KR101134619B1 - Method for purifying 1,3-propanediol - Google Patents

Abstract

The present invention comprises the steps of separating the first solvent layer containing 1,3-propanediol by adding a single or mixed solvent to the microbial culture medium containing 1,3-propanediol; Inducing layer separation by changing a solvent component ratio of the first solvent layer; And separating the 1,3-propanediol-containing layer from the layered first solvent layer with the second solvent layer. According to the present invention, 1,3-propanediol can be separated from the microorganism culture medium containing 1,3-propanediol with excellent efficiency.

1,3-propanediol, separation

Description

Separation method of 1,3-propanediol {Method for purifying 1,3-propanediol}

The present invention relates to a method for separating 1,3-propanediol which can be used in a variety of industrially contained in the microbial culture. More specifically, by adding a single or mixed solvent from the microbial culture medium containing 1,3-propanediol to separate the first solvent layer containing 1,3-propanediol, the solvent of the separated first solvent layer The present invention relates to a method for separating high concentration of 1,3 propanediol by inducing layer separation by changing the component ratio.

Recently, as there is a growing concern about resource depletion and environmental pollution due to excessive use of fossil fuels, strengthened regulations on the use of fossil fuels and renewable energy such as biodiesel to solve the environmental pollution and warming problems in developed countries. Policies are in place to promote the dissemination of chemicals, and are spurring the development of environmentally friendly processes that re-use waste and break away from existing chemical processes.

In such an attempt, a high concentration of glycerol is generated as a by-product in the process of producing biodiesel in an amount of about 10% of the biodiesel production, and a large amount and a large treatment cost have been required to be treated as general waste.

In order to overcome this, deterioration of the economics of the biodiesel production process, the development of a new use of the biodiesel waste by-product glycerol has been made mainly in the United States and Europe.

As the supply of glycerol obtained as a by-product of the biodiesel production process and the price decrease, the possibility of using it as a raw material for various fine chemicals is increasing. The production of 1,3-propanediol through the microbial conversion of glycerol It is an area of great interest in terms of feasibility and potential market size. It is also possible to produce 1,3-propanediol not only in glycerol but also from sugars such as glucose.

1,3-propanediol can be used as a monomer of polytrimethlene terephthalate, which has recently been commercialized and expanded in use, and is currently produced by Shell or Dupont. Polytrimethylene terephthalate is expected to form a large market in 2010, with production exceeding 1 million tons. In order to cope with this, it is necessary to secure eco-friendly production technology and price competitiveness of 1,3-propanediol, a polytrimethylene terephthalate monomer. There is a need for techniques to effectively separate and purify from.

An object according to an embodiment of the present invention is to provide a method for separating 1,3-propanediol, which can be industrially used in various concentrations and with high efficiency.

One embodiment according to the present invention for achieving the above object is to separate the first solvent layer containing 1,3-propanediol by adding a single or mixed solvent to the microbial culture medium containing 1,3-propanediol Making a step; Inducing layer separation by changing a solvent component ratio of the first solvent layer; And separating the 1,3-propanediol-containing layer from the layered first solvent layer with the second solvent layer.

In addition, one embodiment according to the present invention relates to 1,3-propanediol separated by the above separation method.

According to the present invention, since 1,3-propanediol can be efficiently separated at high concentration by a simple method that can be industrially applied, the production and separation technology of 1,3-propanediol, which has recently been spotlighted as a useful compound It can contribute greatly to development.

As a conventional technique for separating 1,3-propanediol from a microbial culture, there are methods such as pervaporation, distillation, or adsorption. However, the above prior art shows a blockage phenomenon in the separator used during pervaporation, very low removal rate of 1,3-propanediol during pervaporation or adsorption, or 1,3- There are disadvantages such as low selectivity to propanediol.

Another method of separating 1,3-propanediol from microbial cultures is to save energy if extraction is used for 1,3-propanediol separation, but selectively select 1,3-propanediol from aqueous solution. The choice of solvent to extract is important.

