KR20200075971A - A method of increasing the content of triglycerides having saturated fatty acids at the sn-2 position in vegetable oil and a method of converting triglyceride having an unsaturated fatty acid at the sn-2 position to triglyceride having a saturated fatty acid at the sn-2 position - Google Patents

Abstract

The present invention relates to a method of increasing a content of triglycerides having saturated fatty acids at an sn-2 position in vegetable oil, and a method of converting triglyceride having unsaturated fatty acid at the sn-2 position to triglyceride having saturated fatty acid at the sn-2 position. The above methods includes: step 1 in which transesterification and acyl migration take place; and step 2 in which transesterification takes place, wherein a triglyceride having an unsaturated fatty acid at the sn-2 position is converted to a triglyceride having a saturated fatty acid at the sn-2 position at a high conversion rate, so that β-palmitin-type breast milk-substituting oil and fat having saturated fatty acids at the sn-2 position can is prepared from vegetable oils with high iodine values.

Description

A method for increasing the content of triglycerides having saturated fatty acids at the sn-2 position in vegetable oil and a method for converting triglycerides having unsaturated fatty acids at the sn-2 position to triglycerides having saturated fatty acids at the sn-2 position{A method of increasing the content of triglycerides having saturated fatty acids at the sn-2 position in vegetable oil and a method of converting triglyceride having an unsaturated fatty acid at the sn-2 position to triglyceride having a saturated fatty acid at the sn-2 position}

A method for increasing the content of triglycerides with saturated fatty acids at the sn-2 position in vegetable oil and a method for converting triglycerides with unsaturated fatty acids at the sn-2 position to triglycerides with saturated fatty acids at the sn-2 position .

Breast milk is the first source of nutrition for infants, and is a complete food product consisting of carbohydrates (6.9-7.2%), lipids (3-5%), proteins (0.8-0.9%), minerals (0.2%) and vitamins. Among them, breast milk fat accounts for more than 50% of the calories provided from breast milk, and thus serves as an important source of nutrition for infants with limited digestibility and absorption.

The main fatty acids in breast milk are unsaturated fatty acids oleic acid (28-43%), linoleic acid (10-25%), saturated fatty acids palmitic acid (15-25%), US Myristic acid (3-12%), stearic acid (4-8%), and other small amounts of heavy chain fatty acids (caprylic acid, capric acid) and polyunsaturated fatty acids (arachidonic acid, eicosapentaenoic acid, docosahexaenoic) acid) is present in small amounts.

In particular, the fatty acid distribution at the position of sn -2 (fatty acid position 2) in triglyceride (TG, triacylglycerol, TAG, triacylglyceride) of breast milk fat is more than 70% saturated fatty acids, and many of them are palmitic acid (C _{16: 0} , P) is occupied, while the unsaturated fatty acids such as oleic acid (C _18:1 , O) and linoleic acid (C _18:2 , L) are mainly in the sn -1 and 3 positions, so the main part of breast milk fat is The TAG types are 1,3-dioleoyl-2-palmitoylglycerol (OPO) and 1-oleoyl-2-palmitoyl-3-linoleoylglycerol (OPL). Thus, human milk fat sn -2 position because the β-palmitin form that contains a large amount of palmitic acid in the sn -2 position the most show a vegetable oil with a notable difference that consists of unsaturated fatty acids.

When the pancreatic lipase reacts with the TAG, the fatty acids at positions sn -1 and 3 in the TAG are hydrolyzed to free fatty acids according to the substrate specificity of the enzyme, thereby producing 2-monoacylglycerol (MAG). In the case of 2-MAG, it forms a micelle structure by bile salts and is absorbed into epithelial cells of the small intestine regardless of the type of fatty acid, and is easily used as an energy source. However, the hydrolyzed free fatty acid shows a different pattern of absorption rate in the body depending on the carbon number length and the degree of unsaturation.

Palmitic acid because hydrolyzed free fatty acid is a saturated fatty acid and palmitic acid having a carbon number length greater than that of the medium chain fatty acid, and forms an insoluble salt by combining with divalent cations such as Ca ²⁺ and Mg ²⁺ under alkaline conditions of pH 7 or higher. And mineral absorption decreases and constipation is caused. Conversely, in the case of β-palmitin, which is the main TAG form of breast milk fat, the fatty acids at positions sn -1 and 3 are hydrolyzed by pancreatic lipase, and 2-MAG palmitic acid is absorbed into the epithelial cells of the small intestine and used as an energy source. Hydrolyzed oleic acid forms salts with divalent cations. The salts produced at this time have a higher absorption rate in the body than the insoluble salts, which increases the utilization rate of fats and minerals, thereby positively affecting the growth of infants.

The method of synthesizing the mother milk substitute can use a variety of acidolysis, esterification, glycerolysis, and alcoholysis reactions based on the enzymatic transesterification reaction. The synthesized mother milk fat maintains digestion and absorption of lipids according to the molecular structure of TAG. And physical properties are determined.

The reaction of lipase in the enzymatic transesterification reaction is a reversible reaction of acylglycerol hydrolysis and ester bond, but under conditions where the amount of water is limited, hydrolysis of acylglycerol is minimized, and transesterification reaction by lipase occurs predominantly. In addition, the immobilized enzyme used has the advantage of immobilizing the enzyme on a hydrophobic or hydrophilic support to increase stability against temperature and pH and to reduce by-products. Here, it is used industrially because of the effect of reducing the production cost due to the reusability of enzymes and the characteristics of easy process control.

In general, the manufacturing process of breast milk substitute maintenance consists of a large amount of palmitic acid, and it has a low iodine value (typically less than 30) and oleic acid or its ester as a substrate to selectively act at the positions of sn- 1 and 3 in the TAG. This enzyme is used as a reaction catalyst. Due to the site specificity of the enzyme, the fatty acid at the sn -2 position in the TAG reactant is mostly preserved to synthesize the β-palmitin form. The substrate mainly used for this is a mixture of PS or palm oil fractions with a high content of tripalmitin (WO 2006114791 A1; EP 0496456 A1; WO 1994026855 A1; Lee et al., New Biotechnology, 27:38-45, 2010; Sahin et. al., J. Agric.Food Chem., 53:5779-5783, 2005)

One object of the present invention is to provide a method for maintaining a milk substitute in the form of β-palmitin having saturated fatty acids at the sn- 2 position.

Another object of the present invention is to provide a method for increasing the content of triglycerides with saturated fatty acids at the sn-2 position in vegetable oil.

Another object of the present invention is to provide a vegetable oil with an increased content of triglycerides having saturated fatty acids at the sn-2 position prepared according to the above method.

Another object of the present invention is to provide a method for converting triglycerides having an unsaturated fatty acid at the sn-2 position to triglycerides having a saturated fatty acid at the sn-2 position.

