KR100903705B1 - Enzymatic interesterification in a continuous packed bed reactor using stepwise in temperature for development of trans fat free and low saturated fat in the fats and oils - Google Patents

Abstract

Continuous enzymatic interesterification in a stepwise temperature changing system is provided to produce oils and fats containing less than 30% of saturated fat without trans fats by reacting to mixed oil of soybean oil-derived extremely hardened oils and high-oleic acid sunflower oils at higher temperature than a melting point of the mixed oil and re-reacting to the mixed oil at lower temperature than the melting point of the mixed oil. A method for preparing oils and fats containing less than 30% of saturated fat comprises the following steps of: mixing soybean oil-derived extremely hardened oils and high-oleic acid sunflower oils; primarily reacting to the mixed oil in a continuous packed-bed reactor with lipase at 70°C for 3-15 minutes; and secondarily reacting to the mixed in the continuous packed-bed reactor with lipase at 60°C for 3-200 minutes. The mixing ratio of the soybean oil-derived extremely hardened oils and high-oleic acid sunflower oil is 1 to 9 or 9 to 1. The lipase is selected from the group consisting of: lipase derived from microorganism containing Rhizopus delemar, Mucor miehei, Alcaligenes sp., Aspergillus niger, Candida Antarctica, Candida cylindracea and Geotrichum candidum; lipase derived from plant containing seeds from soybean, manuka and castor bean; and animal pancreatic lipase.

Description

Enzymatic interesterification in a continuous packed bed reactor using stepwise in temperature for development of trans fat free and low saturated fat in the fats and oils}

 The present invention relates to fats and oils containing 30% or less of saturated fat and a method for producing the same, and more specifically, to 1) soybean oil-derived ultra-cured oil and refined high oleic sunflower oil. Mixing; 2) a first reaction step of reacting the mixed oil in a continuous enzyme reactor (PBR: Packed Bed Reactor) in which lipase is present at 70 ° C. for 3 to 15 minutes; And 3) a second enzymatic transesterification reaction (EI) for reacting the mixed oil passed through step 2) in a continuous enzyme reactor (PBR) in which lipase is present at 60 ° C. for 3 to 200 minutes. : It is produced through Enzymatic Interesterification, and it does not contain trans fats and contains fats and fats containing 30% or less of saturated fat and its manufacturing method.

In nature, trans fats are found in extremely low levels in dairy products from ruminants. Most trans fats are produced in large quantities as vegetable fats and oils undergo hydrogenation, especially in partially hardened oils. Trans fats have been used in a variety of ways because they are convenient to carry, increase the shelf life of the goods, and also have the added effect (plasticity) that makes the sweets more crispy and savory. However, recent studies have reported that trans fat intake is highly correlated with the incidence of cardiovascular disease, and is considered to be a more dangerous risk factor for cardiovascular disease than saturated fatty acids. Therefore, in the food industry, trans fat reduction and development of alternative fats and oils have become the biggest issues.

Of the low-trans fat manufacturing processes under study, the most visible part of industrialization is based on transesterification. Although the transesterification reaction is difficult due to the introduction of new equipment and a lot of technical know-how and production technology, it is most suitable to replace various curing oils and meets the solid fat index according to the temperature required by margarine and shortening. It has the advantage of being able to.

The transesterification reaction includes chemical interest exchange (CI) using a chemical catalyst and enzymatic transesterification (EI) using an enzyme as a catalyst. Among the two methods, the enzymatic method has higher specificity and continuous reaction than the chemical method, the reaction proceeds at a relatively low temperature, and is environmentally friendly. The transesterification reaction using enzymes is to prepare triacylglycerols with desired properties by using trans fat-free fats and oils as substrates and lipases as enzymes to exchange positions of fatty acids 1 and 3 attached to glycerol. . The triacylglycerols thus prepared are trans fat-free, with no trans fat or present below the reference. It can also modify the melting point or crystallization properties of fats under certain conditions. This enzymatic transesterification has the characteristics of producing a desirable product in terms of environmental as well as the functionality of fats and oils. Therefore, the current global trend is trying to produce fats and oils through environmentally friendly enzymatic transesterification which makes it easier to produce high value-added fats and oils and secures food safety.

