KR20040092067A - Novel strain Paenibacillus kribbensis AP-2 producing flocculant polysaccharide material - Google Patents

Abstract

PURPOSE: A microorganism Paenibacillus kribbensis AP-2 producing flocculant polysaccharide materials is provided, thereby environment-friendly purifying the polluted water and removing heavy metals from the water. CONSTITUTION: The microorganism Paenibacillus kribbensis AP-2(KCTC 10421BP) isolated from the polluted soil and water produces flocculant polysaccharide materials. The flocculant polysaccharide materials produced from Paenibacillus kribbensis AP-2(KCTC 10421BP) (i) have molecular weight of 230 kDa, (ii) are composed of glucose, mannose, glucuronic acid and fucose in a mole ratio of 5:1.86:1.4:1, (iii) have infrared spectrum shown in the figure 5, and (iv) have coagulation activity and turbidity removing activity.

Description

Novel strain Paenibacillus kribbensis AP-2 producing flocculant polysaccharide material

The present invention relates to a novel strain producing viscous polysaccharides, namely Paenibacillus kribbensis AP-2; accession number-KCTC10421BP, deposit date-February 10, 2003.

In order to supply clean water resources that are most needed for daily life, it is necessary to remove suspended solids (SS) and various organic and inorganic materials that cause turbidity in the water system in particular. Accordingly, for the removal of the solid suspended substances contained in various water systems (including purified water, sewage and wastewater, etc.), most of them are manufactured by chemical synthesis such as polyacrylamide (PAA, polyacrylamide), aluminum sulfate (Alum), and the like. The compounds to be used are mainly used for the treatment of aqueous systems. In particular, for water treatment, alum, polyaluminum chloride (PAC) or polyaluminum chloride silica (PACS) is mainly used.

However, as the above chemical compounds are continuously used for a long time as a water treatment agent, the problem caused by the accumulation of aluminum, which is a heavy metal in the case of alum, and a strong neurotoxin depending on the degree of use in the case of polyacrylamide (PAA) There are many concerns about the environmental and safety aspects of many chemical synthetic materials that are currently widely used in water treatment formulations such as wastewater and sewage treatment, such as the problem of being able to work. {Dearfield & Ambermathy, 1988, "Acrylamide: its metabolism, developmental and reproductive effects, genotoxicity, and carcinogenicity," Mutant Research. 195; 45-77}.

Therefore, there is a great need for the development of water treatment formulations and water treatment methods that are more environmentally friendly and safe to the human body, and have excellent control of environmental factors (including coagulation activity and turbidity, total nitrogen removal, chemical oxygen demand, and heavy metal removal). have.

The object of the present invention is more environmentally friendly and safer to the human body than the conventional water treatment formulations and water treatment methods using chemical synthetic materials, and environmental factors (coagulation activity and turbidity, total nitrogen removal ability, chemical oxygen demand, heavy metal removal, etc.). It is to provide a water treatment method having a better controllability of).

1 is a newly identified and separated one penny Bacillus Cri Ben cis -2 AP in the present invention, - a (Paenibacillus kribbensis AP-2 Accession No. KCTC10421BP) Scanning electron micrographs of the strains also (× 8,000).

Figure 2 is a schematic diagram showing the systematic relationship with other strains of the genus Penivacillus belonging to the strain of the present invention.

3A and 3B are cell fatty acid analysis (MIDI) results of the strain of the present invention.

Figure 4 is a graph of the HPLC analysis of the components of the viscous material obtained through the culture of the strain of the present invention.

Figure 5 is an infrared spectroscopic graph of the main functional groups of the viscous material obtained through the culture of the strain of the present invention.

Figure 6 is a graph showing the change in final pH, strain growth, viscosity and aggregation activity according to the culture time of the culture of the strain of the present invention.

7 is a view comparing the aggregation activity of the viscous substances obtained through the culture of the strain of the present invention, according to the carbon ratio in the culture conditions.

8 shows turbidity when treated with ferric chloride (FeCl ₃ ) in river water alone and when treated with viscous material obtained from the culture of the strain of the present invention after ferric chloride (FeCl ₃ ). It is a graph comparing (turbidity).

