JPH06113877A - Production of alpha-and beta-glucosides - Google Patents

Abstract

PURPOSE:To obtain a method for producing alpha- and beta-glucosides having physiological actions such as surface-active, plant growth promoting or plant aroma imparting actions. CONSTITUTION:The objective method for producing an alpha-glucoside of formula II is to subject a beta-glucosidase to the catalytic reaction with glucosides of formula I (R is aliphatic alcohol residue, aromatic alcohol residue or alicyclic alcohol residue) which are a mixture of the alpha- with the beta-isomers and hydrolyze the beta-isomer. Furthermore, the objective method for producing a beta-glucoside of formula III comprises subjecting an alpha-glucosidase to the catalytic reaction with the glucosides of formula I to hydrolyze the alpha-isomer.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel method for producing α-glucosides and β-glucosides utilizing the enzymatic reaction of α-glucosidase and β-glucosidase. More specifically, the present invention provides the following formula (B):

[Chemical 5] (In the formula, R represents an aliphatic alcohol residue, an aromatic alcohol residue or an alicyclic alcohol residue), and glucosides of a mixture of α- and β-isomers ( Less than,
(Sometimes simply referred to as glucosides) is contacted with β-glucosidase, and the following formula (A) -1 obtained by hydrolyzing β-glucosides

[Chemical 6] (Wherein R is as defined above), a method for producing α-glucosides represented by the following, and a compound obtained by contacting the compound of the formula (B) with α-glucosidase to hydrolyze α-glucosides: Formula (A) -2

[Chemical 7] (In the formula, R has the same meaning as defined above), and relates to a method for producing β-glucosides.

[0002]

2. Description of the Related Art The α-glucosides of the above formula (A) -1 and the β-glucosides of the above formula (A) -2 have been found as constituents of plants, and α- and β-
The glucosides of the formula (B), which are a mixture of isomers, are conventionally known compounds that are chemically synthesized. However, recently, it was discovered that the glucosides have various useful functions such as surface activity and physiological activity, and these compounds include, for example, surfactants, plant growth promoters, plant aroma imparting agents, etc. Has been developed and used for various consumer products. In particular, the α-glucosides of formula (A) -1 and the β-glucosides of formula (A) -2 are α-glucosides
It is a compound that has been attracting attention in recent years because it exhibits superior physiological activity as compared with the glucosides of the formula (B), which is a mixture of β-isomers with and. Along with this, a method for producing α-glucosides and β-glucosides with good selectivity or α- and β-isomer glucosides with good selectivity for α-glucosides
The development of a method for separating glucosides or β-glucosides is strongly desired.

Conventionally, as for the method for producing the α-glucosides of the formula (A) -1, a general synthetic method having particularly excellent selectivity has not been known. In addition, for the production method of the β-glucosides of the formula (A) -2, for example, Koenigs-K
Synthetic methods using the norr reaction are known [Che
m. Ber. , 34 957 (1901)].

[0004]

However, the above-mentioned conventionally proposed production methods are not always satisfactory, and for example, in the method for synthesizing the compound of formula (A) -2,
It has drawbacks such as requiring complicated steps and using expensive reagents.

[0005]

[Means for Solving the Problems] The inventors of the present invention have conducted extensive studies to solve the above problems. As a result, a reaction using an enzyme that has been synthesized by a chemical method but has never been proposed in the past, that is, the above formula, which is a mixture of α- and β-isomers produced by chemosynthesis in a specific ratio, is used. The conventional problems can be solved all at once by bringing the compound of (B) into contact with β- or α-glucosidase and hydrolyzing it, and in addition, the compounds of formula (A) -1 and formula (A) -2 can be obtained in good purity and good yield. The present invention has been completed by discovering that the compound of 1 can be produced in only one step. Therefore, it is an object of the present invention to contact a compound of formula (B) with β-glucosidase,
Method for producing α-glucosides of formula (A) -1 for hydrolyzing glucosides and β of formula (A) -2 for hydrolyzing α-glucosides by contacting the compound of formula (B) with α-glucosidase -Providing a method for producing glucosides. This reaction can be represented by the following process diagram.

