JPH0712313B2 - How to decompose fats and oils - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a method for decomposing fats and oils, and more specifically, in the presence of a specific amount of water, a microbial high molecular weight alkaline lipase is added to a reaction system containing fats and oils and alcohol. The present invention relates to a method of causing a fatty acid alcohol ester and glycerin to decompose in a short time with a high decomposition rate.

[Conventional technology]

There are various chemical decomposition methods for fats and oils, but the ones that have been put into practical use are the saponification decomposition method with an alkali and the corrugate that decomposes under high temperature and high pressure with water. The main ones are the emery method and the methanolysis method which decomposes by heating in the presence of anhydrous methanol and a catalyst.

In recent years, a method for hydrolyzing fats and oils using lipase has been studied from the viewpoint of energy saving. Regarding the role of water in the hydrolysis reaction of fats and oils by the enzymatic method, it acts as a component directly involved in the reaction, as a solvent for enzymes, or as a dispersion solvent for fats and oils. However, since water can disperse oils and fats but cannot dissolve it, the rate of hydrolysis is mainly determined by the interfacial area during the reaction. Inability to exert sufficient ability, the reaction rate decreases significantly. Therefore, in order to obtain a high hydrolysis rate, there is a problem that it takes a long decomposition time or that a large amount of enzyme which is not originally necessary is required.
The economic effect is not always satisfactory.

Kobayashi et al. Are studying decomposition in a two-phase system of water and a non-polar organic solvent such as isooctane for the purpose of facilitating the dispersion of fats and oils as a method of compensating for such defects (Fermentation Engineering., 63, 439).
(1985)).

As a result, the decomposition rate was considerably improved compared to the system without organic solvent addition, but since it is a two-phase system consisting of an organic solvent phase that dissolves fats and oils and an aqueous phase that dissolves enzymes, The interface area must be maintained, and there are problems in practical use such as required reaction time and effective use of enzymes. As an example of using alcohol in the reaction system, there is known a method characterized by carrying out an alcoholysis reaction using alcohol in the absence of water (JP-A-60-78587). Japanese Patent Laid-Open No. 60
In the official publication of the application of −78587, in the conventional alcoholysis reaction, the alcoholysis reaction of alcohol and fats and oils is carried out in the presence of water, but the presence of water in the reaction system lowers the fatty acid ester production rate and the purity. Ishihara et al. (Chem.Pharm.Bull, 23,3266 (1975)) cites that such problems cannot be prevented, and states that such problems can be prevented only by carrying out the alcoholysis reaction in the absence of water. .

[Problems to be solved by the invention]

However, according to the research conducted by the inventors of the present application, the role of water in the alcoholysil reaction is important, and if the amount of water is too large, the fatty acid ester production rate may decrease, or the reaction efficiency of lipase may decrease due to a two-phase emulsion. However, on the contrary, it was found that even in the absence of the lipase, the expression of lipase activity was inhibited and the alcoholysis reaction was reduced, or the reaction was stopped, and satisfactory results were not always obtained.

An object of the present invention is to solve the above-mentioned drawbacks and to provide a method for decomposing fats and oils into fatty acid esters in a short time with a high decomposition rate.

[Means for solving problems]

Therefore, the present inventors have variously studied the amount of water necessary for the efficient alcoholysis reaction of fats and oils, and have a fat and oil in the presence of 0.02% to 3.0% of water and a substituent. C ₁ -C ₂₂ saturated or unsaturated primary or secondary alcohols and organic solvents other than alcohols (excluding tertiary alcohols) in the presence or absence of organic solvents, microbial high molecular weight alkaline The present invention has been completed by discovering that the above-mentioned drawbacks can be solved by causing lipase to act to decompose fats and oils. That is, the present invention is a range in which a homogeneous system can be maintained, and in addition, a microbial high molecular weight alkaline lipase acts on fats and oils and alcohol in the presence of water necessary for the alcoholysis reaction to proceed most efficiently, and the fats and oils can be efficiently treated in a short time. It is a method that often decomposes into fatty acid ester and glycerin.