Enzymatic transesterification can be carried out in batch and continuous reactions, which have many advantages over batches. The biggest advantage of the continuous reaction is that the time to reach the reaction equilibrium is short, mass production is easy to apply, product recovery is relatively easy, and process control is easy. Therefore, the food industry prefers a continuous enzymatic transesterification reaction which is easy to mass produce rather than a batch reaction.

Accordingly, the present inventors have tried to replace the existing curing process that produces a large amount of trans fat and develop a fat-free and low fat containing fat that is suitable for the physical properties of processed fats. The enzymatic transesterification reaction is carried out by reacting the mixed oil mixed with oleic acid sunflower seed oil at a temperature higher than the melting point of the mixed oil and then reacting at a temperature lower than the melting point of the mixed oil to increase the residual activity of the enzyme and decrease the temperature. This resulted in less oxide production, no trans fat, and less saturated fat. In addition, the continuous reaction has a short time to reach the reaction equilibrium, easy to apply mass production, relatively easy product recovery, easy to control the process compared to the conventional batch reaction, and completed the present invention.

Accordingly, an object of the present invention is to prepare a mass production type continuous enzymatic transesterification reaction by lowering the reaction temperature according to the reaction time to increase the remaining activity of the enzyme, while not containing trans fat and a low saturated fatty acid content. To provide.

In order to achieve the above object, the present invention comprises the steps of 1) mixing soybean oil-derived ultra-cured oil and refined high oleic acid sunflower seed oil; 2) a first reaction step of reacting the mixed oil in a continuous enzyme reactor (PBR) in which lipase is present at 70 ° C. for 3 to 15 minutes; And 3) a second enzymatic transesterification reaction comprising reacting the mixed oil passed through step 2) in a continuous enzyme reactor (PBR) having lipase at 60 ° C. for 3 to 200 minutes. Provided is a method for producing fats and oils containing not more than 30% of saturated fat and no saturated fat.

 In the present invention, the transesterification reaction using the enzyme, which was only reacted at high temperature, was carried out by reducing the temperature to a low temperature, thereby increasing the remaining activity of the enzyme, improving the economics by saving energy, and maintaining the oil at a lower temperature. By reducing the production of free radicals, which are the cause, the production of oxides is reduced, and the reduction of hydrolysis due to the decrease of reaction temperature reduces the byproducts such as free fatty acids, thereby reducing the loss of purification.

In addition, the present invention has a shorter time to reach the reaction equilibrium compared to the conventional batch reaction, easy to apply mass production, relatively easy product recovery and easy process control.

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

The present invention provides a method for preparing a fat or oil comprising the following steps:

1) mixing soybean oil-derived extreme hardened oil with refined high oleic acid sunflower seed oil;

2) a first reaction step of reacting the mixed oil in a continuous enzyme reactor (PBR) in which lipase is present at 70 ° C. for 3 to 15 minutes; And

3) a second reaction step of reacting the mixed oil passed through step 2) in a continuous enzyme reactor (PBR) in which lipase is present at 60 ° C. for 3 to 200 minutes.

As used herein, the term "ultra-cured oil" refers to a fat or oil prepared so that the fatty acid content having a double bond is 0.2% or less through a hydrogenation reaction to the vegetable fat or oil.

The present invention uses, but is not limited to, soybean oil-derived ultra-cured oil as vegetable oil-derived ultra-cured oil, and is used in the stepwise temperature system proposed by the present invention even when other vegetable oil-derived ultra-cured oils are commonly used by those skilled in the art. The same efficacy can be obtained through the formula enzymatic transesterification, and it will be obvious that this also belongs to the scope of the present invention.

In addition, the high oleic acid sunflower seed oil used in the present invention is derived from the sunflower seed improved by raising the oleic acid content can be easily selected by those skilled in the art.