[0006]

[Chemical 8]

The method for producing the compounds of formula (A) -1 and formula (A) -2 will be described in detail below with reference to the above process chart. The glucosides of the formula (B) used in the present invention can be easily synthesized by a known method. For example, about 2 to about 20 parts by weight of alcohols (RO
H) in the presence of about 0.1 to about 10 parts by weight of acids such as hydrochloric acid, sulfuric acid and acidic ion exchange resin, and about -20 ° C to about 1
Reacting in a temperature range of about 00 ° C. for about 0.5 to about 24 hours,
If necessary, the acid can be removed by neutralization or filtration, and excess alcohols can be recovered to facilitate the synthesis. Compounds of formula (B) prepared by synthetic means as described above are:
It is a mixture having an isomer content ratio of α-glucosides and β-glucosides of about 70:30 to about 40:60.

An alcohol containing an R residue of an alcohol in the glucosides of the formula (B), which is the starting material of the present invention,
Examples of the alcohol include methyl alcohol, ethyl alcohol, isopropyl alcohol, propyl alcohol, isobutyl alcohol, butyl alcohol, amyl alcohol, isoamyl alcohol, 3-methyl-1-.
Pentanole, 2-hexanol, 2-heptanol,
Undecyl alcohol, 9-undecenol, cis-3
-Hexenol, 6-nonene-1-ol, 2,6-nonadiene-1-ol, homocitronellol, homogeraniol, rosinol, geraniol, citronellol
Le, nerol, linalool, tetrahydrolinalol,
Myrcenol, dihydromyrcenol, Lavanjuro
Aliphatic alcohols such as phenol, hydroxycitronellol and mugol; menthol, terpineol, piperitol, peryl alcohol, carveol, myrtenol
Fentyl alcohol, farnesol, nerolidol, lavangeol, santalol, cedrole,
Alicyclic alcohols such as patchouli call;
Benzyl alcohol, cumin alcohol, phenyl ethyl alcohol, phenyl propyl alcohol, cinnamyl alcohol, α-amyl cinnamyl alcohol, anise alcohol, vanillyl alcohol, piperonyl alcohol, eugenol And aromatic alcohols such as isol and isoeugenol.

1) Production of a compound of formula (A) -1. The production of the α-glucosides of the formula (A) -1 according to the present invention can be carried out by bringing the compound of the formula (B) into contact with β-glucosidase to hydrolyze the β-glucosides. The contact between the compound of formula (B) and β-glucosidase is preferably performed in the form of an aqueous solution, for example, about 1.0 to
It can be carried out with an aqueous solution containing about 80% of the compound of formula (B). Examples of the type of β-glucosidase used in the present invention include cellulase AP5 “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.) produced by Aspergillus genus, and Tri.
Cellulase T "Amano" produced by genus choderma
Commercial products such as β-glucosidase contained in commercial cellulase enzyme preparations (manufactured by Amano Pharmaceutical Co., Ltd.) and β-glucosidase derived from almond (manufactured by Oriental Yeast Co., Ltd.) can be mentioned. In addition, β-glucosidase which is widely present in other plants and animals and microorganisms can also be used. The β-
The amount of glucosidase used can be appropriately selected and is, for example, in the range of about 0.1 to about 0.0001 times by weight, more preferably about 0.01 to about 0.001 times by weight, of the compound of the formula (B). Can be used as an example. Further, generally, in the enzymatic reaction, reaction conditions such as pH and temperature are important, but in the present invention, the pH and temperature may be set within a range in which the β-glucosidase used has enzymatic activity, for example, It is desirable to adjust the pH of the reaction aqueous solution to a range of 2 to 11, more preferably a range of 4 to 10. Regarding the temperature, the reaction may be performed within a range that does not impair the stability of the enzyme, and for example, about 0 ° C to about 40 ° C is desirable.
However, when a so-called thermostable enzyme, alkaline enzyme or acidic enzyme is used, the range is not limited to the above range. The above hydrolysis reaction is carried out by injecting β-glucosidase directly into an aqueous solution of the compound of formula (B) and contacting it, or by immobilizing β-glucosidase and passing it through an aqueous solution of the compound of formula (B). Can be adopted.

From the reaction mixture aqueous solution obtained as described above, enzymes such as β-glucosidase can be easily removed by filtering and separating with a membrane filter or the like. Then, the alcohol and glucose produced by the reaction and the compound of the formula (A) -1 are separated from each other by water, if necessary, and then subjected to a separation means utilizing an adsorption / desorption difference such as activated carbon, an ion exchange resin or silica gel column chromatography. By using it, it can be easily separated and purified. Further, the alcohol product generated by the reaction can be easily removed by extraction separation with an organic solvent or steam distillation, if necessary. Further, if necessary, the remaining water or the like is concentrated under reduced pressure to obtain the compound of formula (A) -1 in high yield and high purity. Specific examples of the formula (A) -1 compound include α-form glucosides of the exemplified formula (B) compound.

