KR100713860B1 - Cyanobacterium Phormidium parchydematicum TB164 having turbidityremoval activity - Google Patents

Abstract

FIELD OF THE INVENTION The present invention relates to Cyanobacteria formidium pachydematicum TB164 [KCTC 10851BP] having turbidity removal activity, and more particularly to formium pachydematicom TB164 [KCTC 10851BP] from natural water systems. Isolation, and the strain was cultured in turbid water to reveal the turbidity removal ability, relates to the formdium pachydematicom useful as a turbidity remover.

Turbidity-removal, cyanobacteria, Phormidium parchydematicum

Description

Cyanobacterium Phormidium parchydematicum TB164 having turbidity­removal activity}

1 is an optical micrograph of the bacterium Formidium Pakidematicom TB164 [KCTC 10851BP] isolated and cultured in a natural water system.

Figure 2 is a graph showing the change in turbidity according to the culture of the bacterium Formidium Pakidematicom TB164 [KCTC 10851BP].

Figure 3 is a graph showing the change in turbidity according to the concentration of the bacterium Formidium Pakidematicom TB164 [KCTC 10851BP].

FIELD OF THE INVENTION The present invention relates to Cyanobacteria formidium pachydematicum TB164 [KCTC 10851BP] having turbidity removal activity, and more particularly to formium pachydematicom TB164 [KCTC 10851BP] from natural water systems. Isolation, and the strain was cultured in turbid water to reveal the turbidity removal ability, relates to the formdium pachydematicom useful as a turbidity remover.

In the case of water sources, lake water and other streams, turbidity-inducing substances include fine sand particles, fine suspended solids, and solid suspended solids with very small particles. For this purpose, various treatment methods including physicochemical methods have been applied, but due to the variety, properties, and inflow characteristics of the turbidity-inducing substance, it takes a lot of time and money to process.

Increasing turbidity in natural water systems not only leads to an increase in pretreatment costs, but also has led to the need for several subsequent processes for its flocculation and precipitation. In general, various industrial flocculants have been used in various forms to remove turbidity-inducing substances such as suspended solids and other particulates of sand. It is known to show a difference.

In general, flocculants are used to remove fine particles and colloidal particles in water to bind and remove them through precipitation, filtration and flotation. In the past, aluminum sulfate was mainly used, but in recent years, it has more excellent performance. Polyaluminum chloride, polyaluminum silicate silica, polyhydraulic hydrochloride silicate, and the like. Accordingly, countries have designated water treatment agents that can be used for water treatment, such as tap water, to regulate the types and maximum usage of chemicals. In Korea, quality standards for water treatment agents are also defined and managed, and aluminum sulfate is managed.

However, the method using such a flocculant has many problems, such as causing a secondary process due to the residual of the polymer flocculant in the treatment liquid due to its chemical properties, which has limitations in a wide range of applications.

The bacterium is well known as a group of organisms that often occur in water sources, causing off-flavors or producing harmful substances for removal. Particularly, the bacterium Formidium pachydematicomb proliferates while forming a long chain filament, and forms a colony on the mucous membrane and appears as a bacterium that grows on the substrate and appears in various appeals and reservoirs, and is 2-methylisobolneor. (2-MIB) or geosmin is produced, and it is generally known to grow or adhere to water in the water system.

On the other hand, some bacteria are reported to produce various bioactive substances as secondary metabolites. As a representative example, the bacterium such as Spirulina is used to produce useful substances such as feed additives, dietary supplements, and gamma linolenic acid (GLA) using biomass. Korean Patent Laid-Open Publication No. 10-2005-0082713 has reported that the bacterium Osillatoria has antibiotic activity against various pathogenic bacteria such as Salmonella typhimurium . -2005-0082714 reports that the culture extract of Anavena pros-Aqui has excellent inhibitory activity against vaccinia H1-related protein tyrosine phosphatase (VHR), an enzyme involved in the regulation of cancer cell growth and proliferation. There is a bar. Recently, researches on the production of various bioactive substances, which are secondary metabolites from bacteria, have been actively conducted. Application to various fields using the bacterium has been attempted, but the case of applying the bacterium to the removal of turbidity has not been reported.

The present invention is to identify the removal ability for turbidity by separating the bacterium Formidium Pakidematicom TB164 [KCTC 10851BP], which occurs in large quantities in natural water systems, including a water source, and is a polymetal that is a heavy metal of a cationic system generally used in a water source. The present invention provides a method for removing turbidity using Formidium Pakidematicom TB164 [KCTC 10851BP], an environmentally friendly material, focusing on the problems of using flocculants such as aluminum chloride (PAC), aluminum sulfate, ferrous acid, and ferric chloride. .

