KR100459852B1 - The microbial seeds for the sewage/wastewater treatment and the method for it's development - Google Patents

Abstract

The present invention relates to a microbial agent suitable for wastewater treatment and a method for manufacturing the same, and more particularly, finely ground rice bran is used as a carrier for growth substrate and preservation, and can be grown at medium and high temperatures (30 to 45 ° C). Fast growth rate, excellent bactericidal activity of organic matter, in particular Bacillus cereus DSC 1021 [KCTC 10161BP] in combination with the addition of inorganic salt mixture in addition to wastewater treatment rate It further relates to a microbial agent and a method for producing the same, which can reduce the size of the treatment reaction tank and expand the range of the treated wastewater.

Description

The microbial seeds for the sewage / wastewater treatment and the method for it's development}

The present invention relates to a microbial agent suitable for wastewater treatment and a method for manufacturing the same, and more particularly, finely ground rice bran is used as a carrier for growth substrate and preservation, and can be grown at medium and high temperatures (30 to 45 ° C). Fast growth rate, excellent bactericidal activity of organic matter, in particular Bacillus cereus DSC 1021 [KCTC 10161BP] in combination with the addition of inorganic salt mixture in addition to wastewater treatment rate It further relates to a microbial agent and a method for producing the same, which can reduce the size of the treatment reaction tank and expand the range of the treated wastewater.

The environmental pollution source that occurs as a result of industrial development and various daily activities (centralization of the city of the population) has already surpassed the midnight action of the natural world, accelerating and widening the environmental degradation, and has a direct and indirect impact on the health and health of humankind. .

Methods for improving and restoring a contaminated natural environment include physical, chemical, and biological methods, and these methods may be used in combination depending on the contaminated form. Among these methods, biological methods artificially enhance self-cleaning by using the functions of microorganisms (material transformation, accumulation, decomposition, etc.) that play a role in natural self-cleaning. It is a feature of technology.

The activated sludge method, developed as a method for treating industrial wastewater, which has emerged as a social problem since the Industrial Revolution, is a representative example of biological methods. to be. The main members of activated sludge are microorganisms adsorbed by bacteria and algae, and these microbial groups, and the action of these microorganisms causes decomposition and removal of organic matter in the wastewater, thereby contributing to wastewater treatment. In the actual treatment of industrial wastewater using activated sludge method, MLSS (Mixed Liquor Suspended Solids), BOD loading, amount of return sludge, oxygen demand of aeration tank, and various physical and chemical factors are important operation factors. The most important factor to determine the type of microorganisms present in activated sludge and their function.

Therefore, these manipulators can be varied in various ways depending on the ability of the microorganism to be used in the activated sludge process and the wastewater phase to be applied. In other words, by developing excellent activated sludge spawn, it is expected to simplify the treatment process and thereby install, reduce operating costs and improve treatment efficiency.

However, it is difficult to install such a wastewater purification treatment facility in a relatively small farmhouse unit or a livestock farm, and the water quality of a river is worsened by discharge | released without the purification process.

On the other hand, in the wastewater treatment method, the sprinkling filter method is a biochemical treatment method for removing organic matter from wastewater by the action of microorganisms attached to the surface while the sewage water which is sprayed on the filter passes through the filter media. Apparatuses used as media have been devised. In addition, a variety of active sludge spawning agents are developed and marketed by foreign companies (polybac, IRL, etc.), and they are commercialized and marketed mainly in small and medium enterprises in Korea.

However, these prior arts have problems such as exchange of media, clogging caused by fine particles, remarkable degradation of wastewater treatment ability, degradation of microorganisms as well as growth of organic matters, and changes in wastewater phase and basic operating factors. Suitable microbial agents are urgently needed.

In view of the above, the present inventors can search for microorganisms capable of growing even at medium and high temperatures, having a high growth rate, and excellent organic matter decomposition activity by various enzyme activities, and thus, Bacillus cereus DSC1021. [KCTC 10161BP] was discovered, and the present invention was completed by developing a microbial preparation produced by culturing together with microorganisms having excellent organic degradation activity using finely ground rice bran as a carrier for growth substrate and preservation.

