JPH0440995B2 - - Google Patents

Description

[Detailed description of the invention]

"Industrial Application Field" The present invention relates to a method for producing a cocoa butter substitute by transesterification of fats and oils using lipase. The present invention relates to a method for producing a cocoa butter substitute, which is characterized by carrying out a transesterification reaction in a short period of time through a stepwise reaction. "Prior Art and Problems" Hitherto, the transesterification reaction of oils and fats has been widely applied mainly as a means to improve the physical properties of edible oils and fats, such as crystallinity and degree of hardness. However, conventional transesterification methods using alkali metals, alkali metal alcoholates, alkali metal hydroxides, etc. as catalysts have low selectivity for the position of fatty acids in glycerides. On the other hand, by using lipase, which is a lipolytic enzyme, as a catalyst, the non-selectivity that was a drawback of chemical transesterification reactions has been improved, and transesterification methods enable selective transesterification depending on the purpose. has been developed (Japanese Unexamined Patent Publication No. 104506/1983). In the known transesterification method of oils and fats using lipase, lipase is used to esterify the middle melting point of palm oil, fats and oils such as shea butter, and other fats and oils, or the middle melting point of palm oil, fats and oils such as stearic acid, and free fatty acids. An exchange reaction is carried out (Japanese Patent Application Laid-Open No. 1973-
104506). In addition, a transesterification reaction between fractionated palm oil, fats and oils such as methyl stearate, and lower alcohol esters of fatty acids is carried out using lipase (Japanese Patent Publication No. 57-27159). Furthermore, the transesterification reaction of oils and fats has been carried out by a hydrolysis reaction and a continuous synthesis reaction to achieve a diglyceride content of 70% by weight or more in the total glyceride (Japanese Patent Application Laid-Open No. 1983-1991).
−203196). The transesterification reaction of fats and oils using lipase as a catalyst is considered to be a reversible reaction between the reaction of hydrolyzing fats and oils and the resynthesis reaction of the hydrolysis reaction product. In the conventional transesterification method of fats and oils using known lipase, water in the reaction system is
It has the disadvantage that the amount is small, 2.0% by weight or less, and the hydrolysis reaction rate is therefore significantly suppressed. As a result, conventional methods for transesterifying fats and oils using lipases have the disadvantage of requiring a long period of time, from one to three days, or more, to complete the reaction. It is expected that the transesterification reaction will be shortened by hydrolysis to diglyceride as disclosed in JP-A-60-203196. However, when carrying out the transesterification reaction of the 1st and 3rd positions of glyceride using a 1st and 3rd position-specific lipase, it is clear that the time-saving effect is greater if the hydrolysis reaction is carried out up to 2-monoglyceride. It is. It is known that 2-monoglyceride is rearranged to 1- or 3-monoglyceride, and rearrangement to 1- or 3-monoglyceride means that the transesterification reaction of fatty acid residues at positions 1 and 3 of the target glyceride Instead, a transesterification reaction at the 1st and 2nd positions or the 2nd and 3rd positions will be carried out. 2-A method of hydrolyzing fats and oils capable of obtaining a high content of monoglycerides has not been found;
Furthermore, as mentioned above, since the problem of rearrangement of 2-monoglyceride has not been solved, it is difficult to hydrolyze it to 2-monoglyceride.
We had no choice but to stop at hydrolysis to diglycerides like 203196. "Means for Solving the Problem" In view of the fact that the conventional transesterification of fats and oils using a known lipase requires a long time, the present inventors proposed a transesterification reaction of fats and oils using a lipase as a catalyst. As a result of conducting research on the production of cocoa butter substitutes, we conducted a hydrolysis reaction to obtain a high content of 2-monoglyceride by applying a lipase with 1- and 3-position specificity to fats and oils, or mixtures of fats and other fats. Then, lipase is applied to the hydrolyzed product or a mixture of the hydrolyzed product and the free fatty acid and/or the lower alcohol ester of the fatty acid to perform a resynthesis reaction, that is, to perform a transesterification reaction, that is, to perform a transesterification reaction. The present inventors have discovered that the reaction time can be surprisingly shortened by carrying out the transesterification reaction of fats and oils through a two-step reaction in which a decomposition reaction and a resynthesis reaction are carried out separately, leading to the present invention. In the hydrolysis reaction of fats and oils using lipase, which the present inventors previously proposed, it is possible to obtain hydrolysis products with a high monoglyceride content (Japanese Patent Application No. 72324/1982), and (Japanese Patent Application No. 22829/1989). As a result of further research, it was found that 2-monoglyceride among the monoglycerides obtained by this hydrolysis method was 95%
It has become clear that it reaches more than % by weight. However, it is known that 2-monoglyceride rearranges to 1- or 3-monoglyceride, and 2-oleomonoglyceride rearranges into 1- or 3-monoglyceride.
- Rearrangement to monoglyceride means that it becomes impossible to produce a cocoa butter substitute by the transesterification reaction of the present invention. The rearrangement of 2-oleo monoglyceride manufactured by Serdary Research Lab. into 1-oleo monoglyceride in a 40°C hexane solution was as shown in the following table.

