WO2003040275A1

WO2003040275A1 - Fats and oils rich in linear isoprenoid fatty acid esters and process for production thereof

Info

Publication number: WO2003040275A1
Application number: PCT/JP2002/011607
Authority: WO
Inventors: Gou Shinohara; Chiemi Satou; Seiichi Shirasawa
Original assignee: The Nisshin Oillio,Ltd.
Priority date: 2001-11-07
Filing date: 2002-11-07
Publication date: 2003-05-15
Also published as: JPWO2003040275A1

Abstract

Fats and oils rich in linear isoprenoid fatty acid esters, characterized by being produced by using as the raw materials (A) one or more members selected from the group consisting of linear isoprenoid alcohols and(B) one or more members selected from the group consisting of fatty acids, fatty acid methyl esters, fatty acid ethyl esters, monoglycerides, diglycerides and triglycerides or an oil or fat containing one or more of these substances and subjecting a mixture of the components (A) and (B) to treatment with a carboxylic ester hydrolase.

Description

BACKGROUND OF THE INVENTION

The present invention relates to fats and oils containing a large amount of chain isoprenoid fatty acid esters and a process for producing the same, and also relates to chain isoprenoid fatty acid esters obtained by isolating from the fats and oils containing a large amount of chain isoprenoid fatty acid esters. And its manufacturing method.

Isoprenoids are a general term for a group of natural organic compounds whose constituent units are isoprene (C5H8). Monoterpenes (C10), sesquiterpenes (C15), diterpenes (C20), Evening terpenes such as terpenes (C25) and triterpenes (C30), various steroids derived from squalene of C30, carotenoids of C40 各種 various polyprenols ranging from C30 to C120, and the molecular weight It is said that there are more than about 20,000 types of isoprenoids that exist in nature, up to 100,000 natural rubbers. In addition, chlorophyll, which has an isoprenoid in part of its structure, and bisubimine K1, K2, and complex isoprenoids such as ubiquinone and menaquinone are also naturally present in a wide variety of forms, such as in photosynthesis and electron transport. It is responsible for important biological functions in the body. Alternatively, pendecabrenyl phosphate (C55) as a sugar carrier lipid in the biosynthesis of bacterial cell walls and dolichol phosphate (C80-C120) in eukaryotes are N-glycoside-type sugar chains and sugar proteins. It has also been found to be important as a sugar carrier lipid for quality biosynthesis. Furthermore, the so-called prenylated protein modified with a phenylesyl group (C 15) or geranylgeranyl group (C 20) plays an essential role in the cancer gene product Ras and G proteins important for signal transduction. The physiological functions of isoprenoids have been elucidated.

On the other hand, it is well known that sterols and higher alcohols are present in fats and oils in trace amounts mainly in the form of fatty acid esters. In particular, in recent years, it has been found that these components are involved in the quality, physical properties and nutritional physiological functions of fats and oils, and fats and oils containing these have been attracting attention as functional fats and oils. It has been confirmed that chain isoprenoid fatty acid esters are present in trace amounts in fats and oils.For example, in crude vegetable oil, it is known to be present in tens to hundreds of ppm depending on the type, but refined. Since the processed edible oil contains only a few ppm at most (Oil Chemistry, Vol. 4, No. 3, 192-196, 1995), the fatty acid esters of chained isoprenoid fatty acids have so far been actively contained in fats and oils. No attempt has been made to do so. Disclosure of the invention

An object of the present invention is to provide an oil or fat containing a high content of chain isoprenoid fatty acid esters and a method for producing the same.

The present inventors have conducted intensive studies to achieve the above object, and as a result, using a carboxylic acid ester hydrolytic enzyme, a chain isoprenoid alcohol, a fatty acid, a fatty acid methyl ester, a fatty acid ethyl ester, From one or more of the group consisting of monoglycerides, diglycerides, and triglycerides and fats and oils containing these, it is possible to easily obtain fats and oils high in the chain isoprenoid fatty acid ester represented by the following general formula (II). They have found that they can do this and have completed the present invention.

(Π)

That is, the present invention is characterized by being obtained by treating the following components (A) and (B) as raw materials with a carboxylic acid ester hydrolase, and selecting from linear isoprenoid fatty acid esters. Fats or oils containing more than one kind of fats and oils more than ordinary fats and oils, preferably obtained by treating with carboxylesterase and / or triacyl "lycerol lipase as carboxylic ester hydrolase. The present invention relates to a method for producing fats and oils containing one or more kinds selected from chain isoprenoid fatty acid esters in comparison with ordinary fats and oils.

(A) one or more selected from the group consisting of linear isoprenoid alcohols represented by the following general formula (I);

(In the formula, the wavy line means a single bond or a double bond, and n means any one integer selected from 1 to 14.)

(B) One or more of the group consisting of fatty acids, fatty acid methyl esters, fatty acid ethyl esters, monoglycerides, diglycerides, and triglycerides, and fats and oils containing these.

At this time, the oils and fats to be used are preferably vegetable oils which contain a small amount of chain isoprenoid fatty acid esters, and if edible, can be used as they are as edible oils.

Examples of the linear isoprenoid alcohols include geraniol, fuarnesol, geranylgeranol, geranylfurnesol, fuarnesyl fuarnesol, geranylgeranyl fuarnesol geranyl fuarnesyl fuarnesol, and fuarnesol. It is preferable to use one or two or more selected from the group consisting of nesyl phenyl nesyl phenyls, phthals and dihydro phthals. As the fatty acid used, a fatty acid having 2 to 30 carbon atoms is preferable. Similarly, as the fatty acid methyl ester, fatty acid ethyl ester, monoglyceride, diglyceride, or triglyceride used, the fatty acid residue has 2 carbon atoms. It is preferably ~ 30.

The fats and oils containing high content of chain isoprenoid fatty acid esters obtained in this way may be subjected to concentration and Z or purification treatment in order to further increase the concentration thereof, whereby the chain isoprenoid fatty acid ester having a higher concentration is obtained. Fats and oils with a high content can be suitably obtained. Further, by further isolation, chain-like isoprenoid fatty acid esters can be obtained very easily and inexpensively.