Therefore, the present invention is capable of growing at medium and high temperatures (30 to 45 ° C.), including a microorganism capable of growing at a high rate of growth and having excellent organic matter degrading activity by various enzyme activities. The addition of a salt mixture further improves the rate of wastewater treatment and provides a microbial agent that can extend the range of the target wastewater that can be treated.

Another object of the present invention is to provide Bacillus cereus dies. C. 1021 [KCTC 10161BP] having an organic component decomposition activity.

1 is a photograph showing the activity of various enzymes produced by Bacillus cereus DSC 1021 [KCTC 10161BP].

The present invention is characterized by the new strain Bacillus cereus DSC 1021 [KCTC 10161BP] having an organic component decomposition activity.

In addition, using the crushed rice bran as a growth substrate and storage carrier of the microbial strain microbial agent produced by combining one or two or more strains of microorganisms excellent in organic component decomposition activity even at medium and high temperatures (30 ~ 45 ℃) and its It includes a manufacturing method.

The present invention will be described in more detail as follows.

The present invention uses finely crushed rice bran as a carrier for growth substrate and preservation, and can grow at medium and high temperatures (30 to 45 ° C.), have a fast growth rate, and have excellent organic decomposition activity. Cereus DS 1021) [KCTC 10161BP] may be included in an appropriate combination, and inorganic salt mixtures may be added to further improve the wastewater treatment rate, to reduce the size of the treatment reactor and to expand the range of the target wastewater to be treated. It relates to a microbial agent for wastewater treatment and a method of manufacturing the same.

First, microbial strains were purely separated from soil and sludge in the vicinity of a wastewater treatment plant, and finally, strains having a rapid growth rate at 30 to 45 ° C. were identified and identified. As a result, Bacillus cereus DSC 1021 was identified. It was named.

On the other hand, in medium and high temperature (30 ~ 45 ℃) one or more strains of microorganisms excellent in organic component decomposition activity was inoculated in the primary liquid medium and incubated for 24 hours at 25 ~ 30 ℃ and then crushed rice bran Using the growth substrate and preservation carrier as a secondary solid culture for 48 hours at 30 ~ 37 ℃ to create a spawn. At this time, the middle and high temperatures (30 ~ 45 ℃) in strains of the superior microbial organic components decomposing activity is Bacillus brevis (Bacillus brevis), Bacillus core tangerine lance (Bacillus coagulans), bacilli such as Bacillus MEGATHERIUM (Bacillus megaterium) Genus is preferred, with the new strain Bacillus cereus dies 1021 [KCTC 10161BP] being most preferred. These strains can be used 1 type or in combination of 2 or more types.

In addition, inorganic salts ((NH ₄ ) H ₂ PO ₄ , (NH ₄ ) ₂ HPO ₄ , CaCl ₂ ㆍ 2H ₂ O mixed in a ratio of 2: 2: 1) were added to the produced spawn at a weight ratio of 1: 1. After the addition, the microorganisms are finally prepared by air drying at room temperature for 4 hours.

As described above, microorganisms having a high growth rate and excellent organic decomposition activity are used as a seed, and thus, problems caused in the aeration tank (various operation factors, complexity of the processing process, excessive operating cost, and treatment efficiency reduction) can be overcome. In addition, by adding inorganic salts, it supplies nitrogen and phosphorus, which are essential ingredients for the growth of microorganisms, and acts as a cofactor for enzymes. It also has an excellent effect on the treatment of wastewater that is not treated, and high cohesive effect can be expected.

Hereinafter, the present invention will be described in detail based on the following examples, but the present invention is not limited thereto.

