JPH04258291A - Immobilized enzyme and its production - Google Patents

Abstract

PURPOSE:To obtain an immobilized enzyme suitable for synthesis and exchange reaction of fats and oils and ester of fatty acid derivatives, having high activity and excellent durability by bringing an aqueous solution of a lipid hydrolyzing enzyme into contact with an immobilized carrier composed of a specific synthetic adsorbent. CONSTITUTION:An aqueous solution of a lipid hydrolyzing enzyme (e.g. lipase or phospholipase) is brought into contact with an immobilized carrier of a synthetic adsorbent composed of an acrylic resin having pores to give an immobilized enzyme. A methacrylic alkyl ester-based resin having 150-200Angstrom pore diameter is preferable as the acrylic resin. Immobilization of the enzyme is carried out at 25-40 deg.C, pH 3-9. When the synthetic adsorbent is subjected to adsorption treatment with an oil-soluble compound such as oleic acid previously or simultaneously with the immobilization, preferably an immobilized enzyme having high activity and high durability is obtained.

Description

[Detailed description of the invention]

0001

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an immobilized enzyme suitable for the synthesis and exchange reaction of esters of fats and oils and fatty acid derivatives, and a method for producing the same.

Ester synthesis reaction is an important technology for the synthesis of alcohol fatty acid esters from alcohol and fatty acids, the synthesis of polyhydric alcohol fatty acid esters such as monoglycerides, polyglycerol fatty acid esters, and sugar esters, and the production method of fragrances such as geranyl butyrate. be.

On the other hand, transesterification of oils and fats is an important technology along with hydrogenation for the modification of edible processed oils and fats such as margarine and shortening.

0004

[Prior art and problems to be solved by the invention] In recent years,
BACKGROUND ART Research and industrialization of synthesis and exchange reactions of fats and oils and esters using lipase as an enzyme have become active in various fields. This takes advantage of the fact that lipase reacts under mild conditions and has regioselectivity and alkyl selectivity. However, these reactions differ from the hydrolysis reactions inherent to lipase, and can proceed only in a system with limited moisture. On the other hand, in order to increase the ester synthesis and exchange activities of lipase, water is required as an enzyme. For example, in the low-moisture system reaction disclosed in JP-A-55-71797, a sufficient reaction rate cannot be obtained, and if more water than necessary is added to increase the reaction rate, the ester decomposition reaction is There is a problem with the process being prioritized. Furthermore, as disclosed in JP-A-60-19495 and JP-A-60-203190, a method has been proposed in which the reaction is divided into two stages: a step of decomposing a polyhydric system and a synthesis step of removing water. However, the synthesis reaction rate of the latter cannot be said to be sufficient compared to the usual transesterification rate, and the process operation becomes unavoidably complicated.

[0005] In order to solve the above problems and use lipase efficiently, attempts have been made to immobilize lipase. The expected advantages of lipase immobilization are as follows. That is, (i) conventionally, when lipase was used in the form of an aqueous solution, it was difficult to mix and disperse it uniformly in oil; however, by immobilizing lipase on the surface of an insoluble carrier, it can be easily dispersed in oil. In addition, since an appropriate amount of water can be retained in the carrier, ester synthesis and exchange reactions can be easily carried out under low moisture conditions. (ii) Lipase, which is expensive as a catalyst, can be easily recovered and reused, and a reaction apparatus can be easily made continuous even in industrial implementation of ester synthesis reactions or exchange reactions.

However, even with the immobilized enzymes having the above-mentioned advantages, it is necessary to maintain both the water content necessary for increasing lipase synthesis activity and the suppression of hydrolysis, which is the reverse reaction. Not yet reached. For example, Jour
al of American Oil Chemist
'sSociety, Volume 60, 291-29
4 (1983) also points out that the hydrolysis reaction proceeds when a small amount of water is added. Further, when a polyhydric alcohol such as glycerin is added instead of water, the hydrolysis reaction is suppressed to some extent, but the ester synthesis/exchange reaction becomes extremely slow. In addition, in order to retain enzyme moisture, a method using a porous carrier and a superabsorbent resin comprehensively bonded with chitosan and then pulverized (Japanese Unexamined Patent Publication No. 59-213
390) also employs a two-step reaction method (Japanese Patent Laid-Open No. 60-203196) in order to achieve both ester synthesis/exchange reaction and decomposition reaction of the immobilized enzyme. However, such a method has disadvantages in that the operation is complicated and it is difficult to suppress diglyceride as a by-product.

