KR20000028522A - Biological treatment method of wastewater including surface active agent - Google Patents

Abstract

PURPOSE: A treatment method of waste water including a surface active agent is provided, which is characterized in that microorganism, Erwinia sp. CJ-S01, which grows well in aerobic condition and can disintegrate all sorts of surface active agents is used for biological treatment process. CONSTITUTION: To select a specific microorganism group which can disintegrate surface active agent included in synthetic detergent, the most appropriate microorganism group is necessary. As representative surface active agents, linear alkylbenzene sulfonate: LAS, sodium lauryl sulfate: SLS, sodium lauryl sthylene sulfate: SLES, and alpha-olefin sulfonate: AOS are used for the method. Water and soil from waste water from plants for disposing industrial waste water are collected, and the microorganism samples are collected from the water and soil. Collected microorganisms are cultivated in specific culture ground which the surface active agent is exclusive source of carbon. Then, specific microorganism group which grows well in the condition is selected as Erwinia sp. CJ-S01.

Description

Biological method and microorganism for treating waste water containing surfactants

The present invention relates to biological wastewater treatment. More specifically, the present invention relates to a microorganism having good growth under aerobic conditions of biological wastewater treatment, having an excellent ability to decompose various surfactants, a separation method thereof, and a method for treating surfactant-containing wastewater using the same.

Synthetic detergents discharged after being used at home now not only pollute the water, hinder the self-cleaning action, act as a source of BOD (biological oxygen demand) in the stream, but also generate a lot of bubbles, degrading the value of the river water. If it is late, the treatment cost of the sewage treatment plant is increased. It also affects the growth and growth of aquatic organisms and destroys ecosystems. Substances related to environmental pollution among the components of these synthetic detergents are mainly surfactant components and affect aquatic organisms even at low concentrations. Therefore, research is actively underway to utilize microorganisms that decompose such surfactants. Swisher has reported on the biodegradation of surfactants (Surfactant Science Series, 18, 751: 1987), and Yu-Chih Hsu is a bacterium that produces enzymes that break down sodium lauryl sulfate, a type of surfactant. (Nature, 200, 1091: 1963), a lot of research has recently been conducted to screen the microorganisms that decompose the surfactant and to apply to industrial wastewater treatment. However, the selection of microorganisms showing excellent degradability for various kinds of surfactants has been difficult.

In selecting microorganisms excellent in the ability to decompose the surfactant used as the main component of the synthetic detergent, it can be said that it is important to determine the source and target of the separation. In the present invention, four types of typical surfactants, linear alkylbenzene sulfonate (LAS) and sodium lauryl sulfate (SLS), which are mainly used to isolate microorganisms that decompose various kinds of surfactants, are used. Sodium lauryl ethylene sulfate (SLES) and alpha-olefin sulfonate (AOS) were used. In order to obtain microorganisms showing excellent performance, it is necessary to select microorganisms capable of degrading for each.

Therefore, the inventors of the present invention, while the microorganisms are separated from the microorganisms in industrial wastewater treatment plants and municipal sewage containing a large amount of various surfactants and tested for degrading the surfactants, the microorganisms identified as Ewinia CJ-S01 have excellent surfactant degrading ability. It turned out to have completed the present invention.

1 is a graph comparing growth in a minimal medium containing various surfactants against microorganisms purely separated according to the present invention.

Figure 2 is a graph showing the growth rate in a variety of surfactant-containing minimal medium of the novel microorganisms Erwinia sp. CJ-S01 according to the present invention.

Figure 3 is a graph showing the removal tendency of the surfactant in a minimal medium containing a variety of surfactant using the novel microorganisms of the genus Wikia CJ-S01 according to the present invention.

Figure 4 is a graph comparing the treatment rate for decomposing the surfactant in the synthetic wastewater containing various surfactant using the novel microorganisms of the genus Awinea CJ-S01 according to the present invention.

Figure 5 is a graph showing the relationship between the surfactant concentration and the removal efficiency in the surfactant-containing wastewater using the novel microorganisms of the genus Wikia CJ-S01 according to the present invention.

FIG. 6 is an electron micrograph (× 12000) of a novel microorganism of the genus Cacteria CJ-S01.

The present invention provides a novel microorganisms of the genus Wikia CJ-S01 having excellent degradability of various surfactants. This microorganism is a strain isolated and selected directly from industrial wastewater treatment plant and municipal sewage, and is deposited with the KFCC-11501 on October 24, 1998 to the Korean spawn association.

The present inventors separated the microorganisms purely by using the water and soil around the stream from the industrial wastewater treatment plant in the Seoul, Gyeonggi-do, Gangwon-do, Chungcheong-do region, urban sewage or streams flowing out of the wastewater. Adapted by adapting the isolated microorganisms on a minimum medium containing each surfactant as the only carbon source, and then screening a single group of microorganisms showing excellent growth in agar-containing media, each containing microorganisms The microorganisms showing good growth when cultured on one liquid medium were finally selected and identified as C.-S01 in Erwinia after various tests. In addition, the selected microorganisms were found to have good degradability of the surfactant.

