HK1158894A - Cocoa butter equivalents produced by the enzymatic interesterification process and method for preparing the same - Google Patents

Abstract

The present invention relates to a process for preparing hard butter having high SOS content by mixing oil for preparing butter with fatty acid or fatty acid ester, adding 1,3 regio-specific enzymes to the obtained mixture to carry out interesterification, distilling the obtained reactants to remove fatty aicd, ethyl ester, and monoglyceride and diglyceride formed after the reaction and fractionally extracting the obtained reactants to separate a solid phase, and to cocoa butter equivalents prepared by the hard butter and a process for preparing the same in which the cocoa butter equivalents can replace existing import cocoa butter equivalents with 1 : 1 because of its equivalent chemical properties, and have no difference in taste and properties with natural cocoa butter and also have lower trans fatty acid. Hard butter according to the present invention can make desired triglyceride structure in oil based on the reaction conditions and have a improved purity and yield in the whole process by recycling all of byproduct other than major product in the distillation and fractional distillation process and is eco-friendly matter by using the enzymatic interesterification reaction, and also cocoa butter equivalents made by the hard butter is characterized in replacing existing import cocoa butter equivalents with 1 : 1 because of its equivalent chemical composition and properties in the production of chocolate with no difference in taste.

Description

Cocoa butter replacer prepared by enzymatic transesterification and preparation method thereof

Technical Field

The invention relates to cocoa butter replacers prepared by enzymatic transesterification and a preparation method thereof, and particularly provides a preparation method of hard cream with high-concentration SOS content, cocoa butter replacers prepared from the hard cream and a preparation method thereof. The hard butter is prepared by mixing an oil and fat raw material for preparing butter with fatty acid or fatty acid ester to obtain a mixture, then adding 1-and 3-site specific enzyme into the mixture to perform transesterification reaction, distilling the obtained reactant, removing fatty acid, ethyl acetate and monoglyceride and diglyceride generated after the reaction, and then separating out a solid phase through fractionation. The cocoa butter substitute has the same chemical properties as the existing imported cocoa butter substitute, can be replaced by 1: 1, has no difference with natural cocoa butter in aspects of taste, properties and the like, and simultaneously has reduced trans-fatty acid content.

Background

Cocoa butter is a natural fat component of cocoa beans (Theobroma cacao), which is defined by the united states food and drug administration as "edible fat obtained before or after roasting (roasting) ripe cocoa beans", which contain 50-55% cocoa butter.

Cocoa butter is valuable in its specific flavor and specific physical properties not possessed by other fats and oils. Cocoa butter is hard at temperatures below room temperature and dissolves rapidly at body temperature, thus having a fresh mouth-feel. And is extremely stable against oxidation, and therefore, is suitable for the production of foods requiring long-term storage such as chocolate. The melting point of cocoa butter is 32-35 deg.C. The hardness varies according to the place of origin, with cocoa butter produced in Malaysia being the most hard, cocoa butter produced in Brazil being the most soft, and cocoa butter produced in Gardner and Kotedawa being of moderate hardness.

Cocoa butter contains 99% triglycerides, less than 1% mono-and diglycerides, 0.2% sterols, and 150-250ppm tocopherols. The saponification value is 188-195, the iodine value is 34-41, and the unsaponifiable matter is 0.3-0.8%.

In particular, unlike other fats or oils, cocoa butter contains more than 80% of the three main fatty acids, i.e., palmitic, stearic and oleic acids, and more than 75% of a symmetrical triglyceride structure in which oleic acid is linked to the 2-position of the triglyceride and palmitic and stearic acids are linked to the 1, 3-positions of the triglyceride. The main symmetrical oil contains palmitic acid-oleic acid-stearic acid (POS) 34-39%, stearic acid-oleic acid-stearic acid (SOS) 23-30%, palmitic acid-oleic acid-palmitic acid (POP) 14-17%, and oleic acid more than 80% is located at 2-position of triglyceride.

Cocoa butter is used not only for chocolate but also for ice cream, candy, etc., and is a light yellow fat obtained by mechanical compression, has a hard texture at a temperature of 20 deg.C or lower, has a solid fat content of about 80% at room temperature, softens at 30-32 deg.C, and dissolves at 32-35 deg.C. Chocolate has such a sensitive melting point because cocoa butter, as mentioned above, has a symmetrical triglyceride structure.

Because cocoa butter is a relatively simple triglyceride structure, it exhibits a unique variety of morphologies upon cooling and solidification. Chocolate has six crystal forms, which can be divided into I-VI forms, and the melting point of the chocolate is increased from 16 ℃ to 36 ℃. The temperature adjusting process ensures that the chocolate has stable V type (melting point is 34-35 ℃) and VI type (melting point is 36 ℃) as a main component of bloom crystal. Recently, a total of 6 crystal forms of α, γ, and both of β' and β have been reported. V and VI are in beta form.

