KR102729011B1 - Method for preparing triglyceride utilizing fractionation byproduct - Google Patents

Abstract

The present invention relates to a method for producing triglycerides by utilizing fractionation by-products, and more specifically, to a method for producing triglycerides by utilizing an enzymatic esterification reaction in which an acyl group of a triacylglyceride is exchanged with another acyl group, wherein the reaction product is distilled and then fractionally purified, and the obtained fractionation by-product is included in the reaction mixture of the enzymatic esterification reaction, thereby lowering the cost of producing triglycerides and increasing the yield, thereby improving the efficiency and economic feasibility of the overall process.

Description

{Method for preparing triglyceride utilizing fractionation byproduct}

The present invention relates to a method for producing triglycerides by utilizing fractionation by-products, and more specifically, to a method for producing triglycerides by utilizing an enzymatic esterification reaction in which an acyl group of a triacylglyceride is exchanged with another acyl group, wherein the reaction product is distilled and then fractionally purified, and the obtained fractionation by-product is included in the reaction mixture of the enzymatic esterification reaction, thereby lowering the cost of producing triglycerides and increasing the yield, thereby improving the efficiency and economic feasibility of the overall process.

Triacylglyceride fats and oils are substances that are widely used in the food industry, and among them, 1,3-dioleoyl-2-palmitoyl glyceride (OPO) is an important component of breast milk fat and has been extensively studied nutritionally.

The fat composition containing the major fatty acids that make up the fat of human milk is found in many vegetable oils and fats, but almost all vegetable glycerides do not have a saturated fatty acid in the 2-position. In contrast, in human milk fat, a significant amount of the saturated fatty acid palmitic acid occupies the 2-position of the glycerides.

As people's interest in infant health has increased recently, the demand for foods with similar ingredients to breast milk is increasing.

Innis (Adv. Nutr., 2:275-283, 2011) reports on the structural role of triacylglycerols in the infant diet.

However, regular powdered milk is a mixture of powdered milk and vegetable oil, and when the oil content of powdered milk is analyzed, it contains almost no palmitic acid in the 2-position.

Accordingly, the application of high-quality human milk fat substitutes for adding 1,3-dioleoyl-2-palmitoyl glycerides (OPO), which contain palmitic acid in the 2-position and have a composition similar to human milk fat, to infant formula is increasing.

OPO is a type of Unsaturated-Saturated-Unsaturated (USU) triglyceride. Mohamad, et al. (Lipase-catalyzed interesterification reactions for human milk fat substitutes production, 2013, 115, 270-285) showed that research has mainly been conducted to synthesize 1,3-dioleoyl-2-palmitoyl glyceride by substituting the 1,3-position of triacylglyceride using lipase. Due to the 1,3-position-specific reactivity of lipase for triacylglycerides, providing a glyceride backbone containing a saturated fatty acid at the 2-position of triacylglyceride fat is important in synthesizing 1,3-dioleoyl-2-palmitoyl glyceride (OPO).

These prior technologies utilize enzymatic esterification reactions, but as the reaction progresses, enzyme activity decreases, resulting in lower efficiency and economic feasibility of the overall process.

Therefore, there is a demand for the development of a new method that can further improve process efficiency and economic feasibility in producing 1,3-dioleoyl-2-palmitoyl glyceride (OPO) using an enzymatic esterification reaction.

The present invention has as a technical problem the provision of a method for producing triglycerides (e.g., 1,3-dioleoyl-2-palmitoyl glycerides (OPO)) through an enzymatic esterification reaction in which an acyl group of a triacylglyceride (e.g., tripalmitoyl glyceride (PPP)) is exchanged with another acyl group (e.g., an oleyl group), thereby reducing the cost of producing triglycerides and increasing the yield, thereby improving the overall process efficiency and economic feasibility.