Regarding the separation of 1,3-propanediol, Korean Patent Application Publication No. 2006-0020864 discloses a mixture containing 1,3-propanediol, 1,2-propanediol, glycerol and glucose under reduced pressure. It is concentrated, and the concentrate is treated with ethyl acetate, methyl ethyl ketone and a mixed solvent thereof, and then a method of separating 1,3-propanediol and 1,2-propanediol using low pressure chromatography is disclosed. .

However, in the above separation method, 1,3-propanediol, which has been transferred to a solvent, is subjected to low pressure chromatography using a methanol solvent in a column filled with silica, and then treated with a solvent. There is a difference from the present invention in which a solvent layer containing, 3-propanediol is separated and a high concentration of 1,3-propanediol is separated using 1,3-propanediol solubility difference according to the solvent component. In addition, even when separated by the above method, glycerol is only partially removed (see Examples 1 to 3 in the specification of 2006-0020864), so that glycerol remaining together with 1,3-propanediol is affected as an impurity. It can be said that the efficiency is inferior to the separation method of the present invention in that it can be. In particular, in Korean Patent Application Publication No. 2006-0020864, the separable 1,3-propanediol concentration in the solvent layer loaded on low pressure chromatography is limited to 40 g / L or less, so that the separable amount per unit time is low and the final concentration is Low efficiency.

Thus, the present invention comprises the steps of separating the first solvent layer containing 1,3-propanediol by adding a single or mixed solvent to the microbial culture medium containing 1,3-propanediol; Inducing layer separation by changing a solvent component ratio of the first solvent layer; And separating the 1,3-propanediol-containing layer from the layered first solvent layer with the second solvent layer.

First, the present invention includes the step of separating the first solvent layer containing 1,3-propanediol by adding a single or mixed solvent to the microbial culture medium containing 1,3-propanediol.

In one embodiment, the single solvent added to the microbial culture is not particularly limited as long as it is a solvent suitable for extracting 1,3-propanediol from the microbial culture, for example methyl-ethyl- ketone), ethyl acetate, di-ethyl ether and MTBE (methyl tert-butyl ether) may be at least one selected from the group consisting of, specifically methyl-ethyl-ketone may be a single solvent. EXAMPLES As illustrated below, the 1,3-propanediol concentration remaining in the microbial culture was analyzed by gas chromatography to extract 1 by solvent through the difference in concentration before and after performing extraction with the added solvent. The amount of, 3-propanediol can be calculated, and when methyl-ethyl-ketone is added to the microbial culture, the extraction concentration of 1,3-propanediol in the solvent is high and the extraction rate is 35% or more. .

In another embodiment, the mixed solvent is at least one selected from the group consisting of methyl-ethyl-ketone, ethyl acetate, di-ethyl ether and methyl tert-butyl ether (MTBE) (solvent A) And a mixed solvent of a hydrophobic solvent (solvent B). The inventors of the present application show that the hydrophobic solvent is methyl-ethyl-ketone, ethyl acetate, di-ethyl ether and MTBE even when the above solvents are mixed as illustrated in Example 6 below. (methyl tert-butyl ether) was confirmed that does not affect the extraction performance of one or more selected from the group consisting of, 1,3-propanediol may be extracted through a mixed solvent.

The hydrophobic solvent is not particularly limited, but may be, for example, at least one selected from the group consisting of hexane, heptane, decane, dodecane, ether, paraffin solution, and oleyl alcohol.

At this time, the volume ratio of the at least one solvent and hydrophobic solvent selected from the group consisting of methyl-ethyl-ketone, ethyl acetate, di-ethyl ether and MTBE (methyl tert-butyl ether) in the mixed solvent Although it will not restrict | limit especially if it is a grade which does not affect the extraction rate of 1, 3- propanediol, For example, it may be more than 1: 0 and 1: 9 or less.