In addition, the present invention relates to a method for producing a fatty acid / fat having a high content of linear isoprenoid fatty acid esters, which comprises treating the above components (A) and (B) as raw materials with a carboxylic ester hydrolase. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in detail.

The present invention relates to a chain isoprenoid fatty acid ester-rich fat or oil obtained by treating the following components (A) and (B) as raw materials with a carboxylic ester hydrolase.

The term "chain isoprenoid fatty acid ester" refers to a substance having a structure resulting from dehydration condensation from a chain of isoprenoid alcohol and a fatty acid. The chain isoprenoid alcohol generally refers to one having a chain structure in which a plurality of isoprene units having 5 carbon atoms are bonded and having a hydroxyl group. Further, the ester form refers to an ester form which can be formed from a hydroxyl group of a chain isoprenoid alcohol and a carboxyl group of a fatty acid.

The carboxylate hydrolase used in the present invention may be of any origin, such as animals, plants, and microorganisms, and may be extracted and purified from the tissue or culture solution by a conventional method, and may be prepared. It is convenient to use commercially available products. In addition, any of these enzymes which are immobilized, those which are not immobilized, and those which are not immobilized can be suitably used, but immobilized enzymes are preferred from the viewpoint of ease of use. As the carboxylic ester hydrolase used in the present invention, carboxyesterase and / or triacylglycerol lipase are preferable. Carboxylesterases are generally enzymes that hydrolyze fatty acid monoesters into alcohols and fatty acids. Triacylglycerol lipase is an enzyme that generally catalyzes the hydrolysis of fats (triglycerides) into fatty acids and glycerol. It is. The carboxylesterase and / or triacylglycerol lipase used in the present invention may be of any origin such as animals, plants, and microorganisms, and is not limited to the following. Carboxylesterase and / or triacylglycerol lipase derived from malt, castor seed, etc., Aspenole: Zoles Niger, A sperg 111 usiiiger), Candy Evening, Candidacy 1 indracea, Candy Da An Evening — Cutica (C and idaantarctica), Rhizopus delemar (R hizopusde 1 emar), Rhizopus jyanicas (R hizopusj avanicus) Pannare force Alcal igenes esp. (A 1 ca) 1 igenesf aecal is)ヽMucor Mihai (Mucor mi e he i) _s Mucor Jabani dregs (Mu cor jav an ic us) , Yudomonasu Furuoretsusensu (P s eudomonas f 1 uorescens), may be mentioned mono- Momaisesu Ranukinosa scan (T he rm o my ces lanuginosus) carboxy esterase from such and / or tri § sill glycerol lipase one peptidase and the like. These can be prepared by extraction and purification from the tissue or culture solution by a conventional method, but it is convenient to use commercially available products. Any of these enzymes may be suitably used, either immobilized or non-immobilized. However, immobilized enzymes are preferred for ease of use.

The chain isoprenoid alcohol used as a raw material of the present invention is not particularly limited as long as it is represented by the following general formula (I). For example, geraniol, fuarnesol, geranir geraniol, geranyl fuarnesol, fuarnesyl arnesol, geranyl geranyl arnesol, geranyl fuarnesol arnesol, fuar

-, Phytol, dihydrophytol, etc.

(In the formula, a wavy line means a single bond or a double bond, and n means any one integer selected from 1 to 14.)

The raw material of the present invention is not particularly limited as long as it is one or more of fatty acids, fatty acid methyl esters, fatty acid ethyl esters, monoglycerides, diglycerides, triglycerides, and fats and oils containing these. The fats and oils may be any of crude oils, unrefined fats and oils in the middle of refining, refined fats and oils, for example, soybean oil, rapeseed oil, cottonseed oil, castor oil, safflower oil, sesame oil, orip oil, Vegetable oils such as linseed oil, rice oil, palm oil, cocoa butter, and kapok oil; animal fats and oils such as lard, beef tallow, fish oil, etc., but there is no particular limitation. MCT, MLC Ts diglyceride, monoglyceride II, structural fats and oils with designed fatty acid structures, and the like. Of these, vegetable oils such as soybean oil, rapeseed oil, cottonseed oil, castor oil, safflower oil, sesame oil, olive oil, linseed oil, rice oil, palm oil, cocoa butter, and kapok oil are preferred, and most preferred. Contains a very small amount of linear isoprenoid alcohol, soybean oil, rapeseed oil, cottonseed oil, castor oil, safflower oil, sesame oil, olive oil, linseed oil, rice oil, Vegetable oils such as palm oil, cocoa butter and kapok oil are preferred. Furthermore, in order to reduce the number of steps after the enzyme treatment in the present invention, it is preferable to use edible vegetable oil that has already been purified.

The fatty acid used as a raw material of the present invention is not particularly limited as long as it is a fatty acid having a carbon number in the range of 2 to 30; for example, acetic acid, butyric acid, caproic acid, caprylic acid, Strength acid, pendecanoic acid, lauric acid, tridecanoic acid, myristic acid, pendecanoic acid, normitic acid, margaric acid, stearic acid, nonadecanoic acid, arakidic acid, behenic acid, lignoceric acid, serotinic acid, montanic acid, Straight-chain saturated fatty acids such as melicic acid, succinic acid, lindelic acid, pedic acid, olemic oleic acid, oleic acid, elaidic acid, paxenic acid, cis vaccenic acid, petroselinic acid, gadoleic acid, eicosenoic acid, L-force Monounsaturated fatty acids such as acid, cetrenic acid, nervonic acid, xymenic acid, and lamequenic acid, monolinolenic acid, stearidonic acid, eicate traenoic acid, eicosapenic acid, docosapenic acid, docosahexane N-3 unsaturated fatty acids such as acid, linoleic acid, linoleic acid, N-6 unsaturated fatty acids such as monolinolenic acid, bishomoarlinolenic acid, and arachidonic acid; conjugated fatty acids such as conjugated linoleic acid and hystereostearic acid; pyrenonic acid, siadonic acid, juperonic acid; Fatty acids having a double bond at the 5-position such as columbic acid, other unsaturated unsaturated fatty acids such as hiragonic acid, moloctic acid, clupanodonic acid, disinic acid, etc. Branched fatty acids such as butyric acid, isovaleric acid, isoacids, antiisoic acids, etc., hydroxy fatty acids such as polyhydroxy acids, / 5-hydroxy acids, mycolic acid, polyhydroxy acids, epoxy fatty acids, keto fatty acids, cyclic fatty acids, etc. In particular, linear fatty acids are preferable from the viewpoint of natural abundance and the like, and linear unsaturated fatty acids are more preferable. Among them, palmitooleic acid and o Mono-unsaturated fatty acids such as innoic acid, noxenoic acid, and erlic acid, n-6 unsaturated fatty acids such as linoleic acid, arlinolenic acid, bishomo-arenolenic acid, and arachidonic acid; hy-linolenic acid, stearidonic acid; N-3 unsaturated fatty acids such as eicosatetraenoic acid, eicosapenic acid, docosapenic acid, and docosahexanoic acid; conjugated linoleic acid; conjugated S-fatty acid such as hyi-eleostearic acid; preferable.