In order to achieve the above object,

According to one aspect of the invention,

Reacting vegetable oil with saturated fatty acid alkyl ester in the presence of Lipase (step 1); And

Reacting the reaction product of step 1 and the unsaturated fatty acid alkyl ester in the presence of Lipase (step 2),

A method for increasing the content of triglycerides with saturated fatty acids at the sn-2 position in vegetable oil is provided.

According to another aspect of the present invention, there is provided a vegetable oil with an increased content of triglycerides having saturated fatty acids at the sn-2 position prepared according to the above method.

According to another aspect of the invention,

reacting a triglyceride having an unsaturated fatty acid at the sn-2 position with a saturated fatty acid alkyl ester in the presence of Lipase (step 1); And

Reacting the reaction product of step 1 and the unsaturated fatty acid alkyl ester in the presence of Lipase (step 2),

A method for converting a triglyceride having an unsaturated fatty acid at the sn-2 position to a triglyceride having a saturated fatty acid at the sn-2 position is provided.

Method of increasing the content of triglycerides with saturated fatty acids at the sn-2 position in the vegetable oil of the present invention and converting triglycerides with unsaturated fatty acids at the sn-2 position to triglycerides with saturated fatty acids at the sn-2 position Including silver transesterification and acyl migration step 1 and transesterification step 2, the triglyceride having an unsaturated fatty acid at the sn-2 position is converted to a triglyceride having a saturated fatty acid at the sn-2 position at a high conversion rate. By conversion, it is possible to prepare β-palmitin-type breast milk substitutes having saturated fatty acids at the sn-2 position from vegetable oils having a high iodine value.

1 is a fraction of the palm oil of the present invention as a starting material, the step of reacting the fraction and PEE in the presence of Lipase (step 1); And a step of reacting the reaction product of step 1 and OEE in the presence of a lipase (step 2) is a schematic diagram showing the reaction process at an example level.
Figure 2 converts triglycerides with unsaturated fatty acids at the sn-2 position of the invention to triglycerides with saturated fatty acids at the sn-2 position to maintain β-palmitin-type breast milk substitutes from palm oil fraction (POF). It is a chemical reaction formula represented by the chemical structural formula of triglyceride. P: palmitic acid ester, O: oleic acid ester, P/O or O/P: triglyceride in which the corresponding fatty acid position is palmitic acid ester or triglyceride in oleic acid ester is present to be.
Figure 3 shows the chromatogram of the substrate analyzed by Ag-HPLC (S = saturated fatty acids, U = unsaturated fatty acids). A) Palm oil, B) Palm oil fraction (palm oil fraction fractionated in step 1 of Example 1), C) Palmitic ethyl ester, D) Oleic ethyl ester
Figure 4 shows a chromatogram of the substrate analyzed by Ag-HPLC for the reaction products of Examples SL1-1 to SL1-12.
Figure 5 shows a chromatogram of the substrate analyzed by Ag-HPLC for the reaction products of Examples SL2-1 to SL2-4. A) SL2-1 (20 wt% TLIM, 1 hr, 1: 6 mol ratio, 45° C., 200 rpm), B) SL2-2 (20 wt% TLIM, 3 hr, 1: 6 mol ratio, 45° C., 200 rpm), C) SL2-3 (20 wt% RMIM, 1 hr, 1: 6 mol ratio, 45°C, 200 rpm), D) SL2-4 (20 wt% RMIM, 3 hr, 1: 6 mol ratio , 45℃, 200 rpm)

One object of the present invention is to provide a method for maintaining a milk substitute in the form of β-palmitin having saturated fatty acids at the sn- 2 position.

In order to achieve the above object, the present invention provides a method for increasing the content of triglycerides with saturated fatty acids at the sn-2 position in vegetable oil and triglycerides with unsaturated fatty acids at the sn-2 position with saturated fatty acids at the sn-2 position. It provides a method for converting to triglycerides.

Through the above methods, β-palmitin-type breast milk substitutes having saturated fatty acids at the sn- 2 position can be prepared.

Hereinafter, the present invention will be described in detail.

One aspect of the invention,

Reacting vegetable oil with saturated fatty acid alkyl ester in the presence of Lipase (step 1); And

Reacting the reaction product of step 1 and the unsaturated fatty acid alkyl ester in the presence of Lipase (step 2),

It provides a method of increasing the content of triglycerides with saturated fatty acids at the sn-2 position in vegetable oil.

Hereinafter, a method of increasing the content of triglycerides having saturated fatty acids at the sn-2 position in the vegetable oil will be described in detail.

The method converts triglycerides having unsaturated fatty acids at the sn-2 position in vegetable oil to triglycerides with saturated fatty acids at the sn-2 position, thereby increasing the content of triglycerides with saturated fatty acids at the sn-2 position in vegetable oil. .

The vegetable oil has an IV (iodine value) of 45-65, which indicates a high IV value, and is a concept including both raw oil or fractions of crude oil.

또는

형태의 트리글리세라이드를 많이 포함하고 있으며,

를 30-40%,

를 40-50%의 함량으로 포함할 수 있다. 또한, 상기 식물유는

,

,

및

로 이루어지는 군으로부터 선택되는 트리글리세라이드를 하나 이상 더 포함할 수 있다.In addition, the vegetable oil

or

Contains a lot of triglycerides in the form,

30-40%,

It may contain 40-50% of the content. In addition, the vegetable oil

,

,

And

Triglyceride selected from the group consisting of may further include one or more.

In the formula of the triglyceride, S is a saturated fatty acid and U is an unsaturated fatty acid. Further, the saturated fatty acid may be palmitic acid, myristic acid, or stearic acid. The unsaturated fatty acid may be oleic acid or linoleic acid.

The lipase of step 1 is sn-1,3 specific lipase, and may be Lipozyme TLIM (Thermomyces lanuginosus).

Step 1 is a transesterification reaction and an acyl migration reaction, where the fatty acids at the sn-1 and sn-3 positions of triglycerides in vegetable oil undergo transesterification reaction with saturated fatty acids of saturated fatty acid alkyl esters, sn The content of triglycerides with saturated fatty acids in -1 or sn-3 increases, and due to acyl migration, the unsaturated fatty acids at the sn-2 position are converted to saturated fatty acids in sn-1 or sn-3.

After performing the above step 1, with respect to vegetable oil, sn-2 may contain 40% or more of triglyceride, which is a saturated fatty acid.

At this time, the saturated fatty acid of the saturated fatty acid alkyl ester of step 1 may be palmitic acid, myristic acid, or stearic acid. The alkyl may be straight or branched C _1-10 alkyl, and may be ethyl. In one embodiment of the present invention, palmitic ethyl ester (PEE) was used, but this is only an example and is not limited thereto.

In step 1, the saturated fatty acid alkyl ester may be used in a ratio of 1 to 10 moles of saturated fatty acid alkyl ester, 1 to 6 moles, and 6 moles to 1 mole of vegetable oil. have.

The reaction time in step 1 may be 1-24 hours, 1-18 hours, 3-12 hours, or 12 hours.

