KR101734260B1 - Method for preparing galactose using red alae - Google Patents

Abstract

The present invention relates to a method for producing galactose using a red algae, and more particularly, to a method for producing galactose including a saccharification step, a filtration step, a concentration step and a precipitation step, It has an effect of providing a process technology capable of industrially mass-producing galactose, which is used as an important intermediate material in material production.

Description

TECHNICAL FIELD The present invention relates to a method for preparing galactose using red algae,

The present invention relates to a method for producing galactose using red algae, and more particularly, to a method for producing galactose including a saccharification step, a filtration step, a concentration step, and a precipitation step.

Galactose is one of the sugars which is a constituent of red algae contained in seaweeds. It is one of very important candidates which is expected to have useful functionality for development of raw materials for chemical reaction and physiologically active substances and application in pharmaceutical field.

This galactose belongs to aldohexose, rarely present in free form in nature, and widely distributed in polymer state. The molecular formula is C ₆ H ₁₂ O ₆ and the melting point is 167 ° C. It is sweet with white powder. It is soluble in water and contains crystalline water has melting point of 118 ℃. There are D- and L-type optical isomers, but there are many D-forms in nature. Generally, when referring to galactose, it refers to D-galactose.

Such galactose is widely distributed in the biological system and is mainly contained in red algae. Galactose production by seaweed saccharification has not been established globally, and there is no report on complete process development at the laboratory level. After the saccharification treatment, the level of the sugar in the liquid phase is analyzed by analyzing the liquid substance.

Further, in the case of producing galactose through a glycosylation process using algae as a raw material, it is very difficult to obtain galactose, which is a monosaccharide, directly as a particle in a glycosylated state. This is due to the presence of protein components and impurities originally present in seaweeds, as well as the presence of acidic chemicals that are introduced during acid glycosylation. At present, the groups that research and develop algae saccharification are mainly engaged in studies for manufacturing fuel materials such as bioethanol through fermentation process after neutralization treatment after acidification, and the process technology of the granulation manufacturing technology has not been completed .

In order to solve the above problems, the present inventors have completed the present invention by confirming that solid state galactose particles can be produced from glycosylated algae through connection of specific unit processes.

Accordingly, an object of the present invention is to provide a process for producing a sugar mixture from a red algae through a saccharification process; Filtering the sugar mixture to obtain a filtered sugar mixture; Concentrating the filtered sugar mixture to obtain a concentrated sugar mixture; And a precipitating step of adding an alcohol to the concentrated sugar mixture to precipitate galactose.

In order to accomplish the object of the present invention described above, the present invention provides a method for producing a sugar mixture, comprising: saccharifying step of preparing a saccharide mixture from a red algae through a saccharification step; Filtering the sugar mixture to obtain a filtered sugar mixture; Concentrating the filtered sugar mixture to obtain a concentrated sugar mixture; And a precipitation step of precipitating galactose by adding alcohol to the concentrated sugar mixture.

Hereinafter, the present invention will be described in more detail.

The saccharification step may be a step of obtaining a sugar mixture containing galactose produced from a red algae cell wall by subjecting red algae to hydrolysis by contacting the acid substance in a liquid state. The sugar mixture may be in a liquid phase, e.g., in the form of an aqueous solution.

The saccharification step may be performed at 80 to 150 ° C, 100 to 150 ° C, or 120 to 150 ° C, but is not limited thereto.

The hydrolysis of the saccharification step may be carried out in the presence of an acid in an amount of 0.05 to 5% (w / v), 0.1 to 5% (w / v), 0.5 to 5% (w / v), or 1 to 5% Concentration in the reaction mixture.

Further, the acid is sulfuric acid (H ₂ SO _4), hydrochloric acid (HCl), hydrobromic acid (HBr), nitric acid (HNO _3), acetic acid (CH ₃ COOH), formic acid (HCOOH), perchloric acid (HClO _4), phosphoric acid (H ₃ PO ₄ ), and paratoluene sulfonic acid (PTSA). However, the present invention is not limited thereto.

The red algae may be red algae containing galactose or a polymer thereof, for example, Chondrus , Eucheuma , Gigartina , Pterocladia , Hephnia spp. But are not limited to, Hypnea , Iridaea , Kappaphycus , Gellidium , or Gracilaria .

