KR100512287B1 - Novel microbe gordonia sihwaniensis pkl-1 enabling degradation of volatile organic compounds and methods for disposal of waste water, sewage and air using the same - Google Patents

Abstract

A novel microorganism capable of effectively decomposing dimethyl sulfide (DMS) among volatile organic compounds without secondary pollution, and a method for decomposing DMS contained in sewage, wastewater, air or soil using the same are disclosed. . To this end, a novel microorganism Godonia cywannisis piquel (PKL) -1 (Accession No .: KCCM-10443) having a resolution of DMS, a volatile organic compound, is provided. In addition, there is provided a method for treating wastewater, air, and the like containing volatile organic compounds using Godoni and Shiwaniensis PKL-1. By using the novel microorganism according to the present invention, it is possible to effectively remove volatile organic compounds, especially DMS, which are spilled during industrial activities and have a harmful effect on humans and organisms and further cause environmental pollution.

Description

Microorganism Godonia siwanenisis picella-1 having a resolution of volatile organic compounds and treatment method for wastewater and air using the same AND AIR USING THE SAME}

The present invention relates to a novel microorganism Gordonia sihwaniensis (PKL) -1 (Accession No .: KCCM-10443) and a biological treatment method of wastewater and air using the same, in more detail. Is a novel microorganism Godonia cywannisis PKL-1 (KCCM-10443) having excellent resolution of hardly decomposable volatile organic compounds and biological treatment method of wastewater, soil and air containing hardly decomposable volatile organic compounds using the same. It is about.

As civilization develops, interest in ecological environment is increasing. In particular, the health problems of numerous volatile volatile organic compounds produced by organic synthesis methods on various living organisms have emerged as serious social problems.

In general, volatile organic compounds such as dimethyl sulfide (DMS) are produced in large quantities as the industry develops, and are widely used in many industrial processes. As a result, workers in industrial sites are increasingly exposed to volatile organic compounds. However, exposure to these volatile organic compounds has been reported to cause abnormalities in urinary and serum bile acids secretion.

In particular, DMS in volatile organic compounds is known to be toxic to carcinogenesis, nervous system and respiratory system. Increasing interest in environmental pollution and the adverse effects of these substances on the environment and human health has led to the development of methods for purifying volatile organic compounds.

Among the volatile organic compounds present in aquatic ecosystems, bacteria that degrade DMS are Thiobacillus (Kyeoun-Suck Cho, Journal of Fermentation and Bioengineering Vol. 71, No. 6, 384-389. 1991), Hippo. Only the degree of Hyphomicrobium (Liang Zhang, Journal of Fermentation and Bioengineering Vol. 72, No. 5, 392-396. 1991) is known. However, these studies are not comprehensive studies considering various environmental factors such as temperature and pH, so that the maintenance of their resolution by microorganisms is questioned.

In addition, the volatile organic compounds are classified as recalcitrants that are difficult to be decomposed by microorganisms, and the physicochemical treatment method for decomposing them is generally not only likely to cause secondary environmental pollution but also has a disadvantage of high processing cost. have. Therefore, there is an urgent need for the development of effective treatment techniques using highly active microorganisms.

In order to solve this problem, genetically recombined strains were made to completely decompose volatile organic compounds, but stability problems remain when these strains are actually used for treating volatile organic compounds. Therefore, in order to completely decompose volatile organic compounds, it is necessary to secure microorganisms having excellent resolution and to develop efficient and natural methods for removing pollutants.

Accordingly, a first object of the present invention is to provide a novel microorganism capable of effectively decomposing DMS in volatile organic compounds without secondary contamination.

It is a second object of the present invention to provide a method for decomposing DMS contained in sewage, wastewater, air, soil, etc. using the novel microorganisms.

One embodiment of the present invention provides a novel microorganism Godonia cywanissis PKL-1 (PKL) -1 having a resolution of DMS which is a volatile organic compound. The novel microbial Goni and Shiwaniensis PKL-1 were deposited on November 4, 2002 at the Korea Microorganism Conservation Center under Accession No. KCCM-10443.

One embodiment of the present invention in order to achieve the second object of the present invention described above is the treatment of wastewater and air containing volatile organic compounds using a novel microorganism Godi and Siwannisis PKL-1 (KCCM-10443) Provide a method.