Typically chocolate contains less than 50% sugar, about 30-50% cocoa powder, and about 30% pre-fat components including fat. In particular, fats and oils used in chocolate have a great influence on the melting point characteristics (melting in the mouth). The market for chocolate and chocolate-related products is expanding, but natural cocoa butter is available in limited quantities, making it expensive and subject to wide variations in price. Therefore, various alternative fats and oils are being developed according to their uses.

Since cocoa butter is expensive, only a part of the total fat required for chocolate is used, and the remainder is mixed with other vegetable fats, and this cocoa butter substitute is called hard butter. Furthermore, the quality of chocolate varies depending on the quality of cocoa butter replacers, which can be replaced by 1: 1 in nature and chemical composition with natural cocoa butter.

Cocoa Butter alternatives are broadly classified as Cocoa Butter Equivalents (CBE), Cocoa Butter Replacers (CBR), and Cocoa Butter Substitutes (CBS).

Triglyceride composition of Cocoa Butter Equivalent (Cocoa Butter Equivalent: CBE) is similar to that of natural Cocoa Butter, excellent in compatibility, and requires tempering. Palm oil intermediate Fraction (Palm Middle Fraction oil), illipe, shea, veronicastrum, and Garcinia indica lipid, among others, belong to this class. Compared with the addition of cocoa butter, the fat similar to cocoa butter can be obtained by adding cocoa butter equivalent with high stearic acid content into palm oil intermediate fraction rich in palmitic acid and mixing. In the preparation of chocolate, cocoa butter equivalent is in most cases not much different from cocoa butter in terms of taste, properties, etc., and can replace cocoa butter. That is, there is no great difference in texture, flavor, and mouthfeel from natural cocoa butter. The annual demand of European Union countries for the oil is more than 1.5 ten thousand tons, and the oil is widely used for replacing cocoa butter worldwide.

Cocoa Butter Replacer (CBR) is a fat obtained by hydrogenating soybean oil, rapeseed oil and palm oil, and can replace natural Cocoa Butter to some extent without requiring tempering. The hydrogenation process treatment produces more trans fatty acids with high melting point, increased solid fat content, increased slope of SFC curve, and increased oxidation stability, so as to be close to natural cacao butter, but the chewing feeling is soft, and the hard feeling of chocolate is not existed. The substitute cocoa butter can be used in chocolate to replace natural cocoa butter, and the dosage can be about 15%.

Cocoa Butter replacers (CBS) are hydrogenated lauric oils that are not compatible with Cocoa Butter and do not require tempering. Suitable for use in confectionery coatings, typically in the hydrogenated oil or interesterification of palm kernel oil and coconut oil, and also in mixtures with other vegetable hydrogenated oils.

The cocoa butter-substitute fat may be prepared by a method of mixing fats and oils separated from palm, illipe, shea butter, and the like, or a method of performing transesterification reaction with an enzyme.

Transesterification refers to a reaction in which an ester reacts with an alcohol, an acid or other glyceride to produce a new ester, and a reaction between the same species is called an interesterification (interesterification) and a reaction between the different species is called a transesterification (transesterification). These reactions are carried out by exchanging and recombining glycerides and acyl groups with a chemical catalyst or an enzyme as a biological catalyst, thereby changing the composition of glycerides, i.e., the chemical composition, and also changing the physical properties such as the melting point and the solid fat content. The transesterification reaction is directly related to the flavor, heat stability, nutrition and the like of edible oil, and particularly, the edible oil can be improved into oil with proper properties, which is particularly important.

The transesterification reaction is classified into a Chemical method (CIE) and an Enzymatic method (EIE) according to the enzyme used. The EIE method is an improved technique that does not add any chemical agent and does not produce harmful by-products, and thus can be called as an environment-friendly melting point inflection inducing technique. On the contrary, since CIE uses a chemical catalyst, oil is lost in the process of removing residual sodium fatty acid (sodium soap), and in terms of reaction characteristics, discoloration of fats and oils, dag (diacylglycerol) residues, and the like occur, so that a subsequent purification process is required. Further, since EIE reacts at a low temperature as compared with CIE and its reaction is peculiar, natural antioxidant substances such as tocopherol contained in oils and fats can be largely retained, and the fatty acid structure can be changed by the peculiar expression of EIE, which is not achieved by CIE. Therefore, the development trend in the world can be said to be a biological international method "enzymatic transesterification technology" that has high added value oil products, is easy to produce, ensures food safety performance, and is environmentally friendly.

Through research of multiple scientists for years, the technology of the enzymatic ester exchange method is an effective method for adjusting the solid fat index of the grease. However, this technique has not been applied to products other than high-priced products due to the cost of expensive raw materials resulting from curing. With the great improvement of the curing technology, the technology can be used for replacing industrial bulk oil such as cocoa butter and the like, and can produce the best product in terms of functions and health.

The main advantages of the enzymatic transesterification technique are as follows:

1. the process is simple and convenient to adjust; 2. can endow products with various changes; 3. trans fatty acid is not generated; 4. a more natural product can be prepared.