In one embodiment, the volume ratio of the microorganism culture medium containing the single or mixed solvent and 1,3-propanediol may be 40: 1 to 1:40, preferably 10: 1 to 1: 2. . The inventors of the present application confirmed that the extraction amount of 1,3-propanediol increases as the volume ratio of the solvent increases.

The present invention includes the step of inducing a layer separation by changing the solvent component ratio of the first solvent layer.

In one embodiment, the solvent component ratio of the first solvent layer may be changed by adding a hydrophobic solvent.

The hydrophobic solvent may be at least one selected from the group consisting of hexane, heptane, decane, dodecane, ether, paraffin solution and oleyl alcohol as mentioned above, and the volume ratio of the final hydrophobic solvent may be It may be 10 to 90% (v / v) as a reference, preferably 30 to 80% (v / v).

In another embodiment, the solvent component ratio of the first solvent layer may be changed by evaporating the solvent contained in the solvent layer under reduced pressure.

At this time, when the solvent used for separating the first solvent layer is a single solvent, the hydrophobic solvent may be added to change the solvent component ratio to separate the 1,3-propanediol layer, and in some cases, before adding the hydrophobic solvent. Layer separation may be facilitated by evaporating a single solvent contained in the separated first solvent layer to increase the 1,3-propanediol concentration.

In addition, the solvent used to separate the first solvent layer is one or more selected from the group consisting of methyl-ethyl-ketone, ethyl acetate, di-ethyl ether, and methyl tert-butyl ether (MTBE) (solvent In the case of a mixed solvent of A) and a hydrophobic solvent (solvent B), the solvent component ratio may be changed by further adding a hydrophobic solvent B to the separated first solvent layer to separate the 1,3-propanediol layer. In some cases, the separated first solvent layer may be heated to increase the volume ratio of the hydrophobic solvent in the mixed solvent included in the first solvent layer, or increase the concentration of 1,3-propanediol, thereby making layer separation easier. You can also

At this time, when the boiling point of the solvent B (e.g., heptane), which is a hydrophobic solvent, is higher than that of solvent A (e.g., methyl ethyl ketone), the volume of the hydrophobic solvent having a relatively higher boiling point is heated by heating the separated first solvent layer. Since the ratio is higher in the solvent layer, the solvent component ratio is changed at the same time as heating, thereby inducing the layer separation.

In addition, when the boiling point of the solvent B (for example, hexane), which is a hydrophobic solvent, is lower than that of the solvent A (for example, methyl ethyl ketone), an additional hydrophobic solvent is added to the first solvent layer to induce layer separation or the first solvent. The layer may be heated to increase the concentration of 1,3-propanediol in the mixed solvent included in the solvent layer, and then hydrophobic solvent may be added to induce separation of the layers.

Thereafter, a step of separating the 1,3-propanediol-containing layer from the layered first solvent layer with the second solvent layer. The 1,3-propanediol-containing layer may be recovered in the lower layer after the separation of the layer.

In some cases, the separation method includes separating the 1,3-propanediol-containing layer from the second solvent layer, and then evaporating the separated 1,3-propanediol-containing layer to be included in the 1,3-propanediol-containing layer. The method may further include removing water and a solvent, and the water and the solvent may be formed through heat evaporation. This gives a high concentration of pure 1,3-propanediol.

In this regard, US Pat. No. 7,572,376 discloses extracting 1,3-propanediol using a hexanol or tributyl phosphate (TBP) solvent in a concentrated culture, and the number of carbon atoms in the solvent layer. A method of separating 1,3-propanediol by performing back extraction with an alkanes solvent of 6 to 10 or water is presented. However, not only the solvent used is different from the single or mixed solvent used in the present invention, but the final 1,3-propanediol concentration is low after back extraction, so that a large amount of solvent or water must be evaporated to produce pure 1,3-propanedi The ol can be separated (see Examples 1 to 1). In addition, since a small amount of hexanol or tributyl phosphate was dissolved in the finally separated aqueous solution layer containing 1,3-propanediol, hexane was added to remove the layer, which was used to separate the hexanol or tributyl phosphate. Since the boiling point is higher than water, it may remain as an impurity without being removed when the aqueous solution evaporates (see Example 8).