Examples of the fatty acid residue constituting the fatty acid methyl ester used as a raw material of the present invention include the fatty acid residues mentioned in the description of the fatty acid. Examples of the fatty acid residue constituting the fatty acid ethyl ester used as a raw material of the present invention include the fatty acid residues described in the description of the fatty acid.

The monoglyceride used as a raw material in the present invention may be any of 1-monoglyceride and 2-monoglyceride. At this time, examples of the moon fatty acid residue constituting the monoglyceride include the fatty acid residues mentioned in the description of the fatty acid.

The diglyceride used as a raw material of the present invention may be any of 1,2-diglyceride and 1,3-diglyceride. The fatty acid residues constituting the diglyceride at this time include the fatty acid residues mentioned in the description of the fatty acid, and the combination thereof is not particularly limited.

Examples of the fatty acid residue constituting the triglyceride used as a raw material of the present invention include the fatty acid residues mentioned in the description of the fatty acid, and the combination thereof is not particularly limited.

The chain isoprenoid fatty acid ester-rich fats and oils described so far can be mis- and / or purified, if necessary, to obtain a further high-content fat and oil. . Although it is difficult to specify the method of such concentration and / or purification in general, for example, fractional distillation, fractional sublimation, zone melting, solvent extraction, various adsorption methods, foam separation method, membrane separation method, molecular sieve And a method using chromatography.

The oil-and-fat having a high content of chain isoprenoid fatty acid esters of the present invention is characterized by being obtained by treating the above components (A) and (B) as raw materials with a carboxylic ester hydrolase. This is because, for example, the chain obtained by the chemical synthesis method from the component (A) and the component (B) can be obtained by the chemical synthesis method directly from the oil and fat containing high chain isoprenoid fatty acid esters. In the course of the reaction, the isoprenoid fatty acid ester-rich fats and oils are mixed with each other in component (A) or component (B) A non-selective reaction between the two forms a side reaction product, which is not preferable because it may be difficult to suitably obtain the desired linear isoprenide lunar fatty acid esters. In addition, undesirable side reaction products are contained as fats and oils, and a step of producing fats and oils again is required to remove these side reaction products. On the other hand, in the fats and oils containing high content of chain isoprenoid fatty acid esters of the present invention, since an enzyme is used to obtain chain isoprenoid fatty acid esters, component (A) and component (B) are selected in the reaction process. It is preferable because it can be easily treated in the same manner as ordinary fats and oils only by removing the enzyme, because it reacts in a natural manner and hardly generates unnecessary side reaction products. In addition, in the case of a chemical synthesis method, several tens to several hundreds are required to promote the reaction. A temperature of C is required, but if such high heat is applied, the oxidative deterioration of both components (AX (B)) is accelerated, and the fats and oils obtained by such a method are used as ordinary fats and oils. On the other hand, the oils and fats high in the chain isoprenoid fatty acid esters of the present invention can be obtained under very mild reaction conditions because enzymes are used to obtain them. In the course of the reaction, the components (A) and (B) do not undergo oxidative deterioration, etc., so that they can be used in the same manner as ordinary fats and oils. Naturally, since a chemical substance is used, fats and oils obtained through such a process cannot be used as edible oils in particular, but the fats and oils high in the chain isoprenoid fatty acid esters of the present invention are To get From using enzyme, excellent in safety, preferable particularly when Ru is used as edible oil.

The content of the chain isoprenoid fatty acid ester in the oil and fat having a high content of the chain isoprenoid fatty acid ester of the present invention is not particularly limited.However, for example, in consideration of practical use such as edible, medical and cosmetic use, and cosmetic use. When put, it is preferable that the fats and oils are refined, and in such a case, the content is, for example, 0.0001 to 50% by mass, preferably 0.0005 to 45% by mass. , More preferably 0.001 to 40% by mass, more preferably 0.005 to 35% by mass, still more preferably 0.01 to 30% by mass, further preferably 0.05 to 25% by mass, more preferably 0.1 to 20% by mass, and still more preferably Is from 0.5 to 15% by mass, particularly preferably from 1 to 10% by mass. When the chain isoprenoid fatty acid esters are concentrated and / or purified as required, they may be concentrated and / or purified as necessary, and are not particularly limited. For example, 0.01 to 90% by mass, Preferably 0.0

5 to 80% by mass, more preferably 0 :! To 70% by mass, more preferably 0.5

-60 mass%, particularly preferably 1-50 mass%.

Further, as described above, the present invention provides a method for producing a chain-type isoprenoid fatty acid ester-rich fat or oil, comprising treating the following components (A) and (B) as raw materials with a carboxylic ester hydrolase. About the method.

(I)

(In the formula, a wavy portion means a single bond or a double bond, and n means any one integer selected from 1 to 14.)