2) Production of a compound of formula (A) -2. The production of β-glucosides of the formula (A) -2 according to the present invention can be easily carried out by bringing the compound of the formula (B) into contact with α-glucosidase and hydrolyzing the α-glucosides. This reaction can be carried out according to the method for producing α-glucosides except that β-glucosidase in the production of the compound of formula (A) -1 described above is replaced with α-glucosidase. For example, in the hydrolysis reaction α-
The reaction conditions such as the amount of glucosidase used, pH, reaction temperature and reaction time may be the same as those in the production of α-glucosides.

The types of α-glucosidase used in the present invention include, for example, transglucosidase L "Amano" (manufactured by Amano Pharmaceutical Co.), which is an α-glucosidase produced by the genus Aspergillus, α-glucosidase produced by yeast (produced by Oriental Yeast) ), Rice-derived α-glucosidase [Serva-Feinbiochemical
a (Germany)] and the like. In addition, α- which is widely present in flora and fauna and microorganisms
Glucosidase can be used. The compound of formula (A) -2 thus obtained can be purified in high yield and high purity, for example, according to the method for separating the compound of formula (A) -1 described above. Further, specific examples of the compound of formula (A) -2 include β-form glucosides of the exemplified compound of formula (B). Next, the present invention will be described in more detail with reference to Reference Examples and Examples.

[0013]

Reference Example 1 Synthesis of ethylgukoside [compound of formula (B)]. D-Glucose 2 g, 95 ° undenatured ethanol 100
0 ml and 0.5 ml of sulfuric acid were added and the mixture was refluxed for 4 hours. This solution was diluted with 300 ml of distilled water, neutralized with barium carbonate, and then filtered to remove excess barium carbonate.
After applying this filtrate to a column, evaporation and concentration were carried out to obtain ethyl glucoside (a mixture of α-form: β-form = 60: 40).
6.5 g (yield: 72%) was obtained.

[0014]

Reference Example 2 Synthesis of various glucosides [compound of formula (B)]. According to the method of Reference Example 1, various alcohols were reacted with D-glucose instead of ethanol of Reference Example 1 to synthesize corresponding glucosides. Examples of these glucosides include cis-3-hexenyl glucoside, geranyl glucoside, menthyl glucoside, isoamyl glucoside, undecyl glucoside, benzyl glucoside, cinnamyl glucoside, vanillyl glucoside, 6-
Nonenyl glucoside, citronellyl glucoside, terpineryl glucoside, β-phenylethyl glucoside, anisyl glucoside, eugenyl glucoside, neryl glucoside, linalyl glucoside, myrcenyl glucoside,
Examples include peryl glucoside, farnesyl glucoside, santalyl glucoside, and ceryl glucoside.

[0015]

Example 1 Synthesis of α-ethyl glucoside [(A) -1 compound]. Ethyl glucoside (α-form / β-form mixture: 60/4
0) 20 g are dissolved in 180 g water. Commercial enzyme cellulase AP5 "Amano" containing β-glucosidase
(Amano Pharmaceutical Co., Ltd.) 0.1 g is added and the reaction is carried out by stirring for 2 days. After the reaction, the mixture was filtered through a membrane filter, and the mother liquor was concentrated to give α-ethylglucoside [to β-isomer ratio (α / α + β) 98% by C ¹³ NMR] of 17
g was obtained. Furthermore, by removing glucose by silica gel column chromatography, 10.8 g of α-ethyl glucoside
(Yield: 90%) was obtained.

[0016]

Example 2 Synthesis of β-ethylglucoside [(A) -2 compound]. Ethyl glucoside (α-form / β-form mixture: 60/4
0) 40 g are dissolved in 360 g water. 4 g of a commercially available enzyme transglucosidase “Amano” (Amano Pharmaceutical Co., Ltd.) containing α-glucosidase is added, and the mixture is stirred and reacted for 2 days. After the reaction, steam distillation is carried out to deactivate the enzyme and distill off the ethanol produced by decomposition. 3 by collecting water by filtering with a membrane filter
5 g of crude product was obtained. The purity of this product is C ¹³ N
When measured by the MR method, it was confirmed that the purity of β-ethylglucoside [ratio of α-isomer (β / α + β)] was 99% or more. Further, glucose was removed by silica gel column chromatography to obtain 14.7 g (yield: 92%) of purified β-ethylglucoside.