FIG. 9 is a graph comparing turbidity when treated with aluminum chloride (AlCl ₃ ) alone in a river water and treated with a slime obtained from the culture of the strain of the present invention after treatment with aluminum chloride (AlCl ₃ ). It is also.

10 is treated with aluminum sulfate alone (Al ₂ (SO ₄ ) ₃ ; Alum) in river water, and treated with a mixture (M / Alum) of a slime and aluminum sulfate obtained in the culture of the strain of the present invention. It is a graph comparing the turbidity and flocculating activity (FA) in the case.

FIG. 11 shows a case in which river water is treated with polyaluminum chloride (PAC) or polyaluminum chloride silica (PACS) alone, and a mixture of viscous material obtained from the culture of the strain of the present invention and PAC or PACS (M / PAC or It is a graph which compared the turbidity and aggregation activity in the case of M / PACS) treatment.

12 shows environmental factors in the case of treating polyaluminum chloride (PAC) alone in river water and in the case of treating a mixture (M / PAC) of a viscous substance and polyaluminum chloride obtained in the culture of the strain of the present invention. It is a graph which compared and observed the control effect (%) with respect to (turbidity, TOC, COD, and TN).

FIG. 13 shows the results of treatment of polyaluminum chloride (PAC) in river water alone and in the case of treatment of a mixture (M / PAC) of a viscous substance and polyaluminum chloride obtained in the culture of the strain of the present invention. It is a graph which compared and observed the removal effect (%) of aluminum ion ( ^{Al3 +} ) and copper ion (Cu2 ⁺ ).

In the present invention, there is provided a novel strain that produces viscous polysaccharide, ie Paenibacillus kribbensis AP-2 (Accession No. KCTC10421BP).

Preferably, the strains of the present invention can be effectively used for water treatment.

In addition, the present invention provides a viscous polysaccharide obtained by the cultivation of a Penibacillus cribensis AP-2 strain ( Paenibacillus kribbensis AP-2; Accession No. KCTC10421BP) and possessing the following characteristics:

i) the molecular weight is about 230,000 daltons;

ii) contains glucose, mannose, glucuronic acid and fucose as components in a molar ratio of 5: 1.86: 1.4: 1;

iii) the infrared spectroscopy spectrum of FIG. 5;

iv) possesses coagulation activity and turbidity removal activity.

"Viscous polysaccharide" of the present invention means a viscous polysaccharide material that is metabolized intracellularly and released extracellularly in the culture of the strain of the present invention, and the chemical characterization and collection method thereof is specifically described in the following examples. It is described.

Also, preferably, the viscous polysaccharide of the present invention can be effectively used for water treatment.

In one preferred aspect of the present invention, there is provided a viscous polysaccharide obtained by culturing the strain of the present invention under conditions of 3 to 10 in a culture medium condition.

In another preferred embodiment of the invention, the constituents of the culture medium comprise ammonium monophosphate, monopotassium phosphate, dipotassium phosphate, magnesium sulfate, manganese sulfate and calcium carbonate, and glucose and / or dextrin as carbon sources. To provide a viscous polysaccharide obtained by culturing the strain of the present invention in a culture medium. The constituents of the culture medium used to obtain the viscous polysaccharide of the present invention, in addition to the above components may include various kinds of components commonly used in culturing microorganisms such as yeast extract, soyton, tryptone, and the like, This is obvious to those skilled in the art.

As used herein, the term "water treatment" refers to the treatment of all water systems, including contaminated water, heavy water, sewage, and the like.

The novel strain provided by the present invention and the viscous polysaccharide obtained therefrom are more environmentally friendly and safer to the human body than the conventional methods of treating water using chemical synthetic materials, and at the same time environmental factors (coagulation activity and turbidity, total nitrogen). Controllability of removal, chemical oxygen demand, etc.) and removal of heavy metals provide better water treatment methods.

The following examples illustrate the invention in more detail, and these examples should not be construed as limiting the scope of the invention.