The mixing ratio of the soybean oil-derived ultra hardened oil and the high oleic acid sunflower seed oil is preferably 1: 9 to 9: 1 by weight, most preferably 3: 7 by weight.

The lipase used in the present invention can be used either from microorganisms, plants and animals, for example, Rhizopus delemar , Mucor miehei and Alcaligenes sp. .), lipase (lipase having selectivity to 1, 3-position of the glycerides (glyceride) to the origin of the microorganism, and the like); Aspergillus niger , Candida antarctica , Candida cylindracea , and Geotrichum candidum Random lipases derived from microorganisms such as these; Lipases derived from plants such as soybean minuka hima seed; And pancreatic lipases of animals. In general, it is convenient to use commercially available products, and immobilized lipases obtained according to certain rules, such as lipase itself, adsorption ion or covalent bonding method, such as lipase itself, for example, a Novo company lipozyme ( Lipozyme) RM IM ( Rhizomucor miehei ), lipozyme TL IM ( Thermomyces lanuginosus ) or Novozym 435 ( Candida antarctica ); Lipase PS-C ( Burkholderia cepacia ) or lipase PS-D ( Burkholderia cepacia ) from Amano; Or microorganisms such as fungi, yeasts and bacteria that have the ability to produce lipases can be used.

Since the melting point of the mixed oil used in the present invention is 60 ~ 67 ℃ the reaction temperature in the first reaction step is preferably carried out transesterification reaction at 70 ℃ for 3 to 15 minutes, and then in the second reaction step The transesterification is carried out for 3 to 200 minutes at 60 ° C. below the melting point. That is, the melting point of fats and oils is drastically reduced by going through the first reaction step of reacting at 70 ° C. for 3 to 15 minutes, which makes it possible to lower the reaction to 60 ° C. in the second reaction step, such a stepwise temperature system. Through it can maximize the remaining activity of the enzyme lipase.

In addition, the fats and oils prepared by the production method according to the present invention are characterized in that they do not contain trans fat and contain less than 30% of saturated fat.

Hereinafter, the content of the present invention will be described in more detail by examples. However, these examples are only presented to understand the contents of the present invention, but the scope of the present invention is not limited to these embodiments.

Comparative Example 1

After filling a certain amount of enzyme (approximately 0.7 to 0.8 g) in a continuous enzyme reactor (PBR; length = 7.62 cm; inner diameter = 0.48 cm), soybean oil-derived ultra-hardened oil and high oleate sunflower seed oil are mixed at a weight ratio of 3: 7. One sample was flown 22 times the volume of the reactor at a constant rate using a pump at 70 ° C. to reduce the free fatty acid content to less than 0.5%. Thereafter, the reactions were continued for 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 40, 60 and 100 minutes in a continuous reactor.

Example 1

After filling a certain amount (about 0.7 to 0.8 g) of enzyme in a continuous enzyme reactor (PBR; length = 7.62 cm; inner diameter = 0.48 cm), mixing soybean oil-derived hardened oil and high oleic sunflower seed oil in a weight ratio of 3: 7 One sample was flown 22 times the volume of the reactor at a constant rate using a pump at 70 ° C. to reduce the free fatty acid content to less than 0.5%. Samples whose melting point of the reaction oil reacted at 70 ° C was reduced to 60 ° C or lower, that is, for 3 minutes, 6 minutes, and 9 minutes at 60 ° C using a continuous reactor of the same specification for a reaction oil that reacted for 9 minutes or more at 70 ° C , 12 minutes, 15 minutes, 18 minutes, 21 minutes, 31 minutes, 51 minutes and 91 minutes.

Test Example 1 Acid Value Analysis

In order to solve the problem of an increase in acid value which may occur in the initial stage of the continuous enzymatic transesterification process, pretreatment reaction conditions were performed. Before starting the reaction, the mixed sample was flowed into a continuous reactor filled with enzyme at a constant rate. Samples were taken every 4, 10, 16, 22, 29, 35, 41 and 47 times the reactor volume to determine the acid value. At this time, the retention time of the enzyme and the sample was changed to 2, 5, 10 and 20 minutes, respectively, to calculate an acid value (AV: Acid value) according to Equation 1 in the same manner, and the results are shown in Table 1 below.

