified PCR product was commissioned by Macrogen Co., Ltd. to obtain final 5'- and 3'-terminal 16S rDNA sequences, and the sequence of each part was found at http://www.ncbi.nlm.nih.gov. The access was compared and searched in GenBank (National Center for Biotechnology Information, Rockville Pike, Bethesda, MD) through the Advanced BLAST similarity search option (Altschul et. Al., 1997).

As a result, a total of 13 strains were isolated from the two samples (FIG. 2), 5 strains were isolated from sample A, and the strain names were defined as FE 1-FE 5, and 8 strains were separated from sample B to randomly select strain names. FM 1-FM 8 was set. As a result of observing the microscopic and colony form, FM 1 strain and FE 2 strain were identified as microorganisms having the same colonies, excluding FM 1. Also, FE 2 and FE 5 strains were identified as yeast strains of the same type as FM 6 and FM 7, respectively, and these yeasts were named FY 1 and FY 2 separately. Therefore, the total number of microorganisms finally separated is 10 species including 2 yeasts. Identification results of eight microorganisms except yeast are shown in Table 1 below. Identification of yeast will be described in detail below in a separate title.

TABLE 1

The identified strains were Bacillus licheniformis FE1, Bacillus cereus FE3, Paenibacillus sp. H10-02 FE4, Acetobacter pomorum , respectively . FM2, Acetobacter peroxydans FM3, Bacillus subtilis FM4, Lactobacilus casei FM5 and Paenibacillus sp. P33 FM8.

1-3. Yeast Identification

As mentioned above, among the microorganisms isolated from each pure microorganism, the microorganisms estimated as yeast through microscopic observation were separately selected and named FY1 and FY2, and the reaction characteristics according to the composition of each medium by incubation in different yeast media (Table 2) The results were finally identified according to the yeast identification criteria of FIG. 3.

TABLE 2 Yeast Identification Medium Composition (g / 1L)

As a result of microscopic observation, the identification of FY 1 and FY 2 microorganisms presumed to be germ-shaped yeast cells was classified according to the yeast identification criteria in FIG. First, after inoculating FY 1 and FY 2 microorganisms in 0.8% nutrient broth, it was confirmed whether pellicles were formed on the liquid medium in which the bacteria were grown. Since the pellicle was not formed in the 0.8% nutrient broth in both FY1 and FY2, the experiment was conducted according to the identification criteria of FIG. 2, and both colonies and cultures formed by the FY1 and FY2 microorganisms did not have pigmentation (B-2). The experiment was conducted according to the identification criteria. As a result, FY1 and FY2 were grown in lactose medium, and in the case of FY 2, pellicles were formed, and only FY2 was again tested in the order of identification (A).

As a result, it did not grow in 60% glucose yeast extract, but grew in maltose medium. Therefore, FY 2 was classified as Debaryomyces hansenii by the yeast identification criteria of FIG. 2, so it was named Debaryomyces hansenii FY2, and the strain FY1 was identified in (B-2). Therefore, it was classified as Kluyveromyces fragilis according to the identification criteria of FIG. 2 because it was grown in lactose medium, and was named Kluyveromyces fragilis FY1. The K. fragilis is not a pathogenic microorganism, and is known to grow well in the liquor and dairy industry wastes, and is a strain that can be effectively used for wastewater treatment. Therefore, it is judged that the strain can be found in the wastewater treatment reactor with excellent treatment effect.

FY1 and FY2 are fermenting low molecular sugars and converting them into useful amino acids, vitamins, and alcohols, and are considered to be microorganisms involved in organic water treatment of high concentration organic wastewater.

A total of 10 microorganisms including the isolated and identified yeasts were named SejungBio-FA100 as a complex microorganism, and deposited on the 14th of October, 2008 with the accession number KCTC 11400BP to the Biological Resource Center of Korea Research Institute of Bioscience and Biotechnology. Specific contents of the complex microorganisms are shown in Table 3 below. This complex microorganism was stored in a refrigerated state (5 ± 2 ℃).

TABLE 3

microbe Number of bacteria (cfu / g) Content (% by weight) Bacillus licheniformis FE1 9X10 ⁷ 15 Bacillus cereus FE3 7.2 X 10 ⁷ 12 Paenibacillus sp. H10-02 FE4 4.2 X 10 ⁷ 7 Acetobacter pomorum FM2 7.2 X 10 ⁷ 12 Acetobacter peroxydans FM3 7.2 X 10 ⁷ 12 Bacillus subtilis FM4 4.2 X 10 ⁷ 7 Lactobacillus casei FM5 6X10 ⁷ 10 Panibacillus sp. P33 fm8 4.2 X 10 ⁷ 7 Kluyveromyces fragilis FY1 5.4 X 10 ⁷ 9 Debaryomyces hansenii FY2 5.4 X 10 ⁷ 9

Example 2: Preparation of Complex Microbial Formulations

4 parts by weight of the microorganisms in the refrigerated complex of Example 1, 6 parts by weight of sucrose, 2 parts by weight of peptone, 0.1 parts by weight of lactose and 1 part by weight of the yeast extract and mixed in the incubator to 100 parts by weight ( Initial pH 6.8), incubated for 24 hours with a strong air supply to the incubator at 30 ℃. Thereafter, the air supply was stopped, and further cultured for 4 days while stirring to obtain a complex microorganism SejungBio-FA100 culture.