The present invention is characterized by the bacterium Formidium parchydematicum TB164 [KCTC 10851BP] having turbidity removal ability.

In another aspect, the present invention is characterized by a turbidity removal method using the bacterium of the genus Phormidium .

Referring to the present invention in more detail as follows.

The colony formed while cultivating the water sample of natural water in general Allen solid medium is first observed by optical microscopy to isolate Southern bacteria. The primary isolates were maintained as monogal strains through several passages in Allen solid medium, followed by microscopic observation and continuous screening.

As a result of observing size, shape, and pigments, which are the main items for identifying bacteria, by optical microscope, the finally isolated forged cyanobacteria were identified as belonging to formium pachydematicom. It was named Maticcom TB164. In addition, the strain was deposited with the Korea Biotechnology Research Institute Gene Bank on September 28, 2005, the accession number was assigned to KCTC 10851BP.

The isolated formdium pachydematicom TB164 [KCTC 10851BP] proliferates into a chain filament as shown in FIG. 1, and forms a colony on a mucous membrane and enters the substrate.

The isolated formamide pachydematicom TB164 [KCTC 10851BP] was incubated with turbid water and tested for its ability to remove turbidity.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the following Examples are intended to illustrate the present invention and do not limit the present invention.

Example 1 Water Sample Collection

In order to secure new bacteria, 123 water samples were collected from a total of 80 locations, including rivers, rivers and lakes, including Chungcheongnambuk-do, Gyeongsangnambuk-do, and Jeollanam-do.

Example 2: Isolation of Southern Bacteria

The water samples of natural water are plated on a normal allen solid medium and cultured in an algae cultivation room [luminous intensity is 100-130 μmol / m ² / s, temperature is 24-26 ℃, light cycle is = L: D (24: 0)]. Colonies formed while observing with optical microscopy to isolate the first bacteria. The first isolates of bacteria were maintained in monolithic culture by several passages in Allen solid medium through microscopic observation and continuous screening.

In this study, the following two methods were mainly used to purely isolate bacteria. First, 100 μl of a sample collected from nature was plated on a plate medium for bacterial culture. When the plate was incubated for 10 to 14 days in a constant temperature culture room, green or cyan-colored colonies appeared. At this time, it was often contaminated with other microorganisms. The colonies were isolated by repeating subculture on fresh medium. Secondly, the sample was filtered using a sterilized glass microfiber filter (Whatman, Cat. No. 1822 025), and the filter surface of the filter paper was placed on the medium for culture of B. It was cultured in a light stirrer irradiated with light. When gliding motility of filamentous cyanobacteria was observed around 3 days of culture, the glass microfiber filter was removed and the plate medium inoculated with filamentous bacterium was cut with a blade and transferred to a new flat medium. After 3 days from this time, the newly moved tricom (trichome, cell death) was taken and cultured by transferring to a new medium.

Example 3: Identification of Bacteria

Microorganisms grown on microorganisms or solid media contained in water samples transported at 4 ° C. in a refrigerated state without pretreatment were observed with a light microscope (Nikon Microphot FXA) and an inverted microscope (Nikon TMS) (FIG. 1) and the base of the cpcBA-IGS gene region. Molecular identification using sequence (SEQ ID NO: 1) confirmed that the genus was formium, and based on the microscopic observations, it was identified with reference to the bird's book (Prescott, 1973; Jeong, 1993; Canter-Lund and Lund, 1995). Results We named Phormidium parchydematicum TB164 ( Phormidium parchydematicum TB164), which was deposited on September 28, 2005, to the Gene Bank of Korea Research Institute of Bioscience and Biotechnology and was given accession number KCTC 10851BP.

Example 4 Cytobacterial Culture and Determination of Turbidity Removal

In order to cultivate the formdium Pakidematicom TB164 [KCTC 10851BP] of the present invention as a light source was used (cool white fluorescent lamps) and the brightness is 100 ~ 130 μmol / m ² / s, temperature is 24 ~ 26 ℃, Photoperiod was incubated in Allen media of Table 1 in an optical stirrer (Illuminated Shaking Incubator, 150 rpm) adjusted to = L: D (24: 0).