Example

Example 1 Isolation of the Strains of the Invention

Water and soil were collected from polluted water systems such as contaminated soils and wastewater treatment plants in northern Gyeongsangbuk-do and used as samples for the separation of new microorganisms.

First, glucose 10 g / l as a mixed medium, ammonium monophosphate 2 g / l, 0.8 g / l potassium phosphate, 0.6 g / l potassium diphosphate, 0.3 g / l calcium carbonate, 0.1 g / l soyton , Using a plate medium containing 0.1 g / l of tryptone, 0.1 g / l of yeast extract, 0.05 g / l of magnesium sulfate and 0.05 g / l of manganese sulfate, the suspension prepared from the sample using sterile saline solution The dilution method was used. The plated petri dishes were incubated for 1 to 3 days in an incubator at 30 ° C., and 20 kinds of microorganisms were isolated to produce a lot of viscous materials while showing excellent growth on plate medium.

After inoculating the separated 20 kinds of microorganisms in the same complex liquid medium, the culture medium was plated on the solid medium again to select 10 strains having good single colony formation and production of viscous substances. Then, the selected strains were placed in a culture test tube and incubated at 30 ° C. for 48 hours using the complex medium, and then the microorganisms forming a single colony were transferred to a test tube made of a slope medium and stored at 4 ° C. or 25% glycerol. Put and stored at -20 ℃ to use as needed.

10 microorganisms of the primary screened were inoculated by 2% (v / v) by using the above-described mixed liquid medium and shake-rotated at 30 ° C., and the aggregation activity was compared by using kaolin clay. Finally, a novel strain with good growth and good aggregation activity was finally identified and named as Paenibacillus kribbensis AP-2, and assigned to the Genetic Bank of Korea Institute of Science and Technology. Deposited (Accession Number-KCTC10421BP; Deposit Date-February 5, 2003).

Example 2: Microbiological Properties of the Strains of the Invention

(1) appearance analysis

Looking at the photograph taken with a scanning electron microscope for the strain of the present invention cultured in the composite medium of Example 1, the shell-like viscous polysaccharide is metabolized in the cell and produced after being secreted out of the cell It can be seen that it has (see Fig. 1).

In contrast, doubling the growth medium Luria Bertani medium (Luria-Bertani medium, aka LB medium-tryptone 10.0g / L, yeast extract 5.0g / L, sodium chloride 5.0g / L, agar 20.0g / L) When cultured in diluted media, the appearance was smooth.

(2) systematic analysis

In order to investigate the molecular relationship of the strains identified and isolated in the present invention, BLAST after amplification of 16S rDNA by polymer amplification (Polymerase Chain Reaction, PCR) using a method commonly used in the art. The sequence was analyzed using the program.

16S rDNA nucleotide sequence of the strain of the present invention is as follows.

As shown in FIG. 2, as a result of phylogenetic analysis, it was confirmed that the strain of the present invention belongs to the genus of Penivacillus, and the 16S rDNA sequence is different from other strains of the genus reported. For reference, the sequence similarity with the existing strain " Penibacillus cribbensis AM49 " found to be most similar to the 16S rDNA sequence of the strain of the present invention was 99%.

Through these results, it was confirmed that the Penibacillus kribbensis AP-2 strain ( Paenibacillus kribbensis AP-2; Accession No. KCTC10421BP) newly isolated and identified in the present invention is a new strain belonging to the genus Penivacillus.

(3) biochemical and physiological analysis

Biochemical and physiological analyzes of the strains of the present invention were performed according to methods currently widely used by those skilled in the art. See Cappuccino, James G. and Sherman, Natalie, MICROBIOLOGY, A LABORATORY MANUAL, 4ed., THE BENJAMIN / CUMMING PUBLISHING, INC., 1996; YOON et al., Int. Syst. Bacteriology. 48., 833-837, 1998}. The following results are compared with the strains of the present invention and are compared with the other known strain "Penibacillus polymixa DSM36" belonging to the genus Penivacillus (see: Fig. 2).