20% by weight of the composite microbial culture medium, 30% by weight of rice bran, 30% by weight of food waste fermentation (food waste ferment is 100 parts by weight of food waste and 800 parts by weight of sawdust mixed with 100 parts by weight of SejungBio-FA100 and 4 days at 45 ℃ Obtained by stirring and fermentation), 10% by weight of coarse rice husk charcoal and 10% by weight of humus earth and solid fermentation (model name: KJ-3, manufactured by Koena, Korea) at 45 ℃ moisture content between 40 to 50% The composite microbial formulation was prepared by drying with.

Example 3: Denitrification Experiment of Complex Microbial Formulation

3-1 Denitrification Test in Syringe

The aerobic denitrification reaction of the complex microbial preparation of Example 2 was carried out in an aerobic state in which oxygen was supplied using a 50 mL syringe, as shown in FIG. The reaction characteristics were compared. First sterilize 20 mL of liquid medium (PYK medium: 5 g peptone; 3 g yeast extract; 2 g KNO ₃ (initial pH 7.0)) containing 2 g of 5% (w / v) complex microbial preparation of Example 2 10 mL of oxygen was supplied into a 50 mL syringe using a Hamilton gastight syringe for aerobic conditions, and then the 50 mL syringe was placed in a 180 rpm shaking incubator and incubated at 30 ° C. Whenever dissolved oxygen in the medium was insufficient, oxygen was continuously supplied, and the gas generated in the syringe was sampled at regular time and analyzed by GC (Gas Chromatography). Gas Chromatography (GC) analysis was performed to determine the composition of N2 gas produced by denitrification in the gas. Gas measurement was performed by first collecting a gas from a 50 mL syringe using a 50 μL gas-tight syringe. Analysis was performed by injection into GC / TCD (Perkin Elmer Instruments). The concentration of N2 gas produced by the denitrification microorganism was obtained by a standard curve, and the amount of N2 gas was calculated by stoichiometric formula according to the ideal gas law.

As a result, in the case of anaerobic syringe experiment (FIG. 5A), gas bubbles began to be generated after 3 hours of reaction, and 1.5 mL of gas measurement was possible within 8 hours of reaction. As a result of analysis using GC, it was found that N ₂ of 37.0 μmole occurred, and 72.1 μmole generated after 72 hours of the final reaction. At this time, the final gas generated 28.3 μmole, which was analyzed by CO2 gas. It was found that most of this gas was CO2, and it could smell like hydrogen sulfide (Fig. 6).

In the case of aerobic syringe experiment (FIG. 5B), oxygen was almost consumed within 8 hours of reaction to re-inject oxygen three times, and N ₂ gas generated 64.2 μmole in the final 72 hours of reaction. In addition to the N ₂ gas, an unidentified gas generated a final 411.9 μmole, most of which was a CO ₂ gas. Therefore, a little more N ₂ gas was generated in the anaerobic state, but sufficient denitrification occurred in the aerobic state.

3-2 Aerobic Denitrification in a 1L 5-neck Flask

In order to observe the change in the main reaction parameters of the denitrification reaction seen in the syringe reactor of Example 3-1, a liquid medium containing 5% (w / v) of the composite microbial formulation of Example 2 incubated for 3 days (PYK) Medium: 5 g peptone; 3 g yeast extract; 2 g KNO ₃ (initial pH 7)) was inoculated in a 1 L 5-neck flask. 1 L flask was incubated in a 30 ℃ constant temperature water bath. After all experimental equipment was set up for aerobic conditions, aeration was performed for 10 minutes into a 5-neck flask and sampled. Pure O ₂ (80% purity) was fed through a 0.2 μm filter when the DO value dropped below 0.5 mg / L for aerobic conditions.

PH, DO, ORP (Redox Reduction Potential) was measured in real time, and through regular sampling, DSW (Dry Sludge Weight), CODcr, Total Nitrogen (TN) (L), Total Kjeldahl Nitrogen, TKN ) (L + s) and IC (NH4 +, NO3-, NO2-) were analyzed, and TN (s) was determined by theoretical calculation of the N component in the cells using DSW. In addition, the number of viable cells was measured by colony forming individuals (CFU) to finally determine the number of viable cells considering the dilution rate.