In addition, the red algae can be used as a primary dry product, a dried product after washing with water, or a disrupted product thereof, but the present invention is not limited thereto.

Wherein the filtration step comprises:

A primary filtration step in which the sugar mixture obtained in the saccharification step is subjected to silica gel column chromatography to obtain a primary filtered sugar mixture;

A second filtration step of obtaining the first filtered sugar mixture by using a column containing an ion exchange resin to obtain a second filtered sugar mixture; And

And a tertiary filtration step of subjecting the secondary filtered sugar mixture to silica gel column chromatography to obtain a tertiary filtered sugar mixture.

The filtration step removes the protein components in the mixture and neutralizes the acidic components added during the saccharification step, thereby improving the stability in the concentration step and improving the efficiency of the concentration step.

The primary filtration step may be carried out by subjecting the sugar mixture obtained in the saccharification step to silica gel column chromatography one or more repeated cycles, for example, 1 to 10 times, 1 to 5 times, or 1 to 3 repeated cycles, And to remove particulate impurities and protein components contained in the mixture

In addition, the silica particles may have an average diameter of 0.1 to 0.5 mm, 0.1 to 0.4 mm, 0.1 to 0.3 mm, or 0.1 to 0.2 mm, and various materials of neutral components may be used and are not limited to specific silica materials .

The column including the ion exchange resin in the secondary filtration step may be a structure in which an ion exchange resin layer and a silica gel particle layer are alternately laminated. For example, the ion exchange resin layer and the silica gel particle layer may be alternately deposited 2 to 10 times, 2 to 8 times, 2 to 5 times, 2 to 3 times, or 2 times, but the present invention is not limited thereto.

The volume ratio of the ion exchange resin layer and the silica gel particle layer in the column including the ion exchange resin is in the range of 10: 1 to 1:10, 5: 1 to 1: 5, 3: 1 to 1: 3, 2: , Or 2: 1 (ion exchange resin layer volume: silica gel particle layer volume), but is not limited thereto.

The ion exchange resin in the second filtration step may be a basic anion exchange resin, and a weak base or strong base anion exchange resin may be selectively applied depending on the acidity of the mixture.

The secondary filtration step may be to reduce the chloride ion in the sugar mixture through exchange between the chloride ion present in the sugar mixture and the hydroxide ion present in the ion exchange resin, It may be appropriate to treat the ion exchange resin at a ratio of 1/10 volume. The sugar mixture that has passed through the column containing such an ion exchange resin may be one in which the acid component is removed and the pH is converted to a level of 5 to 7.

Commercial products manufactured by domestic and foreign companies can be used for the ion exchange resin. For example, TRILITE AMP16 or TRILITE AMP26, which are commercially available from Samyang Corporation of Korea, can be used. The ion exchange resin is characterized by being spherical particles having a polystyrene resin with trimethylammonium ion and dimethylethanolammonium ion attached to the ends thereof. The specific gravity of the particles is about 1.0 to 1.11 g / cm ³ , and the apparent density is 600 to 700 g / l. And the particle size is in the range of 0.3 to 1.2 mm in diameter.

In the third filtration step, the sugar mixture obtained in the saccharification step is subjected to silica gel column chromatography one or more repeated cycles, for example, 1 to 10 times, 1 to 5 times, or 1 to 3 repeated cycles, It may be to remove some of the fine particles generated during the secondary filtration step once again.

In addition, the silica gel particles may have an average diameter of 0.1 to 0.5 mm, 0.1 to 0.4 mm, 0.1 to 0.3 mm, or 0.1 to 0.2 mm, and various materials of neutral components may be used and are not limited to specific silica materials .

In addition, the volume of the column in the first to third filtration steps is suitably in a ratio of 1/2 to 1/5 times the volume of the mixture per liquid phase. If the ratio is out of the above range, the separation efficiency may be lowered or the content of the impurity particles may be increased to cause a phenomenon of mixing with the galactose particles in the final granulation process.

The flow rate of the saccharide mixture in the column of the primary to tertiary filtration step is 0.1 to 100 ml / min, 0.1 to 80 ml / min, 0.1 to 60 ml / min, 0.1 to 40 ml / min, At a flow rate of 20 ml / min, 0.1 to 10 ml / min, 0.1 to 5 ml / min, or 3.5 ml / min, but not limited thereto, Can be set.