Example 1: Isolation of Microbial Strains

30 strains of microorganisms were purely separated using 10 samples of soil and sludge near the wastewater treatment plant. 0.1 g of the sample was suspended in 9.9 ml of physiological saline (NaCl 0.85%), diluted 105 times, and plated on nutrient agar medium (beef extract 0.3%, peptone 0.5%, agar 1.7%) and incubated at 30 ° C. for 48 hours. The microbial colonies formed after the cultivation were purely separated in a nutrient agar medium by a stroke method.

By inoculating purely isolated strains, 30 strains showing rapid growth of viable cells and viable cells were selected by investigating the growth of bacteria and the presence of transparent ring formation (organic degradation activity).

In order to select strains with high growth rate at 30-45 ℃ among 30 strains, the strains were inoculated in nutrient medium, shaken at 30 ℃ for 24 hours, and absorbance at 660 nm was measured for the best growth. Strains were final selected.

Example 2: Identification of Microorganisms

Types of strains obtained from microbial microorganisms with excellent enzymatic activity and proliferation degree for sewage treatment can be referred to the Manual of Determinative Bacteriology and Manual for the Identification of Medical Bacteria. Strain identification was carried out after examining the physiological, physiological and biochemical properties, and cell fatty acid analysis was performed by extracting fatty acids from 40-50 mg of wet cells in logarithmic growth phase and using a gas by a MIS (Microbial Identification System) [Hewlett-Packard 5890A]. Analysis was performed using liquid chromatography.

As a result, as shown in Table 1, the isolated strain forms a colony of a circle in a solid medium, and the colony color was yellow. The shape was bacterium, the size was 0.4-0.7 μm in diameter, and the length was 1,2-2.5 μm. This strain is a Gram-positive bacterium whose growth pH range is 5.0-10.0, growth temperature range is 20-45 ° C., and is an anaerobic bacterium capable of growing under aerobic and anaerobic conditions. Catalase produced and no oxidase, producing acid using glucose. Gelatin liquefaction, casein and urease and nitrate reducibility were positive, and starch hydrolysis and indole production were negative. Carbohydrates did not produce acids from aerobic conditions from mannitol, xylose and arabinose.

From the above results, this strain was estimated to be Bacillus cereus, but the analysis of the cell fatty acid composition for more accurate identification of the strains revealed that the main components were C 15: 0 and 13: 0. Based on the present invention, the strain shows the most similar cell fatty acid to Bacillus cereus, so the microorganism isolated in the present invention is named Bacillus cereus DSC 1021 and the Korea Biotechnology Research Institute Gene Bank (KCTC), a strain depositing institution. Was deposited on 15 January 2002 and was assigned accession number 10161BP.

In addition, the culture of Bacillus cereus DS 1021 was used for the liquid and solid medium of LB, NB, growth was strong at medium temperature (30 ℃) and high temperature (45 ℃), and colony formation ability was excellent.

Morphological, Culture and Physiological Characteristics of Mycobacteria factor Characteristics Morphological Characteristics Gram Dyeing Size (㎛) Motile Spore Formation Bacillus 0.4-0.7 × 1.2-2.5 ++ Culture Characteristics Aerobic Culture Anaerobic Culture Growth Temperature 30 ℃ 45 ℃ 65 ℃ Growth pH 5.7 Growth 7% Salinity ++++-++ Physiological Properties Catalase Production Oxidase Production Acid Production from Glucose Carbohydrate (O / F ¹ /-) Acid Production from Carbohydrates Test lysozyme fatty acid main ingredient + -d ³ O --- ++++-++ r ² 15: 0 (32%) 13: 0 (11%) 1: oxidation-fermentation test, 2: resistance, 3: incorrect for every species

As shown in Table 2, the Bacillus cereus D. C. 1021 was shown to have high enzyme activity such as amylase, protease, lipase, and the like (FIG. 1).

