KR101497666B1 - Solid substrates immobilized with metal phyllosilicate including amino groups and use thereof - Google Patents

Abstract

A solid substrate on which a metal phyllosilicate containing an amino group is immobilized for use in separating microalgae, a method for efficiently separating microalgae from a sample using the same, a kit for efficiently separating microalgae from a sample containing the same, A filtration system, and a method of manufacturing the same.

Description

TECHNICAL FIELD [0001] The present invention relates to a solid substrate immobilized with a metal phyllosilicate containing an amino group,

A solid substrate on which a metal phyllosilicate containing an amino group is immobilized, a method for efficiently separating microalgae from a sample using the same, a kit and a filtration system for efficiently separating microalgae from a sample containing the same, and a method for manufacturing the same .

Since 2000, biomass, regarded as the third bio-energy associated with microalgae as a transport fuel, has attracted interest not only in schools and research laboratories but also in industry, which has consistently provided biodiesel to replace petroleum-based fuels Production. These fuels are associated with microalgae cultivation, and their advantages are that they can use abandoned land or sea, at least as far as the existing plant biodiesel. Furthermore, it has an effect of reducing carbon dioxide. However, there are many problems in developing the microalgae bio-refinery process so as to be a continuous and economically easy fuel. Generally, microalgae bio-refineries are mainly composed of downstream processes. It extracts proteins, carbohydrates, and lipids from microalgae, which are related to microalgae cultivation, harvesting, drying and value products, and extracts the lipids from fatty acid methyl esters (FAME) (hereinafter also referred to as "biodiesel" . ≪ / RTI > In order to reduce costs in the entire process, many researchers have focused on efficient and economical microalgae harvesting processes, but there are centrifugation, coagulation using inorganic and organic coagulants, sedimentation by gravity, screening, flooding, and electrophoresis There is no obvious practical way to date. Furthermore, extraction of lipids from non-dry biomass through cell wall destruction can eliminate the energy-intensive drying process. Finally, the development of suitable catalysts for the production of high-quality biodiesel and the development of these processes are essential, and there is interest in developing ways to consecutively re-use culture media to reduce additional costs. Therefore, much more efficient technologies must be developed to gain competitiveness at prices comparable to the microalgae biofuels industry and petroleum industry-based diesels.

In order to solve such a problem, there is a demand for a material or a method capable of rapidly and efficiently separating a greenhouse.

One aspect is to provide a solid substrate on which a metal phyllosilicate containing an amino group is immobilized, for use in efficiently separating microalgae.

Another aspect is to provide a method for efficiently separating microalgae from a sample using the solid substrate.

Another aspect is to provide a kit for efficiently separating microalgae from a sample containing the solid substrate.

Another aspect is to provide a filtration system for efficiently separating microalgae from a sample containing the solid substrate.

Another aspect is to provide a method for efficiently producing a solid substrate on which a metal phyllosilicate containing an amino group is immobilized.

One aspect provides a solid substrate on which a metal phyllosilicate comprising an amino group is immobilized, for use in separating microalgae.

In the solid substrate, the metal may be selected from the group consisting of Mg, Ca, Zn, Mn, Cu, Co, Ni, Al, Ce and Fe.

The metal phyllosilicate containing the amino group may be prepared by incubating an aminosilane compound and a metal ion in a solvent to precipitate a metal phyllosilicate containing the amino group. The incubation may be performed at 5 캜 to 30 캜. The incubation may be performed at atmospheric or pressurized conditions. The incubation can be carried out under basic conditions. The basic condition may be achieved by dissolution of the aminosilane itself. The incubation may be performed without stirring or stirring. The precipitated metal phyllosilicate can be separated from the reactants. Separation may be by, for example, sedimentation, centrifugation, filtration, or a combination thereof. The resulting metal phyllosilicate can be immobilized on a solid substrate. The solid substrate can be fixed by physical action, chemical bonding, or a combination thereof. The physical action may be adsorption, capture, or a combination thereof. The chemical bond can be made by covalent bond, coordination bond, Valdervalb bond, hydrogen bond, hydrophobic bond, or a combination thereof. The fixation may be performed simultaneously with the incubation, or may be performed separately. The incubation may be performed at pH 10-12. As used herein, the term "metal phyllosilicate" may be used to form sheets of silicate Tetrahedra containing 2: 5 Si ₂ O ₅ or Si: O containing metal. Phyllosilicates represent sheet-like silicates. The metal phyllosilicate contains an amino group having a positive charge on the surface at a high density, and can bind to a negatively charged microalgae. The coupling may be made, for example, by adsorption or electrostatic coupling. The "metal phyllosilicate containing amino group" is hereinafter also referred to as "aminoclay ".