As shown in FIG. 7A, the concentration of DO in the flask was increased to 4.6 mg / L after 3 hours of reaction by continuously supplied oxygen. However, it decreased to 0.2 mg / L within 1-2 hours after the oxygen supply was stopped. After that, two more oxygens were supplied, but the DO value did not increase but remained low due to active oxygen consumption by the complex microbial preparation of Example 2. At this time, the change in ORP was maintained in the first 3 hours at a high DO concentration, but then decreased to -33.6 mV within 4 hours with a sharp decrease in DO, and gradually increased after 6 hours, starting from 9 hours. The positive value was maintained. However, after 32 hours of reaction, the final ORP value was -19 mV, showing a tendency to decrease again. In the case of pH, it decreased from 6.52 to 5.53 for the first 2 hours, but increased as the reaction time continued, resulting in a final pH of 6.24. Tell them what is happening. In addition, as shown in Figure 7b, the values of CODcr, TN, TKN can be seen to decrease.

As such, it can be seen that the complex microbial preparation of the present invention is actively denitrified even under aerobic conditions.

Example 4 Confirmation of Organic Matter Treatment and Denitrogenation Treatment of Waste and Wastewater in Complex Microbial Formulations

4-1: Confirmation of denitrification of wastewater under aerobic conditions

The wastewater treatment facility of the leather wastewater treatment plant as shown in FIG. 8 was used. The wastewater treatment capacity is about 5,000 m3 / d, and the overflow water treated at the front end is bioreactor [aeration tank capacity 18,000 m3, multi-stage aeration tank (B1, B2, B3, B4, B5, B6, B7, and B8 divided into eight aeration tanks). )] Was the condition to flow into the process. Influent COD concentrations are usually between 100 and 200 mg / l and total nitrogen is between 150 and 200 mg / l. In the treatment tank, the composite microbial preparation of Example 2 was 2 to 3 m3 per day, about 70 to 90% in the first stage B1 aeration tank, and 10 to 30% in the B5 to B6 aeration tank where most of the ammonia nitrogen was converted to nitrate nitrogen. In addition, the processing type of nitrogen was added while fluidly flowing. The aeration tank was continuously charged with 150 L / min (for replenishing insufficient alkalinity and carbon source) into the B6 aeration tank, and the dissolved oxygen (DO) was operated between 0.5 and 2.0 mg / l, which is an aerobic condition. As a result, the ammonia nitrogen (NH ₄ -N) and nitrite nitrogen (NO ₂ -N) were partially reduced or converted, so that the increased nitrogen nitrate (NO ₃ -N) decreased drastically from the rear B5 aeration tank. The results showed that the nitrogen content was reduced (FIG. 9). Therefore, it can be seen that the denitrification phenomenon occurs in the aeration tank under aerobic conditions by the complex microbial preparation of the present invention, so that nitrogen is treated.

4-2: Confirmation of Organics Treatment of Wastewater

As shown in FIG. 10, the chemical oxygen demand (COD), which is a general indicator of organic matter contained in the wastewater, also decreases in the B5 aeration tank where aerobic denitrification occurs. It is believed that COD decreases as some organics are solubilized by other nutrients such as Bacillus.

The effective treatment of nitrogen in aerobic conditions as described above is to take an internal bulkhead in a single aerobic tank to take a multistage structure (such as those in aerobic tanks 1, 2 and 3 of FIG. 11), and to process nitrogen. The method of investigating the type and injecting the complex microbial culture of the present invention to the required area is effective.

4-3: Confirmation of organic matter treatment and denitrification of sewage

In order to apply the composite microbial preparation of Example 2 to sewage, a laboratory level pot test was conducted. 15 L of sewage was added to a 20 L plastic test vessel and the concentration of mixed liquor suspended solids (MLSS) in the reactor was adjusted to 3,000 mg / L, pH was 7.2, DO was 1.0-2.0 mg / L, Example Sewage source water flowing into Janglim sewage treatment plant was tested under 0.1% of the combined microbial preparation of 2.

As shown in FIG. 12, the COD and total nitrogen rapidly decreased from the initial COD of 350 mg / L after the reaction time of 4 hours, and the COD of 17 mg / L and the total nitrogen of 8 hours of reaction time. 12 mg / L. This indicates that the complex microorganism SejungBio-FA100 of the present invention and the complex microbial preparation including the same can perform the high-treatment effect on the sewage treatment plant effluent under a single reactor and aerobic reaction conditions.

Simple modifications or changes of the present invention can be easily carried out by those skilled in the art, and all such modifications or changes can be seen to be included in the scope of the present invention.

1a and 1b schematically show a denitrification reaction according to the prior art.

Figure 2 shows a colony picture of each microorganism purely separated. (A) FE 1; (B) FE 2; (C) FE 3; (D) FE 4; (E) FE 5; (F) FM 1; (G) FM 2; (H) FM 3; (I) FM 4; (J) FM 5; (K) FM 6; (L) FM 7; (M) FM 8.