In contrast, the present invention is lower than the boiling point of the single or mixed solvent used in the separation of the solvent layer containing 1,3-propanediol, it is necessary to undergo a separate layer separation process as in US 7,572,376 Without evaporation, high concentrations of 1,3-propanediol can be efficiently obtained.

In some cases, the second solvent layer solvent other than the 1,3-propanediol-containing layer in the solvent or the separated solvent layer removed above may be separated by distillation and reused as a solvent.

In one embodiment, the culture is not particularly limited as long as it can be obtained from 1,3-propanediol producing microorganisms, for example Klebsiella pneumonae , Klebsiella oxytoca , Enterobacter agglomerans , Clostridium butyricum , Citrobacter freundii , and microorganisms, including recombinant E. coli. .

Since the microorganism ingests a carbon source such as glycerol or glucose and produces a solvent such as ethanol, acetic acid, butanediol, 1,3-propanediol, and the like as a metabolite, the culture medium is 1,3-propanediol, 1 It may comprise one or more mixtures selected from the group consisting of, 2-propanediol, butanediol, glycerol and glucose.

The culture may contain 1,3-propanediol at a concentration of 20 to 100 g / L, preferably 20 to 80 g / L, which means that the microorganism is active at high concentrations of 1,3-propanediol in the culture. This is because it is inhibited. In some cases, the culture solution may be a concentrated culture solution, wherein the culture solution may be concentrated at a concentration of 100 g / L or more, specifically, 100 to 500 g / L.

In some cases, the separation method may further include adjusting the pH by adding an acid or a base to the microbial culture.

The inventors of the present application confirmed that the extraction rate can be increased when the culture solution is adjusted to pH 2 to 3 by adding an acid as described in detail in the following Examples. The acid is not particularly limited as long as it is suitable for controlling the desired pH, but for example, one or more selected from the group consisting of acetic acid, butyric acid, lactic acid and succinic acid, which microorganisms consume glycerol and produce as a by-product, may be added. . In addition, it was confirmed that the extraction rate was increased even when adjusted to pH 10-11 by adding a base such as sodium hydroxide (NaOH).

It is thought that the added acid and base influenced the binding between water and 1,3-propanediol and consequently the 1,3-propanediol extraction rate.

In some cases, the separation method may further include adding water to the first solvent layer having a changed component ratio to induce phase separation. As shown in Example 7 below, for example, when using a solvent in which methyl ethyl ketone and hexane or methyl ethyl ketone and heptane are mixed, even a small amount of water separates the layers, thereby increasing the concentration of 1,3-propane die It was confirmed that the ol could be extracted. Furthermore, in the case of using a solvent in which methyl ethyl ketone and heptane are mixed, evaporating methyl ethyl ketone having a low boiling point by heating, when the volume is reduced to a certain degree, layer separation occurs as a result of cooling to room temperature and high concentration with excellent efficiency. It was confirmed that 1,3-propanediol can be recovered.

At this time, the volume ratio of the first solvent layer and water is not particularly limited as long as it can induce a layer separation, for example, may be 100: 1 to 1:10, preferably 50: 1 to 1: 5. .

Hereinafter, the present invention will be described in more detail with reference to the following examples. The following examples are intended to illustrate the invention and are not intended to limit the scope of the invention.

Example 1 Selection of Effective Extraction Solvent

15 ml of 1,3-propanediol diluted at 50 g / L was added to a 50 mL funnel, followed by extraction solvents methyl-ethyl-ketone (MEK), ethyl acetate, and di-ethyl ether (di- 15 ml of ethyl ether), MTBE (methyl tert-butyl ether), and oleyl alcohol were added, and shaken at 30 minutes intervals at 280 rpm using a shaker for extraction. The concentration of 1,3-propanediol in the aqueous solution before and after extraction was quantified using a gas chromatography equipped with FID (Agilent technology 6890N Network GC system), the column was HP-INNOWAX (30 mx 250㎛ x 0.25㎛, Agilent technology). 1,3-propanediol concentration and extraction rate extracted with each solvent are shown in Table 1.

