KR20200083772A - Method for improving efficiency of enzymatic esterification by adjustment of iodine value of triacylglyceride - Google Patents

Abstract

The present invention relates to a method for improving efficiency of enzymatic esterification. More particularly, the present invention relates to a method capable of producing high quality fats and oils while improving overall process efficiency and economic feasibility by using triacylglycerides having an iodine value (IV) within a range of 12 to 18 in performing a reaction of exchanging an acyl group of triacylglycerides with another acyl group using an enzyme, thereby preventing a decrease in enzyme activity as the reaction proceeds.

Description

Method for improving efficiency of enzymatic esterification by adjustment of iodine value of triacylglyceride}

The present invention relates to a method for improving the efficiency of enzymatic esterification, and more specifically, the iodine number (IV) is 12 to 12 in performing a reaction for exchanging an acyl group of a triacylglyceride with another acyl group using an enzyme. By using a triacylglyceride within the range of 18, it relates to a method capable of producing a high-quality fat and oil while improving the overall process efficiency and economic efficiency by preventing a decrease in enzymatic activity as the reaction progresses.

Triacylglyceride fats and oils are very widely used in the food industry. Among them, 1,3-dioleoyl-2-palmitoyl glyceride (OPO) is breast milk fat. As an important ingredient in the nutritional research has been conducted a lot.

Fat compositions containing the main fatty acids that make up the main milk fat are found in a lot of oils and fats of plant origin, but almost all plant origin glycerides do not have saturated fatty acids in the 2-position. In contrast, in breast milk, a significant amount of saturated fatty acid, palmitic acid, occupies the 2-position of glycerides.

Recently, as people's interest in infant health has increased, the demand for foods having similar ingredients to breast milk is increasing.

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

However, the conventional powdered milk is a mixture of powdered milk and vegetable oil, and contains little palmitic acid in the 2-position when analyzing the oil component of the powdered milk.

Accordingly, the application of high quality breast milk fat substitutes for adding 1,3-dioleoyl-2-palmitoyl glyceride (OPO) containing palmitic acid in the 2-position, which is similar in composition to breast milk fat, to the formula. This is increasing.

OPO is a type of Unsaturated-Saturated-Unsaturated (USU) triglyceride, and Mohamad, et al (Lipase-catalyzed interesterification reactions for human milk fat substitutes production, 2013, 115, 270-285) is a tree when lipase is used. It has been shown that studies on the synthesis of 1,3-dioleoyl-2-palmitoyl glycerides by substituting the 1,3-position of acylglycerides have been mainly conducted. Due to the specific reaction properties of the lipase triacylglyceride in the 1,3-position, the provision of a glyceride backbone containing saturated fatty acids at the 2-position of the triacylglyceride fat is 1,3-dioleoyl-2-palmi It is important in the synthesis of soil glycerides (OPO).

Although these prior art techniques use an enzymatic esterification reaction, the enzyme activity decreases as the reaction progresses, and as a result, the efficiency of the entire process is lowered. Therefore, in the enzymatic esterification reaction, a decrease in enzyme activity due to reaction progress is prevented, and a method capable of producing OPO with higher efficiency than the existing 1,3-dioleoyl-2-palmitoyl glyceride (OPO) production method Development is being requested.

An object of the present invention is to provide a method for preventing a decrease in enzyme activity due to reaction progress in an enzymatic esterification reaction and improving the overall process efficiency and economy.

In order to achieve the above technical problem, the present invention uses a triacylglyceride having an iodine number (IV) within a range of 12 to 18 in performing a reaction of exchanging an acyl group of a triacylglyceride with another acyl group using an enzyme. Enzymatic esterification method is provided.

According to another aspect of the invention, (1) providing an iodine number (IV) of a triacylglyceride having a range of 12 to 18; (2) providing a reaction mixture comprising a triacylglyceride obtained in step (1) and fatty acids having an acyl group different from the acyl group in the triacylglyceride as reaction starting materials; And (3) performing an enzymatic esterification reaction on the reaction mixture obtained in the step (2) to exchange the acyl group of the triacylglyceride with the acyl group of the fatty acid.