The raw material component (A) is as described above, and is not particularly limited as long as it is represented by the general formula (I). Geraniol, farnesol, geranylgeranol, geranyl fuarnesol, Arnesyl arnesol, geranyl geranyl arnesol, geranyl arnesyl arnesol, arnesilfa Renesyl arnesol, phytol and dihydrophytol are preferred, and geranylgeradiol is particularly preferred.

As described above, the raw material component (B) is not particularly limited as long as it is a fatty acid, a fatty acid methyl ester, a fatty acid ethyl ester, a monoglyceride, a diglyceride, a triglyceride, or an oil or fat containing them, and is a crude oil or an unrefined oil. Any of fats and oils, fats and oils in the middle of purification, refined fats and the like may be used, but from the viewpoint of reactivity, triglycerides are particularly preferred, and for example, fats and oils having high triglyceride content are preferred. The fat or oil having a high triglyceride content is not necessarily specified because it may be selected according to the properties of the required fat and the like. For example, the triglyceride content is 90% by mass or more, preferably 95% by mass or more. Is preferably 97% by mass or more. When these triglycerides are mainly used, the reactivity is very high, and the desired product can be obtained in a yield of nearly 100%, which is very preferable.

In producing the chain isoprenoid fatty acid ester-rich fats and oils of the present invention, one or more selected from chain isoprenoid alcohols, and fatty acids, fatty acid methyl esters, fatty acid ethyl esters, monoglycerides, diglycerides, triglycerides The mixing ratio of one or more selected from the following can be set arbitrarily because it depends on the form of the target final product, the reaction efficiency, etc. Taking into account, one or more selected from linear isoprenoid alcohols: one or more selected from fatty acids, fatty acid methyl esters, fatty acid ethyl esters, monoglycerides, diglycerides, and triglycerides (mass: mass ), For example, 10:;! ~ 1: 1 000 000, preferably 5: 1 to 1: 1 000 000, more preferably 2: 1 to 1: 1 000 000.

In particular, as described above, geranylgeranol is used as the raw material component (A), and triglyceride and an oil having a triglyceride content of 90% or more are used as the raw material component (B). When a fat is used, the mixing ratio is as follows: geranylgeranol: triglyceride (mass: mass), for example, 1: 1 to: L: 100000, preferably 1: 1 to L : 1 000 000, more preferably 1: 1 to 1: 1 000 000. In the production, it is preferable to dilute the mixture of the above raw materials in a solvent. Since the carboxylate hydrolase is rapidly deactivated by the presence of water, a hydrophobic organic solvent is preferable as the solvent used in the production of the present invention. Hexane, heptane, octane, isooctane, nonane, decane, cyclohexane, carbon tetrachloride, chloroform, dichloromethane, 1,2-dichloroethane, diethyl ether, isopropyl ether, acetic acid Examples include ethyl, propyl acetate, butyl acetate, benzene, toluene, xylene and the like, and particularly preferred are hexane, heptane, octane and isooctane. Of these, those dehydrated are preferred in terms of reactivity. These solvents are used in an amount of 0 to 100 times (mass), preferably 1 to 100 times (mass), more preferably 1 to 25 times the amount of the above-mentioned raw material mixture 1 (mass). (Mass) may be added.

The amount of the enzyme to be used depends on the type of the enzyme, the reactivity, the content of additives such as the immobilizing agent, etc., and is not specified unconditionally. However, the amount is not limited to the following. 0.01 to 100 times (mass), preferably 0.01 to 100 times (mass), more preferably 0.1 to 25 times (mass) If this happens, the reaction will proceed efficiently.

The time required for the reaction depends on the degree of progress of the reaction and the amount of the target product, and is not unequivocally defined, but is, for example, 0.5 to 72 hours, preferably 1 to 48 hours, more preferably 1 to 48 hours. If it is 24 hours, it can be balanced with the manufacturing cost.

The temperature depends on the degree of progress of the reaction and the amount of the target product, and is not generally defined, but is, for example, 20 to 80 ° C, preferably 25 to 75 ° C, and more preferably 3 to 75 ° C. When the temperature is 0 to 70 ° C, the reaction proceeds efficiently. As other production conditions, it is preferable to carry out the reaction under a stream of nitrogen and / or argon, because it is possible to prevent deterioration of the product due to oxidation, and the like. Stirring or the like is preferable because the reaction can proceed efficiently.

In addition, the chain isoprenoid fatty acid ester contained in the chain isoprenoid fatty acid ester-rich fats and oils described above can be obtained as almost a chain isoprenoid fatty acid ester by isolation. it can. That is, the present invention relates to a chain isoprenoid fatty acid ester obtained by isolating from the above-mentioned chain isoprenoid fatty acid ester-rich oil or fat. Isolation methods for this purpose are generally difficult to specify, but include, for example, fractional distillation, fractional sublimation, zone melting, solvent extraction, various adsorption methods, foam separation, membrane separation, separation using molecular sieves, There is a method using chromatography. From the viewpoints of efficiency, simplicity and yield of the method, chromatography is preferred, and adsorption chromatography is more preferred. As the adsorption chromatography, a method utilizing liquid chromatography is preferred because the linear isoprenoid fatty acid esters in the present invention can be isolated in good yield without decomposing. Specific examples of liquid chromatography include normal-phase liquid chromatography, reversed-phase liquid chromatography, thin-layer chromatography, paper chromatography, and high-performance liquid chromatography (HPLC). However, when isolating the linear isoprenide fatty acid esters in the present invention, any method can be used. In particular, normal phase liquid chromatography, reverse phase liquid chromatography, and high performance liquid chromatography (HPLC) are preferable in consideration of the separation ability, the throughput, the number of steps, and the like.

Here, normal phase liquid chromatography refers to, for example, the following method. That is, for example, a column was prepared using silica gel as a stationary phase, a mixture of hexane-monoethyl acetate, a mixture of chloroform and methyl alcohol as a mobile phase, and the reaction mixture obtained by the above production method was prepared. The compound is supplied at a load ratio of 0.1 to 5% (wt (mass) / v (volume)), and is subjected to a continuous elution method using a single mobile phase or a stepwise elution method in which the solvent polarity is gradually increased. This is a method to elute the fraction.

