KR101859211B1 - Adsorption method of microbes for growth environment of microbes when putting microbes in concrete - Google Patents

Abstract

In order to create a microbial growth environment even after concrete hardening, microorganisms are adsorbed to a material capable of forming a microorganism growth environment without direct introduction of microorganisms into concrete, and the materials are injected into concrete to form microorganisms, A microbial adsorption method capable of preventing and continuously providing an environment for self-sustaining microorganisms is disclosed.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for adsorbing microorganisms to a microorganism,

TECHNICAL FIELD The present invention relates to a technique for introducing a microorganism into concrete, and more particularly, to a method for adsorbing microorganisms for forming a microorganism growth environment upon introduction of microorganisms into concrete.

As the recent economic level has improved, interest in the environment has increased, and discussions on the production of environmentally friendly concrete using microorganisms have become more active in concrete formulation.

Conventionally, in order to inject microorganisms into concrete, it is a simple method to use a method of replacing a certain part or all of the compounding water with a microbial culture liquid during the concrete compounding process. In this method, the moisture content of the cement paste is decreased due to the hydration of the cement paste during the curing process, and the nutrients and growth of the microorganisms are reduced due to the volume expansion. Especially, the concrete is hardened by the hydration heat and the internal temperature becomes 40 ℃ or higher. The strongly alkaline environment (pH 11 ~ 12) caused by the hydration product (Ca (OH) ₂ ) of concrete is the optimum growth environment ) Is the largest inhibitory factor in the composition. These effects inhibit the growth of microorganisms inside the concrete, and after 30 days, all of the microorganisms within the concrete will die.

[Prior Patent Literature]

- Korean Registered Patent No. 10-1273444 (March 31, 2013)

- Korean Registered Patent No. 10-0481287 (Mar. 28, 2005)

The object of the present invention is to provide a microbial growth environment even after concrete hardening. It is an object of the present invention to adsorb a microorganism to a material capable of forming a microbial growth environment without directly inputting microbes into a concrete mixture, To prevent the death of mycobacteria, and to provide an environment for continuous microbial self-growth.

In order to solve the above-mentioned problems, the present invention provides a method for manufacturing a micro-organism comprising the steps of charging a sorbent material into a net-like adsorption pad having a mesh size of 100 to 5 mm and a thickness of 0.5 to 50 mm, The present invention provides a method for adsorbing microorganisms for forming a microorganism growth environment in concrete mixing using microorganisms.

And the material of the adsorption pad is a steel material.

And the adsorbent is at least one selected from the group consisting of expanded vermiculite, diatomaceous earth, and superabsorbent resin.

In addition, the microorganism adsorption using the adsorbent is carried out by immersing the microbial culture broth in an amount of 1 to 30 parts by weight based on 100 parts by weight of the microbial culture broth, maintaining the humidity at 40 to 80% and temperature of 5 to 40 DEG C for 1 to 10 days The method comprising the steps of:

Also, the microbial adsorption using the adsorbent is performed by putting the adsorbent into the mesh-type adsorption pad and then floating the adsorbent pad in the culture solution of the microorganism.

And the weight of the counterweight is determined by the following equation (1): " (1) "

[Equation 1]

(Where d is the immersion depth of the adsorption pad, W _L is the loading force, W _S is the fixed load, L and B are the length and width of the adsorption pad, and γ _w is the unit volume weight of the culture medium).

The microorganism may also be Rhodoblastus acidophilusRhodoblastus acidophilus), Bacillus alcalophilus (Bacillus alcalophilus), Geobacillus caldosilocylticus (Geobacillus caldoxylosilyticus), Anenurina Bacillus Surmoa Elofil (Aneurinibacillus thermoaerophilus), Rhodobacter capsulatus (Rhodobacter capsulatus), Rhodopseudomonas palustris (Rhodoseudomonas palustris), Bacillus thuringiensis and Bacillus subtilis (Bacillus subtilis). ≪ / RTI >

Rhodoblastus acidophilus is prepared by adding 0.05 to 1 g of yeast extract, 0.1 to 5 g of di-sodium succinate, 0.1 to 5 g of ferric citrate solution, 0.1 to 1 g of potassium dihydrogenphosphate (KH ₂ PO ₄ ), 0.1 to 1 g of magnesium sulfate heptahydrate (MgSO ₄ .7H ₂ O), 0.1 to 1 g of sodium chloride (NaCl) 0.1 to 1 g of ammonium chloride (NH ₄ Cl) and 20 to 100 ml of calcium chloride dihydrate (CaCl ₂ .2H ₂ O) at a pH of 5 to 7.