1 is a view showing an example of the structure of a metal phyllosilicate containing an amino group. In Figure 1, a represents the ideal unit structure of the Mg-APTES clay and b represents the Mg-APTES clay unit structure obtained when tetraethyl orthosilicate (TEOS) is added during incubation. And a (CH _{2) 3} NH ₂ moieties cationic metal center that is coupled to a sheet silicate of exposing the tea to a surface (Mg ^2+, or Fe ^{+ 3)} - the resulting clay is through a covalent bond. In the aqueous solution, the amino group is positively charged and can be readily converted to a layered clay sheet. As a result, a high concentration of the transparent solution can be obtained. The Mg-APTES clay shown in FIG. 1 (b) may not be layered in an aqueous solution because the silicate sheet is crosslinked.

The aminosilane compound is a compound having the general formula (R ₁ O) ₃ SiQ ₁ , wherein R ₁ is a C1 to C10 alkyl group and Q ₁ is a group having an amino group. R ₁ can be methyl, ethyl, propyl, isopropyl, butyl, isobutyl, or pentyl. Q ₁ may be a primary amino group, a secondary amino group, a tertiary amino group, a quaternary amino group, or a combination thereof. Q ₁ may contain 1 to 100 carbon atoms, for example, 1 to 80, 1 to 50, 1 to 30, 1 to 150, or 1 to 10 carbon atoms. Group having the group _{- (R 2) NH 2,} R 2 and R ₃ are each independently a C1 to C10 alkyl group and n is an integer of 1 -R ₂ NH- of 1 to _{50 (R 3 NH) n- (} R 4 _{NH) m-, - (R 2} ) N (R 5) (R 6) R 7 , and will be selected from a combination thereof, wherein _{R 2, R 3, R 4} , R 5, R 6, and R ₇ are each with R _2, be selected independently of R alkyl group of _3, R _4, R _5, and R ₆ is a C1 to C10, R ₇ is an alkyl group of C1 to C20, n and m are integers between 0 and 20 be . The aminosilane compound may be, for example, (3- aminopropyl) tetraethyl orthosilicate (APTES), [3- (2-aminoethylamino) propyl] trimethoxysilane, 3- Ethylamino) ethylamino] propyltrimethoxysilane, and combinations thereof.

The incubation may be performed in the presence of a crosslinking agent. The crosslinking agent may be a silane which can be bonded to the amino clay sheet by a condensation reaction. The silane is present in a liquid state and can be precipitated by the reaction. That is, it may be capable of causing a sol-gel reaction. The incubation is carried out in the presence of (3-mercaptopropyl) trimethoxysilane (TTMS), methyl tetraethyl orthosilicate (MTES), phenyl tetraethyl orthosilicate (PTES), tetraethyl orthosilicate (TEOS) Lt; / RTI > in the presence of a compound selected from the group consisting of The crosslinking agent may be one which crosslinks between silicate sheets. The metal phyllosilicate may not be layer separated by crosslinking.

The incubation may be performed in the presence of a solid substrate to allow the metal phyllosilicate to be fixed to the solid substrate.

The incubation may be performed under the condition that the metal ion contains more silicon than that of the aminosilane compound on a molar basis. On a molar basis, the metal ion: silicon may be from 1: 0.5 to 0.9, for example 1: 0.75.

The solvent may be one capable of dissolving at least one of a metal ion and an aminosilane compound. The solvent may be an organic solvent or an aqueous solvent. The solvent may be selected from the group consisting of methanol, ethanol, dimethylformamide (DMF), dimethylsulfoxide (DMSO), and combinations thereof. The aqueous solvent may be a solvent comprising water, for example, water, saline, or an aqueous buffer solution.

The microalgae may be selected from the group consisting of green microalgae, bluegreen microalgae, and combinations thereof. The microalgae may be, for example, Chlorella vulgaris , UTEX-265), Nanochloropsis oculata , KMMCC-16), Synechocystis sp. PCC6803, or a combination thereof.

The metal phyllosilicate containing the amino group may be positively charged at pH 4 to pH 12.

The solid substrate may be made of a porous material. The solid substrate may be made of a porous material having a pore size which allows liquid to pass through but not through microalgae. The average pore size may be in the range of from 0.2 탆 to 1,000 탆, for example, from 10 탆 to 1,000 탆, from 10 탆 to 800 탆, from 10 탆 to 500 탆, from 10 탆 to 300 탆, from 30 탆 to 1,000 탆, 50 袖 m to 500 袖 m, or 50 袖 m to 300 袖 m. The solid substrate may be selected from the group consisting of wood, glass, fabric, plastic, or a combination thereof. The plastic includes polyethylene terephthalate (PET), polyvinylidene fluoride (PVDF), or a combination thereof. The fabric may be selected from the group consisting of cotton, wool, nylon, polyester, and combinations thereof. If the solid substrate is a fabric, the average pore size may be between 1 μm and 1,000 μm. If the solid substrate is wood, glass, plastic, or a combination thereof, excluding fabrics, the average pore size may be 0.2 袖 m to 1 袖 m. The solid substrate may have any shape. For example, in the form of spheres, plates, beads, or membranes.