According to the present invention, when performing the reaction of exchanging an acyl group of a triacylglyceride with another acyl group using an enzyme, it is possible to excellently prevent the enzyme activity from decreasing as the reaction proceeds, thereby improving the overall process efficiency and economy High-quality fat and oil products with high C52 neutral lipid content and high glyceride ratio containing palmitic acid in the 2-position can be prepared. In addition, according to an embodiment of the present invention, dioleoyl monopalmitoyl glyceride can be efficiently and economically produced from tripalmitoyl glyceride through an enzymatic esterification reaction.

Hereinafter, the present invention will be described in detail.

The triacylglyceride used as a starting material for the enzymatic esterification reaction in the present invention may be edible oil.

The edible oil and fat can be used as long as it is suitable for edible use, such as liquid or solid vegetable oil or animal oil, hardened oil or transesterified oil of the oil or fat, liquid oil obtained by fractionating the oil or fat, and solid fat.

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

In one embodiment, the triacylglyceride may be palm fractionation oil that has been subjected to fractionation of palm oil during vegetable maintenance.

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

In one 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 material triacylglyceride is tripalmitoylglyceride (PPP).

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

In one embodiment, the fatty acid is selected from saturated or unsaturated linear fatty acids having 4 to 24 carbon atoms, more specifically 12 to 22 carbon atoms, and more specifically 14 to 20 carbon atoms. One or more can be used. More specifically, the fatty acid may be selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid and combinations thereof, preferably oleic acid, but is not limited thereto.

The mixing ratio of the starting material triacylglyceride to fatty acid may be 1:1 to 1:7 by weight, more specifically 1:1 to 1:5, and more specifically 1:1 to 1: It can be 3.

In the present invention, in order to prevent the enzyme activity from decreasing as the enzymatic esterification reaction proceeds, and at the same time to produce a triglyceride having a high C52 neutral lipid content and a high glyceride ratio containing palmitic acid in the 2-position, As one of the starting materials, triacylglycerides having an iodine number (IV) in the range of 12 to 18 are used. When the iodine value is lower than 12 or higher than 18, the enzyme activity decreases more rapidly, so the enzyme cannot be used for a long time and the C52 neutral lipid synthesis rate is lowered.

More specifically, the iodine value of the triacylglyceride may be 14 to 18, and more specifically, it may be 16 to 18.

In the present invention, the reaction temperature of the enzymatic esterification reaction may be determined according to the type of enzyme used, specifically, 40°C or higher, 45°C or higher, 50°C or higher, 55°C or higher, or 60°C or higher, and 85°C or less, 80°C or less, 75°C or less, 70°C or less, or 65°C or less, but is not limited thereto. In order to optimize the effect of the reaction and the efficiency of the process, the reaction temperature may preferably be 40°C to 60°C or 40°C to 50°C, and when the reaction temperature is relatively low as described above, it has an iodine number within the scope of the present invention. The effect of triacylglycerides may be more pronounced.

There is no particular limitation on the method for adjusting the iodine value of the triacylglyceride among the starting materials included in the reaction mixture, and methods known in the art can be appropriately used within a range capable of achieving the desired effect. In one embodiment, the iodine value of the triacylglyceride may be adjusted by the fractionation temperature and conditions of the raw material maintenance, for example, agitation speed at fractionation, fractionation method and fractionation time, but is not limited thereto. The skilled person can adjust the iodine value of the triacylglyceride to a desired range by changing the fractionation temperature and conditions of the raw material oil by a known method. The raw material oil may be, for example, palm oil, but is not limited thereto.

In one embodiment, the reaction mixture may include alcohol, more specifically, ethanol, and the content of alcohol is about 5% (w/w) or less, about 3% (w/w) or less, or about 2% (w) /w) or less, and preferably about 2% (w/w). When alcohol is added to the reaction mixture as described above, the reaction time of the enzyme is shortened to increase the flow rate and substrate treatment rate, and the C52 neutral lipid and 2-position palmitic acid content in the product may be increased, and the enzyme It can have the effect of extending the life of the enzyme with increasing the half-life of

As the enzyme of the enzymatic esterification reaction in the present invention, known enzymes can be used without particular limitation. Specifically, Candida Antarctica B , Thermomyces Thermomyces lanuginose ), Rhizomucour miehei , Rhizopus delemar , Mucormiehei , Alcaligenes sp . , Aspergillus niger , Candida cylindraces , Candida cylindraces ), Burkholderia cepacia , Geotricum candidum ) and combinations thereof, esterifying enzymes derived from microorganisms selected from the group may be used, but are not limited thereto.