Acid value arrangement 2 minutes 5 minutes 10 minutes 20 minutes 4 5.14 3.95 2.97 5.45 10 1.53 1.54 1.57 1.25 16 0.93 0.93 1.06 0.79 22 0.75 0.88 0.84 0.74 29 0.65 0.60 0.74 0.70 35 0.65 0.60 0.70 0.61 41 0.60 0.60 0.61 0.51 47 0.56 0.51 0.51 0.6

In the results of Table 1, the acid value according to the reaction time of the enzyme and the sample showed an equivalent level. And since the acid value falls below 1 after flowing 22 times of the reactor volume, the above process is necessary as a pretreatment reaction before the continuous enzymatic transesterification reaction. Accordingly, the enzyme and the sample were reacted for 5 minutes in the pretreatment step of all the reactions, and passed through 22 times the reactor volume, followed by continuous enzymatic transesterification.

Test Example 2 Analysis of Transesterification Degree

In order to determine the degree of transesterification by enzymes of the fats and oils prepared in Example 1 and Comparative Example 1, the tristearin content of each fat or oil was analyzed using a liquid chromatography method. Degree of conversion (DC: Degree of Conversion,%) was calculated, and the results are shown in Table 2 below.

Degree of transesterification of fats and oils reacted at reaction temperature I (70 ℃) and reaction temperature II (70 ℃ + 60 ℃) Response time (hr) Reaction temperature I (70 ℃) Reaction temperature II (70 ℃ + 60 ℃) 0 0 3 29 6 78 9 86 84 ^* 12 92 91 ^* 15 94 92 ^* 18 93 96 ^* 21 95 96 ^* 24 96 96 ^* 27 95 96 ^* 30 97 97 ^* 40 99 98 ^* 60 99 99 ^* 100 99 99 ^{* *} Hold at 60 ° C for 3, 6, 9, 12, 15, 18, 21, 31, 51 and 91 minutes

In the results of Table 2, Example 1 showed an equivalent level of transesterification compared to Comparative Example 1. Therefore, it was confirmed that Example 1 is an ideal temperature system for the transesterification of the temperature system step by step.

[Test Example 3] Comparison of solid fat content

The solid fat content (SFC) for Comparative Example 1 and Example 1 was investigated. 3-5 g of each sample was placed in an SFC measuring cell and the solid fat content was measured at 12 temperatures from 5 ° C. to 5 ° C. at an initial temperature of 5 ° C .. At this time, the measurement equipment used was a Bruker solid-state content analyzer (Low Resolution NMR). In the results of Table 1, the transesterification reaction of Comparative Example 1 and Example 1 reached equilibrium at 24 minutes of reaction. Therefore, the SFC measurement was carried out for the transesterification fat which reached the equilibrium after reacting for 24 minutes, and the results are shown in FIG. 1. In the results of FIG. 1, the SFCs of Comparative Example 1 and Example 1 did not show a big difference, and the physical properties of the transesterified oils obtained from Comparative Example 1 and Example 1 were the same.

Test Example 4 Comparison of Melting Point of Oils and Fats

Melting points for Comparative Example 1 and Example 1 were investigated. Each sample was filled in a capillary tube about 1 cm, and one end of the capillary was sealed using a burner. After cooling it in a 4-10 ° C. refrigerator for about 30 minutes, the melting point of the fat was measured while raising the temperature at a rate of 0.5 ° C./min from 30 ° C. in a circulator (LAUDA E200) (AOCS official Method Cc 1-25 ). The temperature at which oil is completely clear is the melting point of the oil. The melting point of fats and oils was measured in three iterations and the average was calculated. The results are shown in Table 3 below.