[Table 1] 1,3-propanediol extraction concentration and extraction rate according to the extraction solvent

Example 2 pH Impact Assessment

This time, sodium hydroxide (NaOH) and acetic acid (acetic acid) produced as a by-product from microbial culture were adjusted to pH 10.8 and pH 2.2, respectively. Add 15 ml of 1,3-propanediol diluted to 50 g / L in a 50 mL funnel, adjust the pH, and add 15 mL of methyl-ethyl-ketone (MEK), an extraction solvent, using a 280 rpm stirrer. After stirring for 30 minutes, the amount of 1,3-propanediol extract was investigated. As shown in Table 2, the extraction rate was higher in the sample adjusted to pH 2.2 or pH 10.8 than the sample without pH adjustment. On the other hand, when sodium acetate, a salt such as acetic acid, was added, the extraction rate was rather reduced. From this result, it can be seen that low pH by acid rather than acetate ion affects extraction.

[Table 2] Extraction pattern of methyl ethyl ketone solvent according to pH

Example 3 Glycerol and Glucose Extraction

To determine whether glycerol is extracted as a solvent for 1,3-propanediol extraction, use the same amount (15 mL) in 15 mL of an aqueous solution containing 46 g / L of glycerol and 30 g / L of 1,3-propanediol. Methyl ethyl ketone was added, followed by stirring.

As a result, it was confirmed that the glycerol of the aqueous solution layer was not extracted to the solvent layer since there is no change in concentration to 46 g / L after extraction, it was confirmed that does not affect the extraction rate of 1,3-propanediol. In the same manner, it was confirmed that glucose was not extracted with methyl ethyl ketone.

Example 4 Extraction Aspect According to Volume of Extraction Solvent

10 mL of 1,3-propanediol was added to each funnel and 5, 10, 20,40 mL of methyl ethyl ketone was stirred at 280 rpm for 30 minutes using a stirrer, followed by extraction. As a result, as shown in Table 3, the higher the methyl ethyl ketone volume ratio, the higher the extraction rate.

[Table 3] Extraction pattern according to extraction solvent volume

Example 5 Extraction Pattern According to 1,3-Propandiol Concentration

15 mL of an aqueous solution containing 20, 40, 60, 100 g / L of 1,3-propanediol was added to each funnel, 15 mL of methyl ethyl ketone was added, and stirred at 280 rpm for 30 minutes using a stirrer. Was observed, and the results are shown in Table 4. The higher the concentration of 1,3-propanediol in the initial aqueous solution layer, the higher the concentration extracted with the solvent. The extraction rate and partition coefficient were the highest at 40 g / L. From the results of this example, it can be seen that even if the aqueous solution layer 1,3-propanediol concentration is high, only up to 27 g / L in methyl ethyl ketone is extracted.

[Table 4] Effect of 1,3-propanediol concentration on extraction

Example 6 Extraction Using Mixed Solvent

15 mL of 1,3-propanediol aqueous solution was added to each funnel, and 15 mL of a mixed solvent in which the volume ratio of methyl ethyl ketone: hexane was adjusted to 10: 0 to 5: 5 was added thereto, followed by 280 rpm. After stirring for 30 minutes, the extraction pattern was observed. As shown in Table 5, the extraction ratio of 1,3-propanediol was the highest at 39% when methyl ethyl ketone: hexane = 10: 0 and the lowest at 19% at 5: 5, but was based on methyl ethyl ketone volume 1,3 -Propanediol extraction concentration is constant at 22 ~ 27 g / L regardless of the volume ratio. It can be seen that even using a solvent mixed with methyl ethyl ketone and hexane does not affect the extraction performance of methyl ethyl ketone, and the amount of 1,3-propanediol extracted is related only to the volume of methyl ethyl ketone. .