The above-mentioned Bacillus alcalophilus may be prepared by adding 0.5 to 5 g of beef extract, 0.5 to 5 g of yeast extract, 1 to 10 g of peptone, 1 to 10 g of sodium chloride, In a medium containing 20 to 200 ml of Na-sesquicarbonate solution (containing 2 to 6 g of sodium hydrogencarbonate (NaHCO ₃ ) and 3 to 7 g of anhydrous sodium carbonate (Na ₂ CO ₃ anhydrous) per 100 ml of purified water) To < RTI ID = 0.0 > 11. ≪ / RTI >

The above-mentioned Geobacillus caldoxylosilyticus and Aneurinibacillus thermoaerophilus may be used in an amount of 0.5 to 5 g / l of beef extract, 0.5 to 5 g of yeast extract, pH 4 to 10 and 37 to 80 ° C in a nutrient agar medium containing 1 to 10 g / l of peptone, 1 to 10 g / l of sodium chloride and 5 to 25 g / l of agar, , ≪ / RTI >

The above-mentioned Rhodobacter capsulatus and Rhodoseudomonas palustris contain 0.1 to 5 g of yeast extract, 0.1 to 5 g of disodium succinate hexa-hydrate, 0.1 to 5 g of absolute ethanol, 0.1 to 1 ml of ferric citrate solution, 0.1 to 5 ml of ferric citrate solution, 0.1 to 1 g of potassium dihydrogenphosphate (KH ₂ PO ₄ ), 0.1 to 1 g of magnesium sulfate heptahydrate (MgSO ₄ .7H ₂ O) Characterized in that it is cultivated at a pH of 5 to 7 in a medium comprising 0.1 to 1 g of ammonium chloride (NaCl), 0.1 to 1 g of ammonium chloride (NH ₄ Cl) and 0.02 to 0.1 g of calcium chloride dihydrate (CaCl ₂ .2H ₂ O) .

In addition, Bacillus thuringiensis and Bacillus subtilis are prepared by adding 1 to 10 g of peptic digest of animal tissue, 0.5 to 3 g of yeast extract, 1 to 10 g of sodium chloride and 0.5 to 3 g of beef extract.

According to the present invention, in the production of concrete using microorganisms, there is an effect of preventing the microorganisms from being killed after curing of concrete and providing a continuous growth environment.

In addition, it is possible to provide a method of making economical and large amount of microorganisms easier and substantially capable of adsorption, as compared with the prior art, such as capsule technology for injecting microorganisms.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view illustrating a culture medium container (b) in which an adsorption pad (a) for adsorbing microorganisms and a adsorption pad are immersed, according to an embodiment of the present invention;
FIG. 2 is a schematic view showing a section of a concrete formed after the adsorption of a microorganism-absorbing material according to an embodiment of the present invention, FIG.
FIG. 3 is a graph showing the distribution of Rhodobacter capsulatus , Rhodoseudomonas < RTI ID = 0.0 > palustris), Bacillus emitter Lindsay N-Sys (Bacillus thuringiensis), and Bacillus subtilis (Bacillus subtilis) a photograph showing a scanning electron microscopic (SEM) analysis results for the adsorption capacity of the adsorbent was evaluated sorbent material (expanded vermiculite),
4 is a photograph showing a result of a scanning electron microscope (SEM) analysis for evaluating the adsorption performance of an adsorbent (superabsorbent resin) adsorbing Rhodobacter capsulatus .

Hereinafter, the present invention will be described in detail with reference to examples. Prior to this, terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms, and the inventor should appropriately interpret the concepts of the terms appropriately The present invention should be construed in accordance with the meaning and concept consistent with the technical idea of the present invention. Accordingly, it is to be understood that the constituent features of the embodiments described herein are merely the most preferred embodiments of the present invention, and are not intended to represent all of the inventive concepts of the present invention, so that various equivalents, And the like.

The inventors of the present invention have conducted intensive studies to prevent the microorganisms from being killed after concrete hardening and to form a continuous growth environment in concrete production using microorganisms. As a result, it has been found that a sorbent material in a mesh- To prevent the microorganisms from being killed and to form a continuous growth environment even after concrete hardening.

Accordingly, the present invention relates to a method for preparing a concrete using microorganisms, which comprises a step of putting a sorbent material into a net-like adsorption pad having a mesh eye size of 100 to 5 mm and a thickness of 0.5 to 50 mm, and then immersing the adsorption pad in a microorganism culture liquid. Disclosed is a microbial adsorption method for forming a microbial growth environment.

In the present invention, as a material for microbial adsorption, a material having a high ability of exchanging a cationic cation can be used, and expanded vermiculite, diatomaceous earth, superabsorbent resin and the like can be used.

When a microorganism is simply injected in the production of concrete using microorganisms, growth is slowed or killed due to lack of moisture after concrete hardening. In order to create an environment in which microorganisms can grow in the interior of hardened concrete, the present invention has a porous structure (effective water content of not less than 40% by volume and porosity of not less than 50%) due to innumerable pores, So that the microorganisms are adsorbed before the concrete is introduced.

The microbial adsorbent materials disclosed in the present invention adsorb organic substances by exchangeable cations (Mg ^{2 +} , Ca ^{2 +,} etc.) present on the surface of the material and absorb the organic nutrients (medium components) necessary for microbial and microbial growth do. In addition, pH is 6 ~ 9, which is the ideal material for optimal environment for microbial growth. On the other hand, porous perlite, cork, and the like have no cation exchange ability and are not preferable as the microbial adsorbent in the present invention.