Composition of the Allen Badge ingredient Content (g / L) NaNO ₃ 1.5 K ₂ HPO ₄ 0.039 MgSO ₄ 7H ₂ O 0.075 Na ₂ CO ₃ 0.021 CaCl ₂ 0.027 Na ₂ SiO _{3 ·} 9H ₂ O 0.058 Ferric citrate 0.006 Citric acid 0.006 EDTA 0.001 Microelement 1 ml-H ₃ BO ₃ : 2.86 g / L-MnCl ₂ 4H ₂ O: 1.86 g / L-ZnSO ₄ 7H ₂ O: 0.22 g / L-Na ₂ MoO ₄ 2H ₂ O: 0.391 g / L- _{CuSO 4 · 5H 2 O: 0.079} g / L - Co (NO 3) 2 · 6H 2 O: 0.0494 g / L

After culturing the homogenized formdium pachydematicom TB164 [KCTC 10851BP] using a tissue crusher (homogenazer), the chlorophyll component or fine particles in the crushing process by filtering with a nylon mesh (nylon mesh, 20 ㎛ pore size) Removed. The strain strained to the mesh was washed with an allene medium and inoculated in a fresh allen medium to prepare a strain for use in the experiment.

The turbidity material to be used for the experiment was prepared in a laboratory, and after the sterilization of the muddy water collected from the bottom of the river for 15 minutes at 121 ° C. for 5 days, the suspended particles were collected and prepared.

The experimental group was inoculated in a 250 ml Erlenmeyer flask with 100 ml of allergen medium inoculated with 5 ml of the prepared strain of Formium Pakidematicom TB164 [KCTC 10851BP] (103 µg / L concentration of chlorophyll a ) and 40 ml of the turbidity prepared. The final turbidity was 50 to 60 NTU (nephelometric turbidity unit). In addition, the control group was not inoculated with the strain, and the turbidity material prepared in 100 ml of allen medium in a 250 ml Erlenmeyer flask was put in a final turbidity of 50 ~ 60 NTU.

Thus prepared flask was measured for turbidity while incubated with a rotary incubator (150 rpm) for 10 days, and measured. The results thus obtained are shown in FIG. 2.

As shown in FIG. 2, the experimental group using formdium pachydematicom TB164 [KCTC 10851BP] was found to remove about 60% of the initial turbidity (about 20 NTU) after 1 day, and the turbidity after 3 days. Has been shown to maintain low turbidity below 3 NTU. Compared with the initial turbidity, the turbidity after 3 days was about 95%, and after 10 days the turbidity removal efficiency was about 98%. In contrast, the control group showed high turbidity of 10 NTU or more after 5 days. Formium Pakidematicom as shown in FIG. In the case of TB164 [KCTC 10851BP] it was confirmed that it has a clear turbidity removal ability, and also showed that it shows a high removal efficiency in a short time.

Example 5: Determination of turbidity removal ability according to strain concentration

The change of turbidity removal ability with the change of the use of formdium pachydematicom TB164 [KCTC 10851BP] was tested. Since more than 60% of the initial turbidity was removed after 1 day in Example 1, changes in turbidity were observed at 1, 3, 5, 7, 22 hours after the start of the experiment.

In the case of the experimental group, the amount of the target strain was changed to 10 ml, 20 ml, 30 ml, 40 ml and 50 ml after filtering the allene medium, and the mixture was mixed with the filtered allene medium so that the total amount was 70 ml. a concentration of each 116.85 ㎍ / L, 166.42 ㎍ / L, 358.61 ㎍ / L, 365.29 ㎍ / L and was 411.51 ㎍ / L. In the mixture of the prepared strain and the filtered allene medium, 10 ml each was injected into the mixed solution to 90 ml of the allene medium and 40 ml of the turbidity-inducing substance, and the concentration of the strain was changed (based on the concentration of chlorophyll- a ). After the turbidity-inducing substance was added to 100 ml of allene medium, 40 ml of turbidity was adjusted to have a turbidity of about 50 NTU, and the turbidity with the experimental group was analyzed.

As shown in FIG. 3, turbidity removal ability was increased as the concentration of formdium pachydematicom TB164 [KCTC 10851BP] was increased, and in particular, the amount of the strain of the present invention in the mixture of the target strain and the filtered allene medium was 50. In the experimental group of ㎖, about 40% of the initial turbidity-inducing substance was removed 1 hour after the start of the experiment, and the turbidity after 22 hours in the entire experimental area showed a low NTU of about 0.5% compared to the initial turbidity value. Dematicom The apparent turbidity removal effect of TB164 [KCTC 10851BP] was confirmed.

Formidium Pakidematicom TB164 [KCTC 10851BP] of the present invention shows very good turbidity removal ability without special treatment, it can be applied as an environmentally friendly biological material turbidity remover in place of the existing chemical flocculant.

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Claims (4)

Phormidium parchydematicum TB164 [KCTC 10851BP] with turbidity removal ability. delete delete A turbidity removal method characterized by injecting Phormidium parchydematicum TB164 [KCTC 10851BP] into turbid water.

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