Reverse phase liquid chromatography refers to, for example, the following method. That is, for example, a column was prepared in which silicic acid (ODS) to which octane decylsilane was bonded was used as a stationary phase, a mixture of water and methanol, a mixture of water and acetonitrile, and a mixture of water and acetone. The reaction mixture obtained by the production method is supplied at a load ratio of 0.1 to 5% (wt (mass) / V (volume)), and is continuously eluted with a single solvent or stepwise elution with a stepwise decrease in solvent polarity Is a method for eluting a predetermined fraction.

High-performance liquid chromatography (HPLC) is, in principle, the same as normal-phase liquid chromatography or reverse-phase liquid chromatography described above, and is used for more rapid and high-resolution isolation. belongs to.

The above-mentioned methods can be used in combination of two or more kinds.In this case, the linear isoprenoid fatty acid esters of the present invention can be highly isolated, and can be obtained in a state in which more impurities are removed. Therefore, it is preferable.

The chain-form isoprenoid fatty acid esters obtained as described above are preferable because they can be obtained inexpensively and in large quantities easily, as compared with those obtained by a method such as chemical synthesis. In particular, when fats and oils are used as raw materials, the fats and oils are very inexpensive. In the production process, for example, the solvent mainly used is an inexpensive solvent such as hexane. Preferred. Of course, the use of chemicals is minimal, which is preferable in terms of safety.

The chain isoprenide S fatty acid esters obtained as described above can be used as they are, but one or more of them may be added to or contained in other fats and oils according to the purpose. Can also. In this case, the chain isoprenoid fatty acid esters are highly isolated, so that unnecessary ones in fats and oils are more removed, This is preferable because it does not add unnecessary color or smell. In this case, it is also preferable in that the content can be easily controlled. In the present invention, the fats and oils thus obtained can also be suitably used. Naturally, after addition of the chain isoprenoid fatty acid esters, if necessary, the oil is refined through a generally performed oil / fat refining step, so that it can be used as a normal oil / fat. In addition, the chain isoprenoid fatty acid esters obtained by the above method may be used for foods, pharmaceuticals and the like.

The present invention relates to fats and oils containing a large amount of chain isoprenoid fatty acid esters and a method for producing the same, and a chain isoprenoid fatty acid ester obtained by isolating from the fats and oils containing a large amount of chain isoprenoid fatty acid esters. And its manufacturing method. ADVANTAGE OF THE INVENTION According to this invention, fats and oils which contain many chain isoprenoid fatty acid esters very simply can be provided. In addition, chain isoprenoid fatty acid esters obtained by isolation from these fats and oils can be safely used in forms such as pharmaceuticals, foods and drinks, and external preparations for skin. Example

Next, the present invention will be further described with reference to examples, but the present invention is not limited to these examples.

Geraniol (manufactured by Wako Pure Chemical Industries, Ltd.), arnesol (manufactured by Sigma), geranylgeraniol (manufactured by Sigma), phytol used as a raw material in the following examples

(Manufactured by Wako Pure Chemical Industries), triolein (manufactured by Wako Pure Chemical Industries), geolein (mixture of 1-diolein and 2-dialeine: Funakoshi), monoolein (mixture of 1-monoolein and 2-monolein: funakoshi) , Methyl oleate (manufactured by Wako Pure Chemical Industries), oleic acid (manufactured by Wako Pure Chemical Industries), triarlinolene (manufactured by Funakoshi), conjugated linoleic acid ethyl ester (manufactured by Funakoshi), eicosapentaenoic acid ethyl ester (Wako Pure Chemical Industries) Made), Tridocosahexaenoin (Funakoshi) was purchased as a reagent. Dihydrophyl was obtained by reducing phytol according to the literature (J. Org. Chem. 58. 5285-5287. 1993).

In the examples described below, 10 zL of each sample was collected, diluted 1000 times with hexane, and analyzed by gas chromatography to determine the content and purity. The measurement conditions of the gas chromatography are shown below.

(One condition of gas chromatography)

Column: DB—lht (J & W, φ 0.32mm x 0.15m 5m) Oven temperature: 100-350 ° C (20. C / min)

Inlet / detector temperature: 300/330 ° C

Inlet pressure: 5p s i

Split ratio: 25: 1 Example 1 Fatty acid containing fatty acid geranylgeranyl ester

Dissolve 0.1 g of geranylgeraniol in 1000 g of refined soybean oil, add 1% of Novozym (Novo) to the total amount, and stir with a propeller stirrer at 60 ° C for 3 hours. did. After the reaction, the lipase was removed by filtration through a filter paper from the mixture diluted by adding 10 times the amount of hexane, and the hexane was completely distilled off by vacuum distillation to obtain a fatty acid geranylgeranyl ester-containing oil and fat. The content of fatty acid geranylgeranyl ester in the obtained fats and oils was 0.0187%. The obtained fats and oils had good taste and flavor, and could be used in the same manner as ordinary refined soybean oil. Example 2 Fatty acids containing fatty acid phulnesyl ester

For 1000 g of refined soybean oil, dissolve 2 g of farnesol, and add 1% to the total amount. Was added and the mixture was stirred with a propeller stirrer at 60 ° C. for 9 hours. After the reaction, the lipase was removed by filtration through a filter paper from the diluted hexane by adding a 10-fold amount of hexane, and the hexane was completely distilled off by vacuum distillation to obtain a fatty acid phthalnesyl ester-containing fat or oil. The content of fatty acid geranylgeranyl ester in the obtained fats and oils was 0.446%. The obtained fats and oils had good taste and flavor, and could be used in the same manner as ordinary refined soybean oil. Example 3 Fatty Acid Containing Fatty Acid Ethyl Ester

To 1000 g of refined soybean oil, 0.5 g of the phytol was dissolved, and Novozyme (manufactured by Novo) was added to 1% of the total amount. . After the reaction, the lipase was removed by filtration through a filter paper from the mixture diluted by adding 10 times the amount of hexane, and the hexane was completely distilled off by vacuum distillation to obtain a fatty acid-containing fatty acid and fat. The content of fatty acid geranylgeranyl ester in the obtained fats and oils was ◦.088%. The obtained fats and oils had good B taste, good flavor and the like, and could be used in the same manner as ordinary refined soybean oil. Example 4 Fatty Acid Containing Fatty Acid Isoprenyl Ester