An immersion process is used for adsorption of microorganisms using the adsorbent. In this case, in the immersion condition for the optimum absorption efficiency of the microorganism, the cultured microorganism culture liquid is added to the microorganism culture liquid in an amount of 1 to 30 parts by weight based on 100 parts by weight of the microorganism culture liquid, after immersion, at a humidity of 40 to 80% More preferably 5 to 15 parts by weight for a period of 2 to 5 days at a temperature of 10 to 30 DEG C and a humidity of 50 to 70% after immersion.

On the other hand, the adsorbent floats when immersed in the culture liquid with a specific gravity of less than 1.0. Therefore, in the present invention, a method of effectively adsorbing microorganisms is considered. FIG. 1 shows an exemplary adsorbent pad (a) for adsorbing microorganisms and a culture tank (b) immersed in adsorbent pads.

1, the adsorbent is introduced and discharged through the opening and closing clip 110. When the plurality of adsorption pads 120 are used, the adsorption pads 120 are connected to each other by using the connecting rod 130, The counterweight 140 is connected to the lowermost adsorption pad 120 so that the pad 120 floats while maintaining a constant spacing. In addition, FIG. 1 shows a screw-type openable lid 150, a threaded opening 160 and an annular fin 170 for preventing contamination by other microorganisms.

The mesh type adsorption pad which can accommodate the adsorbent such as expanded vermiculite is preferably manufactured using a steel material such as aluminum. The size of the mesh is determined in consideration of the particle size of the adsorbent material to be used, The size of the mesh eye is in the range of 100 탆 to 5 mm and the thickness of the mesh type adsorption pad is in the range of 0.5 to 50 mm. Preferably, the mesh eye size is 300 탆 to 3 mm and the thickness of the mesh- May be used in the range of 1 to 30 mm, more preferably 500 to 1 mm in mesh eye size and 2 to 10 mm in mesh adsorption pad thickness.

The length and width of the mesh adsorption pad vary with the diameter of the culture vessel. The mesh-type adsorption pad is provided with an opening-closing clip for freely opening and closing the adsorbent, and an annular pin is provided for fixing. The mesh type adsorption pad is connected to each other by using a steel rod such as aluminum. The weight of the counterweight for the immersion depth design is determined from the following equation (1).

[Equation 1]

In Equation 1, d is the immersion depth of the adsorption pad, W _L is the loading force, W _S is the fixed load, L and B are the length and width of the adsorption pad, and γ _w is the unit volume weight of the culture liquid. The fixed load (W _s ) among the elements for setting the target immersion depth (d) may vary depending on the volume and the specific gravity of the adsorption pad, and accordingly, the loading load (W _L ) Depth can be secured.

As described above, in the present invention, the microbial adsorption using the adsorbent can be performed by adsorbing the adsorbent material on the mesh-type adsorbing pad 120 and floating the adsorbing pad 120 in the culture liquid of the microorganism, Can float in the middle of the culture medium's depth and adsorb microorganisms and organic nutrients (media components) evenly.

FIG. 2 schematically shows a cross-section of a concrete formed after the adsorbent to which microorganisms have been adsorbed, according to an embodiment of the present invention.

Referring to FIG. 2, when the sorbent material to which the microorganisms are adsorbed is put into the form of concrete 210, that is, a mixture containing aggregate, cement, and water, the microorganisms are allowed to grow between the aggregates. The amount of the adsorbent to be added is preferably 1 to 30 parts by weight based on 100 parts by weight of the cement so that the required bending strength of the concrete can be secured. The amount of the adsorbent is preferably 5 to 20 parts by weight.

The microorganism used in the microorganism adsorption method according to the present invention is not particularly limited as long as it is a microorganism suitable for the immersion process using the adsorption pad loaded with the adsorbent material. Preferably, the microorganism is a thermophilic material capable of imparting self- It can grow in high temperature environment (40 ~ 50 ℃) and alkaline environment (pH 4 ~ 10) and can be used as organic fertilizer by water purification and nitrogen fixation. In the present invention, among the isolated microorganisms, Rhodoblastus acidophilus , Bacillus alcalophilus , Geobacillus caldoxylosilyticus , Aneurinibacillus , Bacillus thuringiensis, thermoaerophilus , Rhodobacter capsulatus , Rhodoseudomonas spp. palustris), Bacillus emitter Lindsay N-Sys (Bacillus thuringiensis), and Bacillus subtilis (Bacillus subtilis) if the selected microorganisms used in the soaking process using a suction pad In the particular adsorbent after the concrete mix and cured using the same of microorganisms injected in the concrete It was confirmed that it is possible to prevent the death and to create a sustainable growth environment.

Here, the present invention suggests optimal culture conditions and culture conditions with excellent efficiency in seed culture according to the characteristics of each microorganism based on the result of study on the special medium composition to form the optimum culture environment of the microorganisms.