Figure 3 shows the criteria for identifying yeast.

4 shows a 50 mL syringe photograph used for aerobic / anaerobic denitrification.

5 is a graph showing changes in gas composition in syringe experiments under anaerobic conditions (A) and aerobic conditions (B). (↓) is O ₂ , (●) is O ₂ ; (■) is N ₂ and (▲) is unidentified gas.

Figure 6 shows a photograph of a syringe experiment in anaerobic conditions.

Figure 7a shows the parameter change during the aerobic denitrification experiment in a 1L 5-neck flask according to Example 3-2. (↔) O ₂ is oxygen supply; (●) DO; (▲) is pH; (■) is ORP; And (▼) indicate viable counts.

Figure 7b shows the parameter change during the aerobic denitrification experiment in a 1L 5-neck flask according to Example 3-2. (●) represents CODcr; (▲) is TN; And (■) represents TKN.

8 schematically shows a wastewater treatment plant according to a fourth embodiment of the present invention. P: pump, A1: first setting tank, A2: second setting tank, A3: third setting tank, C1: first liquefaction tank and C2: second liquefaction tank.

9 is a graph showing the relationship between the nitrogen removal type and dissolved oxygen in the aeration tank according to Example 4.

10 is a graph showing the relationship between the COD and DO in a composite microbial preparation aeration tank according to Example 4.

11 is a view illustrating a process for adding a complex microbial preparation and an organic material treatment process and an aerobic denitrification process according to an embodiment of the present invention.

12 is a graph showing the effects of COD and T-N treatment for sewage according to an embodiment of the present invention.