The filtration step may be followed by a concentration step. The concentration method may be a vacuum distillation method. The equipment used in this case is not limited to a specific apparatus as a vacuum distillation apparatus. In the concentration step, it is appropriate to concentrate to a level of about 1/10 to 1/20 of the volume of the sugar mixture filtered in the filtration step.

In addition, the concentration step can be performed at a temperature of 30 to 60 ° C. When the temperature is higher than 60 ° C, there is a disadvantage in that some discoloration occurs in the concentrated galactose component. When the temperature is lower than 30 ° C, have.

Also, the concentration step can be performed at a pressure of 10 to 120 mbar, and if the pressure is more than 120 mbar, the process time becomes longer, and when the pressure is less than 10 mbar, the process stability is deteriorated.

Thereafter, alcohol may be added to the concentrated sugar mixture at a temperature of -10 ° C to 25 ° C, or an alcohol at a temperature of -10 ° C to 25 ° C may be added to induce galactose particle formation (granulation) to precipitate galactose. Precipitated galactose solid particles can be obtained using a vacuum filtration equipment.

The alcohol may be at least one selected from the group consisting of linear or branched alcohols having 1 to 4 carbon atoms such as methanol, ethanol, and propanol (for example, isopropyl alcohol), but the present invention is not limited thereto. The alcohol may be used at a concentration of 10 to 100% (v / v), 20 to 100 (v / v), 30 to 100 (v / v), 40 to 100 (v / , 60 to 100 (v / v), 70 to 100 (v / v), 80 to 100 (v / v), 90 to 100 (v / v), 95 to 100 (v / v), such as 99% (v / v). In an embodiment, the alcohol has a concentration of from 10 to 100% (v / v), from 20 to 100 (v / v), from 30 to 100 (v / v), from 40 to 100 (v / v / v), 60 to 100 (v / v), 70 to 100 (v / v), 80 to 100 (v / v), 90 to 100 (v / Or 98 to 100 (v / v), such as 99% (v / v) methanol, ethanol, and propanol (e.g., isopropyl alcohol).

The amount of the alcohol to be added is preferably 5 to 10 times the volume of the concentrated sugar mixture. If the particle size is less than 5 times, particle precipitation is not smooth, and if it is 10 times or more, there is a disadvantage that cost is increased due to excessive use.

The galactose produced by the method for producing galactose may include D-galactose, L-galactose, or a mixture thereof. In addition, the galactose is white and may be in solid particulate form. When galactose is obtained in the form of solid particles, galactose is advantageous in ease of metering, precise control of dosage, ease of storage, and reduction in volume when starting galactose as a starting material for other chemical reactions. On the other hand, when galactose in a liquid state is used as a starting material of a subsequent chemical reaction material, there is a problem that the above-described advantages are lost and the subsequent process becomes very difficult.

Another example provides the galactose produced by the galactose production method.

The galactose produced by the method for producing galactose may include D-galactose, L-galactose, or a mixture thereof. In addition, the galactose may be in the form of a white, solid particle, but is not limited thereto.

The galactose may have a melting point of 163 to 170 ° C, for example, 167 to 169 ° C. Galactose has a purity of at least 80 wt%, at least 85 wt%, at least 90 wt%, at least 95 wt%, at least 96 wt%, at least 97 wt%, at least 98 wt%, or at least 99 wt% Or more.

The production method of the present invention makes it possible to separate and obtain particulate galactose from the red alga hydrolyzed glycosylated liquid solution, and to provide a new process technology which is industrially economical and highly utilizable. Specifically, it is possible to mass-separate and purify galactose through the enlargement of the silica filled column and the ion exchange resin filled column, and it is possible to mass-produce galactose at a low cost. In addition, galactose can be easily separated and granulated by using the separation and purification method according to the present invention from the saccharified liquid obtained through saccharification of red algae to produce galactose, which is used as an important intermediate material in the production of biochemical materials Process technology that can be

FIG. 1 is a flowchart showing a process for producing galactose according to an embodiment of the present invention.
FIG. 2 is a photograph of a sugar mixture according to an embodiment of the present invention treated with an ion exchange resin and then measured with a pH paper.
3 is a photograph showing particles of galactose prepared according to an embodiment of the present invention.