According to the present invention, it is possible to effectively prevent the enzyme activity from being reduced during the enzymatic esterification reaction, so that a large amount of starting material can be continuously processed without replacing the enzyme. In one embodiment, 100 parts by weight to 3000 parts by weight of the reaction mixture per 1 part by weight of the enzyme, or 100 parts by weight to 2000 parts by weight, or 100 parts by weight to 1000 parts by weight of the enzyme may be continuously treated without replacement of enzyme, but the activity of the enzyme As long as it is maintained, further amounts can continue to be processed. For example, if the enzyme activity is appropriate, the reaction mixture per 1 part by weight of the enzyme may be continuously treated without replacing the enzyme by 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, and the upper limit has no particular limitation. None, and even 10000 parts by weight or more can be processed. According to a preferred embodiment, the reaction mixture throughput per 1 part by weight of enzyme may be 1000 parts by weight to 3000 parts by weight.

According to another aspect of the invention, (1) providing an iodine number (IV) of a triacylglyceride having a range of 12 to 18; (2) providing a reaction mixture comprising a triacylglyceride obtained in step (1) and fatty acids having an acyl group different from the acyl group in the triacylglyceride as reaction starting materials; And (3) performing an enzymatic esterification reaction on the reaction mixture obtained in the step (2) to exchange the acyl group of the triacylglyceride with the acyl group of the fatty acid.

In the above production method, triacylglyceride and fatty acid as reaction starting materials, iodine of triacylglyceride, reaction temperature and enzymatic esterification reaction are as described above.

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

Hereinafter, the present invention will be described in more detail through examples. However, this is provided to aid the understanding of the present invention, and the scope of the present invention is not limited thereto.

[ Example ]

As a starting material, a reaction mixture was prepared by adding 2% ethanol to a 1:2 weight ratio mixture of tripalmitoylglyceride (PPP, superstearin) to oleic acid (OA), and then homogenized with a homogenizer. The iodine number (IV) of tripalmitoyl glyceride was 10, 14, 16, 18 or 22, respectively, depending on the experimental group.

The homogenized reaction mixture was subjected to an enzymatic esterification reaction by continuously introducing a constant flow rate (SV=0.5, 72 ml/h) into a continuous reactor in which the enzyme (Novozym 40086) was packed in a column using a metering pump. . The reaction temperature was maintained at 40°C, 50°C or 60°C, respectively, depending on the experimental group. After the reaction, the product exited the reactor and collected in the product tank.

Every 5 days after the start of the reaction, 1 ml of the upper sample of the reaction mixture was taken and analyzed. The analysis method is as follows.

1) Neutral lipid Carbon number analysis

Neutral lipids are separated by carbon number using gas chromatography, and neutral lipids having C52 (total number of carbons (eg, C18-C16-C18, C16-C18-C18, etc.)), such as SPS, SSP, OPO, OOP, The concentration (%) of LPL, LLP, OPL, etc.) was analyzed. The results of tripalmitoylglyceride iodine value and neutral lipid carbon number analysis over time of samples reacted at reaction temperatures of 40°C, 50°C and 60°C are shown in Tables 1 to 3 below.

2) Fatty acid composition analysis of 2-position of glyceride

The ratio of neutral lipids (eg, PPP, PPO, OPO, etc.) of the type in which palmitic acid is located in the 2-position of glyceride was confirmed by gas chromatography. The results of the analysis of fatty acid composition of tripalmitoylglyceride iodine value and time of samples reacted at reaction temperatures of 40°C, 50°C and 60°C are shown in Tables 4 to 6, respectively.

