JPH0565159B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Application Field] The present invention relates to a method for producing a saccharide fatty acid ester compound, and more particularly to a method for easily producing a saccharide fatty acid ester in high yield using lipase. [Prior Art] As saccharide fatty acid esters based on chemical synthesis methods, sucrose fatty acid esters are industrially produced, in part because sucrose has high purity and can be mass-produced at low cost. Currently, sucrose fatty acid esters are produced using organic synthesis methods, such as the Schünel method, the Nebraska-Schünel method, and the USDA method developed by the United States Department of Agriculture.
Many methods have been proposed today, but the basic method in practical use is the transesterification reaction of sugar and lower alcohol esters of fatty acids at high temperatures. On the other hand, attempts have been made to enzymatically synthesize sugar and fatty acid esters by utilizing the ability of lipase to synthesize esters, which is a reaction opposite to the hydrolysis reaction. That is, Seino, Uchibori et al. used sucrose, glucose, fructose, and sorbitol as sugars and oleic acid and stearic acid as fatty acids as synthetic substrates, and used Rhyzopus, Enterobacterium, and Aspergillus as synthetic substrates. , Pseudomonas, Chrmobacterium, Candida,
reported an attempt to synthesize esters in an aqueous solvent system using lipases produced by Mucor and Penicillium [J.Amer.Oil Chem.Soc., 61, (11), 1761
(1984)]. According to this report, extremely low substrate concentrations, i.e.
It states that 4g/lipase was added to an aqueous solution (PH5.4) containing 0.36-1.7% sugar and 2.26-5.7% oleic acid, and the reaction was carried out at 40°C for 72 hours.
As a result, they found that the lipase from Candida cylindracea (Lipase MY) was good, and to a lesser extent, the lipases from Pseudomonas and Enterobacterium also showed synthetic activity. And in this report,
Enzymatic ester synthesis methods in aqueous systems show significantly low yields. [Problem to be solved by the invention] Schünel method, Nebraska Schünel method,
In chemical synthesis methods such as the USDA method, the transesterification reaction is carried out at high temperatures, which causes structural changes and decomposition of some sugars due to intramolecular dehydration, which can have an unfavorable effect on the functionality of the product. In addition, these methods require a large amount of thermal energy for the reaction, and cannot be said to be a satisfactory method from the viewpoint of energy saving. On the other hand, the enzymatic synthesis method using lipase allows the reaction to be carried out under mild conditions at room temperature and normal pressure, and can produce sugar esters with excellent properties without causing deterioration of sugars or product esters. However, lipase is originally known as a typical hydrolytic enzyme along with protease and amylase, and its general function is to reduce the activity toward synthesizing esters in the presence of a large amount of water. In a reaction system in which water is added to the ester, it is difficult to obtain a satisfactory ester synthesis when lipase is used, and re-decomposition of the produced ester cannot be prevented. In fact, according to a report by Seino et al., an esterification reaction was carried out at a sugar concentration of 0.36 to 1.7% in the presence of about 92 to 97% water using Candida cylindrace lipase, and the This suggests that about 1% of sugar esters were produced. Thus, there has been a desire for a method for easily obtaining saccharide fatty acid ester compounds by an enzymatic method in high yield without the drawbacks of using an aqueous solvent system, and the present invention has solved this problem. [Means for solving the problem] In view of the above-mentioned circumstances, the present inventors have determined that the ester synthesis reaction is Based on the idea that the reaction equilibrium law between hydrolysis reaction and synthesis reaction must be followed, and low synthesis rate and re-decomposition cannot be avoided, we decided to proceed without being bound by conventional concepts. We studied lipases that enable fatty acid ester production. As a result, lipases produced by microorganisms of the genus Achromobacter (Japanese Patent Publication No. 49-32080), lipases produced by microorganisms of the genus Alcaligenes (Japanese Patent Publication No. 58-36953, Kaisho 53-59093