Lipase of step 2 is sn-1,3 specific lipase, and may be Lipozyme TLIM (Thermomyces lanuginosus) or Lipozyme RMIM (Rhizomucor miehei).

The step 2 is a step in which the transesterification reaction takes place, and the fatty acids at the sn-1 and sn-3 positions of the triglycerides in the vegetable oil after performing the step 1 are sn-1 or sn by transesterification with an unsaturated fatty acid alkyl ester. The content of triglycerides having unsaturated fatty acids at -3 is increased, and finally, the content of triglycerides having unsaturated fatty acids at sn-1 or sn-3 and saturated fatty acids at sn-2 is increased.

After performing the above step 2, the content of triglycerides having unsaturated fatty acids in sn-1 and sn-3 and saturated fatty acids in the sn-2 position may be 35% or more with respect to the whole plant oil.

At this time, the unsaturated fatty acid of the unsaturated fatty acid alkyl ester of step 2 may be oleic acid (oleic acid) or linoleic acid (linoleic acid). The alkyl may be straight or branched C _1-10 alkyl, and may be ethyl. In one embodiment of the present invention, oleic ethyl ester (OEE) was used, but this is only an example and is not limited thereto.

The reaction time in step 2 may be 0.5-5 hours, 1-3 hours, or 3 hours.

After performing the above step 2, the vegetable oil having an increased content of triglycerides having saturated fatty acids at the sn-2 position can be used as a milk substitute for β-palmitin having saturated fatty acids at the sn- 2 position.

According to the above method, the conventional oil having a high iodine value (IV) improves the fact that the content of saturated fatty acids, specifically, palmitic acid is low, and thus, a triglyceride in the form of β-palmitin cannot be prepared, thereby maintaining a high iodine value. As a starting material, β-palmitin-type triglyceride may be prepared.

Another aspect of the present invention provides a vegetable oil having an increased content of triglycerides having saturated fatty acids at the sn-2 position prepared according to a method of increasing the content of triglycerides having saturated fatty acids at the sn-2 position in the vegetable oil. do.

The sn-2 of the content of the triglyceride vegetable oil having an increased saturated fatty acids in position may be a human milk replacement maintenance of β-palmitin type with the saturated fatty acids in the sn -2 position.

,

및

로 이루어지는 군으로부터 선택되는 어느 하나 또는 이들의 혼합일 수 있다.Triglyceride having an unsaturated fatty acid at the sn-2 position,

,

And

It may be any one selected from the group consisting of or a mixture thereof.

,

및

로 이루어지는 군으로부터 선택되는 트리글리세라이드를 하나 이상 더 포함할 수 있다.In addition, the vegetable oil,

,

And

Triglyceride selected from the group consisting of may further include one or more.

In the formula of the triglyceride, S is a saturated fatty acid and U is an unsaturated fatty acid. Further, the saturated fatty acid may be palmitic acid, myristic acid, or stearic acid. The unsaturated fatty acid may be oleic acid or linoleic acid.

의 함량이 35%이상일 수 있다.All of the above vegetable oil

The content of may be more than 35%.

Another aspect of the invention,

reacting a triglyceride having an unsaturated fatty acid at the sn-2 position with a saturated fatty acid alkyl ester in the presence of Lipase (step 1); And

Reacting the reaction product of step 1 and the unsaturated fatty acid alkyl ester in the presence of Lipase (step 2),

Provided is a method for converting a triglyceride having an unsaturated fatty acid at the sn-2 position to a triglyceride having a saturated fatty acid at the sn-2 position.

Hereinafter, a method of converting triglycerides having an unsaturated fatty acid at the sn-2 position to triglycerides having a saturated fatty acid at the sn-2 position will be described in detail.

In step 1 of the method,

The triglyceride having an unsaturated fatty acid at the sn-2 position may have an IV (iodine value) of 45-65, 50-60, or 53. In addition, the triglyceride having an unsaturated fatty acid at the sn-2 position may be a fraction fractionated from palm oil. In this case, the triglyceride fraction fractionated from the palm oil may have an IV (iodine value) of 55-75, 55-65, and 63.

또는

형태의 트리글리세라이드 또는 이들의 혼합물 상태일 수 있다.

를 30-40%,

를 40-50%의 함량으로 포함할 수 있다. 또한, 상기 sn-2 위치에 불포화지방산을 갖는 트리글리세라이드물은

,

,

및

로 이루어지는 군으로부터 선택되는 트리글리세라이드를 하나 이상 더 포함된 혼합물 형태일 수 있다.Triglyceride having an unsaturated fatty acid in the sn-2 position of step 1 above

or

In the form of triglycerides or mixtures thereof.

30-40%,

It may contain 40-50% of the content. In addition, the triglyceride product having an unsaturated fatty acid at the sn-2 position

,

,

And

It may be in the form of a mixture containing one or more triglycerides selected from the group consisting of.

In the formula of the triglyceride, S is a saturated fatty acid and U is an unsaturated fatty acid. Further, the saturated fatty acid may be palmitic acid, myristic acid, or stearic acid. The unsaturated fatty acid may be oleic acid or linoleic acid.

In addition, since the triglyceride having an unsaturated fatty acid at the sn-2 position in step 1 of the method may be included in a fraction fraction from palm oil, in step 1, triglyceride having an unsaturated fatty acid at the sn-2 position is obtained from palm oil. It can be used as a fractional fraction.

또는

형태의 트리글리세라이드를 많이 포함하고 있으며,

를 30-40%,

를 40-50%의 함량으로 포함할 수 있다. 또한, 상기 분획물은

,

,

및

로 이루어지는 군으로부터 선택되는 트리글리세라이드를 하나 이상 더 포함할 수 있다.The fraction fractionated from the palm oil

or

Contains a lot of triglycerides in the form,

30-40%,

It may contain 40-50% of the content. In addition, the fraction

,

,

And

Triglyceride selected from the group consisting of may further include one or more.

In the formula of the triglyceride, S is a saturated fatty acid ester, and U is an unsaturated fatty acid ester. Further, the saturated fatty acid may be palmitic acid, myristic acid, or stearic acid. The unsaturated fatty acid may be oleic acid or linoleic acid.

The lipase of step 1 is sn-1,3 specific lipase, and may be Lipozyme TLIM (Thermomyces lanuginosus).

In step 1, a transesterification reaction and an acyl migration reaction occur, and the fatty acids at the sn-1 and sn-3 positions of the triglyceride having an unsaturated fatty acid at the sn-2 position are saturated fatty acids at the saturated fatty acid alkyl ester. By transesterification, the content of triglycerides with saturated fatty acids in sn-1 or sn-3 increases, and the unsaturated fatty acids at the sn-2 position are converted to saturated fatty acids in sn-1 or sn-3 due to acyl migration. .

In step 1, when a fraction fraction from palm oil is used as a triglyceride having an unsaturated fatty acid at the sn-2 position, after performing step 1, the triglyceride having an unsaturated fatty acid at the sn-2 position is sn for the entire fraction. It may contain 40% or more of triglyceride, a divalent saturated fatty acid.