Hereinafter, the present invention will be described in detail.

The method of the present invention comprises a fat and an optionally substituted C ₁
Saturation -C _22, or unsaturated primary or secondary alcohol or optionally in an alcohol (however, tertiary excluding alcohol) these to act a microorganism high molecular weight alkaline lipase in the presence of a non-organic solvent 0.02% or more when decomposing fats and oils into fatty acid esters and glycerin
It is carried out in the presence of water of 3.0% or less. With this amount of water, the reaction system can be maintained in a uniform range, and if some of the raw materials and products become insoluble substances in the reaction system, select an appropriate organic solvent for the reaction system. Can be used to solve the problem. Water in the alcoholysis reaction seems to act as a catalyst for the smooth progress of the reaction, and its role is important, and when the amount of water is 0.02% or less, the fatty acids of oils and alcohols Exchange is significantly hindered. This point will be described with reference to experimental examples.

Experimental Example 1 Olive oil (3 g) and 1-butanol (3 g) were placed in a test tube with a stopper, and lipase which is a microbial high molecular weight alkaline lipase
PL-679 (Meito Sangyo, 70000u / g), Lipase AL (Meito Sangyo, 15000u / g), Microbial High Molecular Weight Alkaline Lipase OF (Meito Sangyo, 360,000u / g), Lipase M
-AP20 (Amano Pharmaceutical, 20000u / g), Lipase AP6 (Amano Pharmaceutical, 6000u / g), Talipase (Tanabe Pharmaceutical, 6000u / g), Pancreatin (Wako Pure Chemicals, 2000u / g) lipase powder 0.1
Add g and dehydrate with Molecular Sieves 3A so that the water content will be 0 to 50% (% based on the total reaction system, measured by Karl Fischer method) or add water and shake react at 37 ° C for 24 hours. The fatty acid 1-butanol ester production rate was measured as described in Example 1. The results are shown in Table 1.

As can be seen from the experimental results of Experimental Example 1 above, in the present invention, the amount of water present in the reaction system is 0.02% to 3.0% as free water in the reaction system, preferably 0.02% or less.
% To 1.0%, particularly preferably 0.02% to 0.6%
The following is preferable. When the amount of water exceeds 3%, the homogeneous phase is broken or the alcoholysis reaction is deteriorated, which is not preferable. On the contrary, even if the amount of water is 0.02% or less, the reaction is deteriorated, which is not preferable.

The lipase used in the present invention is a microbial high molecular weight alkaline lipase, preferably having a molecular weight of 100,000 or more, and an optimum pH of 8.0.
The above-mentioned extracellular lipase, as long as it has any origin, any one can be used, but as such a lipase,
For example, a lipase (Japanese Patent Publication No. Sho 49-32080) (hereinafter referred to as lipase AL) produced by a name sugar AL-865 bacterium belonging to the genus Achromobacter (Ministry of Industrial Science and Technology No. 1213) (hereinafter referred to as Lipase AL), Alcaligenes ( Lipase (Japanese Patent Publication No. Sho 58-36953) (hereinafter referred to as Lipase PL-266) produced by the name sugar PL-266 (Ministry of Industrial Science and Technology No. 3187) belonging to the genus Alcaligenes, also a name belonging to the genus Alcaligenes Sugar PL-679
As specific examples, lipases such as lipase (Japanese Unexamined Patent Publication No. 60-15312) (hereinafter referred to as lipase PL-679) produced by No. 3 (Shin Koken Bacterial Contribution No. 3783) can be mentioned. In addition, Pseudomonas nitroreducens var.thormo
-Tolerans produced lipase (Japanese Patent Publication No. 56-28516)
Issue) could be used as well. These lipases are high-molecular-weight proteins that are thought to have a large amount of intramolecularly bound water, and are characterized by having an optimum pH on the alkaline side. It is presumed that the properties are particularly excellent for efficiently carrying out the alcoholysis reaction in a short time. This is also considered to be related to high stability against organic solvents, and this point will be described below with reference to experimental examples.

