JPH0365950B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for transesterification of oils and fats using lipase. Specifically, the present invention relates to a reaction method in which transesterification of oils and fats using lipase is carried out in two steps: a hydrolysis reaction and an ester synthesis reaction. Transesterification of oils and fats is an important technology along with hydrogenation in the production of processed oils and fats. Conventional transesterification reactions are carried out in the presence of inorganic catalysts such as sodium metal, but such chemical methods have the drawback of low regioselectivity for the binding site of the fatty acid to be exchanged. On the other hand, lipase (EC 3113), which is an enzyme that hydrolyzes fats and oils, is known to catalyze not only hydrolysis reactions but also ester synthesis reactions [M. Iwai, Y. Tsujisaka, J. Fukumoto, J. .Gen.
See App1. Microbiol. 10 , 13, (1964)]. The transesterification reaction of fats and oils using lipase has the advantage of not only the substrate specificity of the enzyme and high selectivity such as positional specificity, but also the fact that the reaction proceeds at room temperature and pressure, so it is also useful from the viewpoint of energy and resource conservation. This is to be expected. It is generally accepted that enzymatic reactions are carried out in aqueous solutions, but in the case of transesterification of fats and oils using lipase, hydrolysis reactions take precedence in a relatively water-rich reaction system, making it difficult to obtain a desirable reaction product. Have difficulty. Therefore, the transesterification reaction of fats and oils using conventionally known lipases is carried out in a reaction system with extremely low moisture content. For example, Tokukai Akira
No. 55-71797 describes a method in which the amount of water present in the reaction system is 0.18% by weight or less based on the substrate. Further, JP-A-52-104506 describes a method in which water is added in the presence of a small amount of water, or in the presence of 0.2 to 1% by weight based on the substrate. However, in a reaction system with extremely low water content, such as in the conventionally known method described above, the enzyme is not sufficiently hydrated and cannot take the optimal structure for reaction, so the enzyme is not fully activated and the reaction does not occur. The speed is also very slow. Furthermore, in preparing the enzyme composition, a complicated drying operation is required to remove excess water from the enzyme composition. For this reason, deactivation of the enzyme due to drying is unavoidable, and adjustments to drying time and moisture content are highly empirical, and stable reaction operations cannot be expected. Further, in repeated use of an enzyme composition, in a system with extremely low water content, the water content in the enzyme composition gradually decreases, so that the enzyme activity gradually declines. For this reason, it is necessary to re-add a very small amount of water before the reaction, but it is extremely difficult to control the amount. As mentioned above, while the transesterification reaction of fats and oils using lipase has advantages over chemical methods using inorganic catalysts, it also has many problems, and these are difficult to achieve for industrial use. It is necessary to solve the problems technically. The purpose of the present invention is to sufficiently activate the lipase in the reaction system, increase the reaction rate to the extent that industrialization is possible, and maintain a stable reaction in order to achieve industrial utilization of the transesterification reaction of oils and fats using lipase. The goal lies in the development of reaction operations to achieve this goal. Furthermore, another object of the present invention is to prevent inactivation of lipase within the reaction system and to enable effective reuse of lipase, thereby increasing the economic efficiency of the transesterification process. . In order to achieve this objective, the present inventors have conducted intensive research into the transesterification reaction of oils and fats using lipase, and as a result, have discovered a reaction operation method that can maximize the functions of lipase. Ta. Regarding lipase, the pioneering research of Tsujisaka, Iwai et al. [for example, ( 1 ) J.Gen.Appl.Microbiol.
