KR0132273B1 - Trichlorophenol assimilating microorganism and process for waste water treatment - Google Patents

Abstract

Wastewater generated at pulp or paper factory is in contact with pseudomonas solancerum strain TCP 114(KCTC 0106BP) including 2,4,6-trichlorophenol magnetization properties to eliminate 2,4,6-trichlorophenol effectively.

Description

Microorganisms having trigonal phenolic magnetizing ability and a method for treating trichlorinol-containing wastewater using the same

FIG. 1 is a graph showing the results of testing TCP resolution by strain Pseudomonas Solanaserum TCP 114 of the present invention in the presence of a secondary carbon source.

The present invention relates to a novel microorganism having trichlorophenol magnetizing ability and a method for treating trichlorophenol-containing wastewater using the same, and more particularly, Pseudomonas solanacearum TCP having trichlorophenol magnetizing ability. 114 and methods of treating trichloride-containing wastewater using the same.

Chlorinated phenols, including trichlorinated phenol (2,4,6-trichlorophenol; weakly referred to as TCP), are used as fungicides and insecticides in addition to bleaching pulp, preservatives in wood, and as dyeing aids in leather or leather industries. It is used as a precursor of pentachloride and tetrachloride and chlorinated chlorinated preservatives (prochloraz).

Chlorinated phenols are concentrated in soil or surface water because they are not easily degraded in nature and show toxicity such as carcinogenesis, and the US Environmental Protection Agency has reported that they are classified as recalcitrants. In addition, when the phenol-containing wastewater generated after the various treatment processes described above flows into the wastewater treatment plant without a bleeding process, the activated sludge is destroyed to cause a bulking phenomenon.

Attempts to treat TCP using microorganisms have been relatively recent and include strains that can be used for TCP treatment, such as, for example, azotobacter sp, Li et al., Appl. Environ. Microbiol., Which are aerobic bacteria. 57, 1920, 1991) and Pseudomonas pickettil (Klyohara et al., Lbld, 58, 1276, 1992).

However, their TCP resolution is quite high, but not yet satisfactory. Also, the microorganisms of azotobacters are soil microorganisms, which can reduce the efficiency of wastewater treatment, and the microorganisms of Plebobacterium, a phenol-magnetizing microorganism, It has been reported that TCP resolution is significantly reduced when there is a carbon source that is easy to decompose. (Sarber, D. L. et., Appl. Environ. Microbiol. 50: 1512-1518, 1987)

Therefore, there has been a demand for the development of a strain capable of maintaining a high TCP resolution even when a very high TCP resolution coexists with an easy magnetization substrate such as glucose or succinic acid.

Under these circumstances, the present inventors have diligently studied to find a new kind of TCP-magnetizing strain, and have found that the present invention can achieve the above object by Pseudomonas solanacearum isolated from plant wastewater. Came to complete.

That is, an object of the present invention is to provide Pseudomonas solanacearum KCTC 0106BP having trichlorophenol magnetization ability.

Another object of the present invention is to provide a method for treating trichloride-containing wastewater using the strain.

Hereinafter, the present invention will be described in more detail.

The present inventors collected the wastewater of Cheongju Industrial Complex to find a strain capable of maintaining high TCP resolution even in the presence of very high TCP resolution and coexistence of easy magnetization such as glucose, phenol or succinic acid. Strains with were first selected. Among the selected strains, particularly, strains capable of degrading 200 mg / l of TCP in 20 hours were selected. This strain is capable of magnetizing TCP at a concentration of 700 mg / l, and furthermore, even if succinic acid, phenol, and glucose are present together with TCP, the rate of TCP degradation remains unchanged or rather increased, and aniline, p-cresol, known as aromatic toxic compounds, It has been observed that phenol, benzoic acid and the like can be magnetized.