When microbial alkaline lipases such as JP-A No. 2003-12212 are allowed to act in the presence of an organic solvent, there is a surprising phenomenon in which highly concentrated esters are produced between a wide range of saccharides and fatty acids, which has never been reported before. I discovered that. The present invention was completed based on this discovery. That is, the present invention provides monosaccharides having at least two or more hydroxyl groups and having a carbon number of _C5 to _C7 (this monosaccharide is
methyl group, phenyl group, nitrophenyl group, phenol group, hydroxymethylphenyl group, mercapto group, halogen, acetylamino group, amino group, uracil group, thymine group, or adenine group), from hexol disaccharide, sugar alcohol with carbon number C ₄ to C ₆ , carbon number
A saccharide selected from the group consisting of C ₆ to C ₇ sugar lactones and a saturated or unsaturated fatty acid with a carbon number of C ₂ to _{C 22} (this fatty acid has a hydroxyl group, a carboxyl group,
or a _C1 to _C4 lower alcohol and the above fatty acid forming an ester are dehydrated in the presence of an organic solvent (excluding primary alcohol solvents). A method for producing saccharide fatty acid ester compounds, which is characterized by allowing alkaline lipase produced by a microorganism belonging to the genus Achromobacter or the genus Alcaligenes to act without dehydration or dehydration. The object of the present invention is to provide a method for easily obtaining a saccharide fatty acid ester compound from a fatty acid in high yield. In the present invention, the saccharide fatty acid ester includes not only esters produced from saccharides and fatty acids by the action of lipase, but also saccharide fatty acid esters produced between lower alcohol esters of fatty acids and saccharides. The present invention will be explained in detail below. The saccharides used in the present invention refer to monosaccharides having a carbon number of _C5 to _C7 and having at least two or more hydroxyl groups (these monosaccharides include methyl groups, phenyl groups, nitrophenyl groups, phenol groups, hydroxymethylphenyl groups, mercapto group, halogen, acetylamino group, amino group, uracil group, thymine group, or adenine group), disaccharide consisting of hexol, sugar alcohol with a carbon number of C ₄ to _{C 6} , carbon number C A saccharide selected from the group consisting of ₆ to _C7 sugar lactones, and in the present invention, saccharides include not only aldoses or ketoses, but also the above-mentioned sugar alcohols and sugar lactones derived from these. As specific examples of monosaccharides without substituents, monosaccharides having _C5 carbon atoms include, for example, arabinose,
Examples include ribose, xylose, lyxose, xylylose, ribulose, 2-deoxyribose, and examples of _C6 monosaccharides include glucose,
Galactose, fructose, mannose, sorbose, talose, 2-deoxyglucose, 6
-Deoxygalactose, 6-deoxymannose, 2-deoxygalactose, etc.
Examples of _C7 monosaccharides include glucoheptose,
Examples include sedoheptulose, mannoheptulose, and glucoheptulose. Further, as specific examples of monosaccharides having a substituent, those having a methyl group as a substituent include α-methyl riboside, β-methyl riboside, β-methyl arabinoside, 2-o-methylxylose, α- Methylxyloside, α-methylglucoside, β-methylglucoside, α-methylmannoside, β-methylmannoside, α-methylgalactoside, β-methylgalactoside, 3-o
Examples of those having a phenyl group as a substituent include phenyl-α-glucoside, phenyl-β-glucoside, phenyl-α-galactoside, and phenyl-β-glucoside.
-galactoside, etc., and those having a nitrophenyl group as a substituent include, for example, o
-Nitrophenyl-α-galactoside, o-nitrophenyl-β-galactoside, m-nitrophenyl-β-galactoside, p-nitrophenyl-
β-galactoside, o-nitrophenyl-β-glucoside, p-nitrophenyl-β-glucoside, p-nitrophenyl-α-glucoside, o-
Nitrophenyl-β-xyloside, p-nitrophenyl-β-xyloside, p-nitrophenyl-
Examples include α-mannoside, p-nitrophenyl-β-mannoside, etc. Examples of those having a phenol group as a substituent include arbutin, and examples of those having a hydroxymethylphenyl group as a substituent include salicin. Examples of those having a mercapto group as a substituent include 1-thioglucose, 1
Examples of those having a halogen as a substituent include clarose, and examples of those having an acetylamino group as a substituent include N-acetylglucosamine, N-acetylgalactosan, and N-thiogalactose. −