Another aspect relates to a method for preparing microspheres, comprising contacting a sample comprising microalgae with a metal phyllosilicate comprising an amino group immobilized on a solid substrate as described above, and binding the microalgae to a metal phyllosilicate comprising an amino group A method for separating microalgae is provided.

In the method, the contacting may comprise passing the sample through the solid substrate. In this case, the solid substrate may be made of a porous material having a pore size which allows liquid to pass through but not through microalgae. The other solid substrates are the same as those described above. As a result, the liquid in the sample is removed through the solid substrate and the microalgae can be bonded to the metal fill silicate of the solid substrate. The bound microalgae can be separated from the solid substrate or further processed as such. The contact may be carried out without stirring or stirring.

Another aspect provides a kit for separating microalgae from a sample, comprising a solid substrate as described above and a reagent used to separate microalgae from the sample. The reagent used to separate the microalgae from the sample may include, for example, a wash liquor, an eluent to separate microalgae bound to the metal phyllosilicate, or a combination thereof. The kit may also include instructions for using the kit to separate microalgae from the sample.

Another aspect provides a filtration system for separating microalgae from a sample, comprising a solid substrate as described above as a filtration media. The filtration system may be a dead-end filter system including a tube or channel through which the fluid at one end of the solid substrate can flow. In addition, the filtration system may be a cross-fluoro filter system in which the solid substrate is installed on a sidewall of a pipe or channel through which a fluid can flow.

In another aspect, there is provided a method for preparing a solid substrate, comprising immobilizing a solid substrate, an amino silane compound and metal ions in a solvent to fix a metal phyllosilicate containing an amino group on a solid substrate surface, The method comprising:

In this method, the metal may be selected from the group consisting of Mg, Ca, Zn, Mn, Cu, Co, Ni, Al, Ce and Fe.

The incubation may be performed at 5 캜 to 30 캜. The incubation may be performed at atmospheric pressure. The incubation may be performed without stirring or stirring. The solid substrate can be separated from the reactants. Separation may be by, for example, sedimentation, centrifugation, filtration, or a combination thereof. The resulting metal phyllosilicate can be immobilized on a solid substrate. The solid substrate can be fixed by physical action, chemical bonding, or a combination thereof. The physical action may be adsorption, capture, or a combination thereof. The chemical bond can be made by covalent bond, coordination bond, Valdervalb bond, hydrogen bond, hydrophobic bond, or a combination thereof. The fixation may be performed concurrently with the incubation. The incubation may be performed at pH 10-12.

The aminosilane compound is a compound having the general formula (R ₁ O) ₃ SiQ ₁ , wherein R ₁ is a C1 to C10 alkyl group and Q ₁ is a group having an amino group. R ₁ can be methyl, ethyl, propyl, isopropyl, butyl, isobutyl, or pentyl. Q ₁ may be a primary amino group, a secondary amino group, a tertiary amino group, a quaternary amino group, or a combination thereof. Q ₁ may contain 1 to 100 carbon atoms, for example, 1 to 80, 1 to 50, 1 to 30, 1 to 150, or 1 to 10 carbon atoms. Group having the group _{- (R 2) NH 2,} R 2 and R ₃ are each independently a C1 to C10 alkyl group and n is an integer of 1 -R ₂ NH- of 1 to _{50 (R 3 NH) n- (} R 4 _{NH) m-, - (R 2} ) N (R 5) (R 6) R 7 , and will be selected from a combination thereof, wherein _{R 2, R 3, R 4} , R 5, R 6, and R ₇ are each with R _2, be selected independently of R alkyl group of _3, R _4, R _5, and R ₆ is a C1 to C10, R ₇ is an alkyl group of C1 to C20, n and m are integers between 0 and 20 be . The aminosilane compound may be, for example, (3- aminopropyl) tetraethyl orthosilicate (APTES), [3- (2-aminoethylamino) propyl] trimethoxysilane, 3- Ethylamino) ethylamino] propyltrimethoxysilane, and combinations thereof.

The incubation may be performed in the presence of a crosslinking agent. The incubation is carried out in the presence of (3-mercaptopropyl) trimethoxysilane (TTMS), methyl tetraethyl orthosilicate (MTES), phenyl tetraethyl orthosilicate (PTES), tetraethyl orthosilicate (TEOS) Lt; / RTI > in the presence of a compound selected from the group consisting of The crosslinking agent may be one which crosslinks between silicate sheets. The metal phyllosilicate may not be layer separated by crosslinking.

The incubation may be performed under the condition that the metal ion contains more silicon than that of the aminosilane compound on a molar basis. On a molar basis, the metal ion: silicon may be from 1: 0.5 to 0.9, for example 1: 0.75.

The solvent may be one capable of dissolving at least one of a metal ion and an aminosilane compound. The solvent may be an organic solvent or an aqueous solvent. The solvent may be selected from the group consisting of methanol, ethanol, dimethylformamide (DMF), dimethylsulfoxide (DMSO), and combinations thereof. The aqueous solvent may be a solvent comprising water, for example, water, saline, or an aqueous buffer solution.