[Table 1] Neutral lipid carbon number analysis according to iodine value at 40℃ reaction temperature

[Table 2] Analysis of neutral lipid carbon number according to iodine value at 50℃ reaction temperature

[Table 3] Analysis of neutral lipid carbon number according to iodine value at 60℃ reaction temperature

[Table 4] Fatty acid composition analysis of sn-2 position according to iodine number at 40℃ reaction temperature

[Table 5] Fatty acid composition analysis of sn-2 position according to iodine number at 50℃ reaction temperature

[Table 6] Fatty acid composition analysis of sn-2 position according to iodine number at 60°C reaction temperature

From the experimental results shown in Tables 1 to 6, when the iodine value of triacylglyceride used as a starting material for the enzymatic esterification reaction is higher than 18, it contains initial C52 neutral lipid and palmitic acid at the 2-position. It can be seen that the synthesis rate of glyceride is low and the substrate oxidation is fast, so the enzyme activity is rapidly reduced. In addition, when the iodine value was as low as 12 or less, the synthesis rate of glycerides containing C52 neutral lipid and palmitic acid in the 2-position was low, and such a tendency was particularly high in a low temperature reaction at 40°C. On the other hand, when the iodine value of the triacylglyceride used as a starting material for the enzymatic esterification reaction was within the range according to the present invention, it was possible to prevent a decrease in the enzyme activity as the reaction proceeded, and the enzyme activity did not decrease sharply even at a low temperature reaction. In addition, it was confirmed that an excellent fat or oil product having a high C52 neutral lipid content and a high glyceride ratio containing palmitic acid in the 2-position could be effectively produced.

Claims (13)

In performing a reaction for exchanging an acyl group of a triacylglyceride with another acyl group using an enzyme, an enzymatic esterification method characterized by using a triacylglyceride having an iodine number (IV) within a range of 12 to 18 . The enzymatic esterification method according to claim 1, wherein the reaction starting material is a triacylglyceride and a fatty acid having an acyl group different from the acyl group in the triacylglyceride. The method according to claim 2, wherein the acyl group present in the triacylglyceride is independently selected from saturated or unsaturated linear aliphatic acyl groups having 4 to 24 carbon atoms, and the fatty acid is saturated or having 4 to 24 carbon atoms. A method of enzymatic esterification, at least one selected from unsaturated linear fatty acids. The acyl group present in the triacylglyceride is independently selected from palmitoyl group, stearoyl group, oleoyl group and linoleoyl group, fatty acids are palmitic acid, stearic acid, oleic acid, linoleic acid and Enzymatic esterification method selected from the group consisting of combinations thereof. The enzymatic esterification method according to claim 2, wherein the triacylglyceride is tripalmitoylglyceride and the fatty acid is oleic acid. The method for enzymatic esterification according to claim 1, wherein the triacylglyceride has an iodine value (IV) within a range of 16 to 18. The enzymatic esterification method according to claim 1, wherein the reaction is performed at 40°C to 60°C. (1) providing a triacylglyceride having an iodine number (IV) within a range of 12 to 18;
(2) providing a reaction mixture comprising a triacylglyceride obtained in step (1) and fatty acids having an acyl group different from the acyl group in the triacylglyceride as reaction starting materials; And
(3) performing an enzymatic esterification reaction on the reaction mixture obtained in step (2) to exchange the acyl group of the triacylglyceride with the acyl group of the fatty acid,
Method of manufacturing triglycerides. 9. The method of claim 8, wherein the acyl group present in the triacylglyceride is independently selected from saturated or unsaturated linear aliphatic acyl groups having 4 to 24 carbon atoms, and the fatty acid is saturated or having 4 to 24 carbon atoms. A method for producing triglycerides, at least one selected from unsaturated linear fatty acids. The acyl group present in the triacylglyceride is independently selected from palmitoyl group, stearoyl group, oleoyl group and linoleoyl group, fatty acids are palmitic acid, stearic acid, oleic acid, linoleic acid and A method for producing an enzymatic triglyceride selected from the group consisting of combinations of these. The method of claim 8, wherein the triacylglyceride is tripalmitoylglyceride and the fatty acid is oleic acid. The method for producing triglycerides according to claim 8, wherein the prepared triglycerides are dioleoyl monopalmitoyl glycerides. The enzymatic esterification method according to claim 8, wherein the enzymatic esterification reaction is performed at 40°C to 60°C.