Examples of those having an amino group as a substituent include glucosamine and galactosamine. Examples of those having an adenine group as a substituent include adenosine, deoxyadenosine, and 8-bromoadenosine. Examples of those having a uracil group as a substituent include uridine and deoxyuridine, and examples of those having a thymine group as a substituent include thymidine. In addition, examples of disaccharides consisting of hexose include maltose, sucrose, etc.
As a sugar alcohol consisting of carbon number C ₄ to C ₆ ,
Examples include erythritol, ribitol, xylitol, arabitol, sorbitol, mannitol, galactitol, etc., and have a carbon number of C ₆
Examples of sugar lactones consisting of ~ _C7 include gluconolactone, glucono-δ-lactone, galactono-γ-lactone, and glucoheptonic acid-γ-lactone. Although the above-mentioned saccharides can be exemplified, the selection of saccharides is not limited in the present invention. Next, the fatty acid used in the present invention is a saturated or unsaturated fatty acid having a carbon number of _C2 to _C22 (this fatty acid may be substituted with a hydroxyl group, a carboxyl group, or a phenyl group), and In the present invention, fatty acid refers not only to acids present in oils and fats, but also includes other organic acids. Examples of unsubstituted fatty acids having a carbon number of _C2 to _C22 include acetic acid, propionic acid, acrylic acid, butanoic acid, crotonic acid, methacrylic acid, valeric acid, caproic acid, 2-methylvaleric acid,
Sorbic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linoleic acid, eicosanoic acid, docosanoic acid,
Examples include arachidonic acid. Among fatty acids with substituents, those with a hydroxyl group include ricinoleic acid, dihydroxystearic acid, etc., and those with a carboxyl group include malonic acid, maleic acid, succinic acid, glutaric acid, adipine, etc. Examples of the acid include pimelic acid, suberic acid, azelaic acid, sebacic acid, and tetradecanedioic acid. Examples of the acid having a phenyl group include phenylacetic acid, cinnamic acid, and 3-phenylpropionic acid. Furthermore, the above fatty acid may form an ester with a _{C 1} to C ₄ lower alcohol;
Examples of _C4 lower alcohols include methanol, ethanol, propanol, propylene glycol, glycerin, isopropanol, butanol, and specific examples of these fatty acid esters include methyl laurate, methyl myristate, and methyl palmitate. , methyl stearate, ethyl laurate, ethyl palmitate, ethyl stearate, ethyl oleate, isopropyl laurate, isopropyl myristate, isopropyl palmitate, butyl laurate, butyl stearate, butyl oleate, propylene glycol monostearate, Propylene glycol dipalmitate, glycerin monolaurate, glycerin monostearate, glycerin monooleate, glycerin dipalmitate,
Examples include glycerin distearate, glycerin diolate, glycerin tristearate, glycerin triolate, coconut oil, soybean oil, cottonseed oil, palm oil, beef tallow, lard, and olive oil. Examples include the fatty acids and lower alcohols thereof as described above; however, in the present invention, there is no restriction on the selection of fatty acids and lower alcohol esters thereof. Next, regarding the microbial alkaline lipase used to carry out the present invention, any microbial alkaline lipase can be used, but for example, Meito-AL- which belongs to the genus Achromobacter Lipase (Special Publication No. 49-32080) (hereinafter referred to as lipase-AL) produced by No. 865 (Feikoken Bibori No. 1213), Meito PL-266 belonging to the genus Alcaligenes [Feikoken Lipase produced by Kenjoyori No. 2985 (Kenjoyori No. 3187)
-36953 Publication) (hereinafter referred to as lipase-PL266), the famous sugar PL-, which also belongs to the genus Alcaligenes.
Lipase (Japanese Patent Application Laid-Open No. 53-59093) produced by No. 679 (Feikoken Bibori No. 3783) (hereinafter referred to as lipase)
PL679) can be cited as a specific example of an effective lipase with particularly excellent solvent resistance and ester synthesis ability. This point will be explained using an experimental example. Experimental example 1 Stability in solvent Lipase-PL679 (Meito Sangyo), Lipase-
Take 25 mg each of PL266 (Meito Sangyo) and Lipase-AL (Meito Sangyo) powders into a 7 ml centrifuge tube with a stopper, and add various solvents such as hexane, acetonitrile, and
Add butyl alcohol, acetone, tertiary amyl alcohol, diacetone alcohol, and 2 ml of water.
The mixture was thoroughly stirred and shaken at 37°C for 24 hours, and the remaining activity was measured by lipase titration. Lipase was measured using the method of Kunio et al. for lipase-PL679 and lipase-PL266 [Agric.Biol.
Chem. 46 (7), p. 1743, 1982], lipase
Regarding AL, the method of Kokusho et al. [Oil Chemistry 23 (2), No. 98]
Page, 1974]. The results are shown in Table 1.