In order to achieve the above technical task, the present invention provides a method for producing a triglyceride, comprising a step of performing an enzymatic esterification reaction on a reactant mixture comprising a triacylglyceride and a fatty acid having an acyl group different from an acyl group in the triacylglyceride to exchange an acyl group of the triacylglyceride with an acyl group of the fatty acid, characterized in that the reactant mixture further includes a fractionation by-product obtained by fractionating a distillation residue of a resultant of the enzymatic esterification reaction.

According to another aspect of the present invention, a triglyceride composition is provided, which is obtained by performing an enzymatic esterification reaction on a reactant mixture including a triacylglyceride and a fatty acid having an acyl group different from an acyl group in the triacylglyceride to exchange an acyl group of the triacylglyceride with an acyl group of the fatty acid, and fractionating a distillation residue of the resultant reaction, wherein the content of a neutral lipid having a total of 52 carbon atoms (C52) is 15 wt% or more.

According to the present invention, in producing a triglyceride (e.g., 1,3-dioleoyl-2-palmitoyl glyceride (OPO)) in which an acyl group of a triacylglyceride is exchanged with another acyl group through an enzymatic transesterification reaction, the production cost can be reduced and the yield increased compared to the prior art, thereby improving the overall process efficiency and economic feasibility.

Hereinafter, the present invention will be described in detail.

The triacylglyceride used as a reactant of the enzymatic esterification reaction in the present invention may be an edible fat.

As the edible fat, any suitable edible fat may be used, including liquid or solid vegetable fat or animal fat, hardened oil or transesterified oil of the fat, liquid oil obtained by fractionating the fat, solid fat, etc.

Specific examples of the vegetable oils include those selected from corn oil, rapeseed oil, soybean oil, cottonseed oil, palm oil, coconut oil, rice bran oil, sesame oil, cocoa fat, olive oil, and combinations thereof, and specific examples of the animal oils include those selected from fish oil, lard, beef tallow, chicken fat, and combinations thereof. These edible oils may be used alone or in combination of two or more.

In one specific example, the triacylglyceride may be fractionated palm oil obtained by fractionating palm oil among vegetable oils.

In one specific embodiment, the acyl group present in the triacylglyceride can be independently selected from a saturated or unsaturated straight-chain aliphatic acyl group having 4 to 24 carbon atoms, more specifically 12 to 22 carbon atoms, and even more specifically 14 to 20 carbon atoms. For example, the acyl group present in the triacylglyceride can be independently selected from a palmitoyl group, a stearoyl group, an oleoyl group, and a linoleoyl group, and is preferably, but not limited to, a palmitoyl group.

In one specific embodiment, at least one of the acyl groups present in the triacylglyceride is a palmitoyl group, preferably two, more preferably all three are palmitoyl groups. That is, in the most preferred embodiment, the starting triacylglyceride is tripalmitoylglyceride (PPP).

The fatty acid used as a reactant for the enzymatic esterification reaction in the present invention has an acyl group different from the acyl group in the triacylglyceride.

In one specific embodiment, the fatty acid may be at least one selected from a saturated or unsaturated straight-chain fatty acid having 4 to 24 carbon atoms, more specifically 12 to 22 carbon atoms, and even more specifically 14 to 20 carbon atoms. More specifically, the fatty acid may be selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid, and combinations thereof, and is preferably, but not limited to, oleic acid.

In the present invention, the reaction temperature of the enzymatic esterification reaction can be determined depending on the type of enzyme used, and specifically, it can be 40°C or higher, 45°C or higher, 50°C or higher, 55°C or higher, or 60°C or higher, and also can be 85°C or lower, 80°C or lower, 75°C or lower, 70°C or lower, or 65°C or lower, but is not limited thereto. In order to optimize the effect of the reaction and the efficiency of the process, the reaction temperature can preferably be 40°C to 80°C, and more preferably can be 45°C to 70°C.