[Table] Additionally, 2-oleomonoglyceride was added to Mc Ilvaine buffer (PH8.0) and stirred at 40°C, and the rearrangement of 1-oleomonoglyceride was measured. The results were as shown in the table below.

[Table] From the above results, the rearrangement of 2-monoglyceride to 1-monoglyceride hardly occurs in hexane solution, and McI-
(Ivaine) It became clear that the rearrangement was faster when in contact with a buffer solution. The present invention involves applying a lipase having 1- and 3-position specificity to fats and oils with a high 2-oleoglyceride content, or mixtures of fats and oils with other fats, to carry out a hydrolysis reaction to obtain a high content of 2-oleomonoglyceride. to obtain a hydrolyzed product of fats and oils having a high content of 2-oleomonoglyceride, and then to the hydrolyzed product of the fats and oils, or to the mixture of the hydrolyzed product of the fats and oils and free fatty acids and/or lower alcohol esters of fatty acids, lipase is added. The gist is the production of a cocoa butter substitute through a two-step reaction in which a transesterification reaction is carried out. The oils and fats used in the present invention include ordinary animal and vegetable oils containing a large amount of oleic acid in the second position, microbial oils and synthetic oils, and specifically, palm oil,
One type or a combination of two or more types of derived oils and fats such as shea butter, olive oil, safflower oil, triolein, diglyceride, and fractionated fats thereof can be used. Preferred is 1,3-saturated 2-oleotriglyceride. The free fatty acids and/or lower alcohol esters of fatty acids used in the present invention include palmitic acid,
These include stearic acid, methyl palmitate, and methyl stearate. The lipase used in the present invention is a normal lipase of animal or plant origin or microbial origin, and any lipase having 1- and 3-position specificity may be used for the hydrolysis reaction. Specifically, these include pancreatic lipase, rice bran lipase, Rhizopus lipase, and Mucor lipase, particularly Mucor javanicus, Rhizopus delemar, Rhizopus japonicus, and Rhizopus chinensis. Microbial lipases such as chinencis) are preferable. The method of the present invention involves the presence of a buffer solution or water in an amount of 20 to 500 times the weight of the substrate oil.
Alternatively, a mixture of fats and oils and other fats is subjected to a hydrolysis reaction by acting on a lipase having 1- and 3-position specificity to obtain a fat-and-oil hydrolysis product with a high content of 2-oleomonoglyceride, and then hexane , an organic solvent that does not inactivate lipase, such as industrial hexane, petroleum ether, or petroleum benzine, is added, and after stirring, the hydrolysis product of fats and oils is recovered by standing and/or centrifugation. If necessary, free fatty acids and/or lower alcohol esters of fatty acids are added to the organic solvent solution of the hydrolyzed product of recovered fats and oils, and lipase is further applied to carry out the resynthesis reaction. The reaction is carried out in two stages. In the hydrolysis reaction, the lipase can be used as a buffer solution or an aqueous solution, but it can also be made to act as an immobilized lipase by a known method such as adsorption onto a support. In the resynthesis reaction, the lipase can be directly dispersed in the reaction system and acted upon, but it is preferable to improve the dispersibility and act as an immobilized lipase by a known method such as adsorption on a support. As the support, known supports such as diatomaceous earth, polysaccharide derivatives, and ion exchange resins can be used. Although the transesterification reaction of the present invention varies depending on the type of lipase, it is usually desirable to carry out the transesterification reaction at a temperature in the range of 30 to 50°C. When using a thermostable lipase, it is possible to perform the treatment at temperatures of 50°C or higher. The reaction time of the hydrolysis reaction to obtain a high content of 2-monoglyceride is preferably within 2 hours. A reaction lasting more than 2 hours is undesirable because the fatty acid at the 2-position is rearranged to the 1- or 3-position, resulting in a decrease in the 2-monoglyceride content. The resynthesis reaction is carried out in a short time, within 10 hours. The production of a cocoa butter substitute by transesterification of fats and oils using the lipase of the present invention includes
