JPS6078587A - Preparation of fatty acid ester - Google Patents

Abstract

PURPOSE:To obtain a high-purity fatty acid ester in high yield, by carrying out alcoholysis reaction of fats and oils or a fatty acid monoalkyl ester and an alcohol in the presence of lipase in a solventless state. CONSTITUTION:In preparing a fatty acid ester by subjecting fats and oils or a fatty acid monoalkyl ester and an alcohol to alcoholysis, lipase produced by a microorganism belonging to the genus Alcaligenus, Arthrobacter, or Pseudomonas, or a culture of the microorganism is used as a catalyst, and the reaction is carried out in a solventless state.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing a fatty acid ester by causing an alcoholysis reaction between an oil or a fatty acid monoalkyl ester and an alcohol.

Conventionally, new fatty acid esters were produced by reacting oils and fats or fatty acid monoalkyl esters with alcohol.
Chemical transesterification reactions using alkali catalysts such as sodium methoxide, sodium ethoxide, sodium carbonate, or metal catalysts such as tin-zeta-zinc have been used. However, these chemical transesterification reactions
Relatively harsh conditions are required, and since the reaction is a commutation reaction, the selectivity to fatty acid esters is poor. Therefore, it has the disadvantage that the yield of the target product is low and, moreover, a large number of impurities are present in the product.

On the other hand, a method for producing fatty acid esters using an enzyme-catalyzed reaction using microorganisms has been proposed, which allows the reaction to proceed even under mild conditions. For example, Ishihara et al. (Chem, Phar
The alcoholysis reaction of fats and oils or fatty acid monoalkyl esters using Mucor or pancreatic lipase as a catalyst is known, according to the method of M., Bullet 23.3266 (1975).

However, this method is carried out in a reaction system containing water. For this reason, it is not possible to suppress the hydrolysis of the raw fats and oils, raw fatty acid esters, and product fatty acid esters in the presence of water in the lipase fraction, so a decrease in the yield and purity of the target fatty acid ester is avoided. It has the disadvantage that it cannot be obtained.

An object of the present invention is to provide a method for producing fatty acid esters using linose, which can produce fatty acid esters with high yield and purity.

The inventors of the present invention have devised an idea to perform the alcoholysis reaction in the absence of water to prevent a decrease in yield and purity due to hydrolysis. However, all 9 p4-ases used in conventional alcoholysis reactions act only in the presence of water. As a result of intensive research, the inventors discovered that alkaline earth metal lipases, produced by bacteria of the genus Alithropacter, or bacteria of the genus Pseudomonas, function satisfactorily even in the absence of water, leading to the completion of this invention.

That is, the present invention provides a method for producing fatty acid esters by causing an alcoholysis reaction between fats and oils or fatty acid monoalkyl esters and alcohol, in which alkaline earth metals, Alithropacter paulownia, or lipase produced by Pseudomonas bacteria are catalyzed. Provided is a method for producing a fatty acid ester, which is used as a fatty acid ester, and is characterized in that the reaction is carried out without a solvent or in a non-aqueous solvent.

The lipase used in this invention includes alkaline earth metals,
It is produced by bacteria of the genus Alithropacter or Sheetmonas. Removal of these bacteria can be carried out in exactly the same manner as in the past. That is,
For example, bacteria are cultured aerobically in a culture solution containing suitable carbon sources, oxygen sources, inorganic salts, and olive oil as a lipase inducer, and ammonium sulfate is added to the culture medium to precipitate the tannin. This can be done by subjecting the lipase to an ion-exchange column such as xemography and collecting the lipase fraction. Alternatively, a bacterial culture can be used as it is as an alcoholysis reaction catalyst.

The amount of lipase added is preferably 0.31 to 20% by weight based on the raw material oil or fat or fatty acid ester.

If this is less than 0.1% by weight, the catalytic action will decrease and 2
If it exceeds 0.00 weight, the recovery rate of the product decreases, which is not preferable.

Furthermore, in the alcoholysis reaction, linose and linase may be used in a free state, but in order to further increase the efficiency, they may be immobilized on a nyosine-active carrier by a conventional method. As the carrier, those that are insoluble in the reaction system of the present invention and do not adversely affect the enzyme activity are used, such as celite, alumina, activated clay, quartz sand, molecular sieves, porous silica, etc. 〇The method of the present invention is carried out in the absence of water.

This prevents the raw materials and products from being hydrolyzed. It is preferable that the reaction system be homogeneous, so if the raw materials and products are liquid and dissolve in each other, a solvent-free system is used, and if at least one component of the raw materials or products is precipitated, the raw materials and products are The reaction is carried out in a non-aqueous solvent that can dissolve the enzyme and does not inactivate the enzyme catalyst. Preferred nonaqueous solvents are, for example, petroleum benzene, n-hexane, petroleum ether, and DMF.

Organic solvents such as DMSO and acetone. These non-aqueous solvents are preferably used in an amount of 1000 parts by weight or less per 1 part by weight of the raw material oil or fat or fatty acid monoalkyl ester.