Strains of the Invention Paenibacillus polymixa DSM36 water 0 0 α-cyclodextrin - - β-cyclodextrin - + dextrin + + Glycogen + + Inulin - - Manan - - Twin 40 - - Twin 80 + - N-acetyl-D-glucoseamine + - N-acetyl-D-manosamine - - Amigdin - - L-Arabinose + + D-Arabitol - - Arbutin + + Cellobiose + + Fructose + + L-fusis - - D-galactose + -

D-galacturic acid - - Gentivios + - D-gluconic acid + - α-D-glucose + + m-inositol - - α-D-lactose + + Lactulose + + Maltose + + Maltorios + + D-mannitol + + D-Manos + + D-Melezitose - - D-Melibiose + + α-methyl-D-galactoside - - β-methyl-D-galactoside + - 3-methyl glucose + - α-methyl-D-glucoside - - β-methyl-D-glucoside + + α-methyl-D-manoside - - Palatine + - D-Sicos + - D-Raffinose + + L-Rhamnose - - D-ribose + + Salinity + + Sedoheptulic acid - - D-sorbitol - - Stachios + + Sucrose + + D-tagatose - - D-trehalose + + Turanos - - Xylitol - - D-Xylose + + Acetic acid + - α-hydroxybutyric acid - - β-hydroxybutyric acid - - γ-hydroxybutyric acid - - ρ-hydroxybutyric acid - - α-ketoglutaric acid - - α-ketovaleric acid - - Lactamide - - D-lactic acid methyl ester - - L-lactic acid - - D-malic acid - -

L-malic acid - - Methyl pyruvate - - Mono-methyl succinate - - Propionic acid - - Pyruvic acid - - Seoknamsan Mountain - - Succinic acid - - N-acetyl-L-glutamic acid - - Alanineamide - - D-alanine - - L-alanine - - L-alanyl-glycine - - L-glutamic acid - - Glysyl-L-Glutamic Acid - - L-pyroglutamic acid - - L-serine - - Putrescine - - 2,3-butanediol - + Glycerol - - Adenosine - - 2'-deoxy adenosine - - Inosine + - Undin - - Adenosine-5'-monophosphate - - Thymidine-5'-monophosphate - - Fructose-6-phosphate - - Glucose-1-phosphate - - D-L-α-glycerol phosphate - - ONPG - - Arginine Dihydrolase - - Lysine decarboxylated - - Ornithine Decoxylace - - Use of citrate - - H ₂ S production - - Eures - - Tryptophan Death Aminees - - Indole production - - VP + + Oxydays + + Galatinaise + - Glucose + + Mannitol inositol +- ++ Sorbitol - - Rhamnos - - Sucrose + + Melibiose + + Amigdalin + + Aebinos - -

In addition, Figure 3a and Figure 3b, after separating the cell fatty acid of the strain of the present invention by gas chromatography, shows the results of fatty acid analysis by the MIDI system.

Example 3 Analysis of Viscous Substances Produced by Strains of the Invention

Pure water formed after pouring the ethanol (98%) three times the volume of the culture solution to the supernatant obtained by appropriately diluting the culture obtained by culturing the strain of the present invention isolated in Example 1 by centrifugation, The white precipitate of viscous material was removed, dissolved again and dried by lyophilization. After the concentration of the sample thus obtained was about 1%, 2N trifluoroacetic acid was added, and hydrolysis was induced at high temperature and high pressure for 30 minutes, after cooling, neutralized, and the salt was removed to obtain a sample.

Sample analysis was predicted as per constituent using an RI analyzer (RID-6A, Shimadzu Co., Japan) through a carbopak NA1 column (flow rate-0.25 ml / min, column chamber temperature-room temperature) by High Pressure Liquid Chromatography (HPLC) As a result of analysis according to the time delay compared to the standard sugar, the viscous substance produced by the strain of the present invention contained glucose, mannose, glucuronic acid and fucose in a molar ratio of 5: 1.86: 1.4: 1. It was found to be a complex polysaccharide (see Figure 4).