As described above, in ester synthesis and exchange reactions, it is desired to develop immobilized enzymes with higher ester synthesis and exchange activities and excellent durability.

Conventionally, inert carriers such as celite, alumina, and diatomaceous earth have been mainly used as adsorbents for immobilization. When transesterification is carried out using immobilized enzymes immobilized on these carriers, the reaction can be carried out without a solvent in a batch reaction, but in the case of a column-packed reactor, the use of a solvent is required due to the high pressure drop that occurs. was necessary. In order to avoid excessive pressure drop and obtain a final product that is optimal for column operation, it is desirable to have an adsorption carrier with a relatively large average particle size, as well as a highly durable support for enzyme immobilization with pressure resistance. was.

The present inventors have previously conducted extensive research on factors that increase the ester synthesis and transesterification activities of lipase. As a result, they added oil to lipase disclosed in JP-A-60-25188 to initiate a hydrolysis reaction. In a method of obtaining a highly active immobilized enzyme by immobilizing the enzyme while the enzyme is immobilized in the presence of fatty acids, fatty acid derivatives, alcohols, or phospholipids, it has been found that ester synthesis and transesterification activities are increased. A patent application was filed (Japanese Unexamined Patent Publication No. 1-153090). However, enzymes immobilized in this manner still have insufficient heat resistance and durability.

0010

[Means for Solving the Problems] The present inventors have solved the above problems, strengthened the bond between the enzyme and the carrier, prevented the desorption of the coexisting fatty acids, and made various efforts to obtain a more durable immobilized enzyme. As a result of investigation, it was discovered that a good immobilized enzyme could be obtained by using a synthetic adsorbent made of an acrylic resin having pores as an immobilization carrier, and the present invention was completed.

Specifically, the present invention provides an immobilization method characterized in that a synthetic adsorbent made of an acrylic resin having pores is used as an immobilization carrier, and an aqueous solution of a lipolytic enzyme is brought into contact with the immobilization carrier. The present invention provides an enzyme and a method for producing the immobilized enzyme.

The acrylic resin used in the present invention includes (meth)acrylic acid-based, (meth)acrylonitrile-based, (meth)acrylic ester-based, (meth)acrylamide-based resins, and the like. Among these, preferred acrylic resins are (meth)acrylic ester resins, with methacrylic ester resins being particularly preferred. As methacrylic acid ester resin,
Methacrylic acid alkyl ester-based resins are preferred, and examples thereof include methacrylic acid methyl ester-based, methacrylic acid ethyl ester-based, methacrylic acid propyl ester-based, methacrylic acid isopropyl ester-based, and methacrylic acid butyl ester-based resins.

The immobilization carrier used in the present invention has various shapes such as powder and granules, and any of them can be used. Regarding the immobilization of enzymes, it is generally said that increasing the number of hydrophilic sites on the carrier and increasing its surface area will increase the amount of enzyme bound per carrier and will often result in highly active immobilized enzymes. Considering this ("Immobilized Biocatalyst", edited by Jiro Chibata, p. 41), a porous material with a large specific surface area is also suitable for the present invention. Therefore, when the average particle size of the carrier is 0.35 to 0.4 mm, 150 to 2
Those having a pore diameter of 0.00 Å are preferred. As such an immobilization carrier, for example, Diaion HP2MG manufactured by Mitsubishi Chemical Corporation is used as a methacrylic acid ester resin.
(registered trademark) etc. are used. The average particle size or pore size of the carrier can be measured by a conventionally known measuring method.

[0014] Lipid degrading enzymes used in the present invention include lipase, phospholipase, cholesterol esterase, sphingomyelinase and various esterases. Among these lipases, Rhizopus and Aspergillus, which react only to the 1st and 3rd positions of glycerides and have excellent regioselectivity, are used as lipases.
rgillus) genus, Mucour genus,
Geotrich with fatty acid specificity
um), genus Candi showing no specificity
da) Genus, Pseudomonas
) Genus Penicillium Genus Chromobacterium
m) Lipases of microbial origin, such as those of the genus, and animal lipases, such as pancreatic lipase. Among these, as lipases whose synthetic activity is particularly likely to increase, lipases originating from the genus Rhizopus, Mucor, and Chromobacterium, which have strong activity against medium-chain or larger alkyl groups, are more preferable. An example of cholesterol esterase is Candida
Examples include those of microbial origin such as the genus Dida). Examples of phospholipases include phospholipase D derived from plants such as cabbage, peanuts, carrots, soybeans, and rapeseed, and moss plants, phospholipase D and phospholipase C derived from microorganisms such as Streptomyces, and phospholipase A derived from yeast. , phospholipase A from viper
2 etc.