For 1000 g of refined soybean oil, dissolve 0.5 g of fornesol, 0.5 g of geranylgeraniol, and 0.5 g of phytol, and make 1% of the total amount of Novozym (Novo ) And stirred with a propeller stirrer at 60 ° C for 24 hours. After the reaction, the lipase was removed by filtration through a filter paper from the diluted hexane by adding a 10-fold amount of hexane, and the hexane was completely distilled off by vacuum distillation to obtain a fat and oil containing a fatty acid ethyl ester. The content of fatty acid isoprenyl ester in the obtained fats and oils was 251% as a whole. The obtained fats and oils had good taste and flavor, and could be used in the same manner as ordinary refined soybean oil. Example 5 Fat and Oil Containing Geranylgeranyl Oleate

0.1 g of geranylgeraniol was dissolved in 10 g of triolein, Novozym (manufactured by Novo) was added to the solution in an amount of 1% based on the total amount, and the mixture was stirred at 60 ° C for 1 hour. After the reaction, the lipase was removed by filtration from the dilution diluted with 10 times the amount of hexane, and the hexane was completely distilled off by vacuum distillation to obtain geranylgeranyl oleate-containing fats and oils. The content of the fatty acid geranyl geranyl ester in the obtained fats and oils was 1.90%. The obtained fats and oils had good taste, flavor and the like, and could be used in the same manner as ordinary triolein. Example 6 Oil and Fat Containing Geranylgeranyl Arlinolenate

1 g of geranylgeraniol was dissolved in 10 g of triarinolene, and Novozym (manufactured by Novo) was added thereto so as to be 1% of the total amount, followed by stirring at 60 ° C for 3 hours. After the reaction, the lipase was removed by filtration from the diluted hexane by adding 10 times the amount of hexane, and the hexane was completely distilled off by vacuum distillation to obtain an oil and fat containing geranylgeranyl monolinolenate. The content of fatty acid geranylgeranyl ester in the obtained fats and oils was 18.2%. The obtained fats and oils had good taste and flavor, and could be used in the same manner as ordinary tria-linolene. Example 7 Geranylgeranyl arlinolenate concentrated oils and fats

10 g of the fat or oil obtained in Example 5 was fractionated by column chromatography packed with about 50 times the amount of silica gel. Using a mixed solution of hexane: ethyl acetate = 10: 1 as the mobile phase, hexane and ethyl acetate were completely distilled off from the second and third fractions among the fractionated fractions of 50 OmL by vacuum distillation. 1.87 g of fats and oils were obtained. Containing geranylgeranyl arlinolenate in the obtained fats and oils The amount was 48.4%. Example 8 Geranylgeranyl stearate

Geranylgeraniol 10 Omg and tristearin 90 Omg were dissolved in 1 g of isooctane, Novosam 435 (manufactured by Novo) was added to 1% of the total amount, and the mixture was stirred at 60 ° C for 3 hours. . After confirming that the reaction reached equilibrium by GC, lipase was removed by filtration from the reaction solution diluted with 10 times the amount of hexane, and hexane was distilled off by vacuum distillation to produce a crude reaction. I got something. Purification by silica gel column chromatography gave 187 mg of geranylgeranyl stearate. Example 9 Geranylgeranyl Oleate

Dissolve 10 mg of geranylgeraniol and 90 mg of triolein in 1 g of isooctane, add 1% lipase QL (Meito Sangyo Co., Ltd.) to the total amount, and add 2 hours at 60 ° C Was disrupted. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding a 10-fold amount of hexane, and lipase was removed by filtration. A reaction product was obtained. Purification by silica gel column chromatography gave 186 mg of geranylgeranyl oleate. Example 10 Geranylgeranyl oleate

Dissolve 10 Omg of geranylgeraniol and 10 Omg of triolein in 50 Omg of isooctane, add lipase QL (manufactured by Meito Sangyo Co., Ltd.) to 1% of the total amount, and add 2 hours at 60 ° C. Stirred. After confirming that the reaction reached equilibrium by GC, dilute the reaction solution by adding 10 times the amount of hexane, Then, hexane was removed by vacuum distillation to obtain a crude reaction product. Purification by silica gel column chromatography gave 62.5 mg of geranylgeranyl oleate. Example 11 Geranylgeranyl oleate

Dissolve 10 Omg of geranylgeraniol and 90 Omg of diolein in 1 g of isooxane, add lipase QL (manufactured by Meito Sangyo Co., Ltd.) to 1% of the total amount, and add 2 g at 60 ° C. Stirred for hours. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding 10 times the amount of hexane.The lipase was removed by filtration, and the hexane was distilled off by vacuum distillation to produce a crude reaction. I got something. Purification by silica gel column chromatography gave geranylgeranyl oleate (164 mg). Example 12 Geranylgeranyl oleate

Dissolve 10 Omg of geranylgeraniol and 90 Omg of monoolein in 1 g of isooctane, add lipase QL (manufactured by Meito Sangyo Co., Ltd.) to 1% of the total amount, and stir at 60 ° C for 2 hours did. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding a 10-fold amount of hexane, and lipase was removed by filtration. A reaction product was obtained. Purification by silica gel column chromatography gave 139 mg of geranylgeranyl oleate. Example 13 Geranylgeranyl oleate

Dissolve 10 Omg of geranylgeraniol and 90 Omg of methyl oleate in 1 g of isooctane, and make 1% of the total amount of Reno, 'Ize QL (Meito Sangyo Co., Ltd.) Was added and stirred at 60 ° C. for 2 hours. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding 10 times the amount of hexane.The lipase was removed by filtration, and the hexane was distilled off under vacuum to remove the crude reaction product. I got Purification by silica gel column chromatography gave 83 mg of geranylgeranyl oleate. Example 14 Geranylgeranyl oleate