In addition, in order to measure the molecular weight of the complex polysaccharide, it was prepared to be 0.3% (w / v) using a PL-GFC 1000Å column, and as a result of using dextran for various molecular weights of standard carbohydrates, the molecular weight of the complex polysaccharide was It turned out to be about 230,000 dalton.

On the other hand, in order to investigate the functional binding of the sugar structure and the structural component of the complex polysaccharide produced in the strain of the present invention, infrared spectroscopy analysis using a KBr pellet method [Model-Infiniti 60MI (Mattson, USA) ); Spectral range-400 to 4000 cm ^-1 ; Beam splitter-KBr] was obtained.

The complex polysaccharide produced by the strain of the present invention, a saccharide and a hydroxy at 3400cm ^-1 general peak (peak) in the protein thiol group (-OH groups), the CH stretching (stretching), 1600cm ^-1 2900cm ^-1 in and glucosamine R-NH ₂ group, specific for the glucan produced possible by the CH ₂ bond absorption peak at 1400cm ^-1 of, by showing a COC bond at 1080cm ^-1, which would normally be on a carbohydrate, a hydroxy activating group It was confirmed that it contained a siloxane group, a carboxyl group, an amino group and the like.

Example 4 Aggregation Activity of Cultures of the Strains of the Invention

A predetermined amount of the culture obtained by culturing the strain at 30 ° C. using the composite liquid medium described in Example 1 in a 5 L-fermentation tank was collected, and the strain growth, final pH, viscosity and aggregation activity according to the culture time were obtained. Investigate.

In this case, the coagulation activity is a control group which is irradiated with the absorbance at 550 nm wavelength and controls the coagulant without adding the coagulant in the reciprocal of the absorbance of the sample cell to which the coagulant (for example, the viscous polysaccharide obtained in the strain culture of the present invention) is added. The inverse of the absorbance of is obtained by subtracting the value. More specific methods are described in the literature (see Yokoi H. et al., 1995, Characteristics of biopolymer flocculant produced by Bacillus sp. PY-90, J. Ferment. Bioeng. 79, 378-380).

As can be seen in Table 2 and Figure 6, the final pH, viscosity, growth and aggregation activity was changed according to the incubation time, especially the cohesive activity of the viscous polysaccharide produced when cultured for about 48 hours to 60 hours This was found to be the highest. In particular, as measured in this experiment, the cohesive activity of viscous polysaccharides produced in the strain of the present invention, Paenibacillus kribbensis AP-2, was observed up to about 110. Has been shown to be superior to the cohesive activity of chemically synthesized materials (eg, in the case of Alum, about 71) that have been widely used as water treatment agents.

Characteristics change according to incubation time of the culture of the present invention Incubation time (hours) Final pH Viscosity (× 10 ³ ) Cell growth (OD, A ₆₆₀ ) Coagulation activity (F.A.) 12 5.92 1.02 0.15 39.66 24 5.75 1.85 0.28 49.65 36 5.64 2.03 1.25 99.65 48 5.45 2.63 2.54 110.77 60 5.40 2.88 3.45 110.77 72 5.52 2.87 2.85 99.65 84 5.60 2.85 2.24 82.99 100 5.78 2.75 2.24 66.32

Example 5 Agglomeration Activity of Viscous Polysaccharides Produced in the Strains of the Invention According to the Carbonity of Culture Medium

In the medium used for culturing the strain of the present invention, in the complex liquid medium described in Example 1, the other conditions are the same, and the strain is grown when the culture is carried out only by different ratios of carbon source (glucose) and nitrogen source (ammonium monophosphate). In addition, the final pH, viscosity and aggregation activity of the viscous polysaccharides obtained in the strain culture of the present invention were investigated, respectively. The results are shown in Table 3 below and FIG. 7.

