KR100713859B1 - Cyanobacterium Anabaena flosaquae BT11 having turbidityremoval activity - Google Patents

Abstract

The present invention relates to Anabaena flos-aqua e BT11 [KCTC 10852BP] having turbidity removal activity, and more particularly, to Anabena pros-aqui BT11 [KCTC 10852BP] from a natural water system. ] And the strain was cultured in turbid water to reveal the turbidity removal ability, and relates to Anavena pros-Aqui useful as a turbidity remover.

Turbidity-removal, cyanobacteria, Anabaena flos-aquae

Description

Cyanobacterium Anabaena flos­aquae BT11 having turbidity­removal activity}

1 is a bacterium Anavena pros-Aqui cultivated by separating from natural water Photomicrograph of BT11 [KCTC 10852BP].

Figure 2 bacterium Anavena prosper-Aqui It is a graph showing the turbidity change according to the culture of BT11 [KCTC 10852BP].

Figure 3 bacterium Anavena Prosper-Aqui It is a graph showing the turbidity change of the initial culture of BT11 [KCTC 10852BP].

The present invention relates to Anabaena flos-aqua e BT11 [KCTC 10852BP] having turbidity removal activity, and more particularly, to Anabena pros-aqui BT11 [KCTC 10852BP] from a natural water system. ] And the strain was cultured in turbid water to reveal turbidity removal ability, and relates to Anabena pros-aqui 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 and the like are 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. 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 reports 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. have. 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 isolate the bacterium Anabena pros-Aqui BT11 [KCTC 10852BP] occurring in large quantities in natural water systems, including a water source, and to remove the turbidity material, polychlorinated heavy metal of the cationic system commonly used in water sources An object of the present invention is to provide a method for removing turbidity using Anavena Pros-Aqui BT11 [KCTC 10852BP], which is an environmentally friendly material, in view of the problems of using flocculants such as aluminum (PAC), aluminum sulfate, ferrous acid and ferric chloride.

The present invention is characterized by Anabaena flos-aquae BT11 [KCTC 10852BP] having turbidity removal ability.

In another aspect, the present invention a turbidity removal method using the vena Ana (Anabaena) in cyanobacteria by another feature.

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, form, and pigment, which are the main items for identifying bacteria, by optical microscope, the finally isolated monotonous bacterium was identified as belonging to Anavena pros-Aqui, and it was identified as Anavena pros-Aqui Named BT11. 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 10852BP.

The isolated Anavena pros-Aqui BT11 [KCTC 10852BP] is proliferated into a chain filament as shown in FIG. 1, and it is generally suspended by using gas vacuole to adsorb a substance that is a substrate. They also adhere, and are often attached in indoor cultures.

When the isolated Anavena pros-Aqui BT11 [KCTC 10852BP] was incubated with turbid water, the turbidity removal ability was examined, and it was confirmed that the turbidity removal ability was excellent.

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 for rivers, rivers and lakes in Korea 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 plate 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

The bacterium grown on microorganisms or solid media contained in water samples delivered at 4 ° C. refrigerated without pretreatment was observed under a light microscope (Nikon Microphot FXA) and an inverted microscope (Nikon TMS) (FIG. 1) and the 16S rDNA gene sequence (SEQ ID NO: Molecular identification using 1) confirmed that it is Pros-Aqui species of the genus Anavena, based on the microscopic observations, it was identified with reference to the algae illustration (Prescott, 1973; Chung, 1993; Canter-Lund and Lund, 1995). Results Anabena Pros-Aqui BT11 ( Anabaena flos-aqua e BT11) was named as a gene bank of Korea Biotechnology Research Institute on September 28, 2005 and was given accession number KCTC 10852BP.

Example 4 Cytobacterial Culture and Determination of Turbidity Removal

For the cultivation of the anavena pros-acqui BT11 [KCTC 10852BP] 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 ℃, Gwangju The group was incubated in Allen media of Table 1 in an 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 homogenizing the cultured Anavena pros-Aqui BT11 [KCTC 10852BP] using a homogenazer, the chlorophyll component or microparticles were removed during the crushing process by filtering with a nylon mesh (20 μm pore size). It was. 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 into a 250 ml Erlenmeyer flask and inoculated with 10 ml of the above-produced Anavena pros-Aqui BT11 [KCTC 10852BP] strain (37 ㎍ / L concentration of chlorophyll- a ) in 100 ml of Allen medium and 40 ml of the prepared turbidity material Final turbidity was set to 50-60 NTU (nephelometric turbidity unit). The flask thus prepared was measured for 30 days of incubation with a rotary incubator (150 rpm) to measure the degree of turbidity removal at 10 day intervals, and chlorophyll a was measured at 10 day intervals while incubating in the same manner as the experimental group to investigate the growth of the cells. It was.

As shown in FIG. 2, the Anavena flossac BT11 [KCTC 10852BP] showed 99.5% turbidity removal efficiency (turbidity 0.2 NTU) at 10 days and most of the turbidity material was removed at 99 days and 98.9 days at 20 days and 30 days, respectively. Similar to the initial in%. At the beginning of the culture with the chlorophyll a concentration of Anavena pros-Aqui BT11 [KCTC 10852BP] of 310 μg / L, most of the turbidity material was removed, showing very good turbidity removal ability.

Example 5 Measurement of Initial Turbidity Removal Capacity

Turbidity removal was determined to be 99% or more at the beginning of the culture of Anavena pros-Aqui BT11 [KCTC 10852BP]. Experiments were prepared in the same manner as in Example 4 and incubated in the same conditions as above for 12 days. In this experiment, however, only Anavena Pros-Aqui BT11 [KCTC 10852BP] was added without the addition of Anabena Pros-Aqui BT11 [KCTC 10852BP] as a control and allergic medium (Control 1) without addition of the Turbidity materials. The experiment was performed by adding one Allen medium (control 2). Turbidity and the value of chlorophyll a were measured daily.

As shown in FIG. 3, the turbidity of 9,5% was removed at 2.75 NTU on day 1 of the culture of Anabena pros-Aqui BT11 [KCTC 10852BP], and the turbidity was further decreased over time to 0.39 NTU on day 7 of the culture. Turbidity removal efficiency was the highest (99.3%). In Control 2 without addition of the turbidity material, turbidity by Anavena Pros-Aqui BT11 [KCTC 10852BP] itself was very low in the range of 1.6 to 0.2 NTU as a whole. On the other hand, in the control group 1, which only added the turbidity material, the turbidity material was agglomerated naturally by rotation during cultivation, forming a lump and sinking to the bottom of the culture vessel, whereby the turbidity gradually decreased. However, the removal of turbidity occurred at a high rate in a short time compared to the natural aggregation of the degree removed by Anavena pros-Aqui BT11 [KCTC 10852BP].

As described above, it can be seen that Anavena pros-Aqui BT11 [KCTC 10852BP] has a very high turbidity removal ability even at low concentrations (initiation of culture).

Anavena pros-Aqui BT11 [KCTC 10852BP] 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)

Anabaena flos-aquae BT11 [KCTC 10852BP] with turbidity removal ability. delete delete The turbidity removal method characterized by adding the Anabaena flos-aquae BT11 [KCTC 10852BP] to turbid water.

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