[Table] Experimental Example 2 Ester synthesis ability of various lipases in solvent Various lipases, namely alkaline lipase Lipase-PL679 (Meito Sangyo), Lipase-PL266
(Meito Sangyo), Lipase-AL (Meito Sangyo), and non-alkaline lipase-MY (Meito Sangyo), Lipase-MAP4 (Amano Pharmaceutical), Lipase
Lipases such as AP10 (Amano Pharmaceutical Co., Ltd.), Oripase 2S (Osaka Seiken Co., Ltd.), and Talypase (Tanabe Seiyaku Co., Ltd.) are used in various solvents, such as hexane, acetonitrile, tertiary butyl alcohol, acetone, tertiary amyl alcohol, diacetone alcohol, etc. In the presence of water, fructose as sugar and oleic acid as fatty acid37
Ester synthesis was performed by reacting at ℃ for 24 hours, and the synthesis of sugar ester was confirmed by thin layer chromatography (TLC). The results are shown in Table 2.
The symbols in Table 2 have the following meanings. +++: Ester synthesis occurred very well ++: Ester synthesis occurred well +: Ester synthesis occurred -: Ester synthesis did not occur

〔Example〕

Examples of the present invention are shown below, but the present invention is not limited thereto. Example 1 0.5 g of oleic acid, 0.5 g of fructose, 0.5 g of Molecular Sieves 3A (dehydrating agent sold by Wako Pure Chemical Industries),
Tertiary butyl alcohol 5ml, lipase-PL679
0.1 g of the powder was placed in a test tube with a stopper, mixed well, and reacted with shaking at 37° C. for 24 hours. After the reaction, transfer 10μ of the reaction solution to a thin layer of silica gel (Silica gel 60TLC plate No. 5721, manufactured by Merck & Co., Ltd., 20×
20 cm) and developed using chloroform-methanol-acetic acid-water (80:10:8:2 V/V) as a developing solvent. For detection of spots, 50% sulfuric acid (detection of organic matter) and naphresorcin phosphate reagent (detection of sugar) were used. That is, when 50% sulfuric acid is used, unreacted fructose, unreacted oleic acid, and two unknown spots are detected, and when naphresorcinol phosphate reagent is used, unreacted fructose and two unknown spots are detected. was detected.
Since the Rf values of these two unknown spots matched by the two detection methods described above, these spots were determined to be fructose oleate, which is an oleic acid ester of fructose. Of the two fructose oleate spots, the spot with an Rf value of 0.33 is designated as spot 1, and the spot with an Rf value of 0.75 is designated as spot 2. Spot 1 was overwhelmingly larger in TLC detection, so the two fructose oleate spots were Of these, we decided to refine Spot 1. Fructose oleate was purified using preparative TLC. 0.5 ml of the reaction solution was spotted in a line on a thin layer of silica gel (Merck Silica gel TLC plate No. 13894, 0.5 mm, 20 x 20 cm), and a mixture of chloroform-methanol-acetic acid-water (80:10:8:
2V/V) was used as a developing solvent. After deployment,
Color was developed with 50% sulfuric acid and naphresorcin phosphate reagent, the fructose oleate was scraped off the TLC plate, and the silica gel was extracted with 10 ml of chloroform-methanol (2:1 V/V). extract liquid
After passing it through No.5C paper and removing the silica gel,
It was washed three times with 3 ml of water, and the chloroform layer was separated. The separated chloroform layer was dried under reduced pressure at 30°C. The yield of fructose oleate obtained was 3
mg was found to be single on TLC. The IR spectrum of fructose oleate was measured by the liquid film method using a JASCO model A202 infrared spectrophotometer. The results are shown in Table 3 (Run No. 9). Furthermore, the same procedure as above was carried out using other saccharides to obtain oleic acid esters of saccharides. The IR spectra are shown in Table 3 (Run Nos. 1 to 8 and Nos. 10 to 44). However, 50% sulfuric acid and benzidine periodate reagent were used to detect sugar alcohol esters, sugar lactone esters, and sugar esters with substituents.