Specifically, Rhodoblastus acidophilus is prepared by adding 0.05 to 1 g of yeast extract, 0.1 to 5 g of di-sodium succinate, 0.1 to 5 g of ferric citrate 0.1 to 1 g of potassium dihydrogenphosphate (KH ₂ PO ₄ ), 0.1 to 1 g of magnesium sulfate heptahydrate (MgSO ₄ .7H ₂ O), 0.1 to 1 g of sodium chloride (NaCl) The yeast extract is preferably cultured at a pH of 5 to 7 in a medium containing 0.1 to 1 g of ammonium chloride (NH ₄ Cl) and 0.1 to 1 g of calcium chloride dihydrate (CaCl ₂ .2H ₂ O) 0.1 to 0.3 g of extract, 0.5 to 2 g of Di-sodium succinate, 3 to 7 ml of ferric citrate solution (including 0.1 g per 100 ml), potassium dihydrogen phosphate (KH ₂ PO ₄₎ 0.3 ~ 0.7g, magnesium sulfate heptahydrate _{(MgSO 4 .7H 2 O) 0.2} ~ 0.6g, sodium chloride (NaCl) 0.2 ~ 0.6g, ammonium chloride (NH ₄ Cl) 0.2 ~ 0.6g and calcium chloride hydrate (CaCl ₂ .2H ₂ O). Lt; RTI ID = 0.0 > pH ~ 5.5 < / RTI >

The above-mentioned Bacillus alcalophilus may be prepared by adding 0.5 to 5 g of beef extract, 0.5 to 5 g of yeast extract, 1 to 10 g of peptone, 1 to 10 g of sodium chloride, In a medium containing 20 to 200 ml of Na-sesquicarbonate solution (containing 2 to 6 g of sodium hydrogencarbonate (NaHCO ₃ ) and 3 to 7 g of anhydrous sodium carbonate (Na ₂ CO ₃ anhydrous) per 100 ml of purified water) Preferably, it is preferably cultivated under the conditions of 1 to 11%, and preferably 0.5 to 2 g of beef extract, 1 to 3 g of yeast extract, 3 to 7 g of peptone, 3 to 7 g of sodium chloride and a sodium sesquicarbonate solution (Na- sesquicarbonate solution (containing 2 to 6 g of sodium hydrogencarbonate (NaHCO ₃ ) and 3-7 g of anhydrous sodium carbonate (Na ₂ CO ₃ anhydrous) per 100 ml of purified water) and cultured at a pH of 9 to 10 in a medium containing 80 to 120 ml Is more preferable.

The above-mentioned Geobacillus caldoxylosilyticus and Aneurinibacillus thermoaerophilus may be used in an amount of 0.5 to 5 g / l of beef extract, 0.5 to 5 g of yeast extract, pH 4 to 10 and 37 to 80 ° C in a nutrient agar medium containing 1 to 10 g / l of peptone, 1 to 10 g / l of sodium chloride and 5 to 25 g / l of agar, , And it is preferable that be cultivated in a medium containing 0.5 to 2 g / l of beef extract, 1 to 3 g / l of yeast extract, 3 to 7 g / l of peptone, 3 to 7 g / l and agar 10 to 20 g / l in a nutrient agar culture medium at pH 5 to 9 and 40 to 60 ° C.

In addition, photosynthetic bacteria such as Rhodobacter capsulatus , Rhodoseudomonas spp. palustris was prepared by adding 0.1 to 5 g of yeast extract, 0.1 to 5 g of disodium succinate hexa-hydrate, 0.1 to 1 ml of absolute ethanol, and an iron citrate solution 0.1 to 1 g of ferric citrate solution, 0.1 to 1 g of potassium dihydrogenphosphate (KH ₂ PO ₄ ), 0.1 to 1 g of magnesium sulfate heptahydrate (MgSO ₄ .7H ₂ O), 0.1 to 1 g of sodium chloride (NaCl) NH ₄ Cl) and 0.02 to 0.1 g of calcium chloride dihydrate (CaCl ₂ .2H ₂ O), preferably 0.5 to 2 g of yeast extract, 0.5 to 2 g of disodium succinate hexa-hydrate, 0.3 to 0.7 ml of absolute ethanol, 0.5 to 1.5 ml of ferric citrate solution, potassium dihydrogen phosphate (KH ₂ PO ₄₎ 0.3 ~ 0.7g, magnesium sulfate heptahydrate _{(MgSO 4 .7H 2 O) 0.2} ~ 0.6g, sodium chloride (NaCl) 0.2 ~ 0.6g, ammonium chloride (NH ₄ Cl) 0.2 ~ 0.6g calcium chloride dihydrate and (CaCl ₂ .2H ₂ O) is more preferably pH 5.5 ~ 6.5 in the culture conditions in a culture medium containing 0.03 ~ 0.07g.

In addition, Bacillus thuringiensis and Bacillus subtilis are prepared by adding 1 to 10 g of peptic digest of animal tissue, 0.5 to 3 g of yeast extract, It is preferably cultured at a pH of 4 to 10 in a medium containing 1 to 10 g of sodium chloride and 0.5 to 3 g of beef extract and preferably 3 to 7 g of peptic digest of animal tissue of animal tissue , 1 to 2 g of yeast extract, 3 to 7 g of sodium chloride, and 1 to 2 g of beef extract, more preferably at a pH of 6 to 8.