Currently, the u.k.et al european union 7 countries stipulate: the chocolate component may contain vegetable oil and fat 5% except for cacao butter. Chocolate products in japan and china also have a rule that chocolate should contain more than 35% cocoa powder and more than 18% cocoa butter according to the fair competition treaty. Further, the use of a vegetable oil or fat in place of cocoa butter is not limited as long as it is within the above-specified range.

Cocoa Butter substitutes for Cocoa Butter Equivalent (CBE) in fats and oils can replace 100% of natural Cocoa Butter, and even experts have difficulty finding differences in the final products. Thus, chocolate is currently prepared by mixing expensive natural cocoa butter and Cocoa Butter Equivalent (CBE). In view of the world trend, the demand for cocoa butter equivalent has recently increased, since the legislation stipulates that up to 5% cocoa butter equivalent can be used in combination with natural cocoa butter, also regarded as chocolate. However, Cocoa Butter Equivalent (CBE) circulating in China is completely imported, so the development of cocoa butter equivalent is particularly important.

"research on the development of cocoa butter replacers according to the reverse micelle-enzyme reaction system" (Korea society of food science, 24(2), p111-116, 1992), published by book et al, was studied to produce cocoa butter replacers by transesterifying palm oil and stearic acid with Rhizopus radiculosus lipase in a reverse micelle-enzyme reaction system, qualitatively and quantitatively analyzing each triglyceride by high performance liquid chromatography, and changing the molar ratio of water to dioctyl sodium sulfosuccinate (Aerosol OT) in a simulated reaction of triolein and stearic acid, showing the maximum turnover rate when 3 times stearic acid is added to 30mM triolein at a ratio of 10, and also obtaining the maximum turnover rate at a pH of 7.5 and a temperature of 50 ℃, when palm oil and stearic acid were used as the base material, the amounts of POPs, POOs, and SOOs in palm oil were reduced, whereas the amounts of POS and SOS produced tended to increase. As a result of the investigation, the composition of triglycerides such as POP, POS and SOS of cocoa butter substitute was closer to that of natural cocoa butter than that of commercial cocoa butter substitute.

Korean patent application No. 10-1992-. Dissolving A.O.T in hexane at 1.5-2.5 wt%, dissolving edible oil and fat at 0.9-6 wt% and fatty acid at 0.1-6 wt%, and adding 1-3 site specific lipase solution to prepare reverse micelle.

Korean patent publication No. 10-1996-0001494 discloses a "method for preparing cocoa butter replacers". The method comprises the following steps: (a) a step of immobilizing the lipase by covalently bonding the hydrophobic carrier to the lipase, wherein the hydrophobic carrier is directly covalently bonded to the lipase or covalently bonded thereto via a hydrophobic spacer; (b) a step of dissolving a mixture of stearic acid and triolein or a mixture of stearic acid and palm oil in hexane; and (c) adding the immobilized lipase obtained in the step (a) to the substrate solution obtained in the step (b) to carry out a reaction.

Korean patent No. 10-0773195 entitled "an oil and fat composition and method for preparing cocoa butter replacer having reduced trans-fatty acid content" discloses an oil and fat composition and method for preparing cocoa butter replacer having reduced trans-fatty acid content using the same. The method comprises the steps of mixing two kinds of oil with proper fatty acid composition, and carrying out transesterification reaction or selective hydrogenation reaction on the oil, so as to prepare the cocoa butter substitute with reduced trans-fatty acid content, wherein the cocoa butter substitute has good taste and heat resistance, and simultaneously has compatibility with the cocoa butter of more than 20 percent, excellent processing performance and reduced possibility of inducing cardiovascular and cerebrovascular diseases.

The present inventors have studied on the production of cocoa butter replacers having a reduced trans-fatty acid content by enzymatic transesterification and a production method thereof, and have completed the present invention. Specifically, no chemical organic solvent such as hexane is added, a continuous reaction which is beneficial to improving yield and purity is adopted in the process operation, the reaction is carried out at 40-50 ℃, a fatty acid raw material with good reaction effect in an ester form is used, a by-product is recycled in the extraction process, and a process which is more convenient than the prior art is adopted, so that the high-purity SOS hard butter is prepared, and therefore, the cocoa butter substitute and the natural cocoa butter have no difference in taste, properties and the like, have the same triglyceride composition and properties as the imported cocoa butter substitute, and can be replaced by 1: 1.

Disclosure of Invention

Technical problem to be solved by the invention

The invention aims to provide hard butter with reduced trans-fatty acid content, which is prepared by transesterification, wherein the SOS content of the hard butter accounts for more than 85% of the triglyceride structure.

The invention also aims to provide cocoa butter replacer prepared from the hard butter and a preparation method thereof.

Means for solving the problems

One embodiment of the present invention relates to a method for producing a trans fatty acid reduced cocoa butter replacer using an enzymatic transesterification process.

