JP2001218592A - Method for producing short-chain fatty acid ester - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing a short-chain fatty acid ester that permits efficiently synthesizing the short-chain fatty acid ester having a benzene ring. SOLUTION: A method for producing a short-chain fatty acid ester is provided, wherein the corresponding short-chain fatty acid shown by the formula R1COOH (R1 is H or a 1C-6C aliphatic hydrocarbon group) is reacted with an aromatic alcohol in the pressure of an immobilized lipase.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for producing a short-chain fatty acid ester having a benzene ring, which is suitably used for a fragrance or the like.

[0002]

2. Description of the Related Art Various reports have hitherto been made on the synthesis of esters using lipase, which is an oil and fat hydrolase. For example, for the synthesis of a long-chain fatty acid ester, Paul A. Claon et al. Used citronellyl acetate, geranyl acetate and the like in an organic solvent using Novozym (registered trademark) 435 (manufactured by Novo Nordisk), which is a kind of the lipase. (Enzyme Microb. Technol., Vol. 16 p8
35-840 (1994), J. Agric. Food chem., Vol. 42 p2349-
2352). Examples of other ester synthesis using lipase include Novozym (registered trademark) 435 by Forro E. et al.
(Nova Nordisk) (Acta Pharm.Hung.69 (3), p155-158).
(1999)). However, there has been no report on the synthesis of a short-chain fatty acid ester having a benzene ring using lipase as an enzyme.

On the other hand, short-chain fatty acid esters having a benzene ring include, for example, β-phenethyl ester and the like.
There are various compounds useful as perfumes. Therefore, a synthesis method capable of obtaining them with high yield is desired.

[0004]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above circumstances, and provides a method for producing a short-chain fatty acid ester capable of synthesizing a short-chain fatty acid ester having a benzene ring in a high yield. The main purpose is to do so.

[0005]

According to the present invention, in order to achieve the above object, R ¹ COO is provided as described in claim ^1.
H (R ¹ represents H or an aliphatic hydrocarbon group having 1 to 6 carbon atoms), and a short-chain fatty acid is reacted with an aromatic alcohol in the presence of immobilized lipase. A method for producing a short-chain fatty acid ester. By using such an immobilized lipase, it is possible to efficiently synthesize a short-chain fatty acid and an aromatic alcohol, and to efficiently obtain a short-chain fatty acid ester having a benzene ring in a simple step. .

[0006] In the first aspect of the present invention,
As described in claim 2, the aromatic alcohol is
Ar-R ² -OH (Ar has a substituent, or represents a benzene ring having no, R ² represents. An aliphatic hydrocarbon group having from 1 to 6 carbon atoms) is preferably, among others billing as described in claim 3 that, the aromatic alcohol is, Ar-R ² -OH (Ar represents a benzene ring, R ² represents. an aliphatic hydrocarbon group having up to 3 carbon atoms of 1) a Is preferred. In view of the usefulness of the obtained short-chain fatty acid ester, such an aromatic alcohol is preferable, and even when such a short-chain alcohol having a benzene ring is used as a raw material, the ester can be synthesized in high yield. This is because the feature of the present invention is that it can be performed.

[0007] Further, in the invention described in any one of claims 1 to 3, claim 4 is provided.
As described in above, the short-chain fatty acid is R ¹ COOH
(R ¹ represents H or an aliphatic hydrocarbon group having 1 to 3 carbon atoms). The present invention is characterized in that a short-chain fatty acid ester having a benzene ring can be synthesized in a high yield in the case of such a short-chain fatty acid, and is also preferable from the viewpoint of use of the obtained fatty acid ester. Because.

[0008] In the invention described in any one of claims 1 to 4, as described in claim 5, the lipase in the immobilized lipase is:
It is preferably a lipase derived from an animal or a microorganism. Particularly, as described in claim 6, the lipase is preferably a lipase derived from Candida antarctica or Mucor miehei . This is because the use of such a lipase can improve the yield of short-chain fatty acid esters.

[0009] In the invention according to any one of claims 1 to 6, as described in claim 7, in the reaction, the aromatic alcohol is used as a solvent, and The concentration of short-chain fatty acids is 1 mo
It is preferably at most 1 / L. This is because the yield of the ester is sharply improved by using the aliphatic alcohol as the solvent and further reducing the concentration of the short-chain fatty acid to 1 mol / L or less.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the method for producing a short-chain fatty acid ester having a benzene ring of the present invention will be described in more detail. As described above, the method for producing a short-chain fatty acid ester of the present invention is characterized by reacting a short-chain fatty acid with an aromatic alcohol in the presence of immobilized lipase.