The microorganisms of the genus Urchinia CJ-S01 according to the present invention forms round and convex colonies on the complex medium, the properties of which are described in Tables 1, 2 and 3 below.

Morphological and Cultural Characteristics of C.-S01 Item Characteristics Gram dyeing - shape Bacillus motility +

Physiological and Biochemical Properties of Urbania CJ-S01 Item Characteristics Hemolytic - Oxidase - Catalase + Oxygen composition Conditional anaerobic Indole formation - Reduction of nitric acid to nitrite + Urease formation - Methyl red test - Starch hydrolysis - Sight rate test + O + F test F

Utilization of Organics in CJ-S01 Item Characteristics Glucose + Fructose + Xylose + Glycerol + Sorbitol - Mannitol + Raffinose - Ramrose + m-inositol -

In addition, the present invention provides a method for biologically treating a surfactant-containing wastewater using the above E. C.-S01 strain. According to the method of the present invention, wastewater containing up to 1000 ppm of surfactant can be treated.

The present invention also uses sodium alkylbenzenesulfonate, sodium lauryl sulfate, sodium lauryl ethylene sulfate and alpha-olefinsulfonate as the only carbon sources based on ammonium sulfate, sodium phosphate, monopotassium phosphate, magnesium sulfate and iron chloride. Provided is a method for separating Swinia CJ-S01 strain using a minimal medium containing a surfactant.

The invention is illustrated in detail in the following examples. However, this embodiment is intended to describe the present invention in more detail, and the purpose thereof is not to be understood as limiting the scope of the present invention.

EXAMPLE

Isolation of Strains

In order to separate microorganisms having excellent degradability and growth ability of various surfactants, samples of industrial wastewater treatment plants and urban sewage in Seoul, Gyeonggi-do, Gangwon-do and Chungcheong-do areas were acclimated and allowed to acclimate for a certain period of time. Luria-Bertani medium (LB medium; tryptone 0.1 g, yeast extract 0.05 g, sodium chloride 0.05 g, glucose 0.01 g, agar 0.2 g) was dissolved in 1 liter of biochemical wastewater (filtered with a sterile filter 0.2 μm). Smeared. The petri dishes were incubated for 1 to 3 days in an incubator at 25 to 30 ° C. to isolate 25 single colonies showing excellent growth on plate media.

Minimal medium containing 40 ppm of LAS, SLS, SLES and AOS, four types of representative surfactants, which are widely used in synthetic detergents, to measure the degradability of surfactants against isolated microorganisms (ammonium sulfate 2.5g, 13.8 g of sodium phosphate, 3 g of potassium phosphate, 12.5 mg of magnesium sulfate, and 17.5 mg of iron chloride were inoculated into one liter of liquid medium) and then inoculated with platinum for one period. Then, when growth was made, the microorganisms showing a single colony were first selected by spreading a minimal medium containing four kinds of surfactants with the above-mentioned composition including agar.

On the other hand, inoculate one platinum nip into a culture test tube containing 5 ml of LB medium of the above composition except for agar, and incubate at 25 to 30 ° C. for 24 hours in a shaking rotation incubator and centrifuge the culture medium. Vision, VS15000CFN) cells were recovered by centrifugation for 5 minutes. Some of the recovered microorganisms were used in subsequent experiments and some were lyophilized and stored.

Screening and processing of strains with surfactant degrading ability

Each microorganism of each of the above-selected 10 kinds of microorganisms was suspended in sterile distilled water, and each microorganism was added to the minimum medium containing the surfactant in which four kinds of surfactants (LAS, SLS, SLES, and AOS) were adjusted to 40 ppm, respectively. Inoculated by 1% (v / v) and incubated at 25 ~ 30 ℃ in shaker incubator. Thereafter, growth was compared to select strains having excellent ability to use each surfactant as the only carbon source. In other words, as a result of observing the growth of the minimum medium containing a surfactant containing 40ppm each of the surfactant, two kinds of microorganisms excellent in growth for each of the four types of surfactants (LAS, SLS, SLES and AOS) are selected secondarily. It was. The results are shown in FIG.

The composition of the minimum medium containing the surfactant used by the present inventors was based on 2.5 g / l ammonium sulfate, 13.8 g / l sodium phosphate, 3 g / l potassium monophosphate, 12.5 mg / l magnesium sulfate, and 17.5 mg / l iron chloride. Each surfactant was used at a concentration of 40 ppm as the only carbon source.

Surfactant degradability was compared using a minimum medium containing a surfactant prepared to contain 40 ppm of the surfactant for each of the secondary screened microorganisms. The secondary cells were suspended in sterile distilled water, and then inoculated by 1% (v / v) for the minimum medium containing a surfactant containing 40 ppm of each surfactant. After inoculation, the growth of the cells was measured while culturing at 25 to 30 ° C. in a shaker incubator. The results are shown in Table 4 and Figure 2, the microorganisms identified in the genus C. J-S01 showed a good growth rate for the minimum medium containing the four types of surfactants, which shows the four types of surfactants All of them were selected as the carbon source, and the growth rate was excellent and the decomposition ability of various surfactants was excellent.