Dissolve 10 Omg of geranylgeraniol and 90 Omg of oleic acid in 1 g of isoquinone, add lipase QL (manufactured by Meito Sangyo Co., Ltd.) to 1% of the total amount, and add to 60 ° C. For 2 hours. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding 10 times the amount of hexane.The lipase was removed by filtration, and the hexane was distilled off by vacuum distillation to produce a crude reaction. I got something. Purification by silica gel column chromatography gave 77 mg of geranylgeranyl oleate. Example 15 Geranylgeranyl arlinolenate

Dissolve 10 Omg of geranylgeraniol and 90 Omg of triarlinolene in 1 g of isooctane, add 1% of the total amount to Novozim (Novo), and stir at 60 ° C for 3 hours . After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding 10 times the amount of hexane.The lipase was removed by filtration, and the hexane was distilled off under vacuum to remove the crude reaction product. I got Purification by silica gel column chromatography gave 183 mg of geranylgeranyl monolinolenate. Example 16 Geranylgeranyl conjugated linoleate

Dissolve 10 Omg of geranylgeraniol and 30 Omg of conjugated linoleic acid ethyl ester in 1 g of isooctane, add 1% of the total amount to Novozym (Novo), and stir at 60 ° C for 24 hours did. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding 10 times the amount of hexane.The lipase was removed by filtration, and the hexane was distilled off by vacuum distillation to produce a crude reaction. I got something. Purification by silica gel column chromatography gave 118 mg of geranilugeranyl conjugated linoleate. Example 17 Geranyl eicosapentaenoate

Dissolve 10 mg of geraniol and 90 mg of ethyl eicosapenenoic acid ester in 1 g of isooctane, add Novozym (manufactured by Novo) to 1% of the total amount, and stir at 60 ° C for 24 hours did. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding 10 times the amount of hexane, the lipase was removed by filtration, and the hexane was distilled off under vacuum to remove the crude reaction. The product was obtained. Purification by silica gel column chromatography gave 102 mg of geranyl eicosapenogenate. Example 18 Fanesyl eicosapentaenoate

Dissolve 10 mg of fornesol and 75 mg of ethyl eicosapenenoic acid ester in 1 g of isooctane, add Novozym (Novo) to 1% of the total amount, add 24 mg at 60 ° C. Stirred for hours. After confirming that the reaction had reached equilibrium by GC, lipase was removed by filtration from the reaction solution diluted with a 10-fold amount of hexane, and the hexane was distilled off by vacuum distillation. A reaction product was obtained. Purified by silica gel column chromatography, eicosapenic acid 107 mg of Fanorenesinore were obtained. Example 19 Eicosapengeranylgeranyl echinate

Dissolve 10 Omg of geranylgeraniol and 60 Omg of eicosapen benzoic acid ester in lg isooctane, add Novozyme (Novo) to 1% of the total amount, and add 60 ° C For 24 hours. GC was used to confirm that the reaction had reached equilibrium.The reaction solution was diluted by adding 10 times the amount of hexane.The lipase was removed by filtration, and the hexane was distilled off under vacuum to remove the crude reaction product. I got Purification by silica gel column chromatography gave 106 mg of geranylgeranyl eicosapenate. Example 20 Fityl eicosapentaenoate

Dissolve 10 Omg of phytol and 60 Omg of eicosapen-u-enic acid ester in 1 g of isooctane, add Novozym (Novo) to 1% of the total amount, add 24 mg at 60 ° C. Stirred for hours. GC was used to confirm that the reaction had reached equilibrium.The reaction solution was diluted by adding 10 volumes of hexane, and lipase was removed by filtration, and the hexane was distilled off under vacuum to remove the crude reaction product. I got Purification by silica gel column chromatography gave 94 mg of phycoyl eicosapenate. Example 21 Eicosapen dihydrophytyl echinate

Dissolve 10 Omg of dihydrophytol and 600 mg of ethyl eicosapenenoic acid ester in 1 g of isooctane, add Novozym (manufactured by Novo) to 1% of the total amount, and stir at 60 ° C for 24 hours did. The reaction was confirmed to have reached equilibrium by GC, and the reaction mixture was diluted with a 10-fold amount of hexane and filtered. The lipase was removed by filtration, and hexane was distilled off by vacuum distillation to obtain a crude reaction product. Purification by silica gel column chromatography gave 74 mg of dihydrophytyl eicosapentaenoate. Example 22 2 Geranylgeranyl hexosahexanoate

Dissolve 100 mg of geranylgeraniol and 90 mg of tridocosahexaenoin in 1 g of isooctane, add Novozym (Novo) to 1% of the total amount, and add 60 ° C. The mixture was stirred at C for 3 hours. After confirming that the reaction reached equilibrium by GC, the reaction solution was diluted by adding 10 times the amount of hexane, lipase was removed by filtration, and the hexane was distilled off by vacuum distillation. A crude reaction product was obtained. Purification by silica gel gel column chromatography gave 193 mg of geranylgeranylgeranyl docosahexaenoate. From the above examples, it is found that the mixing ratio required for the reaction between the linear isoprenoid alcohols, fatty acids, triglycerides, etc., and the fats and oils containing these are particularly 1: 1 to 1: 10000 in order to efficiently react. I understood that. Further, from each of the examples, it was confirmed that any of the chain isoprenoid alcohols can be used. In particular, in Examples 1, 5, 9, 10 and 11, etc., the use of geranylgeranol was effective. It was found that geranylgeranyl fatty acid ester-rich fats and oils could be efficiently obtained. Further, from each of the examples, it was confirmed that any of fatty acids, fatty acid methyl, fatty acid ethyl, monoglyceride, diglyceride, triglyceride, and fats and oils containing these can be used, but in particular, Examples 1 to 6, and , 8, 9, 15 and 22 showed that when triglycerides and fats and oils containing triglycerides were used, fats and oils containing a high content of linear isoprenoid fatty acid esters could be efficiently obtained. Further, from Example 7, the chain isoprenoid fatty acid ester was It was also found that concentrated fats and oils could be suitably obtained. Example 23 Chain Isoprenoid Fatty Acid Ester-rich Fats and Oils