At this time, the saturated fatty acid of the saturated fatty acid alkyl ester of step 1 may be palmitic acid, myristic acid, or stearic acid. The alkyl may be straight or branched C _1-10 alkyl, and may be ethyl. In one embodiment of the present invention, palmitic ethyl ester (PEE) was used, but this is only an example and is not limited thereto.

In step 1, the saturated fatty acid alkyl ester may be used in a ratio of 1-10 moles of saturated fatty acid alkyl esters and 1-6 moles of triglyceride having an unsaturated fatty acid at the sn-2 position. Can be used at a ratio of 6 mol.

The reaction time in step 1 may be 1-24 hours, 1-18 hours, 3-12 hours, or 12 hours.

Lipase of step 2 is sn-1,3 specific lipase, and may be Lipozyme TLIM (Thermomyces lanuginosus) or Lipozyme RMIM (Rhizomucor miehei).

The step 2 is a step in which the transesterification reaction takes place, and the fatty acids at the sn-1 and sn-3 positions of the triglyceride in the primary reaction product after the step 1 is carried out by the transesterification reaction with an unsaturated fatty acid alkyl ester. The content of triglycerides having unsaturated fatty acids in 1 or sn-3 increases, and finally, the content of triglycerides having unsaturated fatty acids in sn-1 or sn-3 and saturated fatty acids in sn-2 is increased. do.

After performing the step 2, the content of triglycerides having unsaturated fatty acids in sn-1 and sn-3 and saturated fatty acids in the sn-2 position may be 35% or more with respect to the entire final reaction product.

At this time, the unsaturated fatty acid of the unsaturated fatty acid alkyl ester of step 2 may be oleic acid (oleic acid) or linoleic acid (linoleic acid). The alkyl may be straight or branched C _1-10 alkyl, and may be ethyl. In one embodiment of the present invention, oleic ethyl ester (OEE) was used, but this is only an example and is not limited thereto.

The reaction time in step 2 may be 0.5-5 hours, 1-3 hours, or 3 hours.

After performing the above step 2, triglycerides having an unsaturated fatty acid in the generated sn-1 and sn-3 and an increased content of triglyceride having a saturated fatty acid in the sn- 2 position are β having saturated fatty acids in the sn- 2 position. It can be used as a milk substitute in the form of palmitin.

The method for converting the triglyceride having an unsaturated fatty acid at the sn-2 position to a triglyceride having a saturated fatty acid at the sn-2 position can be represented by a chemical reaction scheme as shown in FIG. 2.

According to the above method, the conventional oil having a high iodine value (IV) improves the fact that the content of saturated fatty acids, specifically, palmitic acid is low, and thus, a triglyceride in the form of β-palmitin cannot be prepared, thereby maintaining a high iodine value. As a starting material, β-palmitin-type triglyceride may be prepared.

Another aspect of the present invention, the content of triglycerides with saturated fatty acids at the sn-2 position prepared by converting triglycerides with unsaturated fatty acids at the sn-2 position to triglycerides with saturated fatty acids at the sn-2 position This increased β-palmitin form provides breast milk replacement.

,

및

로 이루어지는 군으로부터 선택되는 어느 하나 또는 이들의 혼합일 수 있다.Triglyceride having an unsaturated fatty acid at the sn-2 position,

,

And

It may be any one selected from the group consisting of or a mixture thereof.

,

및

로 이루어지는 군으로부터 선택되는 트리글리세라이드를 하나 이상 더 포함할 수 있다.In addition, the vegetable oil,

,

And

Triglyceride selected from the group consisting of may further include one or more.

In the formula of the triglyceride, S is a saturated fatty acid ester, and U is an unsaturated fatty acid ester. Further, the saturated fatty acid may be palmitic acid, myristic acid, or stearic acid. The unsaturated fatty acid may be oleic acid or linoleic acid.

의 함량이 35%이상일 수 있다.All of the above vegetable oil

The content of may be more than 35%.

Hereinafter, the present invention will be described in detail by examples and experimental examples.

However, the following examples and experimental examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples and experimental examples.

<Production Example> Preparation of palm oil fraction

After dissolving palm oil (IV = 53) in a constant temperature water bath at 80°C, 300 g of palm oil and 900 mL of acetone are quantified in a 2 L beaker in a ratio of 1 to 3 (w/v), stirred with 40°C water, and thoroughly stirred. Did. This was left to stand for 24 hours in an incubator maintained at 4° C., and then separated and the solid layer and the liquid layer were separated using filter paper. The fractionated liquid layer was completely dissolved in a constant temperature water bath at 40°C, and then the solvent was removed through concentration under reduced pressure. The POP and POO contents were 32.84%-33.22%, 44.26%-44.3%, respectively, and the palm oil fraction (POF) with an IV of 63. ).

<Examples SL1-1 to SL1-12> Preparation of primary reaction products

The examples SL1-1 to SL1-12 correspond to step 1 of the methods of the present invention.

POF and palmitic ethyl ester (PEE) obtained in the above preparation examples were respectively quantified in a 1: 1-1: 6 mol ratio in a 250 mL reactor, and then Lipozyme ^® TLIM was added to the reactor at a rate of 20 wt% of the total substrate amount. . The reaction was carried out by changing the reaction time (3 hr-12 hr) at 220 rpm in a shaking water bath at 80°C, and the reaction conditions are shown in Table 1 below. After the reaction was completed, a primary reaction product was prepared by filtering the enzyme using a 0.5 μm PTFE syringe filter (Toyo Roshi Kaisha, Ltd., Japan).

Example Substrate ratio Reaction time Stirring speed Reaction temperature Lipase SL1-1 1:1 3 220 rpm 80 ℃ 20 wt% TLIM SL1-2 1:1 6 220 rpm 80 ℃ 20 wt% TLIM SL1-3 1:1 9 220 rpm 80 ℃ 20 wt% TLIM SL1-4 1:1 12 220 rpm 80 ℃ 20 wt% TLIM SL1-5 1:3 3 220 rpm 80 ℃ 20 wt% TLIM SL1-6 1:3 6 220 rpm 80 ℃ 20 wt% TLIM SL1-7 1:3 9 220 rpm 80 ℃ 20 wt% TLIM SL1-8 1:3 12 220 rpm 80 ℃ 20 wt% TLIM SL1-9 1:6 3 220 rpm 80 ℃ 20 wt% TLIM SL1-10 1:6 6 220 rpm 80 ℃ 20 wt% TLIM SL1-11 1:6 9 220 rpm 80 ℃ 20 wt% TLIM SL1-12 1:6 12 220 rpm 80 ℃ 20 wt% TLIM

In Table 1 above, the substrate ratio is POF:PEE.

<Example SL2-1 to SL2-4> Preparation of secondary reaction product (β-palmitin-type triglyceride (maintaining milk substitute))

The present examples SL2-1 to SL2-4 correspond to step 2 of the methods of the present invention.