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

Example  1. Production of galactose from seaweed

Glycation  step

Red algae collected from Jeonnam coast were dried and crushed. Thereafter, 350 cc of distilled water and 150 cc of 1N HCl solution were added to the 500 cc flask, and the acid concentration of the solution was adjusted to 0.3 N HCl. Then, 25 g of the crushed cucumber was put in the flask and stirred at 120 ° C for 4 hours. Then, stirring was stopped and only the upper liquid phase material was obtained at room temperature (25 캜). The volume of the obtained liquid material was about 250 cc.

Filtration step

A first filtration step of flowing 250 cc of the liquid material obtained above to a silica column filled with 100 cc of neutral silica particles having a size of 0.1 to 0.5 mm at a flow rate of 3.5 ml / min was performed.

Next, a secondary filtration step was performed in which a liquid material passed through a silica column was passed through a column filled with 50 cc of ion exchange resin (TRILITE AMP 26) and 25 cc of silica particles alternately at a rate of 3.5 ml / min.

Then, the liquid material passed through the column was passed through a silica column filled with 100 cc of neutral silica particles having a size of 0.1 to 0.5 mm at a rate of 3.5 ml / min to perform a third filtration step for removing some impurities.

Concentration and Granulation  step( Precipitation step )

The concentrate was then concentrated using a rotary evaporator (IKA® RV 10 rotary evaporator) at a bath temperature of 55 ° C. and a reduced pressure of 90 mbar to a volume of about 1/20 of the pre-condensation volume. Then 99% (v / v) ethanol stored at low temperature was added to the concentrated material to precipitate the particles. At this time, the amount of ethanol added was about 20 times the volume of the concentrated substance. The precipitated particles were recovered by vacuum filtration. The thus obtained galactose is shown in Fig. 3, it can be confirmed that the galactose obtained by the above method is in the form of white solid particles (powder).

Comparative Example  One.

The filtrate was prepared under the same conditions as in Example 1, except that the saccharification step was followed by the concentration and granulation step without filtration step.

Comparative Example  2.

The preparation was carried out under the same conditions as in Example 1, except that only the primary filtration step was performed in the filtration step after the saccharification step, followed by the concentration and granulation step.

Comparative Example  3.

The filtrate was subjected to the first and second filtration steps, followed by concentration and granulation steps.

Comparative Example  4.

The particles were prepared under the same conditions as in Example 1, but concentrated to only one-half of the volume before concentration in the concentration and granulation step.

Comparative Example  5.

(V / v) hexane reagent in place of ethanol in the granulation step. The granulation was carried out in the same manner as in Example 1, Was used to proceed with the granulation step. At this time, the amount of hexane reagent added was about 20 times that of the concentrated material.

Comparative Example  6.

The mixture was concentrated under the same conditions as in Example 1, but concentrated to a half of the volume before concentration. Then, 99% (v / v) ethyl acetate The reagent was used to proceed with the granulation step. At this time, the amount of ethyl acetate reagent added was about 20 times as much as the volume of the concentrated substance.

Comparative Example  7.

(V / v) hexane in place of ethanol in the granulation step, and the mixture was concentrated under the same conditions as in Example 1, but concentrated to a half of the pre-concentration volume in the concentration and granulation step. The granulation step was carried out using a reagent in which 99% (v / v) ethyl acetate was mixed at a volume ratio of 1: 1. At this time, the content of the mixed reagent was about 20 times as much as the volume of the concentrated substance.

Experimental Example  1. Confirmation of galactose particle formation

The production of galactose was confirmed in Example 1 and Comparative Examples 1 to 5.

Whether particles are generated Example 1 Particulate matter generated Comparative Example 1 Sticky material created
No particulate matter generated Comparative Example 2 Sticky material created
No particulate matter generated Comparative Example 3 Sticky material created
No particulate matter generated Comparative Example 4 Sticky material created
No particulate matter generated Comparative Example 5 Not created Comparative Example 6 Not created Comparative Example 7 Not created

As can be seen from the results of Table 1, in Example 1, particulate matter was generated, but in Comparative Examples 1 to 4, only sticky matter was produced, and particulate matter was not produced.