【table】

【table】

[Table] From the results in Table 3, it can be seen that the above product is a saccharide oleic acid ester because absorption of ester bonds is observed near 1740 Kaiser and absorption of sugar is observed near 3400. Example 2 Reaction and purification were carried out in the same manner as in Example 1 using 0.5 g of stearic acid, 0.5 g of fructose, 0.5 g of Molecular Sieves 3A, 5 ml of tertiary butyl alcohol, and 0.1 g of Lipase-PL679 powder. . As a result, 2 mg of fructose stearate was obtained. The IR spectrum of this compound is shown in Table 4 (Run No. 14). Furthermore, the same procedure as above was carried out using other fatty acids to obtain fatty acid esters of fructose. The IR spectra are shown in Table 4 (Run No. 1 to 13 and No. 15
~twenty five). Furthermore, the same procedure as above was carried out using ribose and fatty acid to obtain fatty acid ester of ribose. The IR spectra are shown in Table 4 (Run Nos. 26 to 31).

【table】

[Table] From the results in Table 4, the absorption of ester bonds is seen around 1740 Kaiser and the absorption of sugar is seen around 3400 Kaiser, so it can be seen that the above products are fatty acid esters of fructose and fatty acid esters of ribose. . Example 3 Reaction and purification were carried out in the same manner as in Example 1 using 0.5 g of stearic acid, 0.5 g of sorbitol, 0.5 g of Molecular Sieves 3A, 5 ml of tertiary butyl alcohol, and 0.1 g of lipase-PL679 powder. . As a result, 3 mg of sorbitol stearate was obtained.
The IR spectrum of this compound is shown in Table 5 (Run No. 10). Furthermore, fatty acid esters of sorbitol were obtained in the same manner as above using other fatty acids. Its IR
The spectra are shown in Table 5 (Run No. 1 to 9 and No.
11-18). In addition, fatty acid ester of glucono-δ-lactone was obtained in the same manner as above using glucono-δ-lactone and fatty acid. The 5th IR spectrum
Shown in the table (Run No. 19-23).

【table】

[Table] From the results in Table 5, the absorption of ester bonds is seen around 1740 Kaiser and the absorption of sugar is seen around 3400 Kaiser, so the above product is a fatty acid ester of sorbitol and a fatty acid ester of glucono-δ-lactone. I understand that there is something. Example 4 A reaction was carried out in the same manner as in Example 1 using 0.5 g of methyl oleate, 0.5 g of fructose, 5 ml of tertiary butyl alcohol, and 0.1 g of lipase-PL679 powder.
I did some refining. As a result, 3 mg of fructose oleate was obtained. The 6th IR spectrum of this compound
It is shown in the table (Run No. 1). Furthermore, fructose fatty acid esters were obtained in the same manner as above using other fatty acid esters. Its IR spectrum is shown in Table 6 (Run No. 2~
14).