C / N ratio ¹ (w / w) Final pH Viscosity (× 10 ³ ) Cell growth (A₆₆₀) Cohesive activity ² 1 μl 3 μl 5 μl 7 μl 10 μl 2.5 5.82 0.77 2.43 32.99 41.32 49.66 55.21 45.11 3.3 5.65 2.45 2.52 49.66 76.58 99.66 90.56 82.99 5 5.45 2.54 2.62 66.32 124.66 110.77 99.66 90.56 10 5.46 1.98 2.21 66.32 90.57 99.66 76.58 62.16 20 5.57 0.45 0.89 49.66 66.32 66.32 71.08 49.66

{In the table above,

1: After setting the total amount of glucose and ammonium monophosphate to 12.0 g / L, glucose and ammonium monophosphate were added to the medium according to the C / N ratio described above (for example, when the C / N ratio is 5, glucose Silver 10.0 g / l and ammonium monophosphate 2.0 g / l)

2: shows coagulation activity when treated with different amounts of viscous polysaccharide (ie 1, 3, 5, 7 and 10 μl) each obtained in a strain culture of the invention incubated according to different C / N ratios }.

As can be seen from Table 3 and Figure 7, the C / N ratio in the culture medium conditions 3.3, 5 and 10 was found to be particularly excellent in viscosity and aggregation activity, the C / N ratio is 5 The case was the best.

Example 6 Viscous Polysaccharide and Ferric Chloride (FeCl) Produced in a Strain of the Invention ₃ Agglomeration effect when using

Phenol was treated with ferric chloride (FeCl ₃ ) alone at different concentrations in 1 L of river water with 53.2 NTU (absorbance: 0.085), and then added with ferric chloride (FeCl ₃ ) at different concentrations. Turbidity when treated with 3 ml of viscous polysaccharide obtained in a culture of Baenibacillus kribbensis AP-2 was compared (see FIG. 8).

The characteristics of the river water (1ℓ) used in this experiment are shown in Table 4 below:

Characteristics of river water used in this experiment Turbidity (NTU) TOC (ppm) COD (ppm) T-N (ppm) Alkalinity (ppm) Zeta potential (mV) Al content (ppm) Cu content (ppm) 53.2 9.73 2.52 3.57 48.03 2.53 2.47 1.26

As can be seen in FIG. 8, coagulation activity was higher and turbidity was observed when viscous polysaccharide of Paenibacillus kribbensis AP-2 was used in combination with ferric chloride alone. The removal effect was also excellent. In particular, when about 100-125 ppm of ferric chloride (FeCl ₃ ) was added, then 3 ml of viscous polysaccharide obtained from a culture of Paenibacillus kribbensis AP-2 was added. It showed the highest flocculation activity and turbidity removal effect.

Example 7 Viscous Polysaccharide and Aluminum Chloride (AlCl) Produced by a Strain of the Invention ₃ Agglomeration effect when using

To the same river water 1ℓ as that used in Example 6, in the case where by varying the concentration of single treatment with an aluminum chloride (AlCl ₃₎ and, after at different concentrations was added to aluminum chloride (AlCl ₃₎ Penny Bacillus Cri Ben cis Turbidity when treated with 3 ml of viscous polysaccharide obtained from the culture of Ap-2 ( Paenibacillus kribbensis AP-2) was compared with each other (see Fig. 9). The properties of the river water used in this experiment are shown in Table 4 above.

As can be seen in Figure 9, compared with the case of using aluminum chloride alone, the coagulation activity is increased when the viscous polysaccharide obtained from the culture of Paenibacillus kribbensis AP-2 is used together. It was high and the turbidity removal effect was also excellent. In particular, the highest flocculation activity and turbidity were obtained when 3 ml of viscous polysaccharide obtained in the culture of Paenibacillus kribbensis AP-2 was added after the addition of about 10 to 40 ppm of aluminum chloride. Removal effect was shown.

Example 8: Coagulation Effect of Viscous Polysaccharide / Aluminum Sulfate Mixture (M / Alum) Produced in the Strain of the Present Invention

1 L (53.2 NTU) of the same river water as that used in Example 6 was treated with aluminum sulfate (Al ₂ (SO ₄ ) ₃ , Alum), which has been widely used as a water treatment formulation, alone, and Phenibacillus cribensis Turbidity when treated with a mixture of 3 ml of viscous polysaccharide obtained from the culture of Paenibacillus kribbensis AP-2 and aluminum sulfate (M / Alum; mixing ratios are listed in the table below) Flocculating Activity (FA) was compared with each other (see Fig. 10). The properties of the river water used in this experiment are shown in Table 4 above, and the comparison results are shown in Table 5 below.

