JP2012080872A - Microorganism and method for treating surfactant-containing waste water - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new microorganism, etc., which treats surfactant-containing waste water in high efficiency.SOLUTION: The microorganism is Cupriavidus taiwanensis KR-20R strain (FERM P-22013), Cupriavidus necator KR-26R strain (FERM P-22014), Pseudomonas nitroreducens KR-33R strain (FERM P-22015), Ralstonia pickettii KR-35R strain (FERM P-22016), Ralstonia eutropha KR-41R strain (FERM P-22017) or Ralstonia eutropha KR-43 strain (FERM P-22018). The microorganisms have excellent ability to degrade a surfactant, especially an anionic surfactant. The method for treating surfactant-containing waste water using the microorganisms is provided.

Description

  The present invention relates to a microorganism and a method for treating a surfactant-containing wastewater, and more particularly to a microorganism having a high surfactant resolution and a method for treating a surfactant-containing wastewater using the microorganism.

  Surfactants are used in large quantities in various industries such as the food industry, machine industry, and electronics industry, and are discharged not only as domestic wastewater but also as industrial wastewater. Surfactants are often used for cleaning purposes in particular, and domestic wastewater containing surfactants becomes complex wastewater containing various other pollutants. For example, sodium lauryl sulfate (SDS) is a surfactant widely used as a synthetic detergent, and a reliable treatment of waste water containing SDS is required.

  In general, biological treatment such as an activated sludge method is often applied in wastewater treatment. However, since surfactants are hardly biodegradable and there are surfactants that dissolve microbial membranes, it is difficult to treat with the activated sludge method, which is the mainstay of industrial wastewater treatment. For example, if composite wastewater containing surfactants and general natural organic substances is subjected to biological treatment, easily degradable substances in the wastewater are preferentially decomposed, and surfactants that are hardly decomposable remain in the treated water. The problem that it is easy to occur. As a result, when treated water with residual surfactant is released into the environment such as a river, the decomposition of the surfactant requires a period of several days to several tens of days, resulting in a large environmental burden. ing.

  There are not many microorganisms in nature that can rapidly degrade synthetic surfactants that are not natural products. Therefore, it is difficult to sufficiently treat the waste water containing the surfactant (surfactant-containing waste water) by the conventional biological treatment depending on the indigenous microorganisms. Therefore, it is considered effective to use a specific microorganism having a high ability to decompose a surfactant as a foreign species for biological treatment.

  Under these circumstances, various microorganisms having the ability to decompose surfactants (surfactant resolution) have recently been found (for example, Patent Documents 1 to 3) and applied to the treatment of surfactant-containing wastewater. Has been tried. However, it is difficult to say that it is sufficiently applied to the treatment of surfactant-containing wastewater.

  Further, since the surfactant foams at a slight concentration, a large amount of antifoaming agent is required for biological treatment. Furthermore, when tertiary treatment of treated water is required in a COD total amount regulation area such as a closed water area, physicochemical treatment such as activated carbon adsorption or ozone treatment is performed. However, this process is disadvantageous in terms of both initial cost and running cost.

JP 2004-173697 A JP 2000-354891 A JP 2000-270852 A

  As described above, various problems still remain regarding biological treatment of surfactant-containing wastewater, and a solution is desired. Accordingly, an object of the present invention is to provide a novel microorganism excellent in surfactant resolution and to provide a wastewater treatment technique using the microorganism.

  The present inventors screened microorganisms using soil collected from various environments as a separation source in order to separate novel microorganisms having higher surfactant resolution. As a result, the inventors succeeded in separating a plurality of kinds of novel microorganisms having excellent resolution against surfactants, particularly anionic surfactants such as SDS. And the processing method of the wastewater containing surfactant using the said microorganisms was developed, and this invention was completed. The present invention for solving the above-described problems is as follows.