Therefore, the microorganism of the present invention can be very useful for treating not only TCP but also industrial wastewater containing various organic matters, in particular pulp or paper wastewater. In particular, in the case of treating the plant wastewater by the active sludge method, pretreatment with the microorganism of the present invention before sending the wastewater to the treatment plant decomposes nearly 100% of TCP in the wastewater, thereby destroying the activated sludge of the plant afterwards. It doesn't happen. Pretreatment of TCP-containing industrial wastewater using the microorganism of the present invention can be carried out according to a conventional microbiological method for treating wastewater, for example, pulp or papermaking in a column packed with a high concentration of the microorganism of the present invention. By passing the company's wastewater, TCP can be removed to prevent destruction of the activated sludge to treat these wastewaters.

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1

0.1% dipotassium phosphate per liter (% by weight, same as below), monobasic sodium phosphate 0.06%, ammonium nitrate 0.1%, magnesium sulfate 0.02%, potassium chloride 0.02%, trace elements (copper sulfate 0.2%, sulphate 0.03%, 0.1% cobalt sulfate, 0.1% molybdenum tetraoxide, 0.05% dispersion, and 2% calcium sulfate are dissolved in 0.1 ml of distilled water and diluted 10-fold. 1% of the wastewater of the industrial complex was added and incubated for 1 week at 30 ° C. under aerobic conditions.

0.1 ml of the culture solution was plated on a solid medium prepared by adding 1.8% agar to the screening medium, and exhaled incubation at 30 ° C. for 3 days to obtain about 3000 colonies.

Example 2.

The microorganisms obtained in Example 1 were inoculated in screening medium and incubated at 30 ° C. for 1 week, and then 1 ml of the culture was taken to measure the absorbance at 310 nm using a fluorescence analyzer to quantify TCP. Seven microbial colonies were 100% free of TCP.

In order to identify the microorganisms of these colonies, the physiological, morphological and culture characteristics were examined, and they all showed the same characteristics.

Morphological characteristics

It is negative at gram staining, has a diameter of 0.7 m and a length of 1 m, has a flagellum of mono subpola, and does not form spores.

Biochemical properties

G + C (mol%) in the chromosome is 68.5%, fatty acid is composed of C14: 00, 20H C14: 00, 3OH C14: 00, C16: 19, C16: 00, C17: △, 18:19.

Culture characteristics

Optimum incubation temperature and pH were 27-27 ° C. and 6.8-7.8, respectively, and growth at 41 ° C. and 4 ° C. was not observed.

Physiological characteristics

Oxidase +

Catalase +

Lysine Decarboxylase-

Ornithine Decarboxylase-

Hydrogen Sulfide Generation-

Nitrogen gas release from nitric acid +

Glucose capacity +

Mannitol capacity +

Xylose Ability +

Rhamnose ability +

Sorbitol ability +

Lactose ability +

Sugar utilization +

Arabinose ability +

Adonitol Utilization +

Rafinos ability +

Utilizing Salinity +

o-nitrophenyl-β-D-galactopyranoside-

Indole Generation-

Urea Hydrolysis +

Forges-Pro Scout (V.P) Reaction-

Citric acid use +

Tryptophan Deaminase-

Arginine Hydase-

Gelatin Liquefaction-

Malonsa suppression +

Hydrolysis-

Intracellular Polyhydroxybuntyric Acid Accumulation +

Hydrolysis of Extracellular Polyhydroxybutyl Acid-

From the above data, the microorganism of the present invention was identified as Pseudomonas solanasearum.

The microorganism Pseudomonas solanasearum of the present invention isolated and identified by the above method was named TCP 114 and deposited on March 17, 1994 at the Genetic Engineering Research Institute in Daejeon, and deposited KCTC 0106BP. I am assigned a number.

Example 3

In a 250 ml flask, 560 ml of screening medium and TCP were added at 80, 100, 150, 200, 300, 400, 500 and 700 mg / l, respectively, and shaken at 30 ° C. at 120 rpm.

Specific growth rate was measured for each TCP concentration, and the results are shown in Table 1.

From the above results, it can be seen that the microorganism Pseudomonas solaranaserum KCTC 0106BP of the present invention exhibits optimal growth when the TCP concentration is in the range of 150-200 mg / l. It can also be seen that TCP grows even when present at high concentrations of 700 mg / l.

Example 4.