Enzyme Activity of New Bacteria division Amylase Protease Cellulase Lipase Activity level ++++ +++++ - ++ Note) +++++: very good, ++++: very high, ++: high

Example 3: Microbial preparation for wastewater treatment

The new strain Bacillus cereus D. C. 1021 [KCTC 10161BP] according to the present invention has been shown to have excellent growth at medium and high temperatures, high activity of various enzymes, and excellent organic decomposition activity as described above. Rice bran) adsorption experiment to prepare a microbial agent.

The microbial preparation was prepared by inoculating the seed culture of the new strain Bacillus cereus D.C. 1021 [KCTC 10161BP] into a primary liquid medium, incubating at 30 ° C. for 24 hours, and using the crushed rice bran as a growth substrate and carrier. Seeds were made via secondary solid culture at 48 ° C. for 48 hours. In addition, inorganic salts ((NH ₄ ) H ₂ PO ₄ , (NH ₄ ) ₂ HPO ₄ , CaCl ₂ ㆍ 2H ₂ O mixed in a ratio of 2: 2: 1) were added to the produced spawn at a weight ratio of 1: 1. After mixing, the mixture was air dried at room temperature for 4 hours to prepare a microbial preparation. After culturing for 48 hours at 37 ℃ using the ground skim as a medium, the total number of bacteria was about 3.5 × 10 ⁹ CFU / g, the strain of the present invention was confirmed.

Example 4: Wastewater Treatment Effect

In order to test the effect of the microbial agent for wastewater treatment prepared in Example 3, a sample was taken from a wastewater treatment reactor in Sokcho, Gangwon-do. As shown in Table 3, after treating the wastewater with microbial agents, the influent and effluent were analyzed. As a result, the chemical oxygen demand (COD) was about 90% and the biological oxygen demand (BOD) was 84%.

In particular, the decomposition of nitrogen and phosphorus, which had been neglected in the existing treatment methods, was also confirmed. In addition, the addition of an inorganic salt mixture was shown to further improve the wastewater treatment speed, to reduce the size of the treatment reactor, and to expand the range of the target wastewater that can be treated.

Wastewater Treatment Effect Using Microorganisms Influent (mg / l) Effluent (mg / l) % Removal pH 7.36 6.4 - COD _Mn 82.0 8.5 89.6 TOC 54.0 13.0 75.9 BOD 63.0 10.0 84.1 T-N 60.0 47 21.7 T-P 1.17 0.92 21.4

As described above, the present invention is capable of growing even at medium and high temperatures (30 to 45 ° C.), and has a rapid growth rate, and excellent microorganisms, particularly the bacterium Bacillus cereus dies C. Bacillus cereus DSC 1021) [KCTC 10161BP] containing an appropriate combination, and in addition to the inorganic salt mixture to further improve the rate of wastewater treatment, and to a microbial agent that can expand the range of the treated wastewater and a method for producing the same will be.

Therefore, the microbial agent according to the present invention is very useful for improving and restoring a contaminated natural environment.

Claims (7)

Bacillus cereus DSC 1021 [KCTC 10161BP] with enzymatic activity against amylases, proteases and lipases. Bacillus cereus D. C. 1021 [KCTC 10161BP] strain or a microbial agent for wastewater treatment, characterized in that it contains a culture solution thereof. delete After inoculating Bacillus cereus D.C. 1021 [KCTC 10161BP] strain into a liquid medium and incubating the primary liquid at 25 to 30 ° C., the crushed rice bran is produced as a secondary solid culture at 30 to 37 ° C. using a growth substrate and a storage carrier. Method for producing a microbial agent for wastewater treatment, characterized in that the dried strain or culture medium thereof is dried delete The method of claim 4, wherein the inorganic salt is mixed with the strain produced by the solid culture or the culture solution thereof. 7. The wastewater treatment according to claim 6, wherein the inorganic salts contain (NH ₄ ) H ₂ PO ₄ : (NH ₄ ) ₂ HPO ₄ : CaCl ₂ ㆍ 2H ₂ in a weight ratio of 2: 2: 1. Method for producing a microbial agent for use.