[Table 5] Extraction pattern according to the mixed solvent volume ratio

Example 7 Layer Separation According to Change of Solvent Component Ratio

As shown in Table 6, when 1 mL of water was added to 15 mL of methylethylketone containing 1.8 g of 1,3-propanediol, no layer separation occurred, and when 3 mL of water was added, layer separation occurred, but The volume was less than 1 mL.

As shown in Table 6, 3.8 mL of hexane was added to 15 mL of methylethylketone containing 1.8 g to 2.2 g of 1,3-propanediol (methylethyl ketone: hexane = 8: 2), followed by stirring No separation occurs. However, 15 mL of hexane was added to 15 mL of methyl ethyl ketone containing 1.8 g of 1,3-propanediol to change the solvent component ratio (methyl ethyl ketone: hexane = 5: 5). After 2 minutes it is separated into upper and lower layers. As a result, the volume of the lower layer was 1.2 mL, the concentration of 1,3-propanediol 800 g / L, and thus the weight of 1,3-propanediol separated into the lower layer was 0.96 g, yielding 53.2%. When 1mL of water was added to the upper / lower layer solution and stirred, and observed, the upper layer and the lower layer were immediately separated from the case where no hexane was added. In other words, when methyl ethyl ketone is used as a single solvent, a small amount of water and a layer are not separated, and back extraction is impossible. However, when a solvent mixed with methyl ethyl ketone and hexane is used, a small amount of water is used to separate the layers. This makes it possible to back extract 1,3-propanediol at high concentrations.

As shown in Table 6, when 1 mL of water was added, the concentration of 1,3-propanediol in the lower layer was 450 g / L, and the weight of 1,3-propanediol separated in the lower layer was 1 g and the recovery rate was 55.1%. In the same way, when 3 mL of water is added, it is separated into an upper layer and a lower layer. As shown in Table 6, when 3 mL of water was added, the concentration of 1,3-propanediol in the lower layer was 343 g / L, the weight of 1,3-propanediol separated in the lower layer was 1.44 g, and the recovery rate was 79.7%. Increasing this, it can be seen that the recovery of 1,3-propanediol is also improved.

As shown in Table 6, 5 mL of heptane was added to 5 mL of methyl ethyl ketone containing 0.625 g of 1,3-propanediol to change the solvent component ratio (methyl ethyl ketone: heptane = 5: 5) and stirred. After observation, it immediately separates into upper and lower layers. As a result, the volume of the lower layer was 0.5 mL, the concentration of 1,3-propanediol was 1020 g / L, and thus the weight of 1,3-propanediol separated into the lower layer was 0.53 g, yielding 84.8%. Considering that the 1,3-propanediol density is 1.05 g / mL, it can be confirmed that the 1,3-propanediol separated in the lower layer has a purity of 97% or more.

After confirming that 15 mL of heptane was added to 15 mL of methyl ethyl ketone containing 0.6 g of 1,3-propanediol to form a monolayer, it was heated at 80 degrees to change the solvent component ratio, and then the low boiling ethyl ethyl The ketone was intended to evaporate. When the volume was reduced to 20 mL, cooling to room temperature resulted in layer separation. As a result, the volume of the lower layer was 0.6 mL, the concentration of 1,3-propanediol was 967.1 g / L, and thus the weight of 1,3-propanediol separated into the lower layer was 0.58 g, yielding 96.6%.

Methyl ethyl ketone contained in the lower layer containing the high concentration of 1,3-propanediol separated in the above embodiment can be removed by evaporation with water because the boiling point is lower than water, thus high purity 1,3-propanediol You can get it.

[Table 6] Layer separation and 1,3-propanediol separation according to the solvent component ratio

Those skilled in the art to which the present invention pertains will be able to perform various applications and modifications within the scope of the present invention based on the above contents.