Prior to proceeding with the second reaction, the first reaction product (SL1-12) having the best results and methanol were quantified at 1: 5 (w/v) and sufficiently stirred. The supernatant (methanol) separated through centrifugation (2500 rpm, 5 min) was passed through a pasteur pipet column filled with anhydrous sodium sulfate to remove moisture and impurities, and the result was methanol and 1:1 (w/v). It was quantified once again to proceed in the same way. As the ethyl ester and acid components are dissolved in the supernatant, impurities are removed from the remaining primary reaction product and only the TAG component remains.

After quantifying 0.3 g of the first reaction product (SL1-12) and 0,6 g of OEE, respectively, in a 50 mL reactor with a substrate ratio of 1: 6 mol ratio, the enzyme type (TLIM or RMIM) and the reaction time (1 hr or 3 hr) was performed differently, and the reaction conditions are shown in Table 2 below. The amount of the enzyme was added at a rate of 20 wt% of the total substrate amount, and after the reaction was completed, the enzyme was filtered using a 0.5 μm PTFE syringe filter to obtain β-palmitin type triglyceride (maintaining breast milk substitute) as shown in Table 2. It was prepared.

Example Substrate ratio Reaction time Stirring speed Reaction temperature Lipase SL2-1 1:6 One 220 rpm 45 ℃ 20 wt% TLIM SL2-2 1:6 3 220 rpm 45 ℃ 20 wt% TLIM SL2-3 1:6 One 220 rpm 45 ℃ 20 wt% RMIM SL2-4 1:6 3 220 rpm 45 ℃ 20 wt% RMIM

In Table 2 above, the substrate ratio is the primary reactant: OEE

<Comparative Example 1> Preparation of OPO by acyl migration of POO (dioleopalmitin)

Synthesis of OPO was induced by placing the palmitic acid at the sn-2 position of the glycerol backbone using the acyl migration reaction with the POO-rich oil substrate.

Acyl migration was performed using a liquid fraction of palm oil containing POO of about 60% as a substrate. The reaction was carried out in a shaking water bath set at 80° C. and 220 rpm for 12 hours with Lipozyme TLIM lipase containing 20% of the total substrate.

<Comparative Example 2> Preparation of OPO using the reaction of triolein (OOO) and palmitic ethyl ester (PEE)

By using OOO, transesterification reaction using PEE was performed to prepare OPO (TAG, where the sn-2 position is palmitic acid, and the sn-1 and sn-3 positions are oleic acid).

Step 1: The first reaction was carried out using high oleic sunflower oil and palmitic ethyl ester as substrates containing about 90% of OOO. At this time, the molar ratio of the substrates was 1:0.5 to 1:1 (OOO:PEE, w/w), and the reaction was performed with Lipozyme TLIM lipase for 7 to 12 hours in a shaking water bath set at 80°C and 220 rpm. It proceeded to contain 20%.

Step 2: To the primary reactant (10 g in total), oleic ethyl ester was added so that 15 g to 18 g, and Lipozyme TLIM lipase was added in addition to 0.5 to 2 g, in a shaking water bath set at 45°C and 220 rpm 3 The secondary reaction proceeded for ~ 6 hours.

<Experiment 1> Fatty acid composition analysis using GC 1

In order to confirm the fatty acid content, fatty acids were methylated and converted into a fatty acid methyl ester (FAME) state, and analyzed using gas chromatography (GC, Hewlett-Packard 6890, Avondale, PA, USA). Methylation is a method of replacing fatty acid carboxyl group (-COOH) with methyl ester group (-COOCH ₃ ), after weighing 100 mg of sample in 50 mL vial, add 2 mL of salt catalyst 0.5 N methanolic NaOH, and then stir for 30 seconds. And reacted in a water bath at 85°C for 10 minutes. After the reaction was completed, the mixture was cooled for 20 minutes using water at 20°C, and 3 mL of methanolic BF ₃ was added to perform the experiment under the same conditions. After cooling, 3 mL and 1 mL of iso-octane and saturated NaCl were added, and stirred for 1 minute to extract fatty acids substituted with FAMEs. The supernatant separated through centrifugation (2500 rpm, 3 min) was passed through a pasteur pipet column filled with anhydrous sodium sulfate to remove moisture and impurities, collected in a GC vial and analyzed by GC. SP ^™ -2560 (biscyanopropyl polysiloxane, 100 m × 0.25 mm, 0.2 μm film thickness, Supelco, Bellofonte, PA, USA) was used as the column, and a flame ionization detector (FID) was used as the detector. The conditions for instrument analysis were analyzed by setting the temperature of the detector and injector to 285℃ and 225℃, respectively, and the oven temperature program was started at 100℃ and maintained for 4 minutes, then raised to 240℃ by 3℃/min and held for 15 minutes. Proceeded. High purity He was used as the carrier gas and analyzed at a flow rate of 1.0 mL/min. The sample was injected with 1 uL, and the split ratio was set at a ratio of 200:1. Fatty acid content was calculated using peak area values for FAMEs obtained through GC analysis. The retention time (RT) of each peak was confirmed through the standard material, Supelco 37 Component FAME Mix (Sigma-Aldrich, St. Louis, Mo., USA.).

To confirm the fatty acid composition of the sn -2 position in TAG, hydrolysis was performed using pancreatic lipase, and 2-monoacylglycerol (MAG) was separated using TLC. Sample 10 mg, 1M Tris-HCl buffer (pH 7.6) 7 mL, 0.05% bile salt 1.75 mL, 2.2% CaCl ₂ 0.7 mL, pancreatic lipase 10 mg in a 20 mL glass test tube, stir for 1 minute, and then 37°C The reaction was carried out in a water bath for 3 minutes. After the reaction was completed, the mixture was stirred for 30 seconds, and then reacted for 3 minutes under the same conditions. After stirring for 30 seconds, the mixture was reacted for 2 minutes under the same conditions and stirred for 30 seconds. After the stirring was completed, 4 mL of diethyl ether (DE) was placed in a test tube and stirred for 1 minute to perform centrifugation (2500 rpm, 5 min). Subsequently, the supernatant (DE) was passed through a pasteur pipet column filled with anhydrous sodium sulfate to remove moisture and impurities, and then DE was removed using N ₂ gas. After putting DE 150 uL back into the glass test tube from which DE is removed, stir for 30 seconds, spotting on TLC silica gel 60 F ₂₅₄ (MERCK CO., Darmstadt, Germany), and then mixing solvent (hexane: diethyl ether: acetic acid = It was deployed in a TLC tank filled with 50:50:1, v/v/v). After heating the dry TLC plate, the solvent was blown off, and stained with 5% 2,7-dichlorofluorescein in EtOH to separate the 2-MAG band confirmed through UV-spectrophotometer. The methylation of the separated 2-MAG was performed in the same manner as above, except that the addition amount was changed to 1.5 mL of 0.5 N methanolic NaOH, 2 mL of methanolic BF _{3, and} 2 mL of iso-octane.