<110> LEE, Geon          SEJUNG BIOTECH CO.LTD. <120> Complex strain for the waste water / sewage treatment and Nitrogen          treatment process using the same <130> P08-270 <150> KR10-2008-0107867 <151> 2008-10-31 <160> 8 <170> KopatentIn 1.71 <210> 1 <211> 1402 <212> DNA <213> Bacillus licheniformis <400> 1 ttttaagatt tnnnntcggc tcaggacgaa cgctggcggc gtgcctaata catgcaagtc 60 gagcggacag atgggagctt gctccctgat gttagcggcg gacgggtgag taacacgtgg 120 gtaacctgcc tgtaagactg ggataactcc gggaaaccgg ggctaatacc ggatggttgt 180 ttgaaccgca tggttcaaac ataaaaggtg gcttcggcta ccacttacag atggacccgc 240 ggcgcattag ctagttggtg aggtaacggc tcaccaaggc gacgatgcgt agccgacctg 300 agagggtgat cggccacact gggactgaga cacggcccag actcctacgg gaggcagcag 360 tagggaatct tccgcaatgg acgaaagtct gacggagcaa cgccgcgtga gtgatgaagg 420 ttttcggatc gtaaagctct gttgttaggg aagaacaagt accgttcgaa tagggcggta 480 ccttgacggt acctaaccag aaagccacgg ctaactacgt gccagcagcc gcggtaatac 540 gtaggtggca agcgttgtcc ggaattattg ggcgtaaagg gctcgcaggc ggtttcttaa 600 gtctgatgtg aaagcccccg gctcaaccgg ggagggtcat tggaaactgg ggaacttgag 660 tgcagaagag gagagtggaa ttccacgtgt agcggtgaaa tgcgtagaga tgtggaggaa 720 caccagtggc gaaggcgact ctctggtctg taactgacgc tgaggagcga aagcgtgggg 780 agcgaacagg attagatacc ctggtagtcc acgccgtaaa cgatgagtgc taagtgttag 840 ggggtttccg ccccttagtg ctgcagctaa cgcattaagc actccgcctg gggagtacgg 900 tcgcaagact gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt 960 ttaattcgaa gcaacgcgaa gaaccttacc aggtcttgac atcctctgac aatcctagag 1020 ataggacgtc cccttcgggg gcagagtgac aggtggtgca tggttgtcgt cagctcgtgt 1080 cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct tgatcttagt tgccagcatt 1140 cagttgggca ctctaaggtg actgccggtg acaaaccgga ggaaggtggg gatgacgtca 1200 aatcatcatg ccccttatga cctgggctac acacgtgcta caatggacag aacaaagggc 1260 agcgaaaccg cgaggttaag ccaatcccac aaatctgttc tcagttcgga tcgcagtctg 1320 caactcgact gcgtgaagct ggaatcgcta gtaatcgcgg atcacatgcn gcgggttgaa 1380 tacgttcccg gggcttgtac ac 1402 <210> 2 <211> 1401 <212> DNA <213> Bacillus cereus <400> 2 cactagagtt tgatcatggc tcaggatgaa cgctggcggc gtgcctaata catgcaagtc 60 gagcgaatgg attgagagct tgctctcaag aagttagcgg cggacgggtg agtaacacgt 120 gggtaacctg cccataagac tgggataact ccgggaaacc ggggctaata ccggataata 180 ttttgaactg catggttcga aattgaaagg cggcttcggc tgtcacttat ggatggaccc 240 gcgtcgcatt agctagttgg tgaggtaacg gctcaccaag gcaacgatgc gtagccgacc 300 tgagagggtg atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc 360 agtagggaat cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa 420 ggctttcggg tcgtaaaact ctgttgttag ggaagaacaa gtgctagttg aataagctgg 480 caccttgacg gtacctaacc agaaagccac ggctaactac gtgccagcag ccgcggtaat 540 acgtaggtgg caagcgttat ccggaattat tgggcgtaaa gcgcgcgcag gtggtttctt 600 aagtctgatg tgaaagccca cggctcaacc gtggagggtc attggaaact gggagacttg 660 agtgcagaag aggaaagtgg aattccatgt gtagcggtga aatgcgtaga gatatggagg 720 aacaccagtg gcgaaggcga ctttctggtc tgtaactgac actgaggcgc gaaagcgtgg 780 ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatgagt gctaagtgtt 840 agagggtttc cgccctttag tgctgaagtt aacgcattaa gcactccgcc tggggagtac 900 ggccgcaagg ctgaaactca aaggaattga cgggggcccg cacaagcggt ggagcatgtg 960 gtttaattcg aagcaacgcg aagaacctta ccaggtcttg acatcctctg aaaaccctag 1020 agatagggct tctccttcgg gagcagagtg acaggtggtg catggttgtc gtcagctcgt 1080 gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta gttgccatca 1140 ttaagttggg cactctaagg tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt 1200 caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggac ggtacaaaga 1260 gctgcaagac cgcgaggtgg agctaatctc ataaaaccgt tctcagttcg gattgtaggc 1320 tgcaactcgc ctacatgaag ctggaatcgc tagtaatcgc ggatcacatg cccgggttga 1380 atacgttccc ggggcttgta c 1401 <210> 3 <211> 1312 <212> DNA <213> Paenibacillus sp. H10-02 <400> 3 ttagcggcgg acgggtgagt aacacgtagg caacctgcct ataagactgg gataactatc 60 ggaaacgata gctaagaccg gataactggt tttctcgcat gagagaatca tgaaacacgg 120 agcaatctgt ggcttataga tgggcctgcg gcgcattagc tagttggtga ggtaacggct 180 caccaaggcg acgatgcgta gccgacctga gagggtgaac ggccacactg ggactgagac 240 acggcccaga ctcctacggg aggcagcagt agggaatctt ccgcaatgga cgcaagtctg 300 acggagcaac gccgcgtgag tgatgaaggt tttcggatcg taaagctctg ttgccctaga 360 cgaacagcaa ggcgagtaac tgcgctttgt gtgacggtat aggagaagaa agccccggct 420 aactacgtgc cagcagccgc ggtaatacgt agggggcaag cgttgtccgg aattattggg 480 cgtaaagcgc gcgcaggcgg tcaattaagt tgggtgttta agcccggggc tcaaccccgg 540 ttcgcatcca aaactggttg acttgagtgt aggagaggaa agtggaattc cacgtgtagc 600 ggtgaaatgc gtagagatgt ggaggaacac cagtggcgaa ggcgactttc tggcctataa 660 ctgacgctga ggcgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg 720 