A particular embodiment of the present invention relates to a process for the preparation of a hard butter having an SOS content of more than 85%, comprising the steps of:

(a) a step of mixing an oil and fat raw material for preparing cream with a fatty acid or a fatty acid ester;

(b) a step of adding an enzyme specific to the 1-and 3-positions to the mixture obtained in the step (a) to perform a transesterification reaction;

(c) distilling the reactant obtained in the step (b), and then removing fatty acid, ethyl acetate, and monoglyceride and diglyceride generated after the reaction;

(d) a step of separating a solid phase by fractionating the reaction product obtained in the step (c).

In the method for preparing hard butter according to the present invention, as the oil and fat raw material for preparing butter used in the step (a), there may be used oil and fat or a mixture of two or more kinds of oil and fat known in the art, preferably soybean fully hydrogenated oil, fully hydrogenated tallow, palm oil, palm stearin, palm essential oil, palm kernel essential oil, coconut hydrogenated oil, coconut oil, sal oil, sterculia stearin, indian mangosteen oil, shea butter stearin, cottonseed stearin, soybean oil, corn oil, cottonseed oil, rapeseed oil, canadian rape oil, sunflower seed oil, safflower oil, grape seed oil, or olive oil. More preferably at least one of high oleic sunflower oil, high oleic canola oil, high oleic soybean oil, high oleic corn oil and high oleic safflower oil.

The fat and oil raw material for preparing the cream is mixed with stearic acid fatty acid or fatty acid ester in a mixing ratio of 1: 2-6, and the moisture content in the fat and oil of the raw material is preferably less than 0.02%.

In the method for producing hard butter according to the present invention, the transesterification reaction in the step (b) is carried out in a 1-step continuous process. This is accomplished by a continuous process which provides convenience to the reaction process, does not overload the enzyme, and has high production efficiency and high reaction oil yield as compared to the existing batch process. The enzyme specific to the 1-and 3-positions used in the transesterification reaction is preferably a specific lipase. In this case, the continuous reaction is preferably carried out at 40 to 50 ℃.

In the method for producing hard butter according to the present invention, the step (c) is a step of distilling the reaction oil obtained by the transesterification reaction in a molecular still. By this step, fatty acids, ethyl acetate, and monoglycerides and diglycerides formed after the reaction can be removed. The reaction oil has a monoglyceride and diglyceride content of less than 1% after distillation. And the removed fatty acid and ethyl acetate can be reused in the enzymatic transesterification reaction after being treated by a hydrogenation process.

In the method for producing hard butter according to the present invention, the step (d) is a step of separating a solid portion by a fractionation step after the enzymatic transesterification (step b) and the distillation (step c), whereby a fat or oil having an SOS content of 85% or more in a triglyceride structure can be obtained. In addition, the liquid phase part in the fractionation step can be reused in the transesterification reaction after separation, and at this time, the composition of triglyceride should be controlled to be suitable for the reaction conditions. In the above step, the single solvent used in the wet fractionation is preferably acetone.

Through the extraction steps, triglyceride containing more than 85% of SOS and hard butter with high content of POP triglyceride in the oil can be obtained.

The hard cream prepared by the method of the present invention is completed by an enzymatic transesterification reaction completed in a single step, which can obtain a high yield of reaction oil as compared with the conventional process in which the reaction is performed twice or more, and the process steps are convenient and simple. Moreover, the content of monoglyceride and diglyceride can be reduced to less than 1% by the distillation step. And hard cream having an SOS content of 85% or more can be obtained by the extraction step. And the byproducts generated in the separation step are all recovered and can be reused in the continuous enzymatic transesterification reaction, thereby improving the yield and the purity.

As another embodiment, the invention prepares cocoa butter equivalent by mixing the hard butter prepared by the method and the natural oil and fat containing more than 85% of POP according to a proper proportion, so that the cocoa butter equivalent has properties suitable for application.

A preferred embodiment relates to the cocoa butter equivalent produced by the above-described enzymatic transesterification reaction, having a solid fat index at 10 ℃ of 82 to 90%, at 20 ℃ of 40 to 45%, at 25 ℃ of 9 to 14%, at 30 ℃ of 1 to 4%, and at 35 ℃ of 0 to 0.5%.

Further, since trans fatty acids are not produced during the enzymatic transesterification reaction, the trans fatty acid content of the cocoa butter equivalent prepared by the above method is 0.1% or less.

The cocoa butter equivalent prepared by the preparation method has the triglyceride content of more than 99 percent, the diglyceride and monoglyceride content of less than 1 percent and the lower lauric acid fatty acid content of less than 0.1 percent.

These chemical compositions and properties are the same as those of imported cocoa butter equivalents and natural cocoa butter, and can be replaced by 1: 1, and they are not greatly different from natural cocoa butter in taste and properties when chocolate is prepared, and can be used instead of cocoa butter.

Drawings

FIG. 1 shows the results of HPLC analysis of triglycerides of hard butter after the step of distillation and fractionation after the enzyme transesterification, and the SOS content in this figure is 85% or more.