【table】

[Table] From the results in Table 6, it can be seen that the above product is a fructose fatty acid ester because absorption of ester bonds is observed around 1740 Kaiser and absorption of sugar is seen around 3400 Kaiser. Example 5 A reaction was carried out in the same manner as in Example 1 using 0.5 g of methyl oleate, 0.5 g of sorbitol, 5 ml of tertiary butyl alcohol, and 0.1 g of lipase-PL679 powder.
I did some refining. As a result, 3.5 mg of sorbitol oleate was obtained. The IR spectrum of this compound is shown in Table 7 (Run No. 1). Furthermore, sorbitol fatty acid esters were obtained in the same manner as above using other fatty acid esters. Its IR spectrum is shown in Table 7 (Run No. 2~
14).

【table】

[Table] From the results in Table 7, it can be seen that the above product is a sorbitol fatty acid ester because absorption of ester bonds is observed around 1740 Kaiser and absorption of sugar is seen around 3400 Kaiser. Example 6 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, 15 ml of tertiary butyl alcohol
The mixture was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 30°C for 24 hours. The reaction solution from which insoluble materials were removed by centrifugation was concentrated using a rotary evaporator to remove the solvent. After adding 20 ml of chloroform to this residue and dissolving it, insoluble matter was removed by centrifugation. Iatrobeads 6RS-80100 (manufactured by Yatron Co., Ltd.) were filled with this supernatant and washed with chloroform in advance.
was adsorbed onto a column (2 x 30 cm). After washing this column with 200 ml of chloroform, fructose oleate produced in the above reaction was eluted with 100 ml of acetone. This acetone elution fraction was concentrated using a rotary evaporator and then dried under reduced pressure to obtain 410 mg of fructose oleate. A small amount of this fructose oleate was dissolved in chloroform and subjected to thin layer chromatography (TLC) in the same manner as described in Example 1 and detected with 50% sulfuric acid. Spots were detected, and it was confirmed from the mobility (Rf value) on TLC that these two spots were the same as spots 1 and 2 described in Example 1. Fructose oleate obtained as above: approx.
50mg of preparative TLC (Merck silica gel 60 No.
13895) and developed about 10 cm using a developing solvent of chloroform:methanol:acetic acid:water=80:10:8:2. After drying this plate,
Two components corresponding to spot 1 and spot 2 described in Example 1 were extracted with a solution of chloroform:methanol=2:1 while monitoring with UV light to obtain 32 mg of spot 1 component and 6 mg of spot 2 component. For the two types of samples obtained in this way,
The following operations were performed to determine the molar ratio of fructose and oleic acid constituting the fructose oleate. That is, after dissolving about 5 mg of the sample in 5 ml of chloroform, 1 ml each was taken into two capped test tubes (referred to as A and B). After removing chloroform under reduced pressure for both A and B, 0.1N caustic soda/90
Add 1 ml of % methanol solution, cap, and
Heated at ℃ for 3 minutes. Thereafter, 0.1 ml of 1N hydrochloric acid was added to A to neutralize it. This sample was dried under reduced pressure and then dissolved in water to make 2 ml. The fructose contained in this solution was extracted using an enzymatic method (Hubergmeyer, ed., Methods of
Enzymatic Analysis, 3rd edition, Volume 6, p321,
Verlag Chemie GmbH, Weinheim, 1984). On the other hand, B is contained in the sample in accordance with the method for preparing 2,4,20,2-77 fatty acid methyl esters (boron trifluoride-methanol method) of the Standard Oil and Fat Analysis Test Methods (edited by Japan Oil Chemists' Association). The resulting oleic acid was converted into methyl oleate, and then quantified by gas chromatography using methyl stearate as an internal standard. In this way, the molar ratio of fructose and oleic acid constituting both components of spot 1 and spot 2 was determined, and for spot 1, the ratio was approximately 1:1, which is a monoester of fructose oleate. It turned out to be. Similarly, for spot 2, the ratio was approximately 1:2, and it was found that this was a diester of fructose oleate. In addition, 5 mg of fructose oleate was added to chloroform.