Comparison of Growth Potency of Each Surfactant of S. CJ-S01 Surfactant LAS SLS SLES AOX CJ-S01 +++ +++ +++ +++ CJ-S02 + + +++ ++ CJ-S05 ++ +++ + + CJ-S09 + + + +++

Confirmation of Removal Capability of Surfactants by Surfactant Degrading Strains

In order to determine the degree of removal of the surfactant by using the Erwinia CJ-S01 screened according to the present invention, while culturing at 25 ~ 30 ℃ in a shaker incubator, a certain amount is aseptically collected and diluted in an appropriate amount in a centrifuge Treated. The residual concentration of the surfactant was measured for the treated supernatant. For the analysis of surfactant, shake the solution well by adding a certain amount of methylene blue solution, mix the chloroform solution, centrifuge, leave for 10 minutes at room temperature, remove the aqueous layer, and measure the absorbance at 655nm using absorbance spectrophotometer. Was converted. The results are shown in FIG. 3, and the decomposition of the surfactants was excellent by E. C.-S01 in a surfactant-containing minimal medium containing 20, 50, and 100 ppm of each surfactant mixture.

Determination of Treatability of Surfactant-Containing Wastewater by Surfactant Degrading Strains

In order to confirm the treatability in the mixed wastewater containing surfactant, using a synthetic wastewater containing 5.3% of surfactant, incubate at 25 ~ 30 ℃ in shake rotation incubator, take a certain amount, and take the supernatant to chromium oxidation method. Chemical oxygen demand (CODcr) was measured.

The composition of the synthetic wastewater containing the surfactant used by the present inventors was 110 mg / l of starch, 55 mg / l of lactose, 60 mg / l of glucose, 90 mg / l of potassium monophosphate, 70 mg / l of carboxymethyl cellulose, 315 mg / l of ammonium sulfate, glycerol 50mg / l, juicy 110mg / l, gelatin 55mg / l, peptone 50mg / l, tryptone 50mg / l, sugar 10mg / l, cottonseed oil 40μg / l, butter 55mg / l, beverage waste 1ml / l It contained 5.3% of the active agent. The surfactant-containing mixed wastewater thus prepared was compared with treatability according to the dosage of microorganisms of the present invention CJ-S01 according to the present invention at different loads. The results are shown in Figure 4, when the concentration of the mixed wastewater containing 5.3% of the surfactant was increased to 1000ppm, compared to the control group added activated sludge only CJ-S01 microorganism of the control group added The treatment rate was high in the case (Comparative Group 1: 100 ppm of microorganisms; Comparative Group 2: 200 ppm of microorganisms). Therefore, it was confirmed that the surfactant-decomposing microorganism according to the present invention has excellent ability even in wastewater containing a surfactant.

Confirmation of Removal Efficiency by Surfactant Concentration in Surfactant-containing Wastewater by Surfactant Degrading Strains

In order to confirm the treatability according to the surfactant concentration for the surfactant-containing wastewater, the concentration was adjusted to 1000 ppm by the composition of the synthetic wastewater described above, and the surfactant was added at various concentrations, and the surfactant-containing surfactant at each concentration. The efficacy of the microorganisms of the present invention on activator containing wastewater was confirmed. The results are shown in Fig. 5, when the surfactant was added to 1000 ppm of the surfactant, up to 1000 ppm of the surfactant was added to the control group containing the activated sludge CJ-S01 microorganism compared to the control group (comparative group). The removal efficiency remained high without being affected by the concentration of the active agent. Therefore, it was found that the E. coli SJ-S01 microorganism according to the present invention has excellent degradability to the surfactant.

New Microorganisms of the S.J.S-S01, a microorganism with excellent decomposition of selected and separated surfactants from industrial wastewater treatment plants and municipal sewage, can be used for actual field wastewater treatment, and various biological environmental purification technologies including environmental treatment agents using microorganisms. It is expected to be applicable to.

Claims (5)

S.J.-S01 microorganisms of good growth and aerobic degradability under aerobic conditions (Korean spawn association date: October 24, 1998; accession number: KFCC-11051). The microorganism of claim 1, wherein the surfactant is sodium alkylbenzenesulfonate, sodium lauryl sulfate, sodium lauryl ethylene sulfate or alpha-olefinsulfonate. The biological treatment method of the wastewater containing surfactant characterized by using the Swinia CJ-S01 strain of claim 1. The treatment method according to claim 3, wherein the wastewater contains up to 1000 ppm of surfactant. Minimum medium containing surfactant using sodium alkylbenzenesulfonate, sodium lauryl sulfate, sodium lauryl ethylene sulfate and alpha-olefinsulfonate as the only carbon sources based on ammonium sulfate, sodium phosphate, potassium monophosphate, magnesium sulfate and iron chloride Separation method of claim 1 CJ-S01 strain of claim 1, characterized in that using.