Each of the chain isoprenoid fatty acid esters obtained in the same manner as in Examples 9, 18, and 19 was added to each of the purified soybean oil and the purified rapeseed oil so as to have a mass ratio of 1000 ppm and 1000 Oppm, respectively. By dissolving, a total of 12 types of chain isoprenide fatty acid ester-containing fats and oils were prepared. All of the fats and oils had good taste, flavor and the like, and could be used in the same manner as the fats and oils to which the chain isoprenoid fatty acid ester was not particularly added. Example 24 Fats and oils containing a high content of chained isoprenoid fatty acid esters

Three kinds of chain isoprenoid fatty acid esters obtained in the same manner as in Examples 7, 8, and 22 were added to each of the purified soybean oil and the fat and oil of Example 1 in a mass ratio of 10 ppm, 100 ppm, 1000 ppm, Addition and dissolution were carried out at a rate of 10000 ppm to prepare a total of eight kinds of fats and oils containing the above three kinds of chain isoprenide fatty acid esters in high content. All fats and oils have good taste and flavor, and the above three types of chain-like soprenoid fatty acids! : Can be used with refined soybean oil to which no stell is particularly added and the fat and oil of Example 1. ADVANTAGE OF THE INVENTION According to this invention, chain isoprenoid fatty acid ester-rich fats and oils can be obtained simply and efficiently. In particular, it is very safe in terms of the use of enzymes and can be used as edible fats and oils. In addition, chain isoprenoid fatty acid esters can be isolated from these fats and oils, and these can be used as novel pharmaceuticals and foods.

Claims

The scope of the claims

1. Fats and oils rich in linear isoprenoid fatty acid esters obtained by treating the following components (A) and (B) as raw materials with a carboxylic ester hydrolase:

(In the formula, the wavy line means a single bond or a double bond, and n means any one integer selected from 1 to: L4.)

2. The oil and fat having a high content of linear isoprenide fatty acid esters according to claim 1, wherein the carboxylate ester hydrolase is carboxylesterase and / or triacylglycerol monolulinase.

3. The chain isoprenoid alcohol of component (A) is geraniol, fuarnesol, geranylgeranol, geranylfarnesol, funaresyl fuarsol, geranylgeranyl fuarnesol, geranyl geranyl fuarnesol, geranyl fuarnesyl farnesol. 3. The linear isoprenoid fatty acid according to claim 1, wherein the fatty acid is at least one member selected from the group consisting of funesyl phanesyl pharmacosol, phytol, and dihydrophytol. Fats and oils high in esters.

4. The fats and oils rich in linear isoprenoid fatty acid esters according to claims 1 and 2, wherein the fats and oils of the component (B) are triglycerides and vegetable oils containing the same.

5. The oil and fat having a high content of linear isoprenoid fatty acid esters according to claim 4, wherein the vegetable oil is edible.

6. The fatty acid or fatty acid rich in chain isoprenoid fatty acid esters according to any one of claims 1 to 4, wherein the fatty acid of the component (B) has 2 to 30 carbon atoms.

7. The chain isoprenoid fatty acid ester-rich fat or oil according to claim 1, wherein the fatty acid residue constituting the fatty acid methyl ester of the component (B) has 2 to 30 carbon atoms.

8. The fatty acid / fat rich in chain isoprenoid fatty acid esters according to claim 1, wherein the fatty acid residue constituting the fatty acid ethyl ester of the component (B) has 2 to 30 carbon atoms.

9. The fatty acid or fatty acid high in chain isoprenoid fatty acid esters according to any one of claims 1 to 4, wherein the fatty acid residue constituting the monoglyceride as the component (B) has 2 to 30 carbon atoms.

10. The chain-isoprenoid fatty acid ester-rich fats and oils according to claim 1, wherein the fatty acid residue constituting the diglyceride of the component (B) has 2 to 30 carbon atoms.

11. The chain-isoprenoid fatty acid ester-rich fats and oils according to claim 1, wherein the fatty acid residue constituting the triglyceride as the component (B) has 2 to 30 carbon atoms.

12. A geranylgeranyl-fatty acid ester-rich oil or fat obtained by using geranylgeraniol, triglyceride and an edible vegetable oil containing the same as a raw material and treating with triacylglycerol lipase.

13. An oil and fat having a high content of a chain isoprenoid fatty acid ester, which is obtained by concentrating, Z-purifying or purifying the chain isoprenoid fatty acid ester in the oil or fat according to claim 1 to 12.

14. A chain isoprenoid fatty acid ester obtained by isolating from the fats and oils according to claims 1 to 13.

15. A process for producing an oil containing a high content of linear isoprenoid fatty acid esters, comprising treating the following components (A) and (B) as raw materials with a carboxylic ester hydrolase:

16. The mixing ratio of component (A) and component (B) is (A) :( B) = 10: 1 to 1: 1

16. The method for producing a chain isoprenoid fatty acid ester-rich oil or fat according to claim 15, wherein the amount is 000000.

17. A method for producing a geranylgeranyl fatty acid ester-rich oil or fat, comprising using geranylgeranol, triglyceride and an edible vegetable oil containing the same as raw materials, and treating the mixture with triacylglycerol monolipase.

18. Mixing ratio of geranylgeraniol to triglyceride and edible vegetable oil containing geranylgeraniol: geranylgeraniol: triglyceride and edible vegetable oil containing it = 1: 1 to 1: 1: 0 0 0 0 0 0 0 The method for producing a geranylgeranyl fatty acid ester-rich oil or fat according to claim 17, wherein:

19. An oil-and-fat rich in chain-type isoprenoid fatty acid esters characterized by concentrating and / or purifying chain-type isoprenoid fatty acid esters in the oil or fat obtained by the method according to claim 15 to 18. Manufacturing method.

20. A process for producing a chain isoprenoid fatty acid ester, wherein the fatty acid ester is isolated from the fat or oil obtained by the method according to any one of claims 15 to 19.