By using the novel microorganism according to the present invention, it is possible to effectively remove volatile organic compounds, especially DMS, which are the main culprit, which are spilled during industrial activities and have harmful effects on humans and organisms, and also cause environmental pollution. In addition, when the novel microorganism of the present invention is applied to waste water, sewage, soil, atmosphere, or the like contaminated with volatile organic compounds, the environment can be efficiently purified without secondary pollution by the biodegradation process by the microorganisms.

Therefore, it can be widely used in small businesses such as laundry using volatile organic solvents, chemicals, refineries, waste treatment plants, etc. In particular, the polluted soils around facilities such as gas stations and odorous volatile organic compounds in air and ground water Excellent removal effect.

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

Novel microorganisms of the present invention are separated from solid samples in solid agar medium by using DMS as a carbon source and energy source among samples collected from water and soil sludge around streams where wastewater from chemical wastewater treatment plant is discharged. Get sympathetic to

In order to prepare the strain isolated by the above method as a sample to be used for the decomposition of DMS, a minimum medium (Na2HPO4 4g, KH2PO4, 1.5g, MgSO4 · 7H2O 0.2g, CaCl2 0.01g, FeNH4-citrate 0.005g, agar 15g in 1L To the dissolved liquid medium) and inoculated with the new strain isolated under aerobic conditions of about 25-40 ° C., most preferably about 32 ° C., about pH 6-8, most preferably about pH 7. And cultured by centrifugation to obtain a strain.

The highest rate of decomposition at about 32 ° C. and a significant drop in rate of degradation below about 25 ° C. or above 40 ° C., and in the case of pH as well, the highest rate of degradation at neutral pH of about 7 and an acid below about pH 6 Under conditions or basic conditions of more than about pH 8, there is a problem that the decomposition rate is lowered.

  The novel strains of the invention can be included in known treatments. The novel microorganism Godonia siwanenisis PKL-1 (KCCM-10443) is a microorganism having a resolution of DMS can be usefully used for treatment of wastewater, air and the like.

In particular, when the DMS is applied to waste water containing about 0.001 to 50ppm, air, soil and the like shows excellent DMS resolution. If the DMS exceeds about 50ppm, there is a problem that the degradation capacity is significantly reduced due to the toxicity of the DMS. Decomposition can be performed as long as there is even a small amount of DMS, but especially less than about 0.001 ppm, DMS decomposition is not necessary.

The present invention will be described in more detail with reference to the following examples. However, an Example is for illustrating this invention and is not limited only to these.

Example 1

(Isolation of Strains with DMS Resolution)

To isolate strains that degrade DMS, dissolve the minimum medium (Na2HPO4 4g, KH2PO4, 1.5g, MgSO4 · 7H2O 0.2g, CaCl2 0.01g, FeNH4-citrate 0.005g, 15g agar in 1L) containing the appropriate concentration of DMS. A strain using DMS as a carbon source was added thereto, and then cultured at 32 ° C. using a shake incubator. Thereafter, secondary passage in new minimal medium to confirm the growth of microorganisms. Subsequently, again and again subcultured in the same medium to obtain a stable strain.

The strain was cultured while maintaining the range of 32 ℃ and pH 6-8 with DMS as the only carbon source, and then the culture solution was centrifuged for 4 minutes to recover the cells. Since the recovered strains were suspended in a minimal medium, 2, 5 and 10 ppm of DMS were added thereto. Gas samples were taken at regular intervals during shaking culture at 32 ° C. pH 6-8 to measure the amount of DMS present in the samples by gas chromatography.

On the basis of the amount of the initial DMS used in the invention, the strain with the fastest degradation rate of DMS per unit time was screened as the highest resolution strain.

(Identification of Strains with Volatile Organic Compound Resolution)

The strain isolated in Example 1 is a gram-positive bacterium with motility to form a rounded and coarse colony on the medium, and its characteristics are shown in Table 1.

Item characteristic Gram dyeing + shape Bacillus motility + Colony color Ivory

(+: Positive,-: negative)

In addition, for the identification of strains, fatty acids were separated and analyzed by gas chromatography, and the results are shown in Table 2.

fatty acid Configuration(%) 12: 0 0.14 14: 0 2.41 15: 0 0.67 16: 1 w9c 0.92 16: 0 38.26 17: 1 w8c 1.72 17: 0 0.17 18: 1 w9c 29.87 18: 0 3.41

Referring to Table 2, 16: 0 showed the highest composition as 38.26%. As a result of analyzing the cellular fatty acid composition and 16rDNA sequence in the database, it showed 99% homology with Godonia siwaniensis, and it was confirmed that the new microorganism is a new species of Godonia siwanenisis.