[0017]

Examples 3 to 14 Synthesis of various α-ethyl glucosides [(A) -1 compound]. The following raw material glucosides obtained in Reference Example 2 [formula (B)]
[Compound] except that the glucosidase was appropriately changed, and various corresponding α-ethylglucosides were obtained according to the method of Example 1. The results are shown in Table-1.

Table-1 No Raw material glucosides (α / β ratio) β-Glucosidase type α / α + β (%) after reaction 3 3 (Z) -hexenyl glucosides (55/45) Cellular sese AP5 87.3 4 2 -Phenylethyl glucosido (65/35) 〃 97.8 5 geranyl glucosido (63/37) 〃 89.5 6 Menthyl glucosico (50/50) 〃 89.5 7 Thinner milk glucosido (53/57) Cellular T 82.4 8 cosylate 50) 〃 93.2 9 n-nonyl glucoside (63/37) Cellular AP 5 90.5 10 3,6-nona phenyl alkoxy (65/35) 〃 89.3 11 1-octen-3-yl glucoside (50/50) 〃 95.5 12 Eugenyl Glucose (60/40) Cellular T 93.1 13 Citronellyl Glucocide (50/50) Cellular Gage AP5 94.2 14 n-Dodecanyl Glucocide (55/45) 〃 98.2

[0019]

Examples 15 to 26 Synthesis of various β-ethylglucosides [(A) -2 compound]. The following raw material glucosides obtained in Reference Example 2 [formula (B)]
[Compound] except that the glucosidase was appropriately changed, and corresponding various β-ethylglucosides were obtained according to the method of Example 2. The results are shown in Table-2.

Table-2 No Raw material glucosides (α / β ratio) α-Glucosidase type β / α + β (%) after reaction 15 3 (Z) -hexenyl glucosides (55/45) trans glucosidase L 99.0 16 2-Phenylethyl Glucosid (65/35) 〃 99.5 17 Keranyl Glucosid (63/37) 〃 98.7 18 Menthyl Glucosid (50/50) 〃 99.5 19 Cinnamil Glucosidyl (53/57) α-Glucosidyl 50/50) 〃 97.3 21 n-Nonyl glucoside (63/37) Trans glucoside L 99.3 22 3,6-None phenyl alkoxide (65/35) 〃 98.4 23 1-octen-3-yl glucoside (50/50) 〃 97.3 24 Eugenyl Glucosides (60/40) α-Glucosides 99.1 25 Citronellyl Glucosides (50/50) Trans Glucosides L 94.3 26 n-Nos Gurukoshido (55/45) 〃 98.9

[0021]

INDUSTRIAL APPLICABILITY According to the present invention, a compound of formula (A) -1 can be obtained by catalytically reacting a compound of formula (B) with β-glucosidase, and an α-glucosidase can be added to a compound of formula (B). By the catalytic reaction, the compound of formula (A) -1 or formula (A) -2 can be produced in good yield and high purity. Α-glucosides of formula (A) -1 and formula (A)-
The β-glucosides of 2 are useful as surfactants, plant growth promoters, plant aroma imparting agents and the like.

Claims (2)

[Claims] 1. The following formula (B): In the formula, R represents an aliphatic alcohol residue, an aromatic alcohol residue or an alicyclic alcohol residue,
And a β-isomer mixture glucosides are contacted with β-glucosidase to hydrolyze the β-glucosides, the following formula (A) -1: (Wherein R represents an aliphatic alcohol residue, an aromatic alcohol residue or an alicyclic alcohol residue), and a method for producing an α-glucoside. 2. The following formula (B): In the formula, R represents an aliphatic alcohol residue, an aromatic alcohol residue or an alicyclic alcohol residue,
-And β-isomer mixture glucosides are contacted with α-glucosidase to hydrolyze the α-glucosides, represented by the following formula (A) -2: (In the formula, R represents an aliphatic alcohol residue, an aromatic alcohol residue or an alicyclic alcohol residue), and a method for producing β-glucosides.