1 is a schematic diagram showing a separation method of 1,3-propanediol according to an embodiment of the present invention.

Claims (22)

Separating the first solvent layer containing 1,3-propanediol by adding a single or mixed solvent to the microbial culture medium containing 1,3-propanediol; Inducing layer separation by changing a solvent component ratio of the first solvent layer; And And separating the 1,3-propanediol-containing layer from the layered first solvent layer with a second solvent layer. The single solvent is at least one selected from the group consisting of methyl-ethyl-ketone, ethyl acetate, di-ethyl ether, and methyl tert-butyl ether (MTBE), The mixed solvent is a mixed solvent of a hydrophobic solvent and at least one solvent selected from the group consisting of methyl-ethyl-ketone, ethyl acetate, di-ethyl ether and MTBE (methyl tert-butyl ether) Separation method of 1,3-propanediol. delete The method of claim 1, And wherein said single solvent is methyl-ethyl-ketone. delete The method of claim 1, The hydrophobic solvent is at least one selected from the group consisting of hexane, heptane, decane, dodecane, ether, paraffin solution and oleyl alcohol (oleyl alcohol) separation method of 1,3-propanediol. The method of claim 1, The volume ratio of the hydrophobic solvent with one or more solvents selected from the group consisting of methyl-ethyl-ketone, ethyl acetate, di-ethyl ether, and MTBE (methyl tert-butyl ether) in the mixed solvent is 1: 0. A method of separating 1,3-propanediol that is greater than 1: 9. The method of claim 1, Separation method of 1,3-propanediol is a volume ratio of the microorganism culture medium containing the single or mixed solvent and 1,3-propanediol is 40: 1 to 1:40. The method of claim 7, wherein Separation method of 1,3-propanediol is a volume ratio of the single or mixed solvent and the microbial culture medium containing 1,3-propanediol is 10: 1 to 1: 2. The method of claim 1, The solvent component ratio of the separated first solvent layer is a method of separating 1,3-propanediol is changed by adding a hydrophobic solvent. The method of claim 9, The hydrophobic solvent is a volume ratio of 10 to 90% (v / v) based on the first solvent layer separation method of 1,3-propanediol. The method of claim 1, The solvent component ratio of the first solvent layer is a method for separating 1,3-propanediol is changed by evaporation of the solvent contained in the solvent layer under reduced pressure. The method of claim 11, The solvent is evaporated under reduced pressure is separated by distillation and reuse of 1,3-propanediol. The method of claim 1, The separation method further comprises the step of removing the water and the solvent in the separated 1,3-propanediol-containing layer. The method of claim 13, The removed solvent is a separation method of 1,3-propanediol is separated by distillation and reused. The method of claim 1, The solvent of the second solvent layer is a separation method of 1,3-propanediol separated by distillation and reused. The method of claim 1, The culture medium is a 1,3-propanediol separation method comprising at least one mixture selected from the group consisting of 1,3-propanediol, 1,2-propanediol, butanediol, glycerol and glucose . The method of claim 1, The culture medium is Klebsiella pneumonae , Klebsiella oxytoca , Enterobacter agglomerans , Clostridium butyricum , Citrobacter Freundy ( Clebsiella pneumonae ) Citrobacter freundii ) and a method for separating 1,3-propanediol obtained from one or more 1,3-propanediol producing microorganisms selected from the group consisting of microorganisms including recombinant E. coli. The method of claim 1, The culture medium is a separation method of 1,3-propanediol, characterized in that the culture medium concentrated to a concentration of 100 to 500g / L. The method of claim 1, The separation method of 1,3-propanediol separation method further comprises the step of adjusting the pH by adding an acid or a base to the microbial culture. The method of claim 1 The separation method further comprises the step of inducing a layer separation by adding water to the first solvent layer having a component ratio is changed. The method of claim 20, wherein the volume ratio of the first solvent layer and water is 100: 1 to 1:10. delete

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