Experimental Example 2 Lipase PL- which is a microbial high molecular weight alkaline lipase
679 (Meito Industry, 70000u / g), Lipase AL (Meito Industry, 1
5000u / g), microbial high molecular weight alkaline lipase non-lipase OF (Meito Sangyo, 360,000u / g), Lipase M-AP2
0 (Amano, 20000u / g), Lipase AP6 (Amano, 60
00u / g), Talipase (Tanabe Seiyaku, 6000u / g), Pancreatin (Wako Pure Chemicals, 2000u / g), 25mg each of lipase powder, was placed in a centrifuge tube with a 7ml stopper. Hexane, petroleum ether, acetone, tertiary butanol, and water (2 ml each) were added, thoroughly stirred, and shaken at 37 ° C. for 24 hours, and the residual activity was measured by the Reverse activity measuring method. The lipase activity was determined by the method of Kokusei et al. (Agric.Biol.Chem., 4) for lipase PL-679 and pancreatin.
6, 1159,1982) Kokusho's method for lipase AL (oil chemistry, 23, 98,1974) Yamada et al method for lipase OF (day Noka Journal, 36, 860,1962), and other lipase Fukumoto's method (J.Gen.Appl.Microbiol., 9 , 353,196
I went in 3). The results are shown in Table 2.

The lipase used in the reaction may be a purified product or a crude product, and is used as a powder or granular dry enzyme. Further, it may be a dry enzyme immobilized on an ion exchange resin such as DEAE, celite, bentonite, chitosan or the like. The amount of lipase used is not limited, but for example, about 1,000 to 100,0 per 1 g of oil and fat.
The amount of enzyme may be about 00 units, preferably about 2,000 to 50,000 units.

When the enzyme is immobilized on a carrier and used, the higher the specific activity, the higher the reaction efficiency, which is preferable.
Immobilized enzymes of about 0 to 300,000 units / g can be mentioned.

As a reaction solvent, it is most ideal that the alcohol used in the reaction can be used as a reaction solvent at the same time, but if the substrate or product is insoluble in alcohol and a homogeneous phase cannot be obtained, the alcohol (however, An organic solvent other than the tertiary alcohol is used as a reaction solvent. At this time, any organic solvent may be used as long as an organic solvent which does not inhibit the reaction is selected and used. For example, fatty acid hydrocarbons such as n-heptane, n-pentane, n-hexane, petroleum ether, isooctane, etc. Alicyclic hydrocarbons such as cyclopentane, cyclohexane, cyclobutane, etc .;
Aromatic hydrocarbons such as benzene, toluene, xylene, phenol; ketones such as acetone and methyl isobutyl ketone; nitrogen-containing solvents such as acetonitrile; dimethyl ether, diethyl ether, diisopropyl ether, dioxane, etc. Examples include ethers such as; carbon tetrachloride, chloroform, methylene chloride, and other halogenated hydrocarbons; tertiary butyl alcohol, tertiary amyl alcohol, diacetone alcohol, and other tertiary alcohols. Can be done. You may use a solvent individually or in mixture of 2 or more types. The amount to be used depends on the solvent used, the type and concentration of the substrate, but it is desirable to add it in such a degree that the reaction proceeds well and the fluidized homogeneous phase is maintained.
There is no limit to the amount added, but for example, 10 to 90% (v /
The range of v) can be shown. Depending on the substrate, the reaction may be promoted as compared with the case where no solvent is added due to the addition of the solvent as described above.