ccgtaaacga tgcatactag gtgttgggga ttcgattcct cggtgccgaa gttaacacag 780 taagtatgcc gcctggggag tacgctcgca agagtgaaac tcaaaggaat tgacggggac 840 ccgcacaagc agtggagtat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc 900 ttgacatccc tctgtaagct ctagagatag agccctcctt cggaacatag gagacaggtg 960 gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1020 acccttgatc ttagttgcca gcacttcggg tgggcactct aagatgactg ccggtgacaa 1080 accggaggaa ggtggggatg acgtcaaatc atcatgcccc ttatgacctg ggctacacac 1140 gtactacaat ggtcggtaca acgggaagcg aagccgcgag gcggagccaa tccttataag 1200 ccgatctcag ttcggattgc aggctgcaac tcgcctgcat gaagtcggaa ttgctagtaa 1260 tcgcggatca gcatgccgcg gtgaatacgt tcccgggtct tgtacacacc gc 1312 <210> 4 <211> 1345 <212> DNA Acetobacter pomorum <400> 4 aatagagttt tngactggct cagagcgaac gctggcggca tgcttaacac atgcaagtcg 60 cacgaaggtt tcggccttag tggcggacgg gtgagtaacg cgtaggaatc tatccatggg 120 tgggggataa cactgggaaa ctggtgctaa taccgcatga tacctgaggg tcaaaggcgc 180 aagtcgcctg tggaggagcc tgcgttcgat tagctagttg gtggggtaaa ggcctaccaa 240 ggcgatgatc gatagctggt ttgagaggat gatcagccac actgggactg agacacggcc 300 cagactccta cgggaggcag cagtggggaa tattggacaa tgggggcaac cctgatccag 360 caatgccgcg tgtgtgaaga aggtcttcgg attgtaaagc actttcgacg gggacgatga 420 tgacggtacc cgtagaagaa gccccggcta acttcgtgcc agcagccgcg gtaatacgaa 480 gggggctagc gttgctcgga atgactgggc gtaaagggcg tgtaggcggt tttgacagtc 540 agatgtgaaa tccccgggct taacctggga gctgcatttg agacgttaag actagagtgt 600 gagagagggt tgtggaattc ccagtgtaga ggtgaaattc gtagatattg ggaagaacac 660 cggtggcgaa ggcggcaacc tggctcatta ctgacgctga ggcgcgaaag cgtggggagc 720 aaacaggatt agataccctg gtagtccacg ctgtaaacga tgtgtgctag atgttgggta 780 acttagttac tcagtgtcgc agttaacgcg ttaagcacac cgcctgggga gtacggccgc 840 aaggttgaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa 900 ttcgaagcaa cgcgcagaac cttaccaggg cttgaatgtg gaggctgtag gcagagatgt 960 ctatttcttc ggacctccaa cacaggtgct gcatggctgt cgtcagctcg tgtcgtgaga 1020 tgttgggtta agtcccgcaa cgagcgcaac ccctatcttt agttgccagc atgtttgggt 1080 gggcactcta gagagactgc cggtgacaag ccggaggaag gtggggatga cgtcaagtcc 1140 tcatggccct tatgtcctgg gctacacacg tgctacaatg gcggtgacag tgggaagcta 1200 tgtggtgaca cagtgctgat ctctaaaagc cgtctcagtt cggattgcac tctgcaactc 1260 gagtgcatga aggtggaatc gctagtaatc gcggatcagc atgcccgcgg tgaatacgtt 1320 cccgggcctt gtacacaccg cccgt 1345 <210> 5 <211> 1420 <212> DNA Acetobacter peroxydans <400> 5 tttgacaagg aggacgaatt gtacaacagg atatctatca acacaggaaa gaaaaccggc 60 aggaggcaaa agaaacgaaa caccacccca gaaaaaacac aacactagag ttgaatcatg 120 gctcagagcg aacgctggcg gcatgcttaa cacatgcaag tcgcacgaag gtttcggcct 180 tagtggcgga cgggtgagta acgcgtagga atctatccat gggtggggga taacactggg 240 aaactggtgc taataccgca tgatacctga gggtcaaagg cgcaagtcgc ctgtggagga 300 gcctgcgttc gattagctag ttggtggggt aaaggcctac caaggcgatg atcgatagct 360 ggtttgagag gatgatcagc cacactggga ctgagacacg gcccagactc ctacgggagg 420 cagcagtggg gaatattgga caatgggggc aaccctgatc cagcaatgcc gcgtgtgtga 480 agaaggtctt cggattgtaa agcactttcg acggggacga tgatgacggt acccgtagaa 540 gaagccccgg ctaacttcgt gccagcagcc gcggtaatac gaagggggct agcgttgctc 600 ggaatgactg ggcgtaaagg gcgtgtaggc ggttttgaca gtcagatgtg aaatccccgg 660 gcttaacctg ggagctgcat ttgagacgtt aagactagag tgtgagagag ggttgtggaa 720 ttcccagtgt agaggtgaaa ttcgtagata ttgggaagaa caccggtggc gaaggcggca 780 acctggctca ttactgacgc tgaggcgcga aagcgtgggg agcaaacagg attagatacc 840 ctggtagtcc acgctgtaaa cgatgtgtgc tagatgttgg gtaacttagt tactcagtgt 900 cgcagttaac gcgttaagca caccgcctgg ggagtacggc cgcaaggttg aaactcaaag 960 gaattgacgg gggcccgcac aagcggtgga gcatgtggtt taattcgaag caacgcgcag 1020 aaccttacca gggcttgaat gtggaggctg taggcagaga tgtctatttc ttcggacctc 1080 caacacaggt gctgcatggc tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg 1140 caacgagcgc aacccctatc tttagttgcc agcatgtttg ggtgggcact ctagagagac 1200 tgccggtgac aagccggagg aaggtgggga tgacgtcaag tcctcatggc ccttatgtcc 1260 tgggctaccc acgtgctaca atggcggtga cagttgggaa gcctatgttg gtggaccatt 1320 gtcttatcct ctaaagcccg tcttcatttc gaattgcacc ctgcgaactc caggcggcag 1380 gaaggtggga ttggtttgta atccttggat caccatggcc 1420 <210> 6 <211> 1378 <212> DNA <213> Bacillus subtilis <400> 6 gaaactgggg aacttgagtg cagaagagga gagtggaatt ccacgttgta gcggtgaaat 60 gcgtagagat gtggaggaac accagtggcg aaggcgactc tctggtctgt aactgacgct 120 gaggagcgaa agcgtgggga gcgaacagga ttagataccc tggtagtcca cgccgtaaac 180 gatgagtgct aagtgttagg gggtttccgc cccttagtgc tgcagctaac gcattaagca 240 ctccgcctgg ggagtacggt cgcaagactg aaactcaaag gaattgacgg gggcccgcac 300 aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacca ggtcttgaca 360 tcctctgaca atcctagaga taggacgtcc ccttcggggg cagagtgaca ggtggtgcat 