In the present invention, the reaction time of the enzymatic esterification reaction may be, for example, 0.5 hour or more, 1 hour or more, 2 hours or more, 3 hours or more, or 4 hours or more, and may also be 12 hours or less, 10 hours or less, 8 hours or less, or 6 hours or less, but is not limited thereto.

In the present invention, known enzymes can be used as enzymes for the enzymatic esterification reaction without any particular limitation. Specifically, Candida Antarctica B , Thermomyces ranuginosus lanuginose ), Rhizomucour miehei , Rhizopus delemar , Mucormiehei , Alcaligenes sp . , Aspergillus niger , Candida cylindricales cylindraces ), Burkholderia cepacia , Geotricum candidum ) and combinations thereof. An esterifying enzyme derived from a microorganism selected from the group consisting of, but not limited to, the following may be used.

In one specific embodiment, 100 to 3000 parts by weight, or 100 to 2000 parts by weight, or 100 to 1000 parts by weight, of the reactant mixture per 1 part by weight of the enzyme can be continuously treated without replacing the enzyme, and a higher amount can be continuously treated as long as the activity of the enzyme is maintained. For example, if the enzyme activity is appropriate, 500 parts by weight or more, 1000 parts by weight or more, 2000 parts by weight or more, or 3000 parts by weight or more of the reactant mixture per 1 part by weight of the enzyme can be continuously treated without replacing the enzyme, and there is no particular limitation on the upper limit, and even 10000 parts by weight or more can be treated. According to a preferred specific embodiment, the amount of the reactant mixture treated per 1 part by weight of the enzyme can be 1000 parts by weight to 3000 parts by weight.

In one specific example, the triglyceride manufactured according to the above manufacturing method may be dioleoyl monopalmitoyl glyceride, and more specifically, 1,3-dioleoyl-2-palmitoyl glyceride (OPO).

The method for producing triglyceride of the present invention is characterized in that the reaction mixture of the enzymatic esterification reaction further includes a fractionated by-product obtained by fractionating the distillation residue of the resultant enzymatic esterification reaction.

The distillation of the enzymatic esterification reaction resultant is intended to remove fatty acids remaining in the enzymatic esterification reaction resultant, and in one specific example, this can be performed using a thin-film distillation device. At this time, the distillation can be performed under, for example, pressure conditions of 10 mbar or less, 5 mbar or less, or 2 mbar or less and temperature conditions of 180 to 250°C, 200 to 240°C, or 210 to 230°C, but is not limited thereto. In addition, other conditions during distillation using the thin-film distillation device can be appropriately selected. For example, the cooling temperature of the distillate can be 10 to 30°C, the rotation speed of the still can be 300 to 500 rpm, and the feeding speed of the enzymatic esterification reaction resultant can be 20 to 40 g/h, but is not limited thereto. According to one specific example of the present invention, the distillation of the enzymatic esterification reaction product can be performed using a thin film distillation device under the conditions of a distillation temperature of 225°C, a distillation pressure of 2 mbar or less, a distillation effluent cooling temperature of 20°C, a still rotation speed of 400 rpm, and a feeding speed of 32 g/h.

The distillation residue of the enzymatic esterification reaction product obtained through the distillation process described above is fractionated to obtain a fractionated by-product.

In one specific embodiment, the fractionation of the distillation residue may include dissolving the distillation residue in an organic solvent, cooling the solution to produce a solid, separating the produced solid, and removing the organic solvent from the remaining liquid phase to concentrate the resulting product, the result of the concentration being obtained as a fractionation byproduct.

In one specific embodiment, the organic solvent may be acetone, hexane, ethanol or a combination thereof, and acetone is preferably used. There is no particular limitation on the amount of the organic solvent used for dissolving the distillation residue, and for example, the organic solvent may be used so that the volume ratio of the distillation residue to the organic solvent is 1:5 to 1:30, 1:10 to 1:25 or 1:15 to 1:20, but is not limited thereto. In addition, the cooling of the organic solvent solution of the distillation residue may be performed by, for example, allowing the solution to stand at a temperature of 0° C. or lower, -5° C. or lower or -10° C. or lower for 30 minutes or longer, 60 minutes or longer or 90 minutes or longer. According to one specific embodiment of the present invention, the fractionation of the distillation residue may be performed by dissolving the distillation residue in acetone at a volume ratio of 1:16 and then allowing the solution to stand at -13.4° C. for 93 minutes.