FIG. 2 shows the HPLC analysis results of the triglycerides of cocoa butter equivalents prepared by mixing the synthetic hard butter and natural hard butter according to the present invention. The main peaks in this figure are POP, POS, SOS, which are symmetrical triglycerides.

FIG. 3 shows DSC results of cocoa butter replacers of the present invention and conventional cocoa butter replacers.

FIG. 4 is a graph showing the solid fat content of cocoa butter replacers according to the invention and the solid fat content of the prior art cocoa butter replacer as a function of temperature.

Detailed Description

The present invention is further illustrated by the following examples. These examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.

Example 1 enzyme selection

The lipase TLIM (Lipozyme TLIM; Denmark Novoxil) derived from Thermomyces lanuginosus is immobilized in the porous silica particles and is insoluble in oil.

In this example, the lipase TLIM enzyme was reacted with the same substrate for 24 hours in order to compare the properties of the lipase RMIM (Lipozyme RMIM, Denmark Novovin) which has been widely used.

TABLE 1 (area ratio:%)

	DG	SOO	SOS	SSS
					TLIM	1.4	32.4	49	3.7
RMIM	14.4	34.2	39	2.3

DG: diglyceride, S: stearic acid, O: oleic acid

As a result of the reaction, RMIM activity was lower than TLIM activity, and as shown in Table 1 above, the composition of triglyceride of the reaction product after 24 hours of the reaction was: the high content of diglycerides in RMIM, and therefore, it is understood that the conditions for the enzymatic transesterification reaction in the present invention are economically advantageous in that TLIM is more stable and more effective than RMIM.

Example 2 investigation of whether selected enzymes could undergo transesterification

This example to determine whether the lipase TLIM enzyme selected in example 1 above could be subjected to a transesterification reaction, a high oleic oil and stearic acid were mixed in a ratio of 1: 2 to 6, and then the above mixed oil is subjected to a batch reaction. At this time, the reaction temperature was 45 to 50 ℃, and in order to examine the degree of reaction in the hexane solvent, the reaction temperature was divided into two types, i.e., addition of the solvent and non-addition of the solvent, and the melting point and the structure of triglyceride in the fat and oil were analyzed according to the reaction time. The melting points before and after the transesterification reaction with the enzyme were measured using an automatic melting point measuring apparatus, and the content of the SOS triglyceride structure in the fat or oil was measured using a reversed-phase liquid chromatography-evaporative light scattering detector. The melting point of the mixed oil before reaction is about 25-28 ℃, and the melting point after reaction is 29-32 ℃. The results of the analysis of the content of the triglyceride structure in the fat and oil after 9 hours of the reaction, based on the reaction time, showed that 30 to 35% of SOS was contained (see Table 2). Also, there was no difference in the degree of reaction between the addition of the hexane solvent and the absence of the hexane solvent. The stearic acid-stearic acid (SSS) triglyceride removed from the over-reacted reactants should be below 1%.

TABLE 2

The reaction is carried out for 9 hours	Adding hexane solvent	Without addition of solvent
			OOO	6.1	7.3
SOO	46.2	45.4
			SOS	35.2	34.8
SSS	-	-

Example 3 removal of fatty acids, monoglycerides and diglycerides by distillation

In this example, unreacted fatty acid, ethyl acetate, and monoglyceride and diglyceride formed after the reaction were removed from the reaction oil produced by the enzymatic transesterification reaction using a molecular distillation apparatus. The reaction conditions were 1X 10^-3mbar, 200 ℃, the amount of mono-and diglycerides in the distilled reaction oil does not exceed 1%. And the distilled fatty acid, ethyl acetate, etc. can be reused in the enzymatic transesterification reaction through a hydrogen hydrogenation step.

Example 4 extraction Using fractionation procedure

In the embodiment, acetone is used as a single solvent, and the oil with the SOS content of 85% in the triglyceride structure is prepared by adopting wet fractionation and dry fractionation.

The oil is separated through the separation process, so that the oil is separated into solid and liquid. The solid phase of triglyceride had an SOS content of 85% or more, and the result was analyzed by reversed phase liquid chromatography-evaporative light scattering detection apparatus.

The desired results of the present invention were achieved by separating the solid triglyceride structure separated by the above extraction to have an SOS content of about 85% or more, and analyzing the triglyceride structure of the liquid phase to have more than about 80% of the triglycerides as unreacted stearic acid-oleic acid (SOO) and oleic acid-oleic acid (OOO). Unreacted triglycerides are recovered and can also be reused in the enzymatic transesterification reaction. Such a step can improve the yield and purity of the reaction.

Example 5 preparation of Cocoa Butter Equivalent (CBE) Using SOS synthetic oil

This example describes the preparation of cocoa butter equivalents by mixing a reaction oil having a triglyceride content of 85% or more in the enzymatic transesterification reaction oil with a hard butter having a high palmitic acid-oleic acid-palmitic acid (POP) triglyceride content in fats and oils. The cocoa butter substitute contains more than 85% of SUS (S: Saturate, U: Unsulfate) symmetrical triglyceride structure. This has a great influence on the physical and chemical properties of fats and oils, and this structure also has an influence on the solubility in mouth, which is characteristic of cocoa butter and cocoa butter substitutes.