420 ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt 480 gatcttagtt gccagcattc agttgggcac tctaaggtga ctgccggtga caaaccggag 540 gaaggtgggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca cacgtgctac 600 aatggacaga acaaagggca gcgaaaccgg cgaggttaaa ccaatcccac caatctgttc 660 tcagttccga acgcagtctg caactcgact gcgtgaaact ggaatcccta gtaatcgcgg 720 atccacatgc cgcggtgact agagtttgat catggctcag gacgaacgct ggcggcgtgc 780 ctaatacatg caagtcgagc ggacagatgg gagcttgctc cctgatgtta gcggcggacg 840 ggtgagtaac acgtgggtaa cctgcctgta agactgggat aactccggga aaccggggct 900 aataccggat ggttgtttga accgcatggt tcagacataa aaggtggctt cggctaccac 960 ttacagatgg acccgcggcg cattagctag ttggtgaggt aacggctcac caaggcgacg 1020 atgcgtagcc gacctgagag ggtgatcggc cacactggga ctgagacacg gcccagactc 1080 ctacgggagg cagcagtagg gaatcttccg caatggacga aagtctgacg gagcaacgcc 1140 gcgtgagtga tgaaggtttt cggatcgtaa agctctgttg ttagggaaga acaagtgccg 1200 ttcaaatagg gcggcacctt gacggtacct aaccagaaag ccacggctaa ctacgtgcca 1260 gcagccgcgg taatacgtag gtggcaagcg ttgtccggaa ttattgggcg taaagggctc 1320 gcaggcggtt tcttaagtct gatgtgaaag cccccggctc aaccggggag ggtcattg 1378 <210> 7 <211> 1413 <212> DNA <213> Lactobacillus casei <400> 7 gagtttggat ctggctcagg atgaacgctg gcggcgtgcc taatacatgc aagtcgaacg 60 agttctcgtt gatgatcggt gcttgcaccg agattcaaca tggaacgagt ggcggacggg 120 tgagtaacac gtgggtaacc tgcccttaag tgggggataa catttggaaa cagatgctaa 180 taccgcatag atccaagaac cgcatggttc ttggctgaaa gatggcgtaa gctatcgctt 240 ttggatggac ccgcggcgta ttagctagtt ggtgaggtaa tggctcacca aggcgatgat 300 acgtagccga actgagaggt tgatcggcca cattgggact gagacacggc ccaaactcct 360 acgggaggca gcagtaggga atcttccaca atggacgcaa gtctgatgga gcaacgccgc 420 gtgagtgaag aaggctttcg ggtcgtaaaa ctctgttgtt ggagaagaat ggtcggcaga 480 gtaactgttg tcggcgtgac ggtatccaac cagaaagcca cggctaacta cgtgccagca 540 gccgcggtaa tacgtaggtg gcaagcgtta tccggattta ttgggcgtaa agcgagcgca 600 ggcggttttt taagtctgat gtgaaagccc tcggcttaac cgaggaagcg catcggaaac 660 tgggaaactt gagtgcacaa gaggacagtg agaactccat gtgtagcggt gaaatgcgta 720 gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg acgctgaggc 780 tcgaaagcat gggtagcgaa caggattaga taccctggta gtccatgccg taaacgatga 840 atgctaggtg ttggagggtt tccgcccttc agtgccgcag ctaacgcatt aagcattccg 900 cctggggagt acgaccgcaa ggttgaaact caaaggaatt gacgggggcc cgcacaagcg 960 gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggtct tgacatcttt 1020 tgatcacctg agagatcagg tttccccttc gggggcaaaa tgacaggtgg tgcatggttg 1080 tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa cccttatgac 1140 tagttgccag catttagttg ggcactctag taagactgcc ggtgacaaac cggaggaagg 1200 tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt gctacaatgg 1260 atggtacaac gagttgcgag accgcgaggt caagctaatc tcttaaagcc attctcagtt 1320 cggactgtan ngctgcactc gcctacacga agtcggaatc gctagtaatc gcggatcagc 1380 acgcgcggtg aatacgttcc ccggccttgt aca 1413 <210> 8 <211> 1430 <212> DNA <213> Paenibacillus sp. P33 <400> 8 aaaaaagaca aaacgagcca caacagcgnn nncagggaac ccctagagtt tgatctgggt 60 tcaggacgaa cgctggcggc atgcntaata catgcaagtc gagcggactt gatgagaagc 120 ttgcttctct gatggttagc ggcggacggg tgagtaacac gtaggcaacc tgccctcaag 180 cttgggacaa ctaccggaaa cggtagctaa taccgaatag ttgttttctt ctcctgaaga 240 gaactggaaa gacggagcaa tctgtcactt ggggatgggc ctgcggcgca ttagctagtt 300 ggtggggtaa cggctcacca aggcgacgat gcgtagccga cctgagaggg tgatcggcca 360 cactgggact gagacacggc ccagactcct acgggaggca gcagtaggga atcttccgca 420 atgggcgaaa gcctgacgga gcaatgccgc gtgagtgatg aaggttttcg gatcgtaaag 480 ctctgttgcc agggaagaac gcttgggaga gtaactgctc tcaaggtgac ggtacctgag 540 aagaaagccc cggctaacta cgtgccagca gccgcggtaa tacgtagggg gcaagcgttg 600 tccggaatta ttgggcgtaa agcgcgcgca ggcggtcatt taagtctggt gtttaatccc 660 ggggctcaac cccggatcgc actggaaact gggtgacttg agtgcagaag aggagagtgg 720 aattccacgt gtagcggtga aatgcgtaga tatgtggagg aacaccagtg gcgaaggcga 780 ctctctgggc tgtaactgac gctgaggcgc gaaagcgtgg ggagcaaaca ggattagata 840 ccctggtagt ccacgccgta aacgatgagt gctaggtgtt aggggtttcg atacccttgg 900 tgccgaagtt aacacattaa gcactccgcc tggggagtac ggtcgcaaga ctgaaactca 960 aaggaattga cggggacccg cacaagcagt ggagtatgtg gtttaattcg aagcaacgcg 1020 aagaacctta ccaggtcttg acatccctct gaccggtaca gagatgtacc tttccttcgg 1080 gacagacgag acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa 1140 gtcccgcaac gagcgcaacc cttgatctta gttgccagca tttcggatgg gcactctaag 1200 gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg tcaaatcatc atgcccctta 1260 tgacctgngc tacacacgta ctacaatggc cggtacaacg ggctgtgaag ccgcgaggtg 1320 gaacgaatcc taaaagccgg tctcagttcg gatttgcagc tgcaactcgc ctgcatgaag 1380 tcggaattgc taataatccc ggatcaccat ggcgcggtga atacgttccc 1430  