The microalgae may be selected from the group consisting of green algae, cyanobacteria, and combinations thereof.

The metal phyllosilicate containing the amino group may be positively charged at pH 4 to pH 10.

The solid substrate may be made of a porous material. The solid substrate may be made of a porous material having a pore size which allows liquid to pass through but not through microalgae. The average pore size may be in the range of 1 탆 to 1,000 탆 such as 10 탆 to 1,000 탆, 10 탆 to 800 탆, 10 탆 to 500 탆, 10 탆 to 300 탆, 30 탆 to 1,000 탆, 30 탆 to 800 탆, 30 탆 to 500 탆, 50 [mu] m to 500 [mu] m, or 50 [mu] m to 300 [mu] m. The solid substrate may be selected from the group consisting of wood, glass, fabric, plastic, or a combination thereof. The plastic includes polyethylene terephthalate (PET), polyvinylidene fluoride (PVDF), or a combination thereof. The fabric may be selected from the group consisting of cotton, wool, nylon, polyester, and combinations thereof. If the solid substrate is a fabric, the average pore size may be between 1 μm and 1,000 μm. If the solid substrate is wood, glass, plastic, or a combination thereof, excluding fabrics, the average pore size may be 0.2 袖 m to 1 袖 m. The solid substrate may have any shape. For example, in the form of spheres, plates, beads, or membranes.

According to one aspect of the present invention, a solid substrate on which a metal phyllosilicate containing an amino group is immobilized can be used for efficiently separating microalgae.

According to the method of separating the microalgae from the sample according to another aspect, the microalgae can be efficiently separated from the sample.

The kit for separating the microalgae from the sample according to another aspect can be used for efficiently separating the microalgae.

The filtration system for separating the microalgae from the sample according to another aspect can be used for efficiently separating the microalgae.

According to another aspect of the present invention, there is provided a method for manufacturing a solid substrate on which a metal phyllosilicate containing an amino group is immobilized.

1 is a view showing an example of the structure of a metal phyllosilicate containing an amino group.
Figure 2 shows a photograph of a surface coated twice with an Fe-APTES clay.
FIG. 3 is a view showing the result of chlorella bulgaris harvest using vertical flow filtration using a fabric coated with Fe-APTES clay as a filter. FIG.
Figs. 4 and 5 show the vertical flow (flux, Lm ^-2 hr) and the removal efficiency (%) of microalgae harvest using nylon coated cotton coated with amino clay, respectively.
Figs. 6 and 7 show video microscopy images of micro-algae harvesting (A) and after (B) using two coated surfaces of Fe-APTES clay and nylon.
FIG. 8 is a photograph of a PET film showing the degree of biofouling after a newly formed PET film (left) and a horizontal flow process (right).
9 shows photographs of a newly produced PET film (left side) and a PET film (right side) coated once with a Fe-APTES clay.
FIG. 10 is a photograph of a PET film (left) coated twice with Fe-APTES clay and a PET film showing the degree of biofouling after the horizontal flow process (right).
11 is a photograph of a PET film coated three times with Fe-APTES clay (left) and a PET film showing the degree of biofouling after the horizontal flow process (right).
Fig. 12 is a photograph of a PVDF membrane showing the degree of biofouling after a newly fabricated PVDF membrane (left) and a horizontal flow process (right).
13 shows photographs of a newly fabricated PVDF film (left side) and a PVDF film (right side) coated once with a Fe-APTES clay.
Figure 14 is a photograph of a PVDF membrane (left) coated twice with an Fe-APTES clay and a PVDF membrane showing the degree of biofouling after the horizontal flow process (right).
FIG. 15 is a photograph of a PVDF membrane (left) coated three times with Fe-APTES clay and a PVDF membrane showing the degree of biofouling after the horizontal flow process (right).
Figure 16 is a plot of flux over time in a horizontal flow filtration process through an uncoated PET film and a PET film coated twice and three times with Fe-APTES.
Figure 17 is a plot of flux over time in a horizontal flow filtration process through an uncoated PVDF membrane and a PVDF membrane coated twice and three times with Fe-APTES.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

1. Experimental Method

Unless otherwise stated, the following examples were subjected to the following experimental methods.

(One) Amino clay  Synthetic Amino clay  Method for immobilization on a solid substrate

Each amino clay (Mg-APTES clay and Fe-APTES clay) was prepared. Briefly, 8.4 g (0.0413 mol) of MgCl ₂ .H ₂ O (Junsei) is dissolved in 200 mL ethanol (Samchun Pure Chemical) and sterilized for 10 minutes. After that, 13 mL (0.0587 mol) of (3-aminopropyltriethoxysilane: APTES, Sigma-Aldrich) was pipetted into the MgCl ₂ -oluted ethanol solution and magnetic stirrer was performed for 12 hours do. In this case, the pH was about 10. The precipitated material is collected by centrifugation (2800 rpm, 10 minutes) and then washed twice with ethanol to remove excess MgCl ₂ . Then, the product is put in an oven at 45 ° C, dried for a day, and then grinded in a mortar. Similarly, in the Fe-APTES clay synthesis, FeCl ₃ .H ₂ O (Sigma-Aldrich) is used in place of MgCl ₂ .H ₂ O (quasicrystal).