The raw material fat used in the reaction in the present invention is a natural animal fat,
Vegetable fats and oils, processed fats and oils are used, and examples of natural resins include beef tallow, lard, milk fat, fish oil, coconut oil, palm oil,
Palm kernel oil, olive oil, soybean oil, cottonseed oil, rapeseed oil, corn oil, safflower oil, copra oil, coconut oil, rice oil, sesame oil, linseed oil, castor oil, sunflower oil, peanut oil, etc. , For example, hardened beef tallow, hardened lard,
There is shortening, etc., and waste oil, highly oxidized fats and oils can also be used.

In addition, in the present invention, as the alcohol to be reacted with the fat or oil, primary or secondary alcohols having carbon atoms of C _{1 to} C ₂₂ which may have a substituent are used.

Specific examples of the primary alcohol include methanol, ethanol, 1-propanol, allyl alcohol, propargyl alcohol, 1-butanol, isobutanol, 1
-Pentanol, 2-methyl-1-butanol, isopentyl alcohol, neopentyl alcohol, 1-hexanol, 2-methyl-1-pentanol, 2-ethyl-1-butanol, 1-heptanol, 1-octanol, 2 -Ethyl-1-hexanol, 1-nonanol,
3,5,5-Trimethyl-1-hexanol, pantothenyl alcohol, 1-decanol, geraniol, 1-undecanol, 1-dodecanol, farnesol, phytol, hexadecanol, oleyl alcohol, 1
-Octadecanol, 1-eicosanol, 1-docosanol, and the like, and examples of those having a phenyl group as a substituent include benzyl alcohol, β-phenethyl alcohol, cinnamic alcohol, chlorobenzyl alcohol, p. -Aminophenethyl alcohol, hydroxyethylaniline, 2-naphthalene ethanol having a naphthyl group as a substituent, furfuryl alcohol, tetrahydrofurfuryl alcohol having a furfuryl group as a substituent, and a hydroxy group as a substituent Examples of those having ethylene glycol, 1,4-butanediol, 1,6-hexanediol, 1,10-decanediol,
1,16-hexadecanediol, pentaerythritol,
Polyethylene glycol, those having an amino group as a substituent include 2-aminoethanol, 2- (diethylamino) ethanol, 5-amino-1-pentanol, 6-amino-1-hexanol, those having a halogen as a substituent For example, 3-chloro-1,2
-Propanediol, 6-chloro-1-hexanol,
Examples of compounds having a thienyl group as a substituent include 2
-Thienylethanol, those having a pyridyl group as a substituent, for example, 2-viridine ethanol, 2-pyridinepropanol, pyridoxine, those having a piperazyl group as a substituent, for example, 1-piperazine ethanol, having a pyran group as a substituent Examples thereof include piperonyl alcohol, examples of those having a phthalimido group as a substituent are, for example, phthalimidoethanol, examples of those having a morpholyl group as a substituent are, for example, 1-morpholine ethanol, and examples of those having a pyrroldil group as a substituent are, for example. Examples of 1-pyrrolidine ethanol, 1-pyrrolidone ethanol, and those having an imidazole group as a substituent include thiamine.

Next, specific examples of the secondary alcohol include, for example, 2-propanol, 2-butanol, 2-pentanol and 2-
Hexanol, 3-hexanol, 2-heptanol,
2-octanol, 2-nonanol, 2-decanol,
2-undecanol, 2-tridecanol, 2-tetradecanol, 4-methyl-2-pentanol and the like can be mentioned. Further, as those having a cyclo ring as a substituent, for example, cyclobutanol, cyclohexanol, cyclooctanol, as those having a sterol group as a substituent, for example, cholesterol, ergosterol, as those having a phenyl group as a substituent, For example, 1-phenylethanol and the like can be mentioned, but the kind of alcohol is not limited.