  The invention described in claim 1 includes Cupriavidus taiwanensis KR-20R strain (FERM P-222013), Cupriavidus necator KR-26R strain (FERM P-22014), Pseudomonas nitroreducens KR-33R strain (FERM P-20215), Ralstonia pickettii It is a microorganism that is a KR-35R strain (FERM P-22016), Ralstonia eutropha KR-41R strain (FERM P-20217), or Ralstonia eutropha KR-43R strain (FERM P-22018).

  The microorganism of the present invention has a high resolution for surfactants, particularly anionic surfactants such as SDS. According to the microorganism of the present invention, surfactant-containing wastewater can be treated (purified) with high efficiency. For example, by using the microorganism of the present invention, it is possible to reduce or omit the physicochemical treatment that is performed as a treatment of the surfactant-containing wastewater.

  The invention described in claim 2 is a treatment of surfactant-containing wastewater that contacts at least one microorganism according to claim 1 with wastewater containing a surfactant to decompose the surfactant in the wastewater. Is the method.

  The present invention relates to a method for treating surfactant-containing wastewater, and includes a step of bringing at least one microorganism of the present invention described above into contact with a surfactant-containing wastewater. According to the present invention, surfactant-containing wastewater can be treated with high efficiency.

  In the method for treating surfactant-containing wastewater according to claim 2, it is preferable that the surfactant is an anionic surfactant (claim 3).

  In the method for treating surfactant-containing wastewater according to claim 3, it is preferable that the anionic surfactant is selected from the group consisting of a sulfate ester salt, a sulfonate salt, and a carboxylate salt. Item 4).

  According to the microorganism of the present invention, surfactant-containing wastewater can be treated (purified) with high efficiency. For example, it becomes possible to reduce or omit the physicochemical treatment that is performed as a treatment of the surfactant-containing wastewater.

  The same applies to the method for treating surfactant-containing wastewater of the present invention, and the surfactant-containing wastewater can be treated (purified) with high efficiency.

It is a graph which shows the SDS degradation curve by a novel microorganism (Example). It is a graph which shows another SDS degradation curve by a novel microorganism (Example). It is a graph which shows the SDS decomposition curve by a standard activated sludge (comparative example).

  The microorganisms of the present invention are Cupriavidus taiwanensis KR-20R strain (FERM P-222013), Cupriavidus necator KR-26R strain (FERM P-222014), Pseudomonas nitroreducens KR-33R strain (FERM P-20215), Ralstonia pickettii KR-35R. Strain (FERM P-22016), Ralstonia eutropha KR-41R strain (FERM P-222017), or Ralstonia eutropha KR-43R strain (FERM P-22018). These microorganisms are deposited at the Patent Organism Depositary (IPOD), National Institute of Advanced Industrial Science and Technology. Details of the deposit are shown below.

[KR-20R strain]
Display: Cupriavidus taiwanensis KR-20R strain Accession number: FERM P-22013
Receipt date: September 15, 2010

[KR-26R strain]
Display: Cupriavidus necator KR-26R strain Accession number: FERM P-222014
Receipt date: September 15, 2010

[KR-33R strain]
Display: Pseudomonas nitroreducens KR-33R strain Accession number: FERM P-22015
Receipt date: September 15, 2010

[KR-35R strain]
Display: Ralstonia pickettii KR-35R strain Accession number: FERM P-22016
Receipt date: September 15, 2010

[KR-41R strain]
Display: Ralstonia eutropha KR-41R strain Accession number: FERM P-22017
Receipt date: September 15, 2010

[KR-43R strain]
Display: Ralstonia eutropha KR-43R strain Accession number: FERM P-22018
Receipt date: September 15, 2010

  The mycological properties of each strain are shown in Tables 1-3. Hereinafter, “+” indicates positive and “−” indicates negative.

  Furthermore, the result of the additional test of each strain is shown below.