The microorganisms may be cultured in an incubator for 3 to 12 days, preferably 5 to 9 days, after being inoculated in each culture medium.

As described above, the present invention can solve the microbial death problem, which is a serious problem of the microbial application method when the conventional concrete is mixed through the porous material, the adsorption pad and the culture liquid of the specific composition used as the microbial growth place.

Hereinafter, the present invention will be described in detail with reference to examples.

Microbial Isolation and Culture

TSA medium (15 g of tryptone, 5 g of soytone, 5 g of sodium chloride and 15 g of agar in 15 ml of distilled water was adjusted to pH 7 by taking 50 ml of sterilized distilled water and mixing it with 400 ml of sterilized distilled water. After autoclaving sterilization at 120 ° C for 15 minutes, cooling to 60 ° C and dispensing in a sterilized culture dish) was added and inoculated by the smear method. The inoculated microorganisms were separated and identified by 16S rDNA sequencing. The identified microorganisms were identified as Rhodoblastus acidophilus (Accession No. KEMB (Korea Environmental Microorganisms Bank, 9000-009) , Bacillus alcalophilus (see Accession No. KEMB9000-002), Geobacillus caldoxylosilyticus (Accession No. KEMB563-529), Aneninibacillus thermoairophilus Aneurinibacillus thermoaerophilus (see Accession No. KEMB563-526), Rhodobacter capsulatus (see accession number KEMB1-2), Rhodoseudomonas palustris (see accession number KEMB1-6), Bacillus terrine Ensure that the N-Sys (see accession number KEMB4-218) (Bacillus thuringiensis) (see accession number KEMB4-215) and Bacillus subtilis (Bacillus subtilis) The.

Culture of each of the isolated microorganisms was carried out using media of various compositions. In the case of Rhodoblastus acidophilus and Bacillus alcalophilus , 6.0, 27 ' s media) and Table 2 (pH 9.7, Nutrient Broth), and the best growth was observed in the media composition and culture conditions, and Geobacillus caldoxylosilyticus and Aneurinobacillus thermoairophilus In the case of Aneurinibacillus thermoaerophilus , the highest growth was observed in the medium composition and culture conditions shown in Table 3 (pH 6), and Rhodobacter capsulatus and Rhodoseudomonas < RTI ID = 0.0 > to cases when the following Table 4 (showed the best growth in a culture medium composition and the culture conditions shown to pH 6), Bacillus emitter Lindsay N-Sys (Bacillus thuringiensis), and Bacillus subtilis (Bacillus subtilis) of palustris) Table 5 (pH 6 ) Showed the best proliferation in the medium composition and culture conditions shown in Table 1. In the following experiment, the cultured microorganism culture was used.

composition Yeast extract 1g Disodium succinate hexa - dydrate 1g Absolute ethanol 0.5ml Ferric citrate solution (0.5% (w / v)) 1ml KH2PO4 0.5 g MgSO4 ₄ · 7 H ₂ O 0.4 g NaCl 0.4 g NH ₄ Cl 0.4 g CaCl ₂ · 2 H ₂ O 0.05 g Trace element solution ( SL -6, see below) 1ml Distilled water 1L Trace element solution ZnSO ₄ · 7 H ₂ O 0.1 g MnCl ₂ · 4 H ₂ O 30 mg H ₃ BO ₃ 0.3 g CoCl ₂ · 6 H ₂ O 0.2 g NiCl ₂ · 6 H ₂ O 20 mg CuCl ₂ · 2 H ₂ O 10 mg Na ₂ MoO ₄ · 2 H ₂ O 30 mg Distilled water 1L

Composition Peptic digest of animal tissue 5g Yeast Extract 1.5 g Sodium Chloeide 5g Beef extract 1.5 g Distilled water 1L

Microbial-adsorbed Absorbent material  Produce

In the case of Rhodoblastus acidophilus , Bacillus alcalophilus , Geobacillus caldoxylosilyticus and Aneurinibacillus thermoaerophilus , in the case of Aneurinibacillus thermoaerophilus , 1, each of the prepared microorganism culture solutions was added to the culture solution bottle at a concentration of 10 ³ cells / ml, 10 ⁵ cells / ml and 10 ⁹ cells / ml, and 10 parts by weight Was placed in an adsorption pad (mesh size 700 × 700 μm, pad thickness 5 mm, 5-step connection), and immersed and suspended in the culture solution pail (weight of equilibrium weight was determined according to Equation 1 above) And stored for 72 hours in an environment of 60% humidity and 20 < 0 > C temperature. Then, the adsorption pad was taken out from the culture solution pail, and the adsorbent adsorbed by the microorganism was recovered through the opening and closing clip.