Or 100 parts of a mixture of fats and oils and other fats, 2,000 to 5,000 parts of buffer or water, and 0.05 to 20 parts of a commercially available lipase with 1- and 3-position specificity (a commercially available standard lipase with an activity of about 4,000 to 40,000 units). can be directly added to the reaction system and reacted as described above, but it can also be immobilized by known methods such as adsorption onto a support, or added with lipase and hydrated by stirring at 30 to 50°C. Carry out the decomposition reaction. When the amount of water used in a normal hydrolysis reaction is about 500 parts or less, the amount of 2-monoglyceride accumulated is small, but in a hydrolysis reaction system with a high water content of 2000 parts or more, the amount of 2-monoglyceride accumulated is 50 parts. weight%
reach more than that. To recover hydrolysis products of fats and oils, add an organic solvent that does not inactivate lipase, such as hexane, industrial hexane, petroleum ether, or petroleum benzine, to the reaction system.
This is carried out by stirring and then standing still and/or centrifuging. The resynthesis reaction in the transesterification reaction between fats and oils and other fats and oils is carried out by adding 0.05 to 20 parts of commercially available lipase to an organic solvent solution of the hydrolysis product of fats and oils and stirring the mixture. Although lipase can be directly added to the reaction system and allowed to act, it is preferable to immobilize it by a known method such as adsorption onto a support or allow lipase to act. Alternatively, the resynthesis reaction can also be carried out by passing an organic solvent solution of a hydrolyzed product of fats and oils through a garam filled with immobilized lipase. "Examples" The present invention will be described in more detail below with reference to Examples and Comparative Examples, but it goes without saying that the present invention is not limited thereto in any way. Example 1 100 parts of palm oil medium melting point, 5 parts of commercially available powdered lipase derived from M. javanicus (manufactured by Amano Pharmaceutical), 25000 parts of McIlvaine buffer (PH8.0), IM chloride 50 parts of calcium solution was stirred at 40°C for 90 minutes to carry out a hydrolysis reaction. Immediately after the completion of the hydrolysis reaction
- 200 parts of hexane was added, stirred, and then centrifuged to collect a hexane solution. Next, 100 parts of stearic acid, 200 parts of n-hexane, 30 parts of Celite-adsorbed lipase, and 10 parts of powdered molecular sieve (manufactured by Union Showa) were added to the hexane solution of the hydrolysis product of fats and oils, and the mixture was heated at 40°C for 8 hours. The mixture was stirred and a resynthesis reaction was carried out. After completion of the resynthesis reaction, Celite-adsorbed lipase,
The molecular sieve was filtered off, and hexane was distilled off at 60°C under reduced pressure. Free fatty acids were removed from the resulting transesterification product by steam distillation to obtain transesterified fats and oils. Note that the Celite-adsorbed lipase was obtained by adsorbing commercially available powdered lipase (manufactured by Amano Pharmaceutical Co., Ltd.) originating from M. javanicus to Celite. Unless otherwise specified hereinafter, it is a celite-adsorbed lipase of the same type. Analysis method: Using a differential refractometer (R1) as a detector, tetrahydrofuran as a mobile phase, and high performance liquid chromatography (HPLC) using a gel permeation chromatography (GPC) column, the composition of each glyceride and fatty acid was determined. was analyzed. In addition, triglyceride composition was analyzed by HPLC using an ODS column. The analysis results are shown in Table 1. Values are expressed in weight% (the same applies hereinafter).