[KR-20R strain]
・ Growth under anaerobic conditions:-
・ Growth at 42 ℃: +
・ Lipase activity (Tween 80): +

[KR-26R strain]
・ Growth under anaerobic conditions:-
-Growth under 2% NaCl conditions: +
-Growth under 3% NaCl conditions:-

[KR-33R strain]
・ Growth under anaerobic conditions:-
・ Starch hydrolysis:-
・ Production of fluorescent dyes on King's B agar:-

[KR-35R strain]
・ Growth under anaerobic conditions:-
・ Starch hydrolysis:-
・ Lipase activity (Tween 80): +

[KR-41R strain]
・ Growth under anaerobic conditions:-
・ Lactate utilization: +
-Glutamate assimilation: +

[KR-43R strain]
・ Growth under anaerobic conditions:-
・ Lactate utilization: +
-Glutamate assimilation: +

  As described in detail in Examples below, these microorganisms have been identified by the homology of the 16S rDNA base sequence (Table 5).

  As a method for culturing the microorganism of the present invention, a general method for culturing aerobic microorganisms can be employed as it is. For example, using a liquid medium containing an appropriate carbon source or the like, it can be cultured with aeration and agitation. As culture | cultivation temperature, the range of 5-40 degreeC, for example, Preferably 15-40 degreeC, More preferably, 20-35 degreeC can be selected. When a surfactant is added as a carbon source to the medium, the concentration can be selected, for example, from 750 mg / L or less, preferably 500 mg / L or less, more preferably 100 mg / L or less.

  The microorganism of the present invention has high surfactant degradability. As a method for evaluating the surfactant resolution, for example, it can be evaluated by the rate of decrease of the surfactant in the culture solution when the microorganism of the present invention is cultured in a medium containing the surfactant. The rate of decrease in surfactant is calculated, for example, by using a medium that contains surfactant alone or the main carbon source, and detecting changes in the amount of surfactant in the medium based on changes in total organic carbon (TOC). can do.

  In the method for treating surfactant-containing wastewater of the present invention, at least one of the above-mentioned six types of microorganisms is brought into contact with the surfactant-containing wastewater. Only one type of microorganism may be used, or two or more types may be used in combination.

  In the method for treating surfactant-containing wastewater of the present invention, the surfactant to be treated is not particularly limited, and is an anionic surfactant, a cationic surfactant, an amphoteric surfactant, a nonionic surfactant. Although all the surfactants can be treated, they are particularly effective for the treatment of anionic surfactants. Examples of the anionic surfactant include sulfate ester salts, sulfonate salts, and carboxylate salts. Examples of the sulfate ester salt include sodium lauryl sulfate (SDS), sodium alkyl sulfate ester (AS), and sodium alkyl ether sulfate ester (AES). Examples of the sulfonate include linear alkylbenzene sulfonate (LAS), alkylbenzene sulfonate (ABS), α-olefin sulfonate (AOS), and sodium alkanesulfonate (SAS). Examples of carboxylates include fatty acid soaps made from coconut oil or palm oil, perfluorooctanoic acid (PFOA), and alphasulfo fatty acid methyl ester salts (MES).

  Examples of the cationic surfactant include amine salt type and quaternary ammonium salt type.

  Examples of amphoteric surfactants include betaine type, alkyl betaine type, and glycine type.

  Examples of nonionic surfactants include polyoxyethylene alkyl phenyl ether, polyoxyethylene alkyl ether, glycerin fatty acid ester, and fatty acid polyethylene glycol.

  The surfactant-containing wastewater treatment method of the present invention can be applied to any method of aerobic treatment, for example, the activated sludge method or the biofilm method. When applied to the activated sludge method, the surfactant-containing wastewater can be treated with high efficiency by adding the microorganism of the present invention to the activated sludge and allowing it to coexist with other microorganisms. When applying to the biofilm method, a known method such as a rotating disk method or a trickling filter method can be applied as it is. The microorganisms of the present invention can be immobilized on a carrier and fluidized bed type treatment can be performed. In addition, the method for treating surfactant-containing wastewater of the present invention can be applied to both batch and continuous treatment methods.