[0015] In the present invention, the enzyme is immobilized using the porous synthetic adsorbent described above, preferably modified with fatty acids, fatty acid derivatives, phospholipids, etc. that have affinity for the enzyme. This is carried out by equilibrating the enzyme at a stable pH, bringing it into contact with an aqueous enzyme solution, and adsorbing the enzyme. The enzyme aqueous solution is a solvent of monohydric alcohol or polyhydric alcohol having 1 to 6 carbon atoms, sodium chloride, ammonium sulfate, etc.
It may also be a mixed solution of salts commonly used as enzyme treatment agents.

[0016] In the present invention, the temperature for immobilization may be any temperature that does not cause deactivation of the lipolytic enzyme;
-60°C, preferably 25-40°C. Further, the pH of the lipolytic enzyme aqueous solution may be within a range that does not cause denaturation of the lipolytic enzyme, and preferably pH 3 to 9. In particular, when using a lipase whose optimum pH is acidic, the pH is preferably 4 to 6 in order to obtain maximum activity. In addition, the type of buffer used for the enzyme aqueous solution is not particularly limited, but
Common acetate buffers, phosphate buffers, Tris-HCl buffers, etc. can be used.

[0017] When immobilizing an enzyme according to the present invention, the concentration of the enzyme in the aqueous solution is not particularly specified, but from the viewpoint of immobilization efficiency, it is desirable that the concentration be lower than the solubility of the lipolytic enzyme and be sufficient. Furthermore, if necessary, the insoluble portion may be removed by centrifugation and the supernatant may be used. Further, the proportion (weight ratio) of the enzyme and the immobilization carrier used is preferably 0.01 to 1 part of the enzyme per 1 part of the immobilization carrier, but is not particularly limited thereto.

[0018] Also, before or at the same time as immobilization, the synthetic adsorbent may be mixed with one or more types selected from fatty acids, fatty acid derivatives, phospholipids, alcohols, ethers, carbonyl compounds, and alkyl halides. Highly active and highly durable immobilized enzymes can be obtained by adsorption treatment with oil-soluble compounds. At this time, in order to prevent contamination with impurities, it is preferable to perform pretreatment, that is, dissolve these oil-soluble compounds in a volatile solvent, bring this solution into contact with a synthetic adsorbent, filter it, and then dry it. The appropriate ratio of the oil-soluble compound to the immobilization carrier is 0.001 to 1 part by weight of the oil-soluble compound to 1 part by weight of the immobilization carrier, but is not limited thereto. An excessive amount of the oil-soluble compound is not adsorbed on the immobilization carrier, but is released into the solution and adsorbs the enzyme, resulting in a decrease in the yield of immobilization on the immobilization carrier, and is therefore not effective. A suitable adsorption temperature is 0 to 60°C, preferably 5 to 30°C. A suitable adsorption time is 5 minutes to 2 hours. Temperatures above
The times are not limited to these.

The fatty acids used in the synthetic adsorbent treatment in the present invention include linear saturated fatty acids, unsaturated fatty acids, and branched fatty acids having 4 to 24 carbon atoms. Preferred fatty acids include, for example, oleic acid, ricinoleic acid, and linoleic acid.

Suitable fatty acid derivatives used in the synthetic adsorbent treatment in the present invention include those in which the fatty acid residue has 8 to 24 carbon atoms.
Linear saturated fatty acids, unsaturated fatty acids, or branched fatty acids such as monoglycerides, diglycerides, and their derivatives, triglycerides, or polyhydric alcohol fatty acid esters such as propylene glycol and polyglycerin, sugar esters such as sucrose fatty acid esters, and sorbitan fatty acids. Examples include sugar alcohol esters such as esters.

[0021] The alcohols used in the synthetic adsorbent treatment in the present invention include linear or branched aliphatic monohydric alcohols having 8 to 24 carbon atoms and polyhydric alcohols having 2 to 6 carbon atoms. In addition, phenolic compounds, sterols, terpene alcohols having 10 to 20 carbon atoms, and fat-soluble vitamins are also effective.