On the basis of the above morphological, cultural and biochemical properties, the novel microorganism is named Gordonia sihwaniensis PKL-1 and deposited in Korea Microorganism Conservation Center on November 4, 2002. It was deposited with the number KCCM-10443 and its certificate was attached.

Example 2

(Checking the growth of strains in a minimal medium with volatile organic compounds as the only carbon source)

In order to investigate the resolution of the single volatile organic compound of Godonia siwanenisis PKL-1 (KCCM-10443) separated in Example 1, the minimum medium of Example 1, DMS is a volatile organic compound at 32 ℃ Using a carbon source, and supplied with saturated DMS in a gaseous state in a closed culture vessel, the culture solution was collected, and the absorbance was measured at 600 nm using a spectrophotometer to determine the growth of the strain. The above result is shown in FIG.

The strain showed a stationary phase within 78 hours, and the absorbance after the logarithmic phase was maintained at a constant value. The novel strain of the present invention showed a state of proliferation using DMS as a carbon and an energy source.

(Checking the DMS Resolution of Strains in the Minimum Medium Using DMS as the Only Carbon Source Among Volatile Organic Compounds)

Using the minimum medium of Example 1, DMS, a volatile organic compound at 32 ° C., as the only carbon source, and supplying DMS 2, 5, and 10 ppm, respectively, in a sealed culture vessel, and the amount of DMS remaining in the supernatant over time. As a result of analysis, the removal tendency according to the concentration is shown in FIGS. 2A to 2C.

Figures 2a to 2c shows the concentration of DMS after culturing the new microorganism Godonia cywannisis PKL-1 (KCCM-10443) according to the present invention in a minimal medium containing 2 mg / L, 5 mg / L, and 10 mg / L of DMS. Graphs showing the decrease.

Uncolored portions (one) in FIGS. 2A-2C are the results when strains were not added as a control. After 25 hours of incubation, the removal efficiencies at 2, 5, and 10 ppm showed 60.35, 35.18 and 35.96% removal efficiency, respectively, depending on the concentration. As a result, it was confirmed that the Gothonia siwanenisis PKL-1 microorganism of the present invention had a resolution of DMS in volatile organic compounds.

According to the present invention, the use of the novel microorganisms can effectively remove volatile organic compounds, especially DMS, which are the main culprit, which are spilled during industrial activities and adversely affect the human body and organisms and cause environmental pollution.

In addition, when the novel microorganism of the present invention is applied to waste water, sewage, soil, atmosphere, or the like contaminated with volatile organic compounds, the environment can be efficiently purified without secondary pollution by the biodegradation process by the microorganisms.

Therefore, it can be widely used in small businesses such as laundry using volatile organic solvents, chemicals, refineries, waste treatment plants, etc. In particular, polluted soils around facilities such as gas stations, and odorous volatile organic compounds in the air and groundwater. It will have an excellent effect on the removal of.

Although described above with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and changed within the scope of the invention without departing from the spirit and scope of the invention described in the claims below I can understand that you can.

Figure 1 shows the growth of the novel microorganism Gordonia sihwaniensis PKL-1 (KCCM-10443) according to the present invention after incubation in minimal medium containing dimethyl sulfide (DMS). One graph.

Figure 2a is a graph showing the decrease in the concentration of DMS after culturing the novel microorganism Godonia siwanenisis PKL-1 (KCCM-10443) in a minimal medium containing 2 mg / L DMS according to the present invention.

Figure 2b is a graph showing the decrease in the concentration of DMS after culturing the novel microorganism Godonia cywannisis PKL-1 (KCCM-10443) in a minimal medium containing 5 mg / L of the present invention.

Figure 2c is a graph showing a decrease in the concentration of DMS after culturing the novel microorganism Godonia siwanenisis PKL-1 (KCCM-10443) in a minimal medium containing 10 mg / L DMS according to the present invention.

Claims (6)

Novel microorganism Godonia sihwaniensis PKL-1 (KCCM-10443) with volatile organic compound resolution.   The novel microorganism Godonia siwanenisis PKL-1 (KCCM-10443) according to claim 1, wherein the volatile organic compound is DiMethyl Sulfide (DMS). delete delete delete delete