The screening medium to which TCP was added at a concentration of 100 mg / L was inoculated with Pseudomonas solaranacerum TCP 114 (KCTC 0106BP) of the present invention (dry weight 42 mg / l cell concentration). When the specific degradation rate of TCP was measured by shaking culture at 30 ° C. at 120 rpm, the value was 0.240 per hour.

Example 5

100 mg / l of TCP and 0.5 mM of succinic acid were added to the screening medium and inoculated with Pseudomonas Solanasearum TCP 114 (KCTC 0106BP) of the present invention (dry weight 42 mg / l cell concentration). When the specific decomposition rate of TCP was measured while shaking at 30 ° C. and 120 rpm, it was 0.329 per hour.

Example 6

In the same manner as in Example 5, 0.5 mM of phenol was added to the screening medium instead of succinic acid, and was inoculated with Pseudomonas solaranacerum TCP 114 (KCTC 0106BP) (dry weight 42 mg / l cell concentration). When the specific resolution of TCP was measured under shaking at 30 ° C. at 120 rpm, the value was 0.329 per hour.

Example 7

100 mg / l of TCP and 0.5 mM of glucose were added to the screening medium, and Pseudomonas Solanasearum TCP 114 (KCTC 0106BP) was inoculated at a dry weight of 42 mg / l cell concentration. When the specific resolution of TCP was measured while shaking at 30 ° C. and 120 rpm, the rate was 0.309 per hour.

Example 8

Aniline, p-cresol or benzoic acid, respectively, was added to the screening medium at a concentration of 0.5 mM and Pseudomonas Solanasearum TCP 114 (KCTC 0106BP) was inoculated at a dry weight of 90 mg / l cell concentration. Shake culture was performed for 30 days at 30 ℃ 120 rpm. After incubation using a fluorescence spectrometer, the absorbance change of the substrate and cell growth (absorption at 600 nm) at 250-350 nm were simultaneously measured, and the results are shown in Table 2.

(Note) *: When the substrate was degraded and reddish brown, the medium was changed and cultured.

The results of Table 2 confirm that the strain of the present invention can magnetize phenol, benzoic acid, aniline and p-cresol in addition to trichloride phenol. In addition to these substances, the strains of the present invention can partially magnetize aromatic dichlorophenol (2,6-dichlorophenol) and monochlorobenzoic acid (3-chlorobenzoate), and are substituted with various hydroxyl groups. It was confirmed that the benzoic acid of was completely decomposed and grown.

Example 9

In order to confirm whether the strain of the present invention does not lose the ability to magnetize TCP even in the presence of a secondary carbon source, the same procedure as in Example 4 was carried out in the medium with 0.5 mM glucose, phenol and succinic acid, respectively. Cultures were with or without addition. After incubation, the concentrations of TCP, glucose, phenol and succinic acid were measured, and the residual amount was calculated as a relative value of 100 at each initial concentration, and the results are shown in FIG.

As can be seen in FIG. 1, the strain of the present invention has a glucose addition (-■-), phenol addition (-●-) and succinic acid addition (-) as compared to (-▼-) when no secondary carbon source is added. ▲-) also shows high TCP resolution. For reference, in FIG. 1,-▼-indicates residual TCP concentration when no secondary carbon source is added, and-■-,-●-, and-▲-indicate glucose, phenol and succinic acid, respectively. Residual TCP concentrations are indicated, and-□-,-○-and -Δ- represent the concentrations of glucose, phenol and succinic acid remaining in the medium after incubation, respectively.

Claims (2)

1. Pseudononas solanacearum strain TCP 114 (KCTC 0106BP) with 2, 4, 6-trichlorophenol magnetization ability. 2. Microbiological treatment of wastewater containing 2, 4, 6-trichlorophenol, characterized by contacting the wastewater with Pseudononas solanacearum strain TCP 114 (KCTC 0106BP). How to. A method for microbiologically treating wastewater containing 2, 4, 6-trichlorophenol, wherein the wastewater is contacted with Pseudononas solanacearum strain TCP 114 (KCTC 0106BP). .