The short-chain fatty acid used in the production method of the present invention is a short-chain fatty acid represented by R ¹ COOH.
R ¹ is H or an aliphatic hydrocarbon group having 1 to 6 carbon atoms. The aliphatic hydrocarbon group may be straight-chain or have a side chain, and may be saturated or unsaturated. In the present invention, a short-chain fatty acid in which R ¹ is H or an aliphatic hydrocarbon group having 1 to 3 carbon atoms is particularly preferable. In particular,
Considering the usefulness of the obtained short-chain fatty acid, formic acid, acetic acid, and propionic acid can be regarded as the most preferred short-chain fatty acid.

On the other hand, as the alcohol component used in the method for producing a short-chain fatty acid ester of the present invention, an aromatic alcohol having a hydroxyl group bonded to a side chain of an aromatic hydrocarbon is used. Such an aromatic alcohol is not particularly limited, and may be, for example, one having a plurality of benzene rings, a primary alcohol or a secondary alcohol.

However, considering the usefulness and reactivity of the obtained short-chain fatty acid ester, Ar-R ² -OH
Is preferably an aromatic alcohol represented by Here, Ar in the formula represents a benzene ring, and this benzene ring may be one in which any hydrogen is substituted with a substituent such as an aliphatic hydrocarbon group having 1 to 6 carbon atoms. And may have no substituent.
R ² in the formula represents an aliphatic hydrocarbon group having 1 to 6 carbon atoms. The aliphatic hydrocarbon group may be linear or have a side chain, and may be a saturated hydrocarbon or an unsaturated hydrocarbon.
In addition, the primary alcohol is preferable in terms of the usefulness and reactivity of the obtained short-chain fatty acid ester.

[0014] Aromatic alcohols used in the present invention, from the viewpoint of usefulness of the short-chain aliphatic esters derived, a benzene ring Ar represented by the above formula does not have a substituent, and R ² above Is a primary alcohol having an aliphatic hydrocarbon group having 1 to 3 carbon atoms. Specifically, aromatic alcohols preferably used include β-
Phenethyl alcohol, benzyl alcohol, cinnamyl alcohol and the like can be mentioned.

The present invention is characterized in that in synthesizing the above-mentioned short-chain fatty acid and aromatic alcohol, the synthesis is performed in the presence of immobilized lipase. The immobilized lipase refers to a lipase immobilized on a carrier by subjecting it to a treatment such as adsorption, chemical bonding, or inclusion.

The lipase used in the present invention includes lipase extracted and separated from cells or organs of microorganisms or animals and plants, and purified as necessary, or a lipase to which a stabilizer such as a saccharide is added. There is no particular limitation as long as it is present, but animal-derived lipase and microorganism-derived lipase are particularly preferably used.

Examples of the lipase derived from animals include lipase derived from pig pancreas. Examples of lipases derived from microorganisms include lipases derived from Candida yeast, filamentous fungi such as Aspergillus, Mucor, Rhizopus, Penicillium, Pseudomonas, and Chromobacterium, actinomycetes and bacteria. Alternatively, a lipase produced by a host transformed with DNA encoding the lipase of a microorganism that produces these lipases may be used. The lipase used in the present invention may be used in a form contained in a purified or crudely purified enzyme composition, or dried lipase-producing cells (treated cells, resting cells or quiescent cells). The product can also be used directly.

Specifically, Candida antarctica , Cole Maihei ( Mucor )
miehei , a commercial product of lipase derived from this product (hereinafter the same), "Patalases" manufactured by Novo Nordisk Japan Ltd.), Rhizopus delemar ,
Tanabe Seiyaku Co., Ltd. "Talipase"), Aspergillus niger "lipase Sankyo" manufactured (Aspergillus niger, Sankyo (strain as commercially available products of the lipase derived from Aspergillus spp.)), Candida Shirindorase (Candida cylindr
acea, Meito Sangyo Co., Ltd. "lipase OF"), Jiotorikamu candy dam (Geotricum candidum, Amano Pharmaceutical Co., Ltd. "lipase GC"), Alcaligenes sp. (Alcaligenes sp., Meito Sangyo Co., Ltd. "lipase P
L ") and the like can be mentioned as preferred examples. In the present invention, among others, Candida antarctica or Mu
Lipases from cor miehei are preferred.

In the present invention, the above-mentioned lipase is immobilized and used. That is, the lipase is preferably immobilized on a carrier that is insoluble in the reaction substrate of the short-chain fatty acid ester, for example, a porous substance such as celite, diatomaceous earth, activated carbon, silica gel, calcined body, glass beads, animal bone, and bone powder by an adsorption method. Lipase, alginate, agar, xanthan gum, guar gum, plant seed mucilage, polyacrylamide, etc., immobilized on lipase, hydroxyapatite, cationic or anionic ion exchange resin, etc. by ionic or covalent bonding method Lipase is used.