Examples of ethers used in the synthetic adsorbent treatment in the present invention include ethers having 10 to 18 carbon atoms;
Examples include glyceride-like compounds such as glyceryl ethers having 12 to 18 carbon atoms or glycidyl ethers having 10 to 18 carbon atoms, polyoxy compounds, and silicon compounds of the alcohols.

Examples of carbonyl compounds used in the synthetic adsorbent treatment in the present invention include aliphatic aldehydes having 10 to 18 carbon atoms and aliphatic ketones.

Examples of the alkyl halide used in the synthetic adsorbent treatment in the present invention include alkyl halides having 8 to 18 carbon atoms.

[0025] All of the above oil-soluble compounds are preferably liquid at room temperature from the viewpoint of process operation, but are not limited thereto. In addition, these may be used alone, but
Even better effects may be achieved by appropriate combinations.

Examples of the reaction of lipids using the immobilized enzyme obtained in the present invention include transesterification using immobilized lipase, and examples of such transesterification include:
Examples include acidolysis reactions between esters and fatty acids, alcoholysis reactions between esters and alcohols, and interesterification reactions between esters.

Examples of substrates for transesterification using the immobilized enzyme obtained in the present invention include vegetable oils such as soybean oil, olive oil, and palm oil, and animal fats and oils such as beef tallow, lard, and fish oil. . Although these fats and oils may be used alone, it is preferable to use two or more types of fats and oils, or to perform transesterification between fats and oils and higher fatty acids, or between fats and oils and lower alcohol esters of higher fatty acids. When transesterifying a specific fat and oil with another fat, fatty acid, or a derivative thereof, the ratio of the amount of the other substance to 1 part by weight of the specific fat is 0.05 to 20 parts by weight, preferably 0.1 to 10. If it is not in parts by weight, it is difficult to obtain the effect of modifying fats and oils. Particularly preferred is transesterification of an oil or fat having a large number of oleic acid residues at the 2-position, such as palm oil, with stearic acid. In this reaction, the melting point of stearic acid is high and the viscosity of fats and oils is high, so in order to carry out the continuous transesterification reaction in a column reaction without a solvent, it is necessary to maintain the temperature of the reaction system at 60 to 90°C. The immobilized enzyme of the invention is suitable for this purpose.

Other examples of transesterification using the immobilized phospholipase obtained in the present invention include natural phospholipids and various aliphatic alcohols, polyhydric alcohols, terpene alcohols, sugars, sugar alcohols, sterols, etc. Other examples include transphosphatidylation with bases such as guanine, adenine, thymine, and uracil.

Furthermore, examples of ester synthesis reactions using the immobilized enzyme obtained in the present invention include ordinary monohydric alcohols such as methanol, ethanol, propanol, and oleyl alcohol, propylene glycol, glycerin, sorbitol, and polyglycerin. Examples include esterification reactions between polyhydric alcohols, terpene alcohols such as geraniol, citronellol, and menthol, or sterols such as cholesterol, and fatty acids having 2 to 24 carbon atoms.

The ester synthesis reaction is carried out at 20 to 90°C, more preferably 30 to 80°C, without a solvent or in an inert solvent such as a hydrocarbon or ether. Further, the amounts of alcohol and fatty acid are adjusted as appropriate depending on their valences and the intended product. For example, when the purpose is to synthesize diglyceride, 1 mole of glycerin is reacted with about 2 moles of fatty acid, and when the purpose is to synthesize monoglyceride, 1 mole of glycerin is reacted with about 1 mole of fatty acid.

[0031] In these reactions such as transesterification, esterification, and transphosphatidylation, the amount of water in the reaction system, including the amount of water in the immobilized enzyme, should be kept at 5% by weight or less, preferably It is preferable to keep it at 0.1 to 1% by weight.

[0032] The immobilized lipolytic enzyme obtained in the present invention can also be suitably used in the hydrolysis reaction of oils and fats or various lipids by utilizing the inherent properties of the lipolytic enzyme.

[0033]

[Examples] The present invention will be explained in more detail with reference to Examples below, but the present invention is not limited to these Examples.