In one specific example, the fractionation by-product obtained through the fractionation process as described above may contain C52 (neutral lipid having a total carbon number of 52 (e.g., C18-C16-C18, C16-C18-C18, etc.), such as SPS, SSP, OPO, OOP, LPL, LLP, OPL, etc. (wherein, P=palmitoyl group, S=stearoyl group, O=oleoyl group, L=linoleoyl group)) in an amount of 15 wt% or more, more specifically, 20 wt% or more, even more specifically, 25 wt% or more, and even more specifically, 28 wt% or more, based on 100 wt% of the total fractionation by-product.

In addition, in one specific example, the fractionation by-product may contain C52 neutral lipid in an amount of 60 wt% or less, more specifically 55 wt% or less, even more specifically 50 wt% or less, and even more specifically 45 wt% or less, based on 100 wt% of the total fractionation by-product.

In addition, in one specific example, the fractionation by-product may contain C48 neutral lipid in an amount of 1 to 15 wt%, more specifically 2 to 10 wt%, and even more specifically 5 to 10 wt%, based on 100 wt% of the total fractionation by-product, C50 neutral lipid in an amount of 55 to 80 wt%, more specifically 60 to 75 wt%, and even more specifically 60 to 70 wt%, and C54 neutral lipid in an amount of 0.1 to 5 wt%, more specifically 0.2 to 3 wt%, and even more specifically 0.5 to 1 wt%, but is not limited thereto.

In the method for producing triglyceride of the present invention, the reaction mixture of the enzymatic esterification reaction may contain the fractionated by-product in an amount of 15 wt% or more, 20 wt% or more, 25 wt% or more, or 30 wt% or more, based on 100 wt% of the total of the triacylglyceride and the fractionated by-product.

Additionally, in one specific embodiment, the reaction mixture of the enzymatic esterification reaction may contain the fractionation by-product in an amount of 70 wt% or less, 60 wt% or less, 50 wt% or less, or 40 wt% or less, based on 100 wt% of the total of the triacylglyceride and the fractionation by-product.

When the content of the above-mentioned fractionated by-product in the reaction mixture of the enzymatic esterification reaction satisfies the above-mentioned level, the desired triglyceride production yield and C52 content can be preferably improved, while at the same time excellently reducing the production cost.

In the method for producing triglyceride of the present invention, the mixing ratio of triacylglyceride to fatty acid included in the reaction mixture of the enzymatic esterification reaction may be 1:1.1 or more, 1:1.2 or more, or 1:1.3 or more in weight ratio, and may also be 1:4 or less, 1:3 or less, or 1:2 or less.

When the mixing ratio of triacylglyceride to fatty acid in the reactant mixture of the enzymatic esterification reaction satisfies the above level, the desired triglyceride production yield and C52 content can be desirably improved, while at the same time excellently reducing the production cost.

In a preferred specific embodiment, the reaction mixture of the enzymatic esterification reaction contains the fractionation by-product in an amount of 30 to 40 wt% based on 100 wt% of the total of the triacylglyceride and the fractionation by-product, and the mixing ratio of the triacylglyceride to the fatty acid is 1:1.3 to 1:2 by weight.

According to another aspect of the present invention, a triglyceride composition is provided, which is obtained by performing an enzymatic esterification reaction on a reactant mixture including a triacylglyceride and a fatty acid having an acyl group different from an acyl group in the triacylglyceride to exchange an acyl group of the triacylglyceride with an acyl group of the fatty acid, and fractionating a distillation residue of the resultant reaction, wherein the content of a neutral lipid having a total of 52 carbon atoms (C52) is 15 wt% or more.