Melting point Response time (hr) Reaction temperature I (70 ℃) Reaction temperature II (70 ℃ + 60 ℃) 0 63 - 3 60 - 6 58 - 9 57 57 ^* 12 56 56 ^* 15 55 55 ^* 18 54 54 ^* 21 54 53 ^* 24 54 50 ^* 27 53 52 ^* 30 53 52 ^* 40 54 51 ^* 60 50 50 ^* 100 49 49 ^{* *} Hold at 60 ° C for 3, 6, 9, 12, 15, 18, 21, 31, 51 and 91 minutes

In the results of Table 3, Example 1 was almost similar to the comparative example 1 without showing a large difference in the melting point of fats and oils. That is, the physical properties of the transesterified oils obtained from Comparative Example 1 and Example 1 were the same.

Test Example 5 Residual Activity of Enzyme

From the results of the test example, it can be seen that Example 1 is the optimal condition of the transesterification reaction using a stepwise temperature system, based on this the implementation of the transesterification reaction conditions using Comparative Example 1 and stepwise temperature system of high temperature transesterification reaction conditions. In Example 1, the residual activity of the enzyme was compared. Comparative Example 1 and Example 1 reached an equilibrium after 24 minutes of the transesterification reaction and thus fixed the reaction time to 24 minutes under the conditions of Comparative Example 1 and Example 1. The control group was subjected to transesterification reaction at 70 ° C. for 24 minutes in the same manner as in Comparative Example 1, and the control group was initially transesterified at 70 ° C. for 9 minutes in the same manner as in Example 1, and the remaining transesterification reaction was performed at 60 ° C. 15 minutes was carried out at ° C. Reactions to these controls and treatments were carried out continuously for 14 days, and the remaining activity of the enzyme was measured by measuring the degree of transesterification by taking the reaction maintenance every two days, and the results are shown in Table 4 below.

Remaining activity of the enzyme (degree of transesterification:%) Day Control Treatment 0 49 49 One 35 48 2 28 40 3 22 37 4 19 32 5 18 30 6 18 28 7 15 26 Control: Enzyme treated with reaction cycle at 70 ° C for 24 min. Treatment: Enzyme treated with reaction cycle at 70 ° C for 9 min and the remaining 15 min at 60 ° C.

In the results of Table 4, the treatment treated in the same manner as in Example 1 showed that the residual activity of the enzyme is about 1.6 to 1.8 times higher than the control.

As described above, Example 1 and Comparative Example 1 did not show a great difference in the degree of transesterification and physical properties, while in Example 1, the activity of the enzyme was much higher.

Based on these results, the present invention has developed the optimal conditions of transesterification using a stepwise temperature system in a continuous enzymatic reaction and can produce transesterified oils having the same degree of transesterification and physical properties as in the high temperature transesterification conditions. Could. In addition, it was confirmed that the continuous enzyme reaction reached equilibrium more than 10 times shorter than the batch type, and it was found that the process control was much easier and suitable for mass production type.

1 is a graph showing the solid fat content (SFC) of the fats and oils prepared in Example 1 and Comparative Example 1. FIG.

Claims (4)

1) mixing soybean oil-derived extreme hardened oil with high oleic acid sunflower seed oil; 2) a first reaction step of flowing the mixed oil of step 1) in a Packed Bed Reactor (PBR) in which lipase is present and reacting at 70 ° C. for 3 to 15 minutes; And 3) a second reaction step of reacting the mixed oil passed through step 2) in a continuous enzyme reactor in which lipase is present at 60 ° C. for 3 to 200 minutes; Process for producing fats and oils undergoing a continuous enzymatic transesterification reaction comprising a. The method of claim 1, wherein the soybean oil-derived ultra hardened oil and the high oleic acid sunflower seed oil are mixed in a weight ratio of 1: 9 to 9: 1. The method of claim 1, wherein the lipase is Rhizopus delemar , Mucor miehei , Alcaligenes sp. , Aspergillus niger , Candida antarctica, Candida cylindracea and Geotricum Candidum Lipases derived from microorganisms including Geotrichum candidum ); Lipases derived from plants including soybean minuka hima seed; And pancreatic lipase of an animal. A fat or oil prepared by the method according to any one of claims 1 to 3.

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