In addition, Rhodobacter capsulatus , Rhodoseudomonas < RTI ID = 0.0 > palustris), Bacillus emitter Lindsay N-Sys (Bacillus thuringiensis), and Bacillus subtilis (with reference to Figure 1 for the Bacillus subtilis), into each of said prepared microbial culture to the culture tube with 10 ⁵ cell / ㎖ concentration, each microbial culture 10 parts by weight of expanded vermiculite and superabsorbent resin (powder type, Rhodobacter capsulatus only) of 100 parts by weight were introduced into a suction pad (mesh size 700 × 700 μm, pad thickness 5 mm, connected in 5 steps) And the suspension was immersed and suspended in the culture solution bottle (the weight of the counterweight was determined according to the formula (1)), and then the lid was closed and stored for 72 hours in an environment of a humidity of 60% and a temperature of 20 ° C. Then, the adsorption pad was taken out from the culture solution pail, and the adsorbent adsorbed by the microorganism was recovered through the opening and closing clip.

The average immersion depth (d) is a load weight (W _L) 10kg (5kg counterweight 2), (5 2kg suction pad) Dead load (W _S), 10kg of the absorbent suction pad during manufacture wherein the microorganism adsorbed, adsorption The pad size (L x B = 0.3 m x 0.3 m) and the unit volume weight (y _w ) of the culture medium were determined to be about 0.22 m from 1,000 kg / m 3.

Among the adsorbents adsorbed by the produced microorganisms, Rhodobacter capsulatus , Rhodoseudomonas spp. palustris), Bacillus emitter Lindsay N-Sys (Bacillus thuringiensis), and Bacillus subtilis (Bacillus subtilis) the order to evaluate the adsorption performance of adsorbing adsorbent 3 for analysis by scanning electron microscopy (SEM) results (expansion using vermiculite) and 4 (Use of a superabsorbent resin, observation of inner and surface structure shapes). In FIG. 3, results for the case of not using microorganisms for comparison are also shown. As shown in Fig. 3 and Fig. 4, it can be seen that the microorganism exhibits a good adsorption state (see the circle part).

Standard mixing test of cement mortar

Among the adsorbents prepared above, Rhodoblastus acidophilus , Bacillus alcalophilus , Geobacillus lucilus, caldoxylosilyticus) and Annette us your cement mortar standard formulation for the case with the filler's (Aneurinibacillus thermoaerophilus) with the collected written Bacillus (cement: sand weight ratio of 1: sand volume compared to 0% (control) in one), 5%, 7.5% And 10%, and the compressive strength and pH of the mortar were measured over time after the mortar was produced. In Table 6 and Table 7 for Rhodoblastus acidophilus , Bacillus alcalophilus for Bacillus alcalophilus , and in Table 8 and Table 9 for Rhizoblastus acidophilus , ( Geobacillus caldoxylosilyticus ) are shown in Tables 10 and 11, and in case of Aneurinibacillus thermoaerophilus , the results are shown in Tables 12 and 13 below.

Referring to Tables 6 to 13, in the mortar mixture prepared using the microorganisms according to the present invention, the microorganisms can be continuously grown at a level of pH 10 or less regardless of the concentration of the culture, It is possible to confirm that the growth of the microorganism can be inhibited or killed at pH 11 or higher.

On the other hand, in terms of the compressive strength of the mortar, when the adsorbent is replaced by 10% by volume, the adsorbent may need to be maintained at a proper level.

microbe Absorbent material  Manufacture of concrete spheres by injection

100 parts by weight of ordinary Portland cement, 30 parts by weight of water, and 100 parts by weight of Rhodoblastus acidophilus (10 ⁵ cells / ml) and Bacillus alcalophila (100 parts by weight) Bacillus alcalophilus ) (10 ⁵ cells / ml), 10 parts by weight of the adsorbed adsorbent was compounded, and the compounded mixture was added to the mold and cured to form a concrete sphere. (Production Examples 1 and 2)

On the other hand, for comparison, concrete spheres were prepared in the same manner as in Preparation Examples 1 and 2 (Comparative Preparation Examples 1 and 2), except that the microbial culture solution was directly introduced without adsorbing the microorganisms.

Evaluation of self-purification ability of concrete spheres

In order to evaluate the self - purification ability of the concrete sphere, the wastewater obtained from the livestock wastewater treatment plant (Yeoju) was used as the raw water and the water purification capacity was measured. The reactor type used for the wastewater treatment in this experiment was a batch type reactor in which there was no flow and the liquid was completely mixed in the reactor. The water tank for the water purification experiment was a 25 liter acrylic tank, the volume of the reactor was adjusted to 20 liters, and the water was circulated using a pump. Six reaction vessels were prepared and the concrete spheres of Production Examples 1 and 2 and Comparative Production Examples 1 and 2 were placed in a water bath and the water purification capacity was measured. The measurement was carried out four times for two weeks. The chemical oxygen demand (COD) was determined by the reactor digestion (HS-2300Plus), the biological oxygen demand (BOD) by the BOD sensor system, the total nitrogen (TN) by the chromotropic acid HS-2300Plus) and total phosphorus (TP) were measured using a molybdo vanadate (HS-2300Plus). The results are shown in Tables 14 to 17 below.