Turbidity (NTU) Coagulation activity (F.A.) Alum sole 7.30 71.57 M / Alum (Strain Viscosity: Alum Content Ratio) 1:10 5.58 88.24 1:30 5.87 79.14 1:50 5.72 79.14 1:70 5.61 131.09 1: 100 4.95 131.09

As can be seen in Table 5 and Figure 10, compared to the case of using alum alone, viscous polysaccharides and alum obtained in the culture of Paenibacillus kribbensis AP-2 When the mixture (M / Alum) was used at the above ratio, the flocculation activity and turbidity removal effect were found to be excellent.

Example 9: "Mixed Viscous Polysaccharide / Aluminum Chloride Produced in a Strain of the Present Invention (M / PAC)" or "Mixed Viscous Polysaccharide / Aluminum Chloride Produced in a Strain of the Present Invention (M / PACS Agglomeration effect of

When 1 liter (53.2 NTU, absorbance 0.085) of the same river water used in Example 6 was treated with polyaluminum chloride (PAC) or polyaluminum chloride (PACS), which was previously used as a water treatment formulation, alone (30 ppm each). ); When treated with a mixture of viscous polysaccharides and polyaluminum chloride (M / PAC) obtained in a culture of Penibacillus cribensis epi-2; Turbidity, absorbance and flocculating activity in the treatment of a mixture of viscous polysaccharide and polyaluminum chloride silica (M / PACS) obtained from a culture of Penivacillus cribensis epi-2. FA) was compared with each other. The properties of the river water used in this experiment are shown in Table 4 above, and the comparison results are shown in Table 6 and FIG. 11 below.

Turbidity (NTU) Absorbance (ABS) Coagulation activity (F.A.) PAC alone 4.6 0.006 154.9 PACS only 5.7 0.008 113.2 M / PAC (Strain Viscosity: PAC Content Ratio) ^# 1: 1 6.2 0.011 79.1 1: 3 6.2 0.011 79.1 1: 5 6.1 0.008 113.2 1: 7 5.8 0.008 113.2 1:10 5.4 0.007 131.1 1:30 5.1 0.007 131.1 1:50 4.9 0.006 154.9 1:70 4.1 0.006 154.9 1: 1001: 200 4.54.8 0.0060.007 154.9131.1 M / PACS (strain viscosity: PACS content ratio) ^# 1: 1 10.9 0.018 43.8 1: 3 9.3 0.015 54.9 1: 5 9.2 0.015 54.9 1: 7 8.5 0.013 65.2 1:10 7.5 0.011 79.1 1:30 6.7 0.010 88.2 1:50 6.6 0.009 99.3 1:70 5.4 0.008 113.2 1: 1001: 200 4.75.7 0.0070.009 131.199.3

{In the above table, the total amount of # -M / PAC or M / PACS mixture is 30 μl each, and the viscous polysaccharide obtained from the culture of the strain is mixed with PAC or PACS according to the mixing ratio (for example , PAC content of 1:10 is 27.0ppm, PAC content of 1: 200 is 29.85ppm)}.

As can be seen in Table 6 and Figure 11, in terms of turbidity removal effect and cohesive activity, compared with the case of using polyaluminum chloride (PAC) or polyaluminum chloride (PACS) alone, penicillus cris When used in combination with viscous polysaccharides of Paenibacillus kribbensis AP-2, it showed at least a similar or better effect. Particularly, when the mixing ratio of the viscous polysaccharide and polyaluminum chloride or polyaluminum chloride silica of Paenibacillus kribbensis AP-2 is 1:10 to 1: 200 (weight ratio), the aggregation activity and Turbidity removal effect was found to be better.