[Table] In the table, POP is 1,3-dipalmito-
2-olein, POS represents 1-palmito-2-oleo-3-stearin, and SOS represents 1,3-distearo-2-olein (the same applies below). A significant increase in stearic acid content and a significant decrease in palmitic acid content in triglycerides was confirmed. Table 2 shows the composition of the fatty acid residue at position 2 of triglyceride.

[Table] This transesterified oil showed physical properties and triglyceride composition very similar to those of cocoa butter, and was excellent as a cocoa butter substitute. Table 3 shows the monoglyceride composition contained in the hydrolysis reaction product.

[Table] Example 2 A hydrolysis reaction was carried out according to the method of Example 1 to obtain a hexane solution of the hydrolysis product of fats and oils.
Add 100 parts of stearic acid and hexane to the hexane solution.
A resynthesis reaction was carried out by adding 200 parts of the solution and passing the solution through a jacketed glass column with a diameter of 20 mm and length of 250 mm filled with Celite-adsorbed lipase at a flow rate of 30 ml/H. The column was kept at 40°C. After the transesterification reaction was completed, transesterified fats and oils were obtained according to the method of Example 1. Table 4 shows the results.
Shown below.

[Table] The fat obtained by removing diglycerides from the transesterified fat by treatment with a hydrous florisil column showed physical properties and composition very similar to those of cocoa butter. Example 3 Example 1 except that a commercially available lipase (manufactured by Meito Sangyo) derived from Candida cylindracea adsorbed on celite was used in the resynthesis reaction.
A transesterified oil and fat was obtained according to the method of The results are shown in Table 5.

[Table] Example 4 A transesterified oil and fat was obtained according to the method of Example 1, except that ion-exchanged water was used instead of McIlvaine buffer in the hydrolysis reaction. The results are shown in Table 6.

[Table] The high melting point portion obtained by solvent fractionation of the transesterified fat could be used satisfactorily as a cocoa butter substitute. Example 5 A transesterified oil and fat was obtained according to the method of Example 1, except that olive oil was used instead of the medium melting point part of palm oil and methyl stearate was used instead of stearic acid. The results are shown in Table 7.

[Table] The fat obtained by blending the high melting point part obtained by solvent fractionation of the transesterified fat with the middle melting point part of palm oil could be sufficiently used as a cocoa butter substitute fat. Example 6 Palm oil middle melting point instead of palm oil middle melting point
A hydrolysis reaction was carried out according to Example 1 using 50 parts of shea butter and 50 parts of medium melting point part of shea butter, and further adding 100 parts of n-hexane. n-hexane after completion of hydrolysis reaction
After adding 100 parts, stirring and centrifugation, a hexane solution was collected. Next, according to the method of Example 2, a resynthesis reaction was carried out by passing a hydrolyzed hexane solution of the oil and fat through a column. A transesterified oil and fat was obtained according to the method of Example 1.

[Table] The transesterified fats and oils could be used satisfactorily as cocoa butter substitutes either as they were or when blended with palm oil medium melting point. Comparative Example 1 100 parts of palm oil medium melting point, 100 parts of stearic acid,
30 parts of Celite-adsorbed lipase, 10 parts of powdered molecular sieve (manufactured by Union Showa) and n-hexane
200 parts were added thereto, stirred at 40°C, and transesterification was carried out according to the method of Example 1 to obtain transesterified oil and fat. The results are shown in Table 9.

[Table] Comparative example 2 100 parts of olive oil, 100 parts of methyl stearate,
Add 30 parts of Celite-adsorbed lipase, 10 parts of powdered molecular sieve, and 200 parts of n-hexane, and heat at 40°C.
Transesterification reaction was carried out for 8 hours according to the method of Example 1 to obtain transesterified oil and fat. Display results
Shown in 10.

[Table] "Effects of the Invention" The present invention enables the transesterification of fats and oils using lipase to obtain fats and oils that are extremely excellent as cocoa butter substitutes with high POP, POS, and SOS contents, and which is impossible with conventional methods. Since the transesterification reaction is completed in a short time of 10 hours, this method provides an industrially extremely advantageous method for producing a cocoa butter substitute.

Claims (1)

[Claims] 1. 1, 3 in oil or fat, or a mixture of oil and other fat
A hydrolysis reaction is carried out to obtain a high content of 2-monoglyceride by the action of a lipase having positional specificity,
Next, lipase is applied to the hydrolyzed product of the fat or oil, or a mixture of the hydrolyzed product of the fat and free fatty acid and/or the fatty acid lower alcohol ester to perform a resynthesis reaction. A method for producing a cocoa butter substitute by transesterification. 2. The manufacturing method according to claim 1, wherein the hydrolysis product contains 15 to 50% by weight of 2-oleomonoglyceride based on the total glyceride. 3. The manufacturing method according to claim 1, wherein the hydrolysis product contains 2-oleomonoglyceride in an amount of 50% by weight or more based on the total glyceride. 4. The manufacturing method according to each of claims 1 to 3, using a substrate fat or oil in which the fatty acid residues at the 1st and 3rd positions of the triglyceride are saturated fatty acids and the fatty acid residue at the 2nd position is oleic acid. 5. The manufacturing method according to each of claims 1 to 4, wherein after the hydrolysis reaction is completed, the hydrolysis product is recovered using an organic solvent that does not inactivate the lipase.