Example 1 Commercially available synthetic adsorbent (methacrylic acid ester resin, trade name: Diaion HP2MG, manufactured by Mitsubishi Kasei Corporation,
Pore diameter 150 ~ 200 Å, average particle size 0.35 ~
0.4 mm) was dissolved in 100 ml of 500 mM acetate buffer at pH 5, and stirred for 2 hours. After the resin was filtered off, the same operation was performed with a 50 mM acetate buffer at pH 5 for equilibration, and after drying under reduced pressure, it was used as an immobilization carrier.

Furthermore, 10 g of the resin equilibrated by the above method was further added with 2 g of ricinoleic acid in a pH 550 mM acetate buffer.
The resin was added to a solution dispersed in 0 ml and stirred for 30 minutes, and the resin was filtered from the solution, washed with the buffer, dried under reduced pressure, and used as a support for immobilization.

[0036] 10 g of these resins were added to commercially available lipase (Rhizopus japonicus).
onicus) origin lipase preparation, trade name: Lilipase A10, manufactured by Nagase Seikagaku Kogyo Co., Ltd., 35,000
Unit/g) was added to a solution of 100 ml of 50 mM acetate buffer at pH 5, and stirred for 2 hours. The resin was then filtered from the solution and washed with the buffer. At this time, it was found from the lipase activity of the filtrate that 80% of the lipase was immobilized in the case without treatment with ricinoleic acid, and 98% in the case of treatment with ricinoleic acid.

After washing, drying was performed under reduced pressure to obtain an immobilized enzyme. This immobilized enzyme was evaluated by the following method.

[0038] Immobilized enzyme (moisture 5%) 1 g, palm oil medium melting point (iodine value 32.5, diglyceride content 4.
6%) 8 g and stearic acid (93% purity) 12 g
After reacting at 70°C, the amount of stearic acid contained in the triglyceride was analyzed by gas chromatography, and the reaction rate was calculated using the following formula.

[0039]

[Math 1]

St: Concentration of stearic acid 6.4: Concentration of stearic acid originally contained in the melting point part of palm oil The reaction rate and diglyceride production rate were determined from sampling data at the third hour of the reaction time. The results are shown in Table 1.

Comparative Example 1 In Example 1, a commercially available synthetic adsorbent (Diaion H
Celite 545 (P2MG), an inorganic carrier, was used instead of Celite 545 (P2MG).
An immobilized enzyme was obtained by carrying out the same operation as in Example 1, except that the enzyme (manufactured by Marine Building Co., Ltd.) was used.

[0042] Using the obtained immobilized enzyme, the same transesterification reaction as in Example 1 was carried out. The reaction rate and diglyceride production rate were determined from sampling data obtained at the third hour of the reaction time. The results are shown in Table 1.

[0043]

[Table 1]

Example 2 Using the resin treated with ricinoleic acid shown in Example 1, in addition to Lilipase A10 used in Example 1 as the enzyme, a commercially available lipase (Rhizopus deremer) was used.
using a lipase preparation originating from Rhizopus javanicus (trade name: Lipase D, manufactured by Amano Pharmaceutical Co., Ltd.) and a lipase preparation originating from Rhizopus javanicus (trade name: Lipase F, manufactured by Amano Pharmaceutical Co., Ltd.). Immobilization was carried out in the same manner as in Example 1 (immobilization conditions: 200,000 units/g of resin), and the obtained immobilized enzyme was evaluated by the following method.

That is, once the transesterification reaction has been carried out to the end point of the reaction, it is stored as it is at 70°C for 100 hours, the immobilized enzyme is recovered, washed with hexane, and dried.
The transesterification reaction was carried out again under the same conditions, and the activity at 3 hours of the reaction was evaluated (reaction conditions: 70°C, weight ratio of fatty acid (stearic acid) contained in the reaction system to palm oil middle melting point part FA/OIL = 1 .5). The results are shown in Table 2.

[0046]

[Table 2]

(Upper row: reaction rate, lower row: diglyceride production rate) Example 3 A ricinoleic acid-treated resin was prepared in the same manner as in Example 1, and enzymes were immobilized. A diglyceride synthesis reaction was performed on the obtained immobilized enzyme as follows.

[0048] 300 g of rapeseed fatty acid in a 3-necked flask;
Add 30 g of the above immobilized enzyme (dry weight: enzyme water content 3% or less), and stir at 180 rpm while reducing the pressure (3 mmHg) with a vacuum pump until no foaming occurs to remove excess water. Add 48.8g of glycerin and adjust to 3mmH.
g Start the reaction under vacuum and stirring at 180 rpm. Sampling is performed every hour to measure oxidation (AV) and moisture, and the end point is when the AV becomes 20 or less. The esterification rate and selectivity at 3 hours of reaction were determined using the following formulas, and the immobilized enzyme was evaluated. The results are shown in Table 3.