To coat Fe-APTES clay on fabrics or membranes, the fabrics and membranes were immersed in 200 mL of ethanol solution containing 8.4 g of FeCl ₃ .H ₂ O for 6 hours, taken out, and resuspended in 40 mL APTES and 5 mL tetraethylorthosilicate (TEOS , 0.0240 mol) in 200 mL of ethanol. Polyethylene terephthalate (PET) and polyvinylidene fluoride (PVDF) were used as the membrane material (size 10 cm x 5 cm, size: 110 mm x 65 mm) Effective size 6.3cm x 2.2cm).

This is called coating one time. When the second coating is performed, it is referred to as runs = 2. The coated fabric or membrane is washed in ethanol and dried for 24 hours at room temperature.

(2) Microalga species, culture condition and cultivation amount

Freshwater species microalgae, Chlorella vulgaris, a UTEX-265) was purchase from UTEX birds center of Texas College of standard TAP (in Tris-Acetate-Phosphate) media (final 1L tris (hydroxymethyl) -amino methane 20mM, NH4Cl 7mM, MgSO ₄ and 7H ₂ O 0.83mM, CaCl ₂ and _{2H 2 O 0.45mM, K 2 HPO} 4 1.65 mM, KH 2 PO 4 1.05 mM, Na 2 EDTA and 2H ₂ O 0.134 mM, ZnSO ₄ and 7H ₂ O 0.136 mM, H ₃ BO ₃ 0.184 mM, MnCl ₂ .4H ₂ O 40 μM, FeSO ₄揃 7H ₂ O 32.9 μM, CoCl ₂揃 6H ₂ O 12.3 μM, CuSO ₄揃 5H ₂ O 10 μM, (NH ₄ ) ₆ Mo ₇ O ₂₄揃 4H ₂ O 0.928μM, 1mL of acetic acid, and the balance is water) or standard F / 2 medium (final 1L of NaNO ₃ 0.075g, NaH ₂ PO ₄ and 2H ₂ O 0.00565g, trace element solution 1.0mL Stark, vitamin mix the stock solution 1.0mL, and the remainder water; the stock solution of trace elements final 1L of Na ₂ EDTA 4.16g, FeCl ₃ and 6H ₂ O 3.15g, CuSO ₄ and 5H ₂ O 0.01g, ZnSO ₄ and 7H ₂ O 0.022g, CoCl ₂ · 6H ₂ O 0.01g, MnCl ₂ and _{4H 2 O 0.18g, Na 2 MoO} 4 and 2H ₂ O 0.006g, and the remainder water; vitamin mix Is cyano cobalt amine (vitamin B12), 0.0005g, thiamin HCl (vitamin B1), 0.1g, biotin 0.0005g and the rest water) in the final 1L

Lt; RTI ID = 0.0 > 2000 < / RTI > Seawater species, Nanochloropsis oculata , KMMCC-16) were purchased from the Korean marine culture collection in Busan and cultured in standard F / 2 media with 2,000 CO ₂ at a light intensity of 2000 lux. Cyanobacteria species, Synechocystis sp. PCC6803 were obtained from Chungnam National University and cultured.

All three species of microalgae were cultured in a 500 mL flask under the following conditions. The temperature was 30 占 폚, the stirring speed was 125 rpm. All equipment and media were autoclaved at 121 ° C for 15-20 minutes, then sterilized and incubated. At the end of the log, biomass was used for separation or harvesting experiments. To measure the biomass concentration in the culture, the optical density (OD) was measured at 682 nm, 660 nm, and 730 nm for chlorella bulgurris, nanochloroboscis oculatus, and synechocystis species, respectively. Harvest experiments in batch mode were performed in 100 mL glass cylinders.

(3) Bulk harvesting and vertical flow filtration experiments

Bulk harvesting with Mg-APTES clay was carried out in a 1 L glass cylinder. The Mg-APTES clay was mixed with the microalgae cultured for 1 to 2 minutes and allowed to flocculate. The supernatant samples were taken periodically to determine the optical density of the cultures and to calculate the harvest efficiency (%). The sample was removed from the middle of the glass cylinder and performed with removal according to the height of the glass cylinder. Fabrics coated with Fe-APTES clay were used as filtration filters using a dead-end filter system. 1L of total microalgae was used as a filtration experiment, and the weight of the permeate solution was calculated to calculate the flux through the filter. The effective area size of the filter was 9.60 cm ² , and the harvesting and filtration efficiency was measured using the following equation.