  In the method for treating surfactant-containing wastewater according to the present invention, the amount of microorganisms to be brought into contact with the surfactant-containing wastewater (for example, addition amount or addition cycle) is the type of microorganism to be used, the properties of wastewater to be treated, the wastewater treatment. What is necessary is just to select suitably according to the structure of a facility, etc. The contact time between the surfactant-containing wastewater and the microorganism can also be appropriately selected according to the type and amount of the surfactant contained in the wastewater, the properties of the wastewater to be treated, and the like. The temperature at the time of wastewater treatment may be within the range in which the microorganism of the present invention can grow, for example, the range of 5 to 40 ° C., preferably 15 to 40 ° C., more preferably 20 to 35 ° C. exemplified as the culture temperature. You can choose. Similarly, as the concentration of the surfactant, for example, a range of 750 mg / L or less, preferably 500 mg / L or less, more preferably 100 mg / L or less can be selected.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.

(1) Screening for new microorganisms Soil collected from various environments in nature was used as a source for separating new microorganisms. 0.5 g of soil sample was suspended in 5 mL of sterile physiological saline and allowed to stand for 60 minutes. 100 μL of the supernatant of the suspension was inoculated into 5 mL of a surfactant-containing medium having the composition shown in Table 4, and cultured with shaking at 30 ° C. and 100 rpm. After 5 transplants, multiple samples with degraded SDS were selected. Surfactant-degrading microorganisms were purely isolated from these enriched cultures using an agar medium.

  Among the plurality of isolated microorganisms, 16SrDNA was considered for 6 types (KR-20R strain, KR-26R strain, KR-33R strain, KR-35R strain, KR-41R strain, KR-43R strain) considered to be particularly useful. The V3 region was amplified by PCR and sequence analysis was performed. A homology search with known microorganisms was performed using the BLAST database. The results are shown in Table 5. That is, these six species showed high homology with Cupriavidus taiwanensis, Cupriavidus necator, Pseudomonas nitroreducens, Ralstonia pickettii, Ralstonia eutropha, and Ralstonia eutropha, respectively.

  Six types of purely isolated microorganisms were deposited with the Patent Organism Depositary (IPOD), National Institute of Advanced Industrial Science and Technology.

  A glycerol stock was prepared for each microorganism and used in the following tests.

(2) Evaluation of surfactant decomposition ability [1]
100 mL of the nutrient medium shown in Table 6 was charged into a 300 mL Erlenmeyer flask and sterilized with an autoclave. 1 mL of glycerol stock of each microorganism was inoculated, and culture was started with a rotary shaker (200 rpm, 30 ° C.). After culturing for 36 to 48 hours, cultured cells of each microorganism were collected by centrifugation. The bacterial cells were washed with physiological saline and then made up to 10 mL with physiological saline to obtain a bacterial cell concentrate.

100 mL of the medium containing surfactant shown in Table 6 was charged in a 300 mL Erlenmeyer flask and sterilized by autoclaving. About each microbial cell concentrated liquid, it inoculated to the Erlenmeyer flask which prepared the surfactant containing culture medium so that an inoculum microbial cell density | concentration might be 90 mg / L in dry microbial cell weight conversion. The culture was started with a rotary shaker (200 rpm, 30 ° C.). The culture solution was sampled at 0, 8, 24, and 48 hours from the start of the culture, and OD ₆₆₀ , pH, and TOC were measured. As a control, the same operation was performed using a medium not inoculated with microorganisms. The results are shown in Table 7 and FIG. FIG. 1 is a graph showing an SDS decomposition curve, and the slope of the curve is the SDS decomposition rate (SDS removal rate). For each microorganism, the time when the TOC concentration decreased sharply (KR-20R, KR-26R, KR-33R, and KR-43R were 8-24 hours, KR-35R was 24-48 hours, KR SDS degradation rate (expressed in TOC removal rate, unit: g-TOC / g-MLSS · day) in -41R strain was calculated. As shown in Table 7, each microorganism achieves a high SDS degradation rate of 0.5 (g-TOC / g-MLSS · day) or more, and each microorganism has a high SDS degradation ability. It was shown that