A mode for contacting a fat and oil and an alcohol with a lipase in the presence of water in the above range to decompose the fat and oil into a fatty acid ester and glycerin can be appropriately selected. When a batch-type reaction tank is used, it may be carried out by a method of adding powdery or granular enzyme or immobilized enzyme to the reaction system and stirring. With the amount of water used in the present invention, the enzyme does not dissolve and floats as a solid, so it can be easily recovered from the reaction system and used repeatedly. When a continuous reaction is carried out using a packed tank type reaction tank, it can be carried out by a method of filling the above-mentioned enzyme into the tank and passing or circulating the reaction solution.

The mixing ratio of the oil / fat and alcohol as a substrate in the reaction and the gas concentration can be appropriately selected, but the number of moles of alcohol added to 1 mol of the oil / fat is, for example, 3 to 3,000 mol
It may be carried out preferably at about 3 to 300 mol, and the substrate concentration at that time may be about 10 to 100%. In addition, it is possible to mix and react several kinds of fats and oils and alcohols to be used.

The reaction temperature does not particularly need to be heated because it proceeds even at about room temperature, but it is preferable to carry out the reaction at an appropriate temperature in consideration of the melting point and boiling point of the substrate or solvent used, the action temperature of the enzyme, etc. C., preferably 20 to 60.degree. C.

Also, the reaction time can be appropriately selected, for example, from 3 to
A reaction time such as 48 hours can be indicated. The fatty acid ester produced by the alcoholysis reaction can be separated and purified by means such as distillation and solvent fractionation, if necessary.

Fatty acid esters of various alcohols obtained in the present invention,
Emulsifiers, defoamers, solubilizers in the fields of food industry, enzyme industry, textile industry, pharmaceuticals, paper pulp industry, synthetic rubber, plastics industry, coating industry, cosmetics industry, petrochemical industry, agrochemical industry, machinery industry, etc. It can be used as a lubricant, a separating agent, or the like. In addition, esters composed of higher fatty acids and higher alcohols are also used as waxes.

〔Example〕

Examples of the present invention will be shown below, but the present invention is not limited thereto.

Example 1 3 g of palm oil and 3 g of 1-hexanol were placed in a test tube with a stopper, and the water content in the reaction system was measured by the Karl Fischer method.
After adding water to 0.2%, add 200 mg of lipase PL-679 powder (70000u / g) and react at 37 ℃ for 1,3,6,16,24,48,72 hours to produce fatty acid 1-hexanol ester. The rate was measured. The results are shown in Table 3.

The fatty acid alcohol ester production rate was determined by iatroscan. That is, 0.05 ml of the above reaction solution was dissolved in 3 ml of chloroform, filtered to remove insoluble matter, and 1 μl was spotted on Chromarod S2 (Yatron) to n-hexane-diethyl ether acetic acid (90: 10: 1 v / v). ) Is developed as a developing solvent for about 10 cm and subjected to Iatroscan (Iatroscan TH10 manufactured by Yatron Co., Ltd.) to obtain the weight ratio of the components from the peak area ratio, and the fatty acid alcohol ester production rate is glyceride, fatty acid, fatty acid alcohol ester to fatty acid alcohol ester Expressed as a weight ratio.

From the results in Table 3, it can be seen that the fatty acid 1-hexanol ester formation reaction is completed in a short time of 6 hours.

Example 2 After adding 3 g of oils and fats shown in Table 4 and 3 g of alcohols shown in Table 4 to a test tube with a stopper, the water content of the reaction system was measured by the Karl Fischer method, and water was added so that the water content would be 0.2%. , 200 mg of Lipase PL-679 powder was added, and the mixture was shaken at 37 ° C. for 6 hours to determine the fatty acid alcohol ester production rate. The results are shown in Table 4.

The results in Table 4 show that Lipase PL-679 decomposes oil and fat very well to form fatty acid alcohol ester.

Example 3 3 g of olive oil and 3 g of alcohol shown in Table 5 were placed in a test tube with a stopper, and the water content of the reaction system was measured by the Karl Fischer method. Water was added so that the water content was 0.2%, and then lipase AL was used.
200 mg of powder (15000 u / g) was added and shake reaction was carried out at 37 ° C. for 6 hours to determine the fatty acid alcohol ester production rate. The results are shown in Table 5.