The following experiments were carried out using the hard butter prepared in examples 1 to 5 above.

Experimental example 1 physical property analysis was performed using a differential scanning calorimeter

In the present experimental example, the Melting curve (Melting Profile) before and after transesterification with an enzyme was measured with a differential calorimeter, and the possibility of reaction was known from the measurement. The experimental conditions are shown in table 3 below.

TABLE 3

DSC Instrument name	TA Q20
		Temperature of experiment	-60 ℃ to 80 DEG C
Rate of cooling	10 ℃/min (to-80 ℃ C.)
		Rate of temperature rise	5 ℃/min (to 100 ℃ C.)
Amount of standard sample	15±5mg

The DSC experiment shows that the whole phase change is in the temperature range of-60 to 80 ℃, and the result shows that the invention is almost completely consistent with the existing cocoa butter substitute. The results of the experiment are shown in FIG. 3.

Experimental example 2 analysis of solid fat content by Nuclear Magnetic Resonance (NMR)

In this example, the solid fat content was analyzed by nuclear magnetic resonance under the analysis conditions shown in table 4 below. The solid fat content was analyzed by means of nuclear magnetic resonance, the experimental Method being a Parallel Method (Parallel Method). 6 samples were prepared for each of the enzymatic transesterification reactions, and the pretreatment was carried out by dissolving the oil and fat sufficiently at 100 ℃ and then leaving the solution at 60 ℃ for 5 minutes and then at 0 ℃ for 60 minutes. The pretreatment time was about 80 minutes. The samples were measured by placing them in a previously prepared centigrade bath (Celsius bath) -constant temperature metal bath (metal block thermostat) at 10.0 deg.C, 20.0 deg.C, 25.0 deg.C, 30.0 deg.C and 35.0 deg.C. The measurement of the sample took about 6 seconds.

TABLE 4

NMR Instrument name	BRUKER the minispec
		Frequency of	60MHz
Amount of sample	6ml
		Temperature of pretreatment	100 constant temperature metal bath, 0 DEG C
Temperature of experiment	10.0℃，20.0℃，25.0℃，30.0℃，35.0℃

The results of the natural fats and oils considered as cocoa butter equivalents and the SFC (Solid Fat Contents: Solid Fat content) of the currently used cocoa butter equivalents, as well as of the synthetic hard butter and cocoa butter equivalents of the present invention are shown in table 5 below.

TABLE 5

	10℃	20℃	25℃	30℃	35℃
						Hard butter of the invention	76.6	63.4	46.2	20.4	6.1
The cocoa butter equivalent of the invention	83.2	42.3	12.8	5.5	1
						The existing cocoa butter equivalent	90.5	45.7	9.4	0.7	0.5
Bingbingcao fat	93.7	74.9	26.5	1.3	0.5
						All-grass of Veronicastrum	87	78.4	64.4	21	0.9
Butyrospermum parkii	77.6	62.3	51	7.9	2.5
						Palm oil intermediate fraction	94.2	81	48.9	5.5	0

From the experimental results in said table 5, it is clear that the hard cream of the present invention has very similar properties to the vero fat stearin extract oil. The illipe, veronicastrum, shea butter in table 5 are referred to as natural cocoa butter equivalents. The annual production of these lipids is variable and expensive and uneconomical. The invention prepares hard butter with similar properties to the above lipids through enzyme transesterification, and prepares a cocoa butter equivalent by using the hard butter, wherein the cocoa butter equivalent can replace the existing cocoa butter equivalent by 1: 1.

EXAMPLE 3 melting Point measurement analysis

The melting points were measured and analyzed under the conditions shown in Table 6 below. After mixing the substrates, the melting Point (melting Point) was analyzed by a method capable of rapidly measuring the degree of reaction and physical properties. The melting point was measured by using an EX-871 automatic rising melting point measuring apparatus. Firstly, completely melted grease is filled into a capillary tube about 1cm, then the capillary tube is inserted into a capillary tube supporting table, and then the capillary tube supporting table is placed in a refrigerator for about 10 minutes to obtain a melting point sample. The prepared 10 ℃ distilled water was put into the water tank of the melting point measuring apparatus, and then the prepared sample was inserted on the sensor to measure. The temperature is first raised at a rate of 2 deg.C/min within the temperature range of 10-25 deg.C, and then raised at a rate of 0.5 deg.C/min after 25 deg.C.