Specific examples of such immobilized lipase include Lipozyme (registered trademark) IM (manufactured by Novo), Novo
zym® 435 (Novo Nordisk), KE-100
(Manufactured by Kao Corporation). In the present invention, Novozym (registered trademark) 435 (manufactured by Novo Nordisk) can be mentioned as the most preferred immobilized lipase.

In the method for producing a short-chain aliphatic ester having a benzene ring according to the present invention, the aromatic alcohol,
An esterification reaction is carried out using short-chain fatty acids and immobilized lipase. That is, usually, a predetermined amount of an aromatic alcohol, a short-chain fatty acid, and an immobilized lipase are added to a predetermined solvent, and the ester reaction proceeds under normal pressure or dehydration conditions.

At this time, the input ratio of the aromatic alcohol and the short-chain fatty acid is not particularly limited, but is preferably adjusted so that the aromatic alcohol becomes excessive. In particular, in the present invention, it is preferable to use the above-mentioned aromatic alcohol as a solvent and to use short-chain fatty acids at a concentration of 1 mol / L or less, especially 0.4 mol / L or less. By doing so, the yield of the short-chain aliphatic ester having a benzene ring is remarkably improved. Also,
The immobilized lipase is preferably used in a ratio such that the amount of the immobilized lipase is in the range of 20 mg to 200 mg per 5 ml of the reaction solution.

The esterification reaction in the present invention can be carried out in a batch or continuous manner at room temperature or within a temperature range which does not impair the activity of the immobilized lipase to be used. Specifically, under reduced pressure, 20 to 100 ° C., preferably 25 to 100 ° C.
The reaction is carried out while removing water generated at 80 ° C., and after completion of the reaction,
If necessary, the immobilized lipase is removed / separated by filtration or the like, and excess raw material is distilled off under reduced pressure. Remove here
It is economical and preferred that the separated and recovered immobilized lipase be reused as it is, or washed with a solvent such as hexane which does not impair the activity of the enzyme.

When the esterification reaction is performed under dehydration conditions, the dehydration may be performed by any known method. Preferably, the reaction is carried out by reducing the pressure of the reaction system and distilling off water produced by the esterification under reduced pressure. Dehydration can also be performed by using an insoluble dehydrating agent such as molecular sieve.

The method for producing a short-chain fatty acid ester according to the present invention can be carried out continuously using an enzyme tower filled with immobilized lipase. This is advantageous as an industrial production method because there is no need for this.

The thus obtained short chain aliphatic ester having a benzene ring is used after being separated and purified to the extent necessary. Specific uses include various flavors.

The present invention is not limited to the above embodiment. The above embodiment is an exemplification, and has substantially the same configuration as the technical idea described in the scope of the claims of the present invention. It is included in the technical scope of the invention.

[0028]

EXAMPLES (Example 1: Synthesis of β-phenethyl acetate) 3
0 U / 100 mg of Novozym® 435 (Novo Nor
disk) (immobilized lipase), 0.6 mol / L acetic acid, and 5 ml β-phenethyl alcohol.
-A phenethyl alcohol solution was prepared. The β-phenethyl alcohol solution was placed in a 10-ml test tube with a screw cap, and shaken for 4 hours at a temperature of 40 to 50 ° C and a reciprocal shaking condition of 200 revolutions per minute. The obtained reaction product was measured at 40 ° C. using a high-performance liquid chromatography (HP1100, ODS column, manufactured by Hewlett-Packard Co.) with a 55% (v / v) aqueous methanol solution as an eluent. As a result, 90% molar conversion was obtained. Acetic acid was esterified at a high rate, and β-phenethyl acetate was obtained in high yield.

The esterification was carried out under the above conditions while changing the concentration of acetic acid in the β-phenethyl alcohol solution from 0.2 mol / L to 3.0 mol / L. The results are summarized in FIG. As is clear from FIG. 1, the concentration of acetic acid was 1 mol / mol.
If the concentration is not more than L, esterification is performed at a molar conversion of about 90% or more. Particularly, when the concentration of acetic acid is 0.4 mol / L or less, esterification is performed at a molar conversion of 100%.

A system not using immobilized lipase (acetic acid concentration:
(0.4 mol / L), esterification was carried out in the same manner, but the molar conversion of acetic acid was 1.35%.

Example 2 Synthesis of β-phenethyl formate
Esterification was carried out in the same manner as in Example 1 except that the acetic acid in Example 1 was changed to formic acid of 0.4 mol / L and the temperature during reciprocating shaking (during the reaction) was set to 40 ° C. In the system containing the immobilized lipase, β-phenethyl formate was obtained at a molar conversion of 100%, while in the system not containing the immobilized lipase, the molar conversion was 32.0%.