Harvest efficiency (%) = 1-OD _f / OD _i

Where OD _f and OD _i are the final and first optical density values of the microalgae culture.

The vertical flow through the filter was calculated using the following equation.

Flux (L m ^-2 h ^-1 ) = Permeate volume / (area x time)

Here the area is the effective area of the filter coated with Fe-APTES clay.

(4) microalgae Lipid  Through extraction Fatty acid methyl ester  ( FAME )

Centrifuge the biomass at 4 ° C at 5000 rpm for 10 minutes to discard the supernatant and wash it with distilled water. The biomass obtained by centrifugation was frozen using liquid nitrogen and vacuum freeze-dried at -40 ° C. The total lipid was extracted from 10 mg of dried biomass by a chloroform-methanol (2: 1, v / v) solvent mixture similar to Folch's method.

The extracted lipids were converted to fatty acid methyl esters (FAME) using the transesterification reaction. Briefly, methanol was added to the extracted lipid which contained sulfuric acid as a catalyst, followed by reaction at 100 ° C for 10 minutes. The FAME in the organic layer was analyzed by gas chromatography (GC, HP5890, Agilent, USA) with an INNOWAX capillary column (30 m x 0.32 mm x 0.5 μm, Agilent, USA) with a flame ionization detector (FID). The gas chromatographic column temperature was programmed as follows. (1) The initial column temperature is stopped at 50 ° C for 1 minute. (2) Raise to 200 DEG C at a rate of 15 DEG C per minute. (3) Raise to 250 ° C at a rate of 2 minutes per minute and hold for 2 minutes. Each FAME component was analyzed qualitatively and quantitatively using the standard solution for the residual time and peak area.

(5) Characteristic evaluation and equipment

The concentration of microalgae biomass (g / L) was calculated by measuring the optical density (OD) using a UV-visible spectrophotometer (Beckman Coulter, DU 730).

The morphology of microalgae and coagulated microalgae was observed using an optical microscope (Leica, model DM2500) and field emission scanning electron microscopy (FE-SEM, Magellan 400, FEI). The fabric coated with air-dried amino clay and amino clay was examined by transmission electron microscope (TEM, JEM-2100F), scanning electron microscope (SEM) equipped with energy dispersive x-ray spectroscopy (EDX, Bruker), video microscope -13M), respectively. All photos were captured with an iPhone digital camera. The powder X-ray diffraction (XRD) pattern was evaluated with D / MAX-RB (Rigaku, 12 kW). Fourier transform infrared (FT-IR) spectroscopy was scanned from 4000 cm ^-1 to 400 cm ^-1 in KBr pellet mode. The zeta potential was measured using a Zetasizer Nano-ZS particle analyzer (Malvern). Quantitative determination of silicon concentration in aqueous solution was performed using an inductively coupled plasma spectroscopy (ICP-AES, Optima 7300 DV). The pH change was monitored using a pH / ion meter (D-53, Horiba).

Example  1: metal fastened to the fabric Phyllosilicate

Fg-APTES clay was fixed on the cotton or nylon as described above. The surface or nylon has an average pore size of 1 μm to 1,000 μm.

Figure 2 shows a photograph of a surface coated twice with an Fe-APTES clay. The size of the surface is 110 mm x 65 mm, which shows that the surface is uniformly coated and shows a compactness that the microalgae can not escape. As a result of SEM-energy dispersive X-ray analysis (EDX) analysis of Fe-APTES clay-coated fabric, Fe-APTES clay components Fe, Si, Cl, C, O, and N The ingredients were identified. Where Cl is a < RTI ID = 0.0 _> Is a counter ion for stabilizing the group. C and O can originate from both amino clay and fabric. It was confirmed that the Fe-APTES clay particles were well adsorbed on the surface of the cotton, and the coating was made so that microalgae could be harvested well.

The Fe-APTES clay coated surface was performed using vertical flow filtration with scale-up as a filter. The vertical flow filtration system is made up of a face coated with Fe-APTES clay at the end of a 1.2 L volume vessel, allowing liquid to permeate through the face. Vertical flow filtration systems are capable of handling 1.2 L volume of microalgae samples. Vertical flow filtration systems have a much better permeate flow compared to a cross-flow system, but can quickly biofouling the membrane. Therefore, it is thought that a vertical flow filtration system will be implemented, which can reduce the problem of releasing the amino clay particles immobilized on the surface membrane in advance.

FIG. 3 is a view showing the result of chlorella bulgaris harvest using vertical flow filtration using a fabric coated with Fe-APTES clay as a filter. FIG. The vertical flow filtration system is as described above. (Coating = 1), Nylon (Coating = 1), Cotton (Coating = 1), Nylon, 2), nylon (coating = 2), cotton (coating = 3), and nylon (coating = 3), respectively. In the case of using the metal phyllosilicate fixed surface compared to the surfaces (BC, BN) on which the metal phyllosilicate was not fixed, the harvest efficiency of the microalgae was remarkably high. In particular, the harvest efficiency (%) of the two coatings of amino clay in both cotton and nylon was about 95%.