The above triacylglycerides, fatty acids, enzymatic esterification reaction, distillation of the reaction product, and fractionation of the distillation residue are as described above.

In one specific embodiment, the triglyceride composition may include C52 (neutral lipid having a total carbon number of 52 (e.g., C18-C16-C18, C16-C18-C18, etc.), such as SPS, SSP, OPO, OOP, LPL, LLP, OPL, etc. (wherein, P=palmitoyl group, S=stearoyl group, O=oleoyl group, L=linoleoyl group)) in an amount of 25 wt% or more, more specifically 30 wt% or more, even more specifically 35 wt% or more, and even more specifically 40 wt% or more, based on 100 wt% of the total composition.

In addition, in one specific embodiment, the triglyceride composition may include C52 neutral lipid in an amount of 80 wt% or less, more specifically 75 wt% or less, even more specifically 70 wt% or less, and even more specifically 65 wt% or less, based on 100 wt% of the total composition.

In addition, in one specific embodiment, the triglyceride composition may contain C48 neutral lipid in an amount of 1 to 15 wt%, more specifically 1 to 10 wt%, and even more specifically 1 to 5 wt%, based on 100 wt% of the total composition, C50 neutral lipid in an amount of 25 to 55 wt%, more specifically 30 to 50 wt%, and even more specifically 35 to 45 wt%, and C54 neutral lipid in an amount of 0.1 to 20 wt%, more specifically 0.1 to 15 wt%, and even more specifically 3 to 10 wt%, but is not limited thereto.

Hereinafter, the present invention will be described more specifically through examples. However, these are provided only to help understanding of the present invention, and the scope of the present invention is not limited thereby.

[ Example ]

Manufacturing example

A 1:2 weight ratio mixture of tripalmitoyl glyceride (PPP, super stearin (SS)) and oleic acid (OA) was prepared.

25 g of the above mixture was placed in a 100 mL Erlenmeyer flask, and 2.5 g (10 wt% based on 100 wt% of the reactant mixture) of a separately weighed enzyme (Novozym 40086, an enzyme derived from Rhizomucour miehei) was added thereto, and an enzymatic esterification reaction was performed at 60°C with a stirring speed of 150 rpm for 6 hours.

Thereafter, the reaction product was distilled (using a thin film distillation device, distillation temperature 225℃, cooling temperature 20℃, 400 rpm, feeding speed 32 g/h) to remove residual fatty acid (OA), and the distillation residue was fractionated (distillation residue: acetone = 1:16 (v/v), cooling temperature -13.4℃, standing time 93 minutes, acetone removed under reduced pressure from the filtrate after filtration) to obtain a fractionated by-product.

The contents of each type of neutral lipid in the reactant tripalmitoyl glyceride (SS) and the obtained fractionated by-product were as follows.

Example

Using tripalmitoylglyceride (SS), oleic acid (OA) used as reactants in the above manufacturing example and the fractionated by-product obtained in the above manufacturing example, an enzymatic esterification reaction was performed under the conditions shown in Table 1 and Table 2 below. Novozym 40086 was used as an enzyme in an amount of 10 wt% relative to 100 wt% of the reactant mixture, and the enzymatic esterification reaction was performed at 60°C and a stirring speed of 150 rpm.

Each reaction result was analyzed using the following method, and the results are shown in Tables 1 and 2 below.

For Table 2, distillation and fractional purification were performed for each reaction product as in the manufacturing example above.

1) Neutral lipids Carbon number analyze

Neutral lipids were separated by carbon number using gas chromatography, and the concentration of C52 (neutral lipids with a total carbon number of 52 (e.g., C18-C16-C18, C16-C18-C18, etc.), such as SPS, SSP, OPO, OOP, LPL, LLP, OPL, etc.) was analyzed. The results are shown in Table 1 and Fig. 1 below.