Claims (5)

Bacillus licheniformis FE1 9X10 ⁷ cfu / g, Bacillus cereus FE3 7.2X10 ⁷ cfu / g, Paenibacillus sp. H10-02 FE4 4.2X10 ⁷ cfu / g, Acetobacter pomorum FM2 7.2X10 ⁷ cfu / g, Acetobacter peroxydans FM3 7.2X10 ⁷ cfu / g, Bacillus subtilis FM4 4.2X10 ⁷ cfu / g, Lactobacilus casei FM5 6X10 ⁷ cfu / g, Paenibacillus sp. P33 FM8 4.2X10 ⁷ cfu / g, Kluyveromyces fragilis FY1 5.4X10 ⁷ cfu / g and Debaryomyces hansenii FY2 complex microorganism SejungBio-FA100 (microbial accession number: KCTC 11400BP) for organic matter treatment and denitrogenation treatment of sewage and wastewater consisting of 5.4X10 ⁷ cfu / g. 0.5 to 20% by weight of the composite microorganism SejungBio-FA100 (Microorganism Accession No .: KCTC 11400BP) culture medium of claim 1; Rice bran 20-50% by weight; Food waste fermented 20 to 50% by weight; A composite microbial preparation for organic matter treatment and denitrification of sewage and wastewater, characterized by consisting of 10-30% by weight of coarse rice husk charcoal and 5-10% by weight of humus soil. The method of claim 2, The complex microorganism SejungBio-FA100 (Microorganism Accession No .: KCTC 11400BP) culture medium is 1 to 5 parts by weight of the SejungBio-FA100, 4 to 15 parts by weight of sucrose, 1 to 5 parts by weight of peptone, 0.05 to 1 parts by weight of lactose and yeast extract Mixed microorganism preparation for organic matter treatment and denitrification of sewage and waste water, characterized in that obtained by mixing the remaining water in 0.5 to 10 parts by weight to 100 parts by weight and then incubated at 20-35 ℃ for 6-48 hours. delete The denitrification treatment method of wastewater and wastewater, wherein the complex microbial agent of claim 2 or 3 is added to an aerobic tank or aeration tank.

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