FIGS. 4 and 5 show the vertical flow (flux, Lm ^-2 hr) and removal efficiency (%) of microalgae harvest using nylon coated cotton coated with amino clay, respectively. The permeation flux (flux, Lm ^-2 h) gradually decreased to reach approximately 55.

Figs. 6 and 7 show video microscopy images of micro-algae harvesting (A) and after (B) using two coated surfaces of Fe-APTES clay and nylon. As shown in Fig. 6 and Fig. 7, there was a biofouling film formation after algae harvest.

This permeate flow is the result of proceeding only by gravity without external pressure using a vacuum pump. Therefore, additional vibrations or vacuum modes can be used to enhance and maintain the permeate flow. As a matrix coating the amino clay, the cotton fabric in the cotton and nylon is much more suitable and easy to handle due to its mechanical flexibility, which helps to uniformly coat the cotton surface due to the cotton fibers with various functional groups.

As shown in this example, aminoclay was immobilized on fabrics and applied to vertical flow filtration, allowing both freshwater and seawater microalgae to be harvested. Therefore, this result can be a practical way to reduce the cost of downstream processing in microalgae biorefinery, with microcryogenic harvesting systems based on amino clays. The use of textiles will help to lower investment costs and solve some of the challenges of challenging projects in microalgae biofinery.

Example  2: metal fixed to plastic film Phyllosilicate

As described above, the Fe-APTES clay was fixed to the plastic film, that is, the PET film and the PVDF film. The PET film and the PVDF film have an average pore size of about 4 占 퐉, and about 0.45 占 퐉. Next, these membranes were placed in a cell made of acryl and connected to a cross-flow filtration apparatus. The horizontal flow filtration device uses a pump to flow the algae sample in a feed tank to a circular tube (3/8 inch diameter) and to filter the sample as it passes perpendicularly through the cell.

FIG. 8 is a photograph of a PET film showing the degree of biofouling after a newly formed PET film (left) and a horizontal flow process (right). In FIG. 8, the horizontal flow process was performed by flowing a culture solution of Chlorella vulgaris algae at 30 DEG C pH 6.5 for 7 days in BG11 medium for 3 hours at a flow rate of 4.2 L / min.

9 shows photographs of a newly produced PET film (left side) and a PET film (right side) coated once with a Fe-APTES clay.

FIG. 10 is a photograph of a PET film (left) coated twice with Fe-APTES clay and a PET film showing the degree of biofouling after the horizontal flow process (right). The horizontal flow filtration process was performed in the same manner as in Fig.

11 is a photograph of a PET film coated three times with Fe-APTES clay (left) and a PET film showing the degree of biofouling after the horizontal flow process (right). The horizontal flow filtration process was performed in the same manner as in Fig.

Fig. 12 is a photograph of a PVDF membrane showing the degree of biofouling after a newly fabricated PVDF membrane (left) and a horizontal flow process (right). The horizontal flow filtration process was performed in the same manner as in Fig.

13 shows photographs of a newly fabricated PVDF film (left side) and a PVDF film (right side) coated once with a Fe-APTES clay.

Figure 14 is a photograph of a PVDF membrane (left) coated twice with an Fe-APTES clay and a PVDF membrane showing the degree of biofouling after the horizontal flow process (right). The horizontal flow filtration process was performed in the same manner as in Fig.

FIG. 15 is a photograph of a PVDF membrane (left) coated three times with Fe-APTES clay and a PVDF membrane showing the degree of biofouling after the horizontal flow process (right). The horizontal flow filtration process was performed in the same manner as in Fig.

As shown in FIG. 8 to FIG. 15, it can be seen that the higher the degree of Fe-APTES clay coating is, the less biofouling occurs. The clay coating in this case could be expected to prevent biofouling rather than the concept of coagulation that affects microalgae harvest. The harvesting efficiency was 49.9% for PET, 99.6% for PET (runs = 2) and 99.9% for PET (runs = 3).

In the horizontal flow filtration process, the rejection rate calculation method is as follows.

Rej = 1- C _P / C _F

Here, Rej is rejection, dimensionless, expressed as a fraction,

C _P is the concentration in the permeate, mole / L

C _F is the concentration in feed water, mole / L. The larger the Rej, the less the fine species pass through the membrane. The algae harvested are located in the retentate.

Figure 16 is a plot of flux over time in a horizontal flow filtration process through an uncoated PET film and a PET film coated twice and three times with Fe-APTES. The horizontal flow filtration process was performed in the same manner as in Fig. The rejection rate for each process is shown in Table 1.