Dissolve in 0.5 ml, charge 1 microliter of the solution to Chromatrot SII (silica gel rod, manufactured by Yatron Co., Ltd.) of Iatro Scan (manufactured by Yatron Co., Ltd.), and mix chloroform: methanol: acetic acid:
It was developed for about 10 cm using a developing solvent of water = 80:10:8:2. Thereafter, this lot was subjected to Iatroscan, and two peaks, fructose monooleate and fructose dioleate, were obtained, and the weight ratio of these peaks was about 5:1 based on the ratio of peak areas. Example 7 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, and 20 ml of acetonitrile were placed in a 50 ml Erlenmeyer flask and shaken back and forth at 30° C. for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 427 mg of fructose oleate. Example 8 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, 2 g of molecular sieves 3A,
30 ml of tertiary amyl alcohol was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. From then on,
Purification was carried out in the same manner as in Example 6 to obtain 382 mg of fructose oleate. Example 9 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, and 25 ml of acetone were placed in a 50 ml Erlenmeyer flask and shaken back and forth at 45° C. for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 396 mg of fructose oleate. Example 10 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, 2 g of molecular sieves 3A,
25 ml of diacetone alcohol was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 45°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 280 mg of fructose oleate. Example 11 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, 20 ml of tertiary butyl alcohol,
5 ml of N,N-dimethylformamide was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 50°C for 24 hours.
Thereafter, the product was purified in the same manner as in Example 6 to obtain 287 mg of fructose oleate. Example 12 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, 10 ml of acetonitrile, and 5 ml of dimethyl sulfoxide were placed in a 50 ml Erlenmeyer flask.
It was shaken back and forth at 60°C for 24 hours. Thereafter, it was purified in the same manner as in Example 6 to obtain 371mg of fructose oleate.
I got it. Example 13 Fructose 1g, Sebacic acid 5g, Lipase-PL266 powder 100mg, Molecular sieves 3A2
g. 15 ml of tertiary butyl alcohol was placed in a 50 ml Erlenmeyer flask, and the mixture was shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 50 mg of fructose sebaccinate. Example 14 1 g of fructose, 2 g of oleic acid, 50 mg of lipase-PL266 powder, molecular sieves 3A2
g. 15 ml of tertiary butyl alcohol was placed in a 50 ml Erlenmeyer flask, and the mixture was shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 385 mg of fructose oleate. Example 15 1 g of fructose, 6 g of methyl oleate, 150 mg of lipase-PL266 powder, and 15 ml of tertiary butyl alcohol were placed in a 50 ml Erlenmeyer flask and heated at 40°C for 24 hours.
I shook it back and forth for hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 537 mg of fructose oleate. Example 16 1 g of fructose, 3 g of stearic acid, 200 mg of lipase-PL266 powder, molecular sieves
2 g of 3A and 15 ml of tertiary butyl alcohol were placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 372 mg of fructose stearate. Example 17 1 g of fructose, 5 g of palmitic acid, 100 mg of lipase-PL266 powder, molecular sieves
2 g of 3A and 15 ml of tertiary butyl alcohol were placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 223 mg of fructose palmitate. Example 18 1 g of fructose, 4 g of lauric acid, 100 mg of lipase-PL266 powder, 15 tertiary butyl alcohol
ml was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 272 mg of fructose laurate. Example 19 1 g of fructose, 3 g of ricinoleic acid, 100 mg of lipase-PL266 powder, molecular sieves
2 g of 3A and 15 ml of tertiary butyl alcohol were placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 352 mg of fructose ricinoleate. Example 20 1g ribose, 6g oleic acid, lipase
PL679 powder 100mg, tertiary butyl alcohol 15ml
The mixture was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 12 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 250 mg of ribose oleate. Example 21 1g glucose, 6g oleic acid, lipase
100g of PL679 powder, 2g of molecular sieves 3A,