Example 10 Effect on Environmental Factors of a Viscous Polysaccharide / Aluminum Mixture (M / PAC) Produced in a Strain of the Invention

When 1 liter of the same river water used in Example 6 was treated with 30 ppm of polyaluminum chloride (PAC), which is widely used as a water treatment formulation, alone, and Paenibacillus kribbensis AP-2 30 μl of a mixture of viscous polysaccharides and polyaluminum chloride (M / PAC) obtained in the culture of (wherein the ratio of strain viscous: PAC is 1:70 and PAC content is 29.57 ppm). The effect of control on environmental factors was compared and observed.

Specific properties of the river water used in this experiment are shown in Table 4 above. In addition, in this experiment, four environmental factors, such as chemical oxygen demand (COD), total organic carbon (TOC), turbidity, and total nitrogen (TN), were compared and observed. Same as 12.

Turbidity (NTU) TOC COD T-N River water 53.2 9.73 2.52 3.6 PAC 2.44 (control rate-95.4%) 6.60 (control rate-32.1%) 0.94 (control rate-70.5%) 1.98 (control rate-34.3%) M / PAC mixture (strain viscous: PAC = 1:70) 2.13 (control rate-96.0%) 4.48 (control rate-53.9%) 0.74 (control rate-79.5%) 1.69 (control rate-40.3%)

As can be seen in Table 7 and Figure 12, compared to the case of using the polyaluminum chloride (PAC) that is most commonly used as a conventional water treatment formulation of the Penibacillus cribensis AP-2 When mixed with viscous polysaccharide obtained from the culture, it was more environmentally friendly and showed excellent water treatment effect.

Example 11 Heavy Metal Removal Effect of a Viscous Polysaccharide / Poly Aluminum Chloride Mixture (M / PAC) Produced in a Strain of the Invention

When 1 liter of the same river water used in Example 6 was treated with 30 ppm of polyaluminum chloride (PAC), which is widely used as a water treatment formulation, alone, and Paenibacillus kribbensis AP-2 30 μl of a mixture of viscous polysaccharides and polyaluminum chloride (M / PAC) obtained in the culture of the present invention, wherein the ratio of the strain culture: PAC is 1:70, and the PAC content is 29.57 ppm. The removal effect of aluminum and copper ions contained in the river water was observed.

As can be seen in Table 8 and FIG. 13 below, Paenibacillus kribbensis AP-2 compared to the case of using polyaluminum chloride (PAC), which is most commonly used as a water treatment formulation, alone. When mixed with viscous polysaccharides obtained from the culture of the heavy metals including aluminum and copper was shown to have a great effect.

Al ⁺³ concentration (ppm) Cu ⁺² concentration (ppm) River water 2.472 1.264 PAC 0.558 (Removal rate-77.4%) 1.258 (Removal rate-0.5%) M / PAC mixture (strain viscosity: PAC = 1:70) 0.369 (Removal rate-85.1%) 1.206 (Removal rate-4.6%)

Claims (6)

Paenibacillus kribbensis AP-2; Accession No. KCTC10421BP to produce viscous polysaccharides. 2. The Penibacillus cribensis epi-2 strain according to claim 1, which can be used for water treatment. As a viscous polysaccharide obtained by culturing Paenibacillus kribbensis AP-2 (Accession No. KCTC10421BP), i) the molecular weight is about 230,000 daltons; ii) contains glucose, mannose, glucuronic acid and fucose as components in a molar ratio of 5: 1.86: 1.4: 1; iii) the infrared spectroscopy spectrum of FIG. 5; iv) viscous polysaccharides having coagulation activity and turbidity removal activity. 4. The viscous polysaccharide according to claim 3, which can be used for water treatment. The viscous polysaccharide according to claim 3 or 4, wherein the C / N ratio of the culture medium in the culture is 3 to 10. The method according to claim 3 or 4, wherein the constituents of the culture medium include ammonium monophosphate, monopotassium phosphate, dipotassium phosphate, magnesium sulfate, manganese sulfate and calcium carbonate, and glucose and / or dextrin as carbon sources. Viscous polysaccharide obtained by culturing in a culture medium comprising.