[0049]

[Math 2]

[0050]

[Table 3]

[0051]

Effects of the Invention In the present invention, by using a synthetic adsorbent made of an acrylic resin having pores as an immobilization carrier, it is desired that the adsorbent be removed from the enzyme solution due to its affinity with the lipolytic enzyme. The impurities that were present are not adsorbed by the adsorbent, and only the lipolytic enzymes are specifically adsorbed. Therefore, it is possible to immobilize directly from a crude enzyme solution, and moreover, it has a high enzyme adsorption rate.

[0052] Furthermore, after adsorption of the enzyme, the lipolytic enzyme is very stable on the adsorbent, does not deactivate, does not desorb during the reaction, and has a markedly improved durability.

Since the synthetic adsorbent of the present invention has a relatively large average particle size compared to conventional adsorbents, for example, in the case of a column-packed reactor, there is no excessive pressure drop and the column can be operated smoothly. .

Furthermore, the immobilized enzyme of the present invention has excellent heat resistance, so that the reaction can be carried out at a temperature of 50 to 80°C. Furthermore, in the case of transesterification, for example, by adjusting the water content, continuous transesterification can be carried out in an economically easy manner without using a solvent.

[0055] In particular, the immobilized lipase obtained by immobilizing the regioselective lipase by the method of the present invention has remarkable activity, and has a high activity due to the combination of fats and oils containing a large amount of oleic acid at the 2-position of glyceride and saturated fatty acids. When the aim is to produce a symmetrical fat with a structure similar to natural cacao butter by transesterification, it is possible to reduce the by-product of diglyceride and the transition to an asymmetric type, and the accompanying reduction of the by-product of trisaturated glyceride. It becomes possible.

Furthermore, in the synthesis of esters, in the conventional enzymatic method, the reaction reaches equilibrium due to water produced as the reaction progresses, so that esterification does not proceed. Therefore, attempts were made to further advance the esterification by dehydration operations such as reducing the pressure of the reaction system, but such operations inevitably resulted in a decrease in the ester synthesis activity of the enzyme. In such cases, when the immobilized lipase according to the method of the present invention is used, it retains sufficient ester synthesis activity even under low moisture conditions, so a high esterification rate can be achieved in a short period of time, and the reaction can last for a long time. It has the advantage that there is no deterioration in quality due to side reactions such as coloring due to chemical reaction and generation of off-flavors.

Claims (7)

[Claims] 1. An immobilized enzyme, characterized in that a synthetic adsorbent made of an acrylic resin having pores is used as an immobilization carrier, and an aqueous solution of a lipolytic enzyme is brought into contact with the immobilization carrier. 2. The immobilized enzyme according to claim 1, wherein the acrylic resin is a (meth)acrylic acid ester resin. 3. The immobilized enzyme according to claim 2, wherein the (meth)acrylate resin is a methacrylate resin. 4. The immobilized enzyme according to claim 1, wherein the lipolytic enzyme is selected from lipase, phospholipase, cholesterol ethylase, and sphingomyelinase. 5. An aqueous solution of a lipolytic enzyme,
It has a pore size of 200 Å and an average particle size of 0.35-0.4
1. A method for producing an immobilized enzyme, which comprises bringing the immobilized enzyme into contact with a synthetic adsorbent made of a methacrylic acid ester resin having a diameter of 1 mm. [Claim 6] In immobilizing a lipolytic enzyme,
Claim 5, characterized in that the immobilization is carried out in the presence of one or more compounds selected from fatty acids, fatty acid derivatives, phospholipids, alcohols, ethers, carbonyl compounds, and alkyl halides. Manufacturing method described. 7. A synthetic adsorbent made of a methacrylic acid ester resin having a pore diameter of 150 to 200 Å and an average particle diameter of 0.35 to 0.4 mm is coated with fatty acids, fatty acid derivatives, or phospholipids in advance. 7. The production method according to claim 5, wherein the insoluble carrier obtained by adsorption treatment and the lipolytic enzyme are adsorbed and immobilized in a water-insoluble medium or in a water-soluble or water-insoluble organic solvent.