The fats and oils or fatty acid monoalkyl esters and alcohols used as raw materials in the method of this invention may be the same as conventional ones depending on the purpose. That is, natural vegetable or animal fats or synthetic fats and oils are used as the raw material fats and oils. As a natural oil,
Vegetable oils and fats such as Norm oil, Noyam kernel oil, olive oil, soybean oil, rapeseed oil, corn oil, cottonseed oil, and safflower oil; animal fats and oils such as beef tallow, pig intestine, fish oil, squid oil, and whale oil. As U fatty acid monoalkyl ester, carbon number is 6
Alkyl esters having 1 to 4 carbon atoms of aliphatic monocarboxylic acids having 1 to 22 carbon atoms are preferred. If the number of carbon atoms in the fatty acid and alkyl group is outside the above range, the alcoholysis reaction will be difficult to proceed. Specific examples of raw fatty acid monoalkyl esters include methyl cabroate, ethyl pelargonate, globyl undecanoate \zotel laurate, ethyl thyarumitate, araki>m propyl, methyl behenate, etc.@the above raw material oils and fats or fatty acids The raw material alcohol to be reacted with the monoalkyl ester is the same as the alkoxy group of the target fatty acid ester, but is naturally different from the alkoxy group of the raw material fat and fatty acid ester. As the raw alcohol, preferably a monohydric or dihydric aliphatic alcohol having 1 to 20 carbon atoms is used. If the number of carbon atoms is 21 or more, the alcoholysis reaction will be difficult to proceed, which is not preferable. these are,
It may be linear or branched, and may be a primary alcohol or a secondary alcohol. Specific examples of raw alcohol include methyl alcohol, 2-ethyl alcohol, amyl alcohol, 1-hexanol, l-decanol, lauryl alcohol, cetyl alcohol, oleyl alcohol, l-
Eicosanol, inclovir alcohol, 2-hexanol, 2-/nanol, 2-pentadecanol, 2-
Eicosanol, l-methyl-1-undecanol, 1
-Methyl-1-tetradecanol, 2-hexyl-1-
hexanol, 2-octyl-1-octatool, 2-
7J syl-1-decanol, cholesterin, ergosterin, β-sitosterin, glycerin, ethylene glycol, propylene glycol, pentaerythritol,
Examples include glycerin monolaurate and globylene glycol mono'palmitate. The raw alcohol is
At least 1 part by weight is used per 1 layer of raw material oil or fat or fatty acid monoalkyl ester.

Preferably, the reaction is carried out at a temperature of 1O to 90°C. At temperatures as low as 10°C, the system becomes non-uniform and quickly.
If the temperature exceeds 90°C, lipase is likely to be inactivated.

According to the method of this invention, desired fatty acid esters can be obtained with high purity and high yield. Therefore, the method of this invention can be widely applied to the production of soaps, cosmetics, fragrances, gelastec additives, etc.

Practical row 1 After mixing 1 part by weight of lipase extracted from various microorganisms shown in Table 1 by a conventional method, 2.5 parts by weight of Celite, and 4 parts of distilled water, the mixture was heated to 40°C or less under reduced pressure. The distilled water was evaporated to immobilize the lipase. This immobilized IJ/4'
-ze30 In9 (500 U) was mixed with 151n9 of trimyristin and 0.3 IIL of methanol or ethanol, and reacted at 45°C for 3 days. After the reaction was completed, 1 ml of a hexane solution of methyl behenate, an internal standard substance, was added, and the supernatant was analyzed by gas chromatography.

Column: 5% 5p2300, 60-8o mesh, 2
meters, 135℃~230℃/10℃/min
Met. The production rate of fatty acid ester was calculated with trimyristin hydrolyzed with sodium hydroxide as 100. The results are shown in Table 1.

*, ** Origin of lipase used by Ishihara et al. Example 2 In the same manner as in Example 1, glue derived from Alcaligenes faecalis I? -ze was immobilized on various carriers shown in Table 2.

Mix 30 m9 (500 U) of this immobilized lipase, 15 mf/palm oil, and 0.3 ml methanol-k,
The reaction was carried out at 0°C for 24 hours. The product was analyzed as in Example 1. The results are shown in Table 2.

Table 2 Example 3 In the same manner as in Example 1, lipase derived from Alcaligenes faecalis was immobilized on various carriers shown in Table 3. Mix 3 ml of this immobilized A-ze (500 U) with 15 ml of palm oil and 0.3 ml of methanol, and heat at 50°C for 24 hours.
Allowed time to react. The product was analyzed as in Example 1.

The results are shown in Table 3.

Table 3 Example 4 In the same manner as in Example 1, lipase derived from Alcaligenes faecalis was immobilized on each glue carrier shown in Table 4.

Mix 30 m9 (500 U) of this immobilized Reno-ya-ze, 15 IRg of palm kernel oil, and 0.3 ml of methanol,
The reaction was carried out at ℃ for 24 hours.

The product was analyzed as in Example 1. The results are shown in Table 4.