The fatty acid composition of the crude palm oil, palm oil fraction, and PEE according to the above Experimental Example 1 is shown in Table 3, and the total fatty acids and% of fatty acids in the sn-2 position are evaluated in Table 4 with respect to the palm oil fraction.

Fatty acid composition Primary reaction substrate fatty acid Palm oil Palm oil
fraction(4℃) Palmitic
ethyl ester C12: 0 0.98±0.02 1.22±0.01 ND C14:0 1.62±0.02 1.66±0.01 0.72±0.02 C16: 0 44.94±0.17 38.15±0.00 99.28±0.02 C18: 0 4.75±0.04 4.10±0.08 ND C18:1 (n-9) 36.85±0.12 41.54±0.10 ND C18:1 (n-7) 0.91±0.03 1.01±0.01 ND C18:2 (n-6) 9.94±0.03 12.31±0.00 ND

The results are expressed as the mean ± SD (n=2)

ND :　Not detected

Unit: area%

Fatty acid composition Palm oil fraction(4℃) fatty acid Total fatty acids sn -2 position C12: 0 1.22±0.01 1.29±0.04 C14:0 1.66±0.01 0.56±0.10 C16: 0 38.15±0.00 9.24±0.54 C18: 0 4.10±0.08 0.93±0.20 C18:1 (n-9) 41.54±0.10 64.38±0.78 C18:1 (n-7) 1.01±0.01 0.99±0.05 C18:2 (n-6) 12.31±0.00 22.62±0.06

The results are expressed as the mean ± SD (n=2)

ND :　Not detected

Unit: area%

As shown in Tables 3 and 4, the main fatty acids of the substrate were palmitic acid (38.15 area%) and oleic acid (41.54 area%). Sn- 2 was composed mostly of oleic acid (64.38 area%).

Table 5 shows the evaluation of the total fatty acid and% of the fatty acid in the sn-2 position relative to the primary reaction product according to the Experimental Example 1.

Fatty acid composition 1st reaction product fatty acid Total fatty acids sn -2 position C12: 0 0.37±0.06 ND C14:0 0.86±0.02 ND C16: 0 73.47±0.00 61.57±1.86 C18: 0 2.03±0.02 ND C18:1 (n-9) 17.95±0.05 29.43±2.05 C18:1 (n-7) 0.46±0.04 ND C18:2 (n-6) 4.88±0.04 9.00±0.19

The results are expressed as the mean ± SD (n=2)

ND :　Not detected

Unit: area%

The main fatty acids of the primary reaction products were palmitic acid (73.47 area%) and oleic acid (17.95 area%). Most of sn -2 consisted of palmitic acid (61.57 area%) and oleic acid (29.43 area%). In particular, when compared to the fatty acid composition of the substrate, the palmitic acid content increased from 38.15 area% to 73.47 area%, and the oleic acid content decreased from 41.54 area% to 17.95 area%. This is because a significant amount of oleic acid bound to the sn -1,3 position was replaced by palmitic acid by an enzyme that specifically reacts with the sn -1,3 position in reaction with palmitic ethyl ester. In addition, acyl migration occurred and the palmitic acid at the sn -1,3 position moved to the sn -2 position, increasing the amount from 9.24 area% to 61.57 area%.

Table 2 shows the evaluation of the total fatty acid and% of the fatty acid in the sn-2 position relative to the secondary reaction product (β-palmitin (maintaining milk)) according to the Experimental Example 1.

Total fatty acid composition SL2-2 SL2-4 C10: 0 0.56±0.01 0.57±0.01 C12: 0 2.14±0.02 2.26±0.06 C14:0 0.38±0.08 0.33±0.00 C16: 0 30.82±0.11 30.29±0.24 C18: 0 2.25±0.00 2.30±0.04 C18:1 (trans) 0.68±0.01 0.75±0.02 C18:1 (n-9) 53.37±0.16 53.52±0.11 C18:1 (n-7) 0.63±0.01 0.61±0.03 C10:2 (n-6) 9.20±0.02 9.37±0.01 sn- 2 fatty acids C16: 0 76.05±1.36 70.77±0.23 C18: 0 ND ND C18:1 (n-9) 23.95±1.36 29.23±0.23 C18:2 (n-6) ND ND

The results are expressed as the mean ± SD (n=2)

ND :　Not detected

Unit: area%

As shown in Table 6 above,

The fatty acids with the highest content of β-palmitin (maintaining milk substitute) were oleic acid (53.37-53.52 area%) and palmitic acid (30.29-30.82 area%). Palmitic acid content in breast milk is known to be 15.43-24.26 wt%. In particular, when compared with the fatty acid composition of the primary reaction product, the oleic acid content increased from 17.95 area% to 53.37-53.52 area%, and the palmitic acid content decreased from 73.47 area% to 30.29-30.82 area%. This is because a substantial amount of palmitic acid were coupled in the sn -1,3 positions by an enzyme acting specifically to the sn -1,3 positions was substituted with oleic acid.

The palmitic acid content at the sn -2 position of β-palmitin (maintaining breast milk substitute) was found to be 70.77-76.05 area%. The highest palmitic acid content at sn-2 was found in SL2-2 (76.05 area%), and the lowest value was found in SL2-4 (70.77 area%).

The content of palmitic acid at the sn-2 position in breast milk is It is known to be 50.6-58.0%, and it can be seen that β-palmitin (maintaining breast milk substitute) prepared by the method according to the present invention has an excess range beyond this.

Therefore, the method according to the present invention can be usefully used in the preparation of a breast milk substitute in the form of β-palmitin having saturated fatty acids at the sn- 2 position.

<Experiment 2> TAG analysis using Ag-HPLC

To separate the TAG isomers (symmetric structure and asymmetric structure), analysis was performed using Ag-HPLC (Younglin, Anyang, Korea). The column used a silver ion column (Chromspher 5 lipids 250 mm × 4.6 mm id, Varian, Netherlands), and a detector used a Sedex 75 evaporative light scattering detector (ELSD, Dedere, Alfortville, France). The temperature and pressure conditions of the ELSD were set at 40°C and 2.2 bar, and N ₂ gas was used. The flow rate was set to 1.5 mL/min, and the mobile phase solvent was solvent A (hexane: iso-propanol: acetonitrile = 100:0.1:0.1, v/v/v) and solvent B (hexane: iso-propanol: acetonitrile = 100:1:1, v/v/v) was used, and samples were dissolved in a mixed solvent (chloroform:hexane = 1:4, v/v) at a concentration of 1 uL/1 mL and analyzed by injecting 20 uL. . When the analysis sample was injected, the solvents A and B were set at a ratio of 100:0, flowed for 5 minutes, and then changed at a ratio of 80:20 until 50 minutes. The analysis was then completed by changing to a 50:50 ratio up to 60 minutes and holding for 1 minute, changing to a 100:0 ratio up to 62 minutes and holding up to 70 minutes. The retention time (RT) of each peak was checked through the OPO standard (Samyang, Seoul, Korea).