TABLE 6

Name of instrument	EX-871 AUTOMATIC ELEVATING MELTING POINT DETERMINATION APPARATUS
		Temperature rise	0.5℃/min
Number of samples	8 are provided with
		Pretreatment conditions	-5 ℃ (10 minutes)
Detection method	Detection method of special photoelectric sensor
		Heating device	Special coilHeater 400W
Stirring device	Variable speed vibrating device

The melting point is closely related to the mouth solubility of the oil and fat, and particularly, the oil and fat used for chocolate is required to be stable in the process of circulation at normal temperature and to have a melting point which is just melted in the mouth. And the melting point is the same as the solid fat content in the above experimental example 2 as the weight factor in the physical properties of the fat or oil. The melting point of the cocoa butter substitute generally used is about 34 ℃. The melting point of the hard butter synthetically prepared according to the invention is 39-40 ℃. The melting point of cocoa butter substitute prepared by mixing palm oil intermediate fraction and the like is 35 ℃. The results of the melting point test are as follows.

TABLE 7

	Melting Point (. degree.C.)
		Hard butter	39.3
The invention relates to cocoa butter substitute	35
		The existing cocoa butter substitute	34
Bingbingcao fat	28.6
		Butyrospermum parkii	32.9
All-grass of Veronicastrum	34.6
		Palm oil intermediate fraction	31.7

Experimental example 4 analysis of fatty acids by GC

In this experimental example, fatty acids were analyzed by gas chromatography under the conditions shown in table 8 below. Trans fatty acids and fatty acid compositions were analyzed under the following conditions. The Standard was a Lipid Standard (Lipid Standard) of SIGMA (SIGMA), and the reagents used for the analysis were all special grade reagents. In order to methylate each sample subjected to the enzymatic transesterification, 0.025mg of each sample was taken, 1.5ml of a 0.5N NaOH-methanol solution was added thereto, and after heating on a heating block for about 5 minutes, the mixture was cooled in a thermostatic bath at 30 to 40 ℃. Then 2ml of BF 3-methanol solution was added, heated on a heating block and cooled in a thermostat at 30-40 ℃. Then, 1-2ml of isooctane (iso-octane) and a saturated NaCl solution were added to the sample, mixed and left to stand. After separating the upper layer, the sample was analyzed by removing water with sodium sulfate dehydrate (sodium sulfate dehydrate).

TABLE 8

In order to investigate the fatty acid content of hard butter, cocoa butter replacers, natural cocoa butter replacers and existing cocoa butter replacers prepared by the enzyme transesterification reaction, gas chromatography was used for analysis. The analysis results are shown in table 9 below. From the results of the analysis, it was found that lauric acid, which is a lower fatty acid, was not present, and trans fatty acid was not 0.1%, and that trans fatty acid was not produced in the enzymatic transesterification reaction of the present invention.

TABLE 9

Experimental example 5 triglyceride Structure analysis

This experimental example analyzes the structure of triglyceride under the conditions shown in the following table 10. The structure of triglyceride in fat and oil was analyzed by the following experiment. The triglyceride structure of cocoa butter equivalent was analyzed using a reversed-phase hplc-evaporative light scattering detector system. After 30. mu.l of the sample and 10ml of hexane were put, the mixture was filtered by a Teflon syringe filter (25mm, 0.2 μm), and then the mixture was put into a 2mm vial, followed by injecting 100. mu.l of the sample by an auto sampler. The solvent used was acetonitrile (solvent A), hexane/isopropanol (solvent B) and the flow rate was 1 ml/min. Gradient elution of solvent (A: B, v: v) was carried out as follows: the ratio was 80: 20 for 45 minutes, 54: 6 for 60 minutes, 80: 20 for 60 minutes to 70 minutes, and 70 minutes for total application time.

Watch 10

The structural analysis of the triglycerides of the present invention is very important. The position of fatty acid triglyceride is changed according to the enzyme transesterification, and the triglyceride structure determines the physical and chemical properties of the grease. In particular, cocoa butter or cocoa butter equivalent is characterized by containing 90% or more of a specific triglyceride having a SUS (S: Saturate, U: Unsulfate) symmetric structure.

In addition, the present invention has carried out an analysis experiment on the triglyceride structure after the synthesis, separation and extraction steps.

TABLE 11

Experimental example 6 analysis according to TLC-FID

In this experimental example, TLC-FID analysis was performed in order to determine the content of diglycerides, triglycerides and the like under the conditions shown in Table 12 below.

The content of diglycerides, monoglycerides and triglycerides can be determined by TLC-FID. TLC-FID is an instrument that can perform quantitative and qualitative analysis of organic mixed substances separated on Thin Layer Chromatography (TLC). A chromatographic rod (quartz rod coated with a thin layer of silica or alumina at high temperature to separate and spread the sample) was designed specifically for TLC-FID analysis. The solvent used for the analysis was a special grade reagent from SIGMA (SIGMA) corporation. After dissolving the sample in the solvent, about 1. mu.l of the sample was spotted on the above-mentioned chromatographic bar. After the sample was developed in the developing tank containing the developing solvent for 20 minutes, it was thoroughly dried in a drying oven and then analyzed by TLC-FID.