Table 4 Practical Example 5 In the same manner as in Practical Example 1, Alcaligenes faecalis-derived glue i4-ase was immobilized on various carriers shown in Table 5. 30 mg (500 U) of this immobilized lipase, 15 Iv of palm oil, and 0.3 a of ethanol were mixed and reacted at 50° C. for 24 hours. The product was analyzed as in Example 1. The results were summarized in Example 5. In the same manner as in Example 111, Alithropacter ureafaciens-derived glue/motherase was immobilized on various carriers shown in Table 6. This immobilized mother of 30 mgp (
sou) and 9-mu oil 15In9 and ethanol 0.3
tnl and subjected to fuj reaction at 5o cr: 24 hours. The product was analyzed as in Run 1+IJ 1.

The results are shown in Table 6.

Table 6 Example 7 Lipase derived from Sheetmonas fragi was immobilized on Celite in the same manner as in M9U1.

This immobilized lipase 4rn9 (500U) and palm oil 1
Mix 5~ and 0.3 d of ethanol and heat at 45℃ for 24 hours.
Allowed time to react. The product was analyzed as in Example 1.

The results are shown in Table 7.

Table 7 Ethyl palmitate 69.3% Ethyl oleate 44.9% Example 8 In the same manner as in Example UL, lipase derived from Alcaligenes faecalis was immobilized on various carriers shown in Table 8. 30η (500 U) of this immobilized lipase, 15 liters of barm cedar oil, and 0.3 ml of ethanol were mixed and reacted at 50° C. for 24 hours.

The product was analyzed in the same manner as in the run. The results are shown in Table 8.

Table 8, Example Column 9 In the same manner as Example 1, glue and yase derived from Alcaligenes faecalis was immobilized on various carriers shown in Table 9. This immobilized lipase 30■ (500U) and beef tallow 15
Mix m9 and Ku1/-L0.3 rtLl, 5
The reaction was carried out at 0°C for 24 hours. The product was run and analyzed as in 111. The results are shown in Table 9. In the same manner as in Experiment 1, lipases derived from Alcaligenes faecalis, Mucor pruscis, and porcine pancreas 11 were immobilized on Seven Light. This immobilized lipase 30 I
Q (500U), Noyam oil 30m9 and ethanol 0
．． 3 ml and 0.3 ml of n-hexane or petroleum pendine were mixed and reacted at 50°C for 24 hours. The product was analyzed as in Example 1.

The results are shown in Table 1O.

In the same manner as in Table 10, Example 11, FlJl, lipase derived from Alcaligenes faecalis was immobilized on Celite. 30 μ (500 U) of this immobilized lipase, 15 μ of coconut oil, and 0.3 d of ethanol were mixed and reacted at 50° C. for 24 hours. The product was analyzed as in Example 1. Results first
It is shown in Table 1.

Table 11 Ethyl laurate 91.6 Ethyl myristate 94.0 Ethyl palmitate 100 Ether stearate 86.3 Example row 12 In the same manner as IMIIThlJ1, Alcaligenes
Lipase derived from Faecalis was immobilized on Celite.

Mix 30 mg (500 U) of this immobilized lipase with 15 η of olive oil and 0.3 ml of ethanol,
The reaction was carried out at ℃ for 24 hours. The product was analyzed as in Example 1. The results are shown in Table 12.

Table 12 Ether lumitate 99.1 Ethyl oleate 100 Example 13 2.5 m9 of trilaurin and 4.7 g of cetyl alcohol were dissolved in 0.3 Inl of petroleum benzine. To this, Alithropacter ureafaciens 1Llj: Alcaligenes
Add 30 da of lipase derived from Faecalis or 12 mg of lipase derived from Sheetomonas fragi, and
Allowed time to react. The production rate of cetyl ester was determined by gas chromatography analysis using an internal standard method using behenic acid methyl ester as a standard substance. The results are shown in Table 13.

413 table fruiting I1 14 Mix 15 mg of trilaurin, ethanol Q, 3 ml and lipase 3 In9 derived from Alcaligenes faecalis or Alyslohactor ureafaciens or Pseudomonas flagi,
The reaction was carried out at 45°C for 24 hours. The produced ethyl laurate was analyzed in the same manner as in Example 1. The results are shown in Table 14.

Table 14 Solid line 1+l15 Lipase 12 from Pseudomonas fragi immobilized on Celite in the same manner as in Example 1, casillin modified methyl 3 da, methyl palmitate 3 or stearin methyl 3 and ethanol 0.3 rnl Mix 45
The reaction was carried out at ℃ for 24 hours. Execute the product 1+! Analyzed in the same manner as I1. The yield of the produced ethyl ester is
It is shown in Table 5.

Claims (1)

[Scope of Claims] 0) A method for producing a fatty acid ester by causing an alcoholysis reaction between an oil or fat or a fatty acid monoalkyl ester and an alcohol, in which alkaline earth metals, Alithropacter or Pseudomonas bacteria produce A? - A method for producing fatty acid esters, characterized in that the reaction is carried out without a solvent or in a non-aqueous solvent using enzyme as a catalyst. A method for producing a fatty acid ester by using an alkaline earth metal, a culture of Alithropacter or Pseudomonas bacteria as a catalyst, and carrying out the reaction without a solvent or in a non-aqueous solvent. Method for producing esters.