(3) Evaluation of surfactant decomposition ability [2]
The experiment was conducted in the same manner as in the above (2) except that “medium B” of the surfactant-containing medium shown in Table 6 was used as the surfactant-containing medium. Sampling of the culture solution was performed at 0, 8, 12, 16, 20, 24, and 48 hours from the start of the culture. The results are shown in Table 8 and FIG. For each microorganism, the time when the TOC concentration rapidly decreased (24 to 48 hours for KR-20R and KR-26R, 0 to 8 hours for KR-33R, 16 to 20 hours for KR-35R, KR-41R SDS and KR-43R were calculated for SDS degradation rate in 12 to 16 hours. As shown in Table 8, in any microorganism, an SDS degradation rate of 0.3 (g-TOC / g-MLSS · day) or more was achieved, but in particular, KR-33R strain, KR-35R strain, The KR-41R strain and the KR-43R strain have achieved a high SDS degradation rate of 1.2 (g-TOC / g-MLSS · day) or more, and each microorganism has a high SDS degradation ability. It has been shown.

[Comparative Example]
SDS decomposition rate evaluation by activated sludge method

  1 L of synthetic wastewater (100 mg / L SDS, 2.0 g / L inorganic salts, 3 mg / L silicon-based antifoaming agent) using SDS as a sole carbon source was placed in a 2 L aeration tank. Concentrated activated sludge was added to this synthetic wastewater so that the MLSS would have a concentration of 1000 mg / L, and aeration was started. After starting aeration, sampling was performed every predetermined time, and TOC and pH were measured. The water to be treated was changed by the fill & draw method with an aeration time of 24 hours as one cycle. The activated sludge was continuously conditioned and sampled until the TOC removal rate was stabilized. The TOC removal rate after stabilization of the treatment rate was defined as the TOS removal rate by standard activated sludge.

  The activated sludge was acclimatized five times. Table 9 shows the TOC measurement results, and Table 10 shows the MLSS concentration and the TOC decomposition rate (corresponding to the SDS decomposition rate, unit: g-TOC / g-MLSS · day) at the time of preparation. FIG. 3 shows an SDS decomposition curve. The slope of the curve in FIG. 3 is the SDS decomposition rate (removal rate). As a result, an SDS decomposition rate of 0.17 (g-TOC / g-MLSS · day) was shown as an average of the maximum decomposition rates after the 4th acclimatization with a stable processing rate. This value is considerably lower than the values for the above six types of microorganisms (Tables 7 and 8), indicating that SDS is difficult to be decomposed by a normal activated sludge method.

  From the above, it was shown that all of the above six types of microorganisms have a high SDS degradation ability.

Claims (4)

  Cupriavidus taiwanensis KR-20R strain (FERM P-22013), Cupriavidus necator KR-26R strain (FERM P-22014), Pseudomonas nitroreducens KR-33R strain (FERM P-20215), Ralstonia pickettii KR-35R strain (FERM P-22016) ), Ralstonia eutropha KR-41R strain (FERM P-20217), or Ralstonia eutropha KR-43R strain (FERM P-22018).   A method for treating surfactant-containing wastewater, comprising bringing the at least one microorganism according to claim 1 into contact with wastewater containing a surfactant to decompose the surfactant in the wastewater.   The method for treating a surfactant-containing wastewater according to claim 2, wherein the surfactant is an anionic surfactant.   The method for treating a surfactant-containing wastewater according to claim 3, wherein the anionic surfactant is selected from the group consisting of a sulfate ester salt, a sulfonate salt, and a carboxylate salt.

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