From the results in Table 5, it can be seen that lipase AL decomposes oil and fat very well to form fatty acid alcohol ester.

Example 4 3 g of coconut oil, 3 g of 1-docosanol and the organic solvent shown in Table 6
10 ml was placed in an Erlenmeyer flask with a ground stopper and the water content of the reaction system was measured by the Karl Fischer method. Water was added so that the water content would be 0.2%, 200 mg of Lipase PL-679 powder was added, and the mixture was mixed at 37 ° C for 6 hours.
The reaction with time shaking was performed to obtain the fatty acid 1-docosanol ester production rate. The results are shown in Table 6.

Next, the production amount of fatty acid 1-docosanol ester was determined in the same manner as above using 3 g of palm oil instead of palm oil.
The results are shown in Table 6.

From the results shown in Table 6, it can be seen that the fatty acid 1-docosanol ester production rate is increased by adding an organic solvent such as n-hexane or diethyl ether.

Example 5 5 g of lipase PL-679 powder (70000u / g) was dissolved in 100 ml of water, 15 g of DEOH Toyopearl (adjusted to pH 9.0) activated to OH type was added thereto, and the mixture was stirred at 4 ° C for 3 hours. Lipase PL-679 was immobilized on Toyopearl. DEAE Toyopearl was recovered by centrifugation and freeze-dried to obtain 15 g (20000u / g) of DEAE Toyopearl-immobilized lipase PL-679.

Take 3 g of olive oil and 3 g of oleyl alcohol in a test tube with a stopper, and measure the water content in the reaction system by Karl Fischer's method.Add water so that the water content will be 0.2%, then add 500 mg of DEAE Toyopearl immobilized lipase PL-679. Shaking reaction was carried out at 37 ° C. for 6 hours to determine the fatty acid oleyl alcohol ester production rate. After reaction, centrifuge and DEAE Toyopearl immobilized lipase
Table 7 shows the results obtained by collecting PL-679, adding it to the same new reaction system as above, and repeating the reaction.

From the results in Table 7, DEAE Toyopearl immobilized lipase PL-67
It can be seen that 9 can be used repeatedly to produce fatty acid alcohol esters.

〔The invention's effect〕

According to the present invention, in the Alcolysyl reaction using microbial high molecular weight alkaline lipase, the reaction system contains 0.02% to 3.0%.
By the presence of water, various alcohol esters can be obtained from fats and oils with a high reaction rate in an extremely short time. Further, since the enzyme is not dissolved even in the presence of water in this range, the enzyme can be continuously and repeatedly used for a long time, and the utilization efficiency of the enzyme is enhanced.

Further, in the present invention, since a reaction occurs under mild conditions without requiring a toxic reaction catalyst and high energy as used in a chemical method, various alcohol esters of fatty acids produced are excellent in quality. A highly safe ester can be obtained.

Claims (3)

[Claims] 1. A fat or oil in the presence of 0.02% or more and 3.0% or less of water, and a saturated C _{1 to} C ₂₂ carbon atom which may have a substituent,
Alternatively, unsaturated primary or secondary alcohols are allowed to act on a microbial high molecular weight alkaline lipase in the presence or absence of an organic solvent other than alcohols (excluding tertiary alcohols) to give oils and fats fatty acid esters and glycerin. A method for decomposing fats and oils, which comprises decomposing into oil. 2. A microbial high molecular weight alkaline lipase having a molecular weight of 100,000 or more and an optimum pH on the alkaline side of 8.0.
Method described in section. 3. The method according to claim 1 or 2, wherein the microbial high molecular weight alkaline lipase is an extracellular lipase produced by a bacterium of the genus Alcaligenes or the genus Achromobacter.