Tables 14 to 17 show that the concrete spheres according to the preparation examples have higher values of chemical oxygen demand (COD), biological oxygen demand (BOD), total nitrogen (TN) and total phosphorus (TP) removal efficiency is remarkably excellent.

While the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, Such modifications and changes are to be considered as falling within the scope of the following claims.

110: retractable clip 120: suction pad
130: Steel rod for connection 140:
150: openable lid 160: threaded opening
170: annular pin 210: concrete sphere

Environmental microbial bank KEMB9000-009 20160509 Environmental microbial bank KEMB9000-002 20110608 Environmental microbial bank KEMB563-529 20160204 Environmental microbial bank KEMB563-526 20160204 Environmental microbial bank KEMB1-2 20101124 Environmental microbial bank KEMB1-6 20101124 Environmental microbial bank KEMB4-215 20080228 Environmental microbial bank KEMB4-218 20080228

Claims (17)

The method comprising the steps of: injecting an adsorbent into a mesh-type adsorption pad having a size of mesh mesh of 100 μm to 5 mm and a thickness of 0.5 to 50 mm, and then immersing the adsorption pad in a microbial culture solution; A method for adsorbing microorganisms,
The microbial adsorption using the adsorbent is carried out by immersing the microbial culture broth in an amount of 1 to 30 parts by weight based on 100 parts by weight of the microbial culture broth and maintaining the humidity at 40 to 80% and temperature of 5 to 40 DEG C for 1 to 10 days Wherein the microorganism adsorbing method comprises the steps of: The method comprising the steps of: injecting an adsorbent into a mesh-type adsorption pad having a size of mesh mesh of 100 μm to 5 mm and a thickness of 0.5 to 50 mm, and then immersing the adsorption pad in a microbial culture solution; A method for adsorbing microorganisms,
The microbial adsorption using the adsorbent is performed by putting the adsorbent on the mesh-type adsorption pad and then floating the adsorption pad in the culture medium of the microorganism,
Wherein the adsorption pad is suspended by an equilateral weight connected to a lower end of the adsorption pad, and the weight of the equilibrium weight is determined by the following equation (1).
[Equation 1]

(Where d is the immersion depth of the adsorption pad, W _L is the loading force, W _S is the fixed load, L and B are the length and width of the adsorption pad, and γ _w is the unit volume weight of the culture medium). The method comprising the steps of: injecting an adsorbent into a mesh-type adsorption pad having a size of mesh mesh of 100 μm to 5 mm and a thickness of 0.5 to 50 mm, and then immersing the adsorption pad in a microbial culture solution; A method for adsorbing microorganisms,
The microorganisms may be used in an amount of 0.05 to 1 g of yeast extract, 0.1 to 5 g of di-sodium succinate, 2 to 10 g of ferric citrate solution (including 0.1 g per 100 ml) ㎖, potassium dihydrogen phosphate _{(KH 2 PO 4) 0.1 ~} 1g, magnesium sulfate heptahydrate _{(MgSO 4 .7H 2 O) 0.1} ~ 1g, sodium chloride (NaCl) 0.1 ~ 1g, ammonium chloride (NH ₄ Cl) 0.1 ~ 1g And Rhodoblastus acidophilus cultured in a medium containing 20 to 100 ml of calcium chloride dihydrate (CaCl ₂ .2H ₂ O) at a pH of 5 to 7. The method comprising the steps of: injecting an adsorbent into a mesh-type adsorption pad having a size of mesh mesh of 100 μm to 5 mm and a thickness of 0.5 to 50 mm, and then immersing the adsorption pad in a microbial culture solution; A method for adsorbing microorganisms,
0.5 to 5 g of beef extract, 0.5 to 5 g of yeast extract, 1 to 10 g of peptone, 1 to 10 g of sodium chloride and 1 to 10 g of Na-sesquicarbonate solution on the basis of 1 liter of purified water, (Containing 2 to 6 g of sodium hydrogencarbonate (NaHCO ₃ ) and 3 to 7 g of anhydrous sodium carbonate (Na ₂ CO ₃ anhydrous) per 100 ml of purified water) in a medium containing 20 to 200 ml of Bacillus alcalio Wherein the microorganism is Bacillus alcalophilus . The method comprising the steps of: injecting an adsorbent into a mesh-type adsorption pad having a size of mesh mesh of 100 μm to 5 mm and a thickness of 0.5 to 50 mm, and then immersing the adsorption pad in a microbial culture solution; A method for adsorbing microorganisms,
The microorganism may be selected from the group consisting of Beef extract 0.5-5 g / l, yeast extract 0.5-5 g / l, peptone 1-10 g / l, sodium chloride 1-10 g / ( Geobacillus caldoxylosilyticus ) cultivated at a pH of 4 to 10 and 37 to 80 ° C in a nutrient agar medium containing 5 to 25 g / l of agar ( agar ), or an anurinia bacillus thermoairophilus Aneurinibacillus thermoaerophilus ). The method comprising the steps of: injecting an adsorbent into a mesh-type adsorption pad having a size of mesh mesh of 100 μm to 5 mm and a thickness of 0.5 to 50 mm, and then immersing the adsorption pad in a microbial culture solution; A method for adsorbing microorganisms,
The microorganism is selected from the group consisting of 1 to 10 g of peptic digest of animal tissue, 0.5 to 3 g of yeast extract, 1 to 10 g of sodium chloride and 0.5 to 10 g of beef extract, (Bacillus thuringiensis) or Bacillus subtilis ( Bacillus subtilis ) cultured at a pH of 4 to 10 in a culture medium containing 3 g of a microorganism. 7. The method according to any one of claims 1 to 6,
Wherein the material of the adsorption pad is a steel material. 7. The method according to any one of claims 1 to 6,
Wherein the adsorbent is at least one selected from the group consisting of expanded vermiculite, diatomaceous earth, and superabsorbent resin. 7. The method according to any one of claims 2 to 6,
The microbial adsorption using the adsorbent is carried out by immersing the microbial culture broth in an amount of 1 to 30 parts by weight based on 100 parts by weight of the microbial culture broth and maintaining the humidity at 40 to 80% and temperature of 5 to 40 DEG C for 1 to 10 days ≪ / RTI > 7. The method according to any one of claims 1 to 6,
Wherein the microbial adsorption using the adsorbent is carried out by introducing the adsorbent into the mesh-type adsorption pad and then floating the adsorption pad in the culture medium of the microorganism. 11. The method of claim 10,
Wherein the adsorption pad is floated by a counterweight connected to the lower end of the adsorption pad, and the weight of the counterweight is determined by the following equation (1).
[Equation 1]