2) sn - Fatty acid composition analysis at position 2

The proportion of neutral lipids (PPP, PPO, OPO, etc.) with palmitic acid at the sn-2 position was determined using gas chromatography.

3) Isomeric analysis

Neutral lipids in the unsaturated-saturated-unsaturated (USU) form were separated using liquid chromatography, and their concentrations were analyzed. USU is mainly composed of OPO, and analysis was performed to distinguish it from SUU, which contains POO.

[Table 1]

[Table 2]

From the above Table 2, it can be seen that when utilizing the fractionation by-product as a starting material for the enzymatic esterification reaction according to the present invention, the C52 purity can be increased while maintaining the sn-2 PA at 55% or higher, and both an increase in yield and a reduction in cost can be achieved while maintaining the C52 at 50% or higher. Therefore, it was confirmed that according to the present invention, dioleoyl monopalmitoyl glyceride can be produced with high efficiency and economically through the enzymatic esterification reaction.

Claims (12)

A method for producing triglycerides,
A step of performing an enzymatic esterification reaction on a reactant mixture comprising a triacylglyceride and a fatty acid having an acyl group different from an acyl group in the triacylglyceride to exchange an acyl group of the triacylglyceride with an acyl group of the fatty acid,
The above reaction mixture further comprises a fractionated by-product obtained by fractionating the distillation residue of the enzymatic esterification reaction result,
The above fractionation by-products include neutral lipids having a total number of carbons of 48, neutral lipids having a total number of carbons of 50, neutral lipids having a total number of carbons of 52, and neutral lipids having a total number of carbons of 54, but based on 100 wt% of the total fractionation by-products, the neutral lipids having a total number of carbons of 52 are included in an amount of 15 wt% or more.
A method for producing triglycerides. A method for producing a triglyceride in claim 1, wherein the acyl group present in the triacylglyceride is independently selected from a saturated or unsaturated straight-chain aliphatic acyl group having 4 to 24 carbon atoms, and the fatty acid is at least one selected from a saturated or unsaturated straight-chain fatty acid having 4 to 24 carbon atoms. A method for producing a triglyceride in claim 1, wherein the acyl group present in the triacylglyceride is independently selected from a palmitoyl group, a stearoyl group, an oleoyl group, and a linoleoyl group, and the fatty acid is selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid, and combinations thereof. A method for producing a triglyceride in claim 1, wherein the triacylglyceride is tripalmitoylglyceride and the fatty acid is oleic acid. A method for producing a triglyceride, wherein the produced triglyceride in claim 1 is dioleoyl monopalmitoyl glyceride. A method for producing triglyceride according to any one of claims 1 to 5, wherein distillation of the enzymatic esterification reaction result is performed using a thin film distillation apparatus. A method for producing a triglyceride according to any one of claims 1 to 5, wherein the fractionation of the distillation residue of the enzymatic esterification reaction product comprises dissolving the distillation residue in an organic solvent, cooling the solution to produce a solid, separating the produced solid, removing the organic solvent from the remaining liquid phase, and concentrating the same. delete A method for producing a triglyceride according to any one of claims 1 to 5, wherein the fractionation by-product is contained in an amount of 15 to 70 wt% based on 100 wt% of the total of triacylglyceride and fractionation by-product in the reaction mixture of the enzymatic esterification reaction. A method for producing a triglyceride according to any one of claims 1 to 5, wherein the mixing ratio of triacylglyceride to fatty acid included in the reaction mixture of the enzymatic esterification reaction is 1:1.1 to 1:4 or less by weight. A method for producing a triglyceride according to any one of claims 1 to 5, wherein the fractionation by-product is contained in an amount of 30 to 40 wt% based on 100 wt% of the total of triacylglyceride and fractionation by-product in the reaction mixture of the enzymatic esterification reaction, and the mixing ratio of triacylglyceride to fatty acid is 1:1.3 to 1:2 by weight. delete