The composition of TAG (triglyceride) of the substrate analyzed by Ag-HPLC is shown in Table 7 below, and the chromatogram is shown in FIG. 3.

TAG type Palm oil Palm oil fraction(4℃) SSS 6.81±0.36 ND SUS 48.84±0.27 40.78±0.09 SSU 4.31±0.11 2.57±0.16 SUU 37.79±0.24 54.03±0.08 USU 0.20±0.01 ND UUU 2.04±0.03 2.63±0.15

The results are expressed as the mean ± SD (n=2)

ND :　Not detected

SSS: TAG which is a saturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

SUS: sn-1 and sn-3 positions are saturated fatty acids, and sn-2 positions are unsaturated fatty acids TAG

SSU: sn-1 and sn-2 positions are saturated fatty acids, and sn-3 positions are unsaturated fatty acids TAG

SUU: Sn-1 position is saturated fatty acid, sn-2, sn-3 position is unsaturated fatty acid TAG

USU: sn-2 position is saturated fatty acid, sn-1, sn-3 position is unsaturated fatty acid TAG

UUU: TAG which is an unsaturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

Unit: area%

Figure 3 shows the chromatogram of the substrate analyzed by Ag-HPLC (S = saturated fatty acids, U = unsaturated fatty acids). A) Palm oil, B) Palm oil fraction (palm oil fraction fractionated in step 1 of Example 1), C) Palmitic ethyl ester, D) Oleic ethyl ester

As shown in Table 7 and Figure 3, it can be seen that the palm oil fraction has a content of SUS and SUU of 90% or more.

The composition of TAG (triglyceride) of the reaction products of Examples SL1-1 to SL1-12 analyzed by Ag-HPLC is shown in Table 8 below, and the chromatogram is shown in FIG. 4.

Example SSS SUS SSU SUU USU UUU SL1-1 3.70 42.18 5.28 42.96 1.12 4.78 SL1-2 9.19 33.99 12.72 39.08 1.67 3.36 SL1-3 14.13 26.74 20.36 32.52 2.66 3.58 SL1-4 16.79 23.40 24.48 28.57 3.29 3.47 SL1-5 6.79 44.77 5.78 40.59 ND 2.07 SL1-6 19.43 34.37 14.65 29.39 ND 2.17 SL1-7 32.16 24.10 21.09 19.97 1.34 1.33 SL1-8 40.76 18.13 26.43 12.85 1.02 0.81 SL1-9 8.76 60.16 5.33 25.75 ND ND SL1-10 32.90 43.09 9.64 14.36 ND ND SL1-11 53.89 24.44 13.62 8.04 ND ND SL1-12 67.29 13.89 14.90 3.92 ND ND

ND :　Not detected

SSS: TAG which is a saturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

SUS: sn-1 and sn-3 positions are saturated fatty acids, and sn-2 positions are unsaturated fatty acids TAG

SSU: sn-1 and sn-2 positions are saturated fatty acids, and sn-3 positions are unsaturated fatty acids TAG

SUU: Sn-1 position is saturated fatty acid, sn-2, sn-3 position is unsaturated fatty acid TAG

USU: sn-2 position is saturated fatty acid, sn-1, sn-3 position is unsaturated fatty acid TAG

UUU: TAG which is an unsaturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

Unit: area%

Figure 4 shows a chromatogram of the substrate analyzed by Ag-HPLC for the reaction products of Examples SL1-1 to SL1-12.

As shown in Table 8 and Figure 4,

The longer the reaction time, the more acyl migration occurs and the saturated fatty acid content in the sn -2 position increases. SL1-12 (TLIM, 1:6, 12 hr, 80°C) was the primary product with the highest content of SSS and SSU containing saturated fatty acids at the sn -2 position, and the sum was 82.19 area%. Using SL1-12 as a substrate, the second reaction, β-palmitin (milk fat replacement paper), was synthesized.

The composition of TAG (triglyceride) of the reaction products of Examples SL2-1 to SL2-4 analyzed by Ag-HPLC is shown in Table 9 below, and the chromatogram is shown in FIG. 5.

TAG type SL2-1 SL2-2 SL2-3 SL2-4 SSS 24 9.5 10.6 3.6 SUS 6.7 ND ND ND SSU 48.8 51 51.4 49.6 SUU 9.6 11.2 10.1 10.6 USU 10.9 28.3 27.8 36.2 UUU ND ND ND ND

ND :　Not detected

SSS: TAG which is a saturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

SUS: sn-1 and sn-3 positions are saturated fatty acids, and sn-2 positions are unsaturated fatty acids TAG

SSU: sn-1 and sn-2 positions are saturated fatty acids, and sn-3 positions are unsaturated fatty acids TAG

SUU: Sn-1 position is saturated fatty acid, sn-2, sn-3 position is unsaturated fatty acid TAG

USU: sn-2 position is saturated fatty acid, sn-1, sn-3 position is unsaturated fatty acid TAG

UUU: TAG which is an unsaturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

Unit: area%

Figure 5 shows the chromatogram of the substrate analyzed by Ag-HPLC for the reaction products of Examples SL2-1 to SL2-4.

As shown in Table 9 and Figure 5,

One of the major components of β-palmitin, OPO (sn-2 position is palmitic acid, sn-1, sn-3 position is oleic acid TAG) represents the TAG form of USU. The shape of the USU ranged from 10.9% (SL2-1) to 36.2% (SL2-4). The TAG of the SSU form (PPO) is also β-palmitin with palmitic acid at the sn -2 position, so it can be said to be of nutritional value along with USU.

In terms of the type of enzyme and the reaction time, when TLIM and RMIM were used as catalysts, both of the USU synthesis occurred when the reaction time was 3 hr rather than the short reaction time (1 hr), but more when RMIM was used. A large amount of USU synthesis took place.

In addition, when comparing the TAG compositions of SL2-2 (TLIM, 3 hr, 45°C) and SL2-4 (RMIM, 3 hr, 45°C) with excellent synthesis amounts of USU and SSU in each enzyme, they were saturated at the sn -2 position. The sum of the fatty acid-containing SSU and USU showed that SL2-2 and SL2-4 were 79.3 area% and 85.8 area%, respectively, and both conditions showed a high content of 75% or more.

In Comparative Example 1 analyzed by Ag-HPLC, the results of Ag-HPLC analysis after 12-hour acyl migration reaction of maintaining high POO content are shown in Table 10 below.