TABLE 12

The composition of glycerides was analyzed based on the water content in the mixed oil by using the fact that the triglyceride content and the diglyceride and monoglyceride content after the enzyme transesterification change depending on the water content in the oil and fat. The analysis results are shown in the following Table 13. As a result of the analysis, when the water content in the fat or oil was 0.02% or less before the enzyme transesterification, a hard cream having a triglyceride content of 99% or more and a diglyceride and monoglyceride content of 1% or less was obtained.

Watch 13

Moisture content in oil (%)	Triglyceride (%)	Monoglyceride, diglyceride (%)
			0.01	99.3	0.7
0.02	99.2	0.8
			0.05	98.7	1.3

The results of analyzing the contents of triglyceride, diglyceride, and monoglyceride of the hard butter and cocoa butter substitute synthesized according to the present invention are shown in table 14 below.

TABLE 14

	Triglyceride (%)	Monoglyceride, diglyceride (%)
			Hard butter	99.2	0.8
The invention relates to cocoa butter substitute	99.4	0.6
			The existing cocoa butter substitute	99.4	0.6
Palm oil intermediate fraction	99.5	0.5

Industrial applicability

The preparation method of the cocoa butter equivalent firstly adopts continuous enzymatic transesterification when preparing the fat containing rich SOS, the process is more convenient than the prior batch process, triglyceride is separated by molecular distillation, so that the yield and purity of the reaction are improved, the operation is convenient, and the hard butter with the triglyceride structure, which has high purity and high yield, is prepared by selective extraction by using solvents and the like. The hard butter obtained by the above process is mixed with various fats and oils to prepare cocoa butter equivalent having a composition similar to that of triglyceride of natural cocoa butter. The chemical composition and properties of the cocoa butter equivalent prepared by the method are the same as those of imported cocoa butter equivalent and natural cocoa butter, and can be replaced by 1: 1, so that the cocoa butter equivalent has no great difference with the natural cocoa butter in the aspects of taste, properties and the like when being used for preparing chocolate, and can replace the natural cocoa butter.

Claims (12)

1. A method for preparing hard butter with SOS content of more than 85% comprises the following steps:

a step of mixing an oil raw material for preparing cream with a fatty acid or a fatty acid ester;

b, adding 1 and 3 site specific enzyme into the mixture obtained in the step a to perform ester exchange reaction;

c, distilling the reactant obtained in the step b, and then removing fatty acid, ethyl acetate, and monoglyceride and diglyceride generated after reaction;

d a step of separating the solid phase by separating the reactant obtained in the step c.

2. The method for preparing hard butter according to claim 1, wherein the fat and oil raw material for preparing butter in step a is one selected from soybean fully hydrogenated oil, fully hydrogenated tallow, palm oil, palm stearin, palm essential oil, palm kernel essential oil, coconut hydrogenated oil, coconut oil, salmon stearin oil, indian mangosteen oil, shea butter stearin, cotton seed stearin, soybean oil, corn oil, cotton seed oil, rapeseed oil, canadian rape oil, high oleic canadian rape oil, sunflower seed oil, high oleic sunflower seed oil, safflower oil, grape seed oil or olive oil, or a mixture of two or more thereof.

3. The method for producing hard butter according to claim 1, wherein a mixing ratio of the fat and oil raw material and the fatty acid or fatty acid ester for producing butter is 1.0: 2.0 to 6.0.

4. The method of producing hard butter according to claim 1, wherein the oil and fat raw material for producing butter in step a has a moisture content of 0.02% or less and a reaction temperature of 40 to 50 ℃.

5. The method for producing hard butter according to claim 1, wherein in the step b, the enzyme specific to the 1-and 3-positions is a lipase TLIM (lipozyme TLIM).

6. The method of producing hard butter according to claim 1, wherein the content of diglycerides and monoglycerides is 1% or less and the content of triglycerides is 99% or more in step c.

7. The method of producing hard butter according to claim 1, wherein the fatty acid or fatty acid ester removed in the step c is recovered and reused in an enzymatic transesterification reaction by a hydrogenation step.

8. The method of producing hard butter according to claim 1, wherein the reaction product obtained in step d contains solid triglyceride having an SOS content of 85% or more and an SSS content of 1% or less.

9. The method of preparing a hard butter according to claim 8 wherein the liquid phase triglyceride obtained in step d has an SOS content of less than 5%.

10. The method of producing hard butter according to claim 9, wherein the oil or fat having less than 5% of SOS in the triglyceride obtained in the step d is recovered and then reused in the enzyme transesterification reaction to perform a continuous reaction.

11. A hard butter having an SOS content of 85% or more prepared according to the process of any of claims 1-10.

12. A cocoa butter equivalent prepared from a hard butter obtained by the process according to claim 8, wherein the cocoa butter equivalent has a solid fat content of 82-90% at 10 ℃, 40-45% at 20 ℃, 9-14% at 25 ℃, 1-4% at 30 ℃ and 0-0.5% at 35 ℃.