Experimental Example  2. Galactose through melting point measurement Whether to generate  Confirm

To determine whether galactose was produced in Example 1 and Comparative Examples 1 to 7, melting points were measured using a differential scanning calorimeter (DSC) (TA instruments, USA).

Specifically, the melting point of the galactose reagent was measured to be 167 to 169 占 폚, and the melting points of Example 1 and Comparative Examples 1 to 7 were measured and shown in Table 1 below.

Melting point
(° C) Example 1 167 Comparative Example 1 65 Comparative Example 2 66 Comparative Example 3 50 Comparative Example 4 45 Comparative Example 5 Not measurable Comparative Example 6 Not measurable Comparative Example 7 Not measurable

As can be seen from the results of Table 2, in Example 1, the melting point was measured at 167 ° C and contained in the range of 167 to 169 ° C, which is the melting point of the galactose reagent. As a result, it was confirmed that galactose was produced, To 4 were found to have a melting point of 45 to 65 ° C and were not included in the melting point range of the galactose reagent. As a result, it was confirmed that galactose was not produced. In Comparative Examples 5 to 7, no sticky substance was produced, I could not measure it.

Claims (18)

A saccharification step of producing a saccharide mixture containing galactose from a red algae through a saccharification process;
Filtering the sugar mixture to obtain a filtered sugar mixture;
Concentrating the filtered sugar mixture to a level of 1/10 to 1/20 of the volume of the filtered sugar mixture to obtain a concentrated sugar mixture; And
And a precipitation step of precipitating the concentrated sugar mixture by adding an alcohol,
Wherein the filtration step comprises: a primary filtration step in which the sugar mixture obtained in the saccharification step is subjected to silica gel column chromatography to obtain a primary filtered sugar mixture;
A secondary filtration step of treating the primary filtered sugar mixture with a column containing an ion exchange resin to obtain a secondary filtered sugar mixture having a pH of 5 to 7 converted to a level; And
Wherein the second filtered sugar mixture is subjected to a third filtration step to obtain a third filtered sugar mixture by silica gel column chromatography,
Wherein the column of the secondary filtration step has a structure in which an ion exchange resin layer and a silica gel particle layer are alternately laminated 2 to 10 times, and a volume ratio of the single ion exchange resin layer and the single silica gel particle layer is 10: 1 to 1:10. ≪ / RTI > delete The process according to claim 1, wherein the saccharification step is carried out at 80 to 150 ° C. The process according to claim 1, wherein the saccharification step hydrolyzes the red algae to produce a sugar mixture. The process according to claim 4, wherein the hydrolysis is carried out by adding an acid at a concentration of 0.1% (w / v) to 15% (w / v). The method of claim 5, wherein the acid is sulfuric acid (H ₂ SO _4), hydrochloric acid (HCl), hydrobromic acid (HBr), nitric acid (HNO _3), acetic acid (CH ₃ COOH), formic acid (HCOOH), perchloric acid (HClO ₄₎ , Phosphoric acid (H ₃ PO ₄ ), and paratoluene sulfonic acid (PTSA). The method of claim 1, wherein the red algae are selected from the group consisting of Chondrus , Eucheuma , Gigartina , Pterocladia , Hypnea , Iridaea , Wherein the plant is at least one selected from the group consisting of Kappaphycus , Gellidium , and Gracilaria . The process according to claim 1, wherein the filtration step is carried out at a flow rate of 0.1 to 20 ml / min. delete 2. The method according to claim 1, wherein the silica gel of the silica gel column chromatography used in at least one of the first filtering step and the third filtering step has an average diameter of 0.1 to 0.5 mm. The method according to claim 1, wherein the ion exchange resin in the secondary filtration step is a basic anion exchange resin. delete delete The method according to claim 1, wherein the concentration step comprises concentrating the filtered sugar mixture by distillation under reduced pressure. The process according to claim 1, wherein the concentration step is carried out at 30 to 60 占 폚. The process according to claim 1, wherein the concentration step is carried out under a pressure of 10 to 120 mbar. The process according to claim 1, wherein the precipitation is carried out at a temperature of from -10 ° C to 25 ° C. The production method according to claim 1, wherein the alcohol in the precipitation step is at least one selected from the group consisting of methanol, ethanol, and propanol.

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