(Where d is the immersion depth of the adsorption pad, W _L is the loading force, W _S is the fixed load, L and B are the length and width of the adsorption pad, and γ _w is the unit volume weight of the culture medium). The method according to claim 1,
The microorganism may be selected from the group consisting of Rhodoblastus acidophilus, Bacillus alcalophilus, Geobacillus caldoxylosilyticus, Aneurinibacillus thermoaerophilus ), Rhodobacter capsulatus, Rhodoseudomonas palustris, Bacillus thuringiensis, and Bacillus subtilis. In addition, the present invention relates to How to. 13. The method of claim 12,
Rhodoblastus acidophilus is prepared by adding 0.05 to 1 g of yeast extract, 0.1 to 5 g of di-sodium succinate, and 1 to 5 g of ferric citrate solution 0.1 to 1 g of potassium dihydrogenphosphate (KH ₂ PO ₄ ), 0.1 to 1 g of magnesium sulfate heptahydrate (MgSO ₄ .7H ₂ O), 0.1 to 1 g of sodium chloride (NaCl) 0.1 to 1 g of ammonium chloride (NH ₄ Cl) and 20 to 100 ml of calcium chloride dihydrate (CaCl ₂ .2H ₂ O) at a pH of 5 to 7. 13. The method of claim 12,
The above-mentioned Bacillus alcalophilus is prepared by adding 0.5 to 5 g of beef extract, 0.5 to 5 g of yeast extract, 1 to 10 g of peptone, 1 to 10 g of sodium chloride, In a medium containing 20 to 200 ml of Na-sesquicarbonate solution (containing 2 to 6 g of sodium hydrogencarbonate (NaHCO ₃ ) and 3 to 7 g of anhydrous sodium carbonate (Na ₂ CO ₃ anhydrous) per 100 ml of purified water) 11 < / RTI > conditions. 13. The method of claim 12,
The Geobacillus caldoxylosilyticus and Aneurinibacillus thermoaerophilus may be used in an amount of 0.5 to 5 g / l of beef extract and 0.5 to 5 g / l of yeast extract, l of peptone, 1 to 10 g / l of peptone, 1 to 10 g / l of sodium chloride and 5 to 25 g / l of agar at pH 4 to 10 and 37 to 80? ≪ / RTI > 13. The method of claim 12,
The Rhodobacter capsulatus and Rhodoseudomonas palustris may be prepared by mixing 0.1 to 5 g of yeast extract and 0.1 to 5 g of disodium succinate hexa-hydrate per liter of purified water, 0.1 to 1 ml of absolute ethanol, 0.1 to 5 ml of ferric citrate solution, 0.1 to 1 g of potassium dihydrogenphosphate (KH ₂ PO ₄ ), magnesium sulfate heptahydrate (MgSO ₄ .7H ₂ O) 0.1 to 1 g, 0.1 to 1 g of sodium chloride (NaCl), 0.1 to 1 g of ammonium chloride (NH ₄ Cl) and 0.02 to 0.1 g of calcium chloride dihydrate (CaCl ₂ .2H ₂ O) . ≪ / RTI > 13. The method of claim 12,
Bacillus thuringiensis and Bacillus subtilis are prepared by adding 1 to 10 g of peptic digest of animal tissue, 0.5 to 3 g of yeast extract, 1 to 10 g of sodium chloride and 0.5 to 3 g of beef extract.

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