TAG type POO-rich oil acyl migration SSS 6.5 SUS 5.4 SSU 19.6 SUU 37.0 USU 12.0 UUU 19.6

SSS: TAG which is a saturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

SUS: sn-1 and sn-3 positions are saturated fatty acids, and sn-2 positions are unsaturated fatty acids TAG

SSU: sn-1 and sn-2 positions are saturated fatty acids, and sn-3 positions are unsaturated fatty acids TAG

SUU: Sn-1 position is saturated fatty acid, sn-2, sn-3 position is unsaturated fatty acid TAG

USU: sn-2 position is saturated fatty acid, sn-1, sn-3 position is unsaturated fatty acid TAG

UUU: TAG which is an unsaturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

Unit: area%

As shown in Table 10 above,

As in Comparative Example 1, when performing only acyl migration using a high POO content, it can be seen that the content of triglycerides having saturated fatty acids at the desired sn-2 position is very low.

In Comparative Example 2 analyzed by Ag-HPLC, after the first reaction (step 1) and the second reaction (step 2) by substrate ratio (OOO:PEE=1:0.75 to 1:1 mol), the Ag-HPLC analysis results were obtained. It is shown in Table 11 below.

1st reaction result 2nd reaction result TAG type OOO:PEE
=1: 0.75 mole OOO:PEE
=1:1 mole OOO:PEE
=1: 0.75 mole OOO:PEE
=1:1 mole SSS One 2 0 0 SUS 3 4 One One SSU 7 12 3 5 SUU 32 35 14 17 USU 10 12 14 18 UUU 48 35 67 58

SSS: TAG which is a saturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

SUS: sn-1 and sn-3 positions are saturated fatty acids, and sn-2 positions are unsaturated fatty acids TAG

SSU: sn-1 and sn-2 positions are saturated fatty acids, and sn-3 positions are unsaturated fatty acids TAG

SUU: Sn-1 position is saturated fatty acid, sn-2, sn-3 position is unsaturated fatty acid TAG

USU: sn-2 position is saturated fatty acid, sn-1, sn-3 position is unsaturated fatty acid TAG

UUU: TAG which is an unsaturated fatty acid in all of the sn-1, sn-2, and sn-3 positions

Unit: area%

As shown in Table 11 above,

When the triglyceride having oleic acid as the starting material for all of the sn-1, sn-2, and sn-3 positions, after the first reaction, the sum of the contents of SSS, SSU, and USU is remarkably low compared to the method according to the present invention. , After the second reaction, it can be seen that even in the finally produced material, the contents of USU and SSU show a lower content of 1/3 or less compared to the present invention.

As described above, β-palmitin prepared by the method according to the present invention has a content of SSU such as USU and PPO of 65% or more, and 80% or more of SL2-4, which is very high. It can be seen that the content. In addition, even when the starting material is a vegetable oil or palm oil fraction having a high iodine value as in the present invention, it shows a production rate of β-palmitin (maintaining milk substitute), which is equivalent to a conventional manufacturing method using a starting material having a low iodine value.

Therefore, β-palmitin (maintaining a breast milk substitute) prepared by the method according to the present invention can be usefully used as a fat or oil substitute, and the method according to the present invention can be usefully used to prepare a breast milk substitute.

Claims (23)

Reacting vegetable oil with saturated fatty acid alkyl ester in the presence of Lipase (step 1); And
Reacting the reaction product of step 1 and the unsaturated fatty acid alkyl ester in the presence of Lipase (step 2),
Method for increasing the content of triglycerides with saturated fatty acids at the sn-2 position in vegetable oil.
According to claim 1,
The method comprises converting triglyceride having an unsaturated fatty acid at the sn-2 position in vegetable oil to triglyceride having a saturated fatty acid at the sn-2 position.
According to claim 1,
Lipase of step 1 is a method characterized in that the sn-1,3 specific lipase, Lipozyme TLIM (Thermomyces lanuginosus).
According to claim 1,
Lipase of step 2 is a sn-1,3 specific lipase, characterized in that Lipozyme TLIM (Thermomyces lanuginosus) or Lipozyme RMIM (Rhizomucor miehei).
According to claim 1,
The vegetable oil is a manufacturing method characterized in that the IV (iodine value) is 45-65.
According to claim 1,
The step 1 is a method characterized in that the transesterification reaction and acyl migration (acyl migration) reaction occurs.
The method of claim 6,
A method characterized in that the unsaturated fatty acid at the sn-2 position of triglyceride in vegetable oil is converted to saturated fatty acid by the acyl migration.
According to claim 1,
Step 2 is a method characterized in that the transesterification reaction occurs.
According to claim 1,
The saturated fatty acid of the saturated fatty acid alkyl ester of step 1 is characterized in that the palmitic acid (palmitic acid), myristic acid (myristic acid) or stearic acid (stearic acid).
According to claim 1,
The method of claim 2, wherein the unsaturated fatty acid of the unsaturated fatty acid alkyl ester of step 2 is oleic acid or linoleic acid.
A vegetable oil having an increased content of triglycerides with saturated fatty acids at the sn-2 position prepared according to the method of claim 1.
The method of claim 11,
Triglyceride having an unsaturated fatty acid at the sn-2 position,

,

And

Vegetable oil, characterized in that any one or a mixture thereof selected from the group consisting of:
(In the above formula, S is a saturated fatty acid, U is an unsaturated fatty acid).
The method of claim 11,
The vegetable oil,

,

And

Vegetable oil, characterized in that it may further comprise at least one triglyceride selected from the group consisting of:
(In the above formula, S is a saturated fatty acid, U is an unsaturated fatty acid).
reacting a triglyceride having an unsaturated fatty acid at the sn-2 position with a saturated fatty acid alkyl ester in the presence of Lipase (step 1); And
Reacting the reaction product of step 1 and the unsaturated fatty acid alkyl ester in the presence of Lipase (step 2),
A method for converting a triglyceride having an unsaturated fatty acid at the sn-2 position to a triglyceride having a saturated fatty acid at the sn-2 position.
The method of claim 14,
Triglyceride having an unsaturated fatty acid in the sn-2 position of step 1 is characterized in that the fraction fraction from palm oil.
The method of claim 14,
Lipase of step 1 is a method characterized in that the sn-1,3 specific lipase, Lipozyme TLIM (Thermomyces lanuginosus).
The method of claim 14,
Lipase of step 2 is a sn-1,3 specific lipase, characterized in that Lipozyme TLIM (Thermomyces lanuginosus) or Lipozyme RMIM (Rhizomucor miehei).
The method of claim 14,
Triglyceride having an unsaturated fatty acid in the sn-2 position of step 1 is characterized in that the IV (iodine value) is 55-75.
The method of claim 14,
The step 1 is a method characterized in that the transesterification reaction and acyl migration (acyl migration) reaction occurs.
The method of claim 19,
A method characterized in that the unsaturated fatty acid at the sn-2 position of triglyceride is converted to saturated fatty acid by the acyl migration.
The method of claim 14,
Step 2 is a method characterized in that the transesterification reaction occurs.
The method of claim 14,
The saturated fatty acid of the saturated fatty acid alkyl ester of step 1 is characterized in that the palmitic acid (palmitic acid).
The method of claim 14,
The method of claim 2, wherein the unsaturated fatty acid of the unsaturated fatty acid alkyl ester of step 2 is oleic acid.

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