15 ml of tertiary butyl alcohol was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. From then on,
Purification was carried out in the same manner as in Example 6 to obtain 212 mg of glucose oleate. Example 22 1 g of glucoheptose, 6 g of oleic acid, 100 g of lipase-PL679 powder, and 15 ml of tertiary butyl alcohol were placed in a 50 ml Erlenmeyer flask, and heated to 48°C at 40°C.
I shook it back and forth for hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 62 mg of glucoheptose oleate. Example 23 Maltose 1g, oleic acid 6g, lipase
PL679 powder 100mg, tertiary butyl alcohol 15ml
was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 72 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 41 mg of maltose oleate. Example 24 Methyl-α-glucoside 1g, oleic acid 6
g, Lipase-PL679 powder 100mg, Molecular Sieves 3A 2g, tertiary butyl alcohol 15ml.
The mixture was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 258 mg of methyl-α-glucoside oleate. Example 25 Glucono-δ-lactone 1g, oleic acid 6
Take 100 mg of lipase-PL679 powder and 15 ml of tertiary butyl alcohol into a 50 ml Erlenmeyer flask, and add 40 g.
The mixture was shaken back and forth at ℃ for 36 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 134 mg of glucono-δ-lactone oleate. Example 26 Sorbitol 1g, oleic acid 6g, Lipase-PL679 powder 100mg, tertiary butyl alcohol 15
ml and 2 g of Molecular Sieves 3A were placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 620 mg of sorbitol oleate. Example 27 Mannitol 1g, oleic acid 6g, Lipase-PL679 powder 100mg, tertiary butyl alcohol 15
ml was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 325 mg of mannitol oleate. Example 28 1 g of fructose, 6 g of oleic acid, 70 mg of lipase-AL powder, 15 ml of tertiary butyl alcohol,
2 g of Molecular Sieves 3A was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 30°C for 24 hours. From then on,
Purification was carried out in the same manner as in Example 6 to obtain 480 mg of fructose oleate. Example 29 Fructose 1g, oleic acid 6g, Lipase-PL679 powder 100mg, tertiary butyl alcohol 15
ml and 2 g of Molecular Sieves 3A were placed in a 50 ml Erlenmeyer flask and shaken back and forth at 30°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 620 mg of fructose oleate. Example 30 1 g of fructose, 6 g of oleic acid, 100 mg of lipase-PL679 powder, 15 tertiary butyl alcohol
ml was placed in a 50 ml Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 416 mg of fructose oleate. Example 31 1 g of lipase-PL679 powder was placed in a 100 ml Erlenmeyer flask, 20 ml of distilled water was added to dissolve it, 1 g of bentonite was added, and the mixture was stirred at room temperature for 30 minutes. Bentonite was collected by centrifugation, 30 ml of acetone was added thereto, stirred and suspended, and bentonite was collected again by centrifugation. Thereafter, this operation was repeated twice and then dried under reduced pressure. 50ml of 100mg of Lipase PL679 adsorbed and immobilized on bentonite using the above method, 1g of fructose, 6g of oleic acid, and 15ml of tertiary butyl alcohol.
The mixture was placed in an Erlenmeyer flask and shaken back and forth at 40°C for 24 hours. Thereafter, the product was purified in the same manner as in Example 6 to obtain 526 mg of fructose oleate.

Claims (1)

[Claims] 1 Number of carbon atoms having at least 2 or more hydroxyl groups
_C5 - _C7 monosaccharides (these monosaccharides include methyl, phenyl, nitrophenyl, phenol, hydroxymethylphenyl, mercapto, halogen, acetylamino, amino, uracil, thymine, or a disaccharide consisting of hexose (which may be substituted with an adenine group), carbon number C ₄
A saccharide selected from the group consisting of ~ _C6 sugar alcohols, _C6 - _C7 sugar lactones, and saturated or unsaturated _C2 - _C22 fatty acids (these fatty acids are
(which may be substituted with a hydroxyl group, a carboxyl group, or a phenyl group) or a C ₁ to C ₄ lower alcohol and the above fatty acid forming an ester, an organic solvent (excluding primary alcohol solvents) 1. A method for producing a saccharide fatty acid ester compound, which comprises allowing alkaline lipase produced by a microorganism belonging to the genus Achromobacter or the genus Alcaligenes to act in the presence of the compound, with or without dehydration.