(Example 3: Synthesis of β-phenethyl propionate) Esterification was carried out in the same manner as in Example 2 except that formic acid in Example 2 was replaced with propionic acid. In the system containing the immobilized lipase, β-phenethyl propionate was obtained at a molar conversion of 100%, while in the system not containing the immobilized lipase, the molar conversion was 1.18%.

Example 4 Synthesis of Cinnamyl Acetate Esterification was carried out in the same manner as in Example 2 except that formic acid was changed to acetic acid and β-phenethyl alcohol was changed to cinnamyl alcohol. In the system containing the immobilized lipase, cinnamyl acetate was obtained at a molar conversion of 100%, whereas in the system not containing the immobilized lipase, the molar conversion was:
It was 3.60%.

Example 5 Synthesis of Benzyl Acetate Esterification was carried out in the same manner as in Example 4 except that cinnamyl alcohol in Example 4 was changed to benzyl alcohol. In the system containing immobilized lipase, benzyl acetate was obtained at a molar conversion of 100%, whereas in the system not containing immobilized lipase, the molar conversion was 8.55%.

(Example 6: Synthesis of benzyl propionate) Esterification was carried out in the same manner as in Example 5, except that acetic acid in Example 5 was changed to propionic acid. In the system containing immobilized lipase, benzyl propionate was obtained at a molar conversion of 100%, whereas in the system not containing immobilized lipase, the molar conversion was 20.3%.

The results of Examples 1 to 6 are shown in Table 1.
Put together. As is clear from Table 1, short-chain fatty acid esters containing a benzene ring were obtained at a molar conversion of 100% in all the systems containing the immobilized lipase. Therefore, it became clear that the production method of the present invention has an extremely high yield.

[0037]

[Table 1]

[0038]

According to the present invention, a short-chain fatty acid represented by R ¹ COOH (R ¹ is H or an aliphatic hydrocarbon group having 1 to 6 carbon atoms) is fixed to an aromatic alcohol. A method for producing a short-chain fatty acid ester, characterized in that the reaction is carried out in the presence of an immobilized lipase. By using such an immobilized lipase, it is possible to efficiently synthesize a short-chain fatty acid and an aromatic alcohol, and to efficiently obtain a short-chain fatty acid ester having a benzene ring in a simple step. This has the effect.

[Brief description of the drawings]

FIG. 1 is a graph showing the relationship between the concentration of acetic acid and the molar conversion.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12R 1: 785) C12R 1: 785) (72) Inventor Shoji Usami 3-4-1 Okubo, Shinjuku-ku, Tokyo Waseda University Faculty of Science and Engineering F-term (reference) 4B064 AD64 CA31 CB26 CD06 CD07 DA20 4H059 BA35 BB14 BB45 DA09

Claims (7)

[Claims] 1. A lipase in which a short-chain fatty acid represented by R ¹ COOH (R ¹ represents H or an aliphatic hydrocarbon group having 1 to 6 carbon atoms) and an aromatic alcohol are immobilized. A method for producing a short-chain fatty acid ester, wherein the reaction is performed under the following conditions. 2. The method according to claim 1, wherein the aromatic alcohol is Ar—R ² —
^2. The short-circuit according to claim 1, wherein OH is OH (Ar represents a benzene ring having or not having a substituent, and R ² represents an aliphatic hydrocarbon group having 1 to 6 carbon atoms). Method for producing chain fatty acid ester. 3. The method according to claim 1, wherein the aromatic alcohol is Ar—R ² —.
OH (Ar represents a benzene ring; R ² has 1 to 3 carbon atoms;
Up to the aliphatic hydrocarbon group. 3. The method for producing a short-chain fatty acid ester according to claim 2, wherein 4. The method according to claim 1, wherein the short-chain fatty acid is R ¹ COOH (R
¹ represents H or an aliphatic hydrocarbon group having 1 to 3 carbon atoms. The method for producing a short-chain fatty acid ester according to any one of claims 1 to 3, characterized in that: 5. The method for producing a short-chain fatty acid ester according to claim 1, wherein the lipase in the immobilized lipase is a lipase derived from an animal or a microorganism. Method. 6. The method for producing a short-chain fatty acid ester according to claim 5, wherein the lipase is a lipase derived from Candida antarctica or Mucor miehei . 7. The reaction according to claim 1, wherein in the reaction, the aromatic alcohol is used as a solvent, and the concentration of the short-chain fatty acid is 1 mol / L or less. A method for producing the short-chain fatty acid ester according to claim.