 Uncoated PET PET (runs = 2) PET (runs = 3) Rejection rate (%) 26.7 69.9 97.3

When the coating was applied three times, the rejection rate was 97.3%. In the case of flux, the flux of the PET film with two coatings showed a flux value of about 30, and the coating with three times showed a flux value of about 15. When the coating is applied three times to improve the rejection rate, the flux has a decreasing correlation.

Table 2 shows the rejection rates after 3 hours in a horizontal flow filtration process through uncoated PVDF membranes and PVDF membranes coated twice and three times with Fe-APTES. The horizontal flow filtration process was performed in the same manner as in Fig.

 Uncoated PVDF PVDF (runs = 2) PVDF (runs = 3) Rejection rate (%) 99.7 100 100

When the Fe-APTES clay was coated 2 times and 3 times, the rejection rate was 100%. In the case of flux, the flux was maintained at around 50 for both the 2 and 3 coatings. Therefore, in the case of the uncoated PVDF film, the flux rapidly decreased with time, and was markedly lower than when the coating was applied two times and three times. Therefore, in case of PVDF film, 2 times coating with Fe-APTES clay is suitable. In Table 1 and Table 2, the rejection rate is the rejection rate at 3 hours after the start of filtration.

Claims (20)

A metal phyllosilicate containing an amino group is immobilized on a metal phyllosilicate containing an amino group for use in separating microalgae. The metal phyllosilicate containing the amino group is prepared by incubating an aminosilane compound and a metal ion in a solvent to form a metal filament Wherein said immobilization is performed simultaneously with said incubation. ≪ RTI ID = 0.0 > 11. < / RTI > The solid substrate of claim 1, wherein the metal is selected from the group consisting of Mg, Ca, Zn, Mn, Cu, Co, Ni, Al, Ce and Fe. delete The solid substrate according to claim 1, wherein the aminosilane compound is a compound having the general formula (R ₁ O) ₃ SiQ ₁ , wherein R ₁ is a C1 to C10 alkyl group and Q ₁ is a group having an amino group. The method according to claim 4, a group having an amino group _{- (R 2) NH 2,} R 2 and R ₃ are independently a C1 to C10 alkyl group, and n is (R ₃ NH) -R ₂ NH- integer of 1-50 for each n - (R ₄ NH) m-, - (R ₂ ) N (R ₅ ) (R ₆ ) R ₇ and combinations thereof, wherein R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , And R ₇ are each independently selected from the group consisting of R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are C 1 to C 10 alkyl groups, R ₇ is a C 1 to C 20 alkyl group, Lt; RTI ID = 0.0 > 20. ≪ / RTI > [2] The aminosilane compound according to claim 1, wherein the aminosilane compound is at least one selected from the group consisting of (3-aminopropyl) -tetraethylorthosilicate (APTES), [3- Aminoethylamino) ethylamino] propyltrimethoxysilane, and combinations thereof. The solid substrate of claim 1, wherein the incubation comprises performing in the presence of a crosslinking agent. 2. The method of claim 1, wherein the incubation is conducted in the presence of (3-mercaptopropyl) trimethoxysilane (TTMS), methyl tetraethyl orthosilicate (MTES), phenyl tetraethyl orthosilicate (PTES) In the presence of a compound selected from the group consisting of combinations thereof. The solid substrate according to claim 1, wherein the incubation is performed under the condition that the metal ion contains more silicon than the aminosilane compound silicon on a molar basis. The solid substrate of claim 1, wherein the solvent is selected from the group consisting of methanol, ethanol, dimethylformamide (DMF), dimethylsulfoxide (DMSO), and combinations thereof. The solid substrate of claim 1, wherein the microalgae are selected from the group consisting of green microalgae, bluegreen microalgae, and combinations thereof. The solid substrate according to claim 1, wherein the metal phyllosilicate containing an amino group is positively charged at a pH of from 4 to 10. The solid substrate of claim 1, wherein the solid substrate is of a porous material having a pore size that allows liquid to pass but not micro-algae. The solid substrate according to claim 1, wherein the solid substrate has a film shape. Contacting a sample comprising microalgae with a metal phyllosilicate comprising an amino group immobilized on a solid substrate according to any one of claims 1, 2 and 4 to 14 to bind the microalgae to a metal phyllosilicate comprising an amino group; And separating the microalgae from the sample. 16. The method of claim 15, wherein the contacting comprises passing a sample through a solid substrate. A kit for separating microalgae from a sample, comprising a solid substrate according to any one of claims 1, 2 and 4 to 14 and a reagent used for separating microalgae from the sample. A filtration apparatus for separating microalgae from a sample, comprising a solid substrate according to any one of claims 1, 2 and 4 to 14 as a filtration medium. A step of immobilizing a solid substrate, an aminosilane compound and a metal ion in a solvent to fix a metal phyllosilicate containing an amino group on the surface of the solid substrate; and a method for producing a solid substrate on which a metal phyllosilicate containing an amino group is immobilized . 21. The method of claim 19, wherein the incubation is performed in the presence of a crosslinking agent.

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