(1964), (2)Biochem.Biophys.Acta. 489 , 415
(1977), (3) ibid, 575 , 156, (1979), and (4)
Agric.Biol.Chem. 40 , 655, (1976), etc.], it has been demonstrated that it has regiospecificity and can be used as a catalyst for ester synthesis reaction, which is the reverse reaction of hydrolysis. Among them, the position specificity of glycerol in ester synthesis is consistent with the position specificity in hydrolysis, and in glyceride synthesis from glycerol and fatty acids by lipase, the water content in the reaction system controls the final synthesis rate. It has been experimentally proven that Based on these facts, the present inventors analyzed the transesterification reaction of oils and fats from a reaction industrial perspective, and found that the transesterification reaction rate r of oils and fats is basically expressed by the following equation. I found it. r=k[DG][FA] Here, k is the overall reaction rate constant, [DG] is the diglyceride concentration, and [FA] is the fatty acid concentration. Further, k largely depends on the water content in the reaction system. Regarding the transesterification reaction rate of fats and oils, there have been few kinetic research reports to date. The present inventors conducted basic research and examination on the rate of transesterification of oils and fats using a simplified system, that is, a system consisting of trilaurin and capric acid. As a result of computer-based analysis using reaction rate equations based on all possible reaction paths that correspond to changes in composition over time, it was found that a reaction in which triglyceride and fatty acid directly exchange fatty acid groups and generate new triglyceride cannot occur. It was concluded that no.
On the other hand, assuming a reaction pathway in which a new triglyceride is generated by esterification of diglyceride and fatty acid, that is, assuming that diglyceride is an intermediate in the transesterification reaction, the experimental values and calculated values agree very well, and the above basic We were able to derive a reaction rate equation. Furthermore, as a result of detailed studies on the esterification reaction of fatty acids and alcohols by lipase, they discovered that although this esterification reaction progresses extremely quickly, once the ester is produced, it is hardly hydrolyzed. The method for transesterification of fats and oils using lipase of the present invention is based on the knowledge that diglyceride is an intermediate in the transesterification reaction of fats and oils. The basic idea is to incorporate it into the reaction and artificially control the reaction equilibrium. The method for transesterification of fats and oils using lipase of the present invention is characterized in that the transesterification reaction of fats and oils is carried out in two stages, and alcohol is added to the reaction system. This reaction is mainly a hydrolysis reaction of fats and oils by lipase, and the second stage reaction is mainly a glyceride ester synthesis reaction by lipase.
Incidentally, the two-stage reaction described above can be performed in a continuous operation. Moreover, in order to increase the reaction yield, a multi-stage tank type operation can also be adopted. The alcohol used in the present invention is preferably an aliphatic alcohol having 4 to 18 carbon atoms, and particularly preferred among these are butyl alcohol, hexyl alcohol, octyl alcohol, decyl alcohol, and the like. In the present invention, alcohol may be added only to the first reaction stage or only to the second reaction stage, but it is preferably added to both the first reaction stage and the second reaction stage. In the first stage reaction, that is, the reaction stage mainly consisting of hydrolysis reaction, alcohol is preferably added from the beginning of the reaction, and by adding such alcohol, free fatty acids generated by hydrolysis become alcohol esters. Since the alcohol ester produced is hardly hydrolyzed, the hydrolysis of glycerides is greatly accelerated. The amount of alcohol added in the first stage reaction is preferably equal to or less than the amount of free fatty acid produced by hydrolysis, preferably 50 to 90 mol% of the amount of free fat expected to be produced. . In addition, in the second stage reaction, that is, the reaction stage mainly consisting of ester synthesis reaction, a fatty acid was added to esterify with a partial glyceride mainly consisting of diglyceride, to generate triglyceride, and obtain the desired triglyceride composition. Alcohol is then added to convert most of the remaining free fatty acids into alcohol esters. At this time, the amount of alcohol added in the second stage reaction may be equal to or less than the amount of free fatty acids remaining after triglyceride production, and is preferably 50 to 90 mol% of the amount of free fatty acids. Converting free fatty acids into alcohol esters in this way has the advantage that triglycerides can be very easily recovered after the reaction. In order to stabilize and improve dispersibility of the lipase used in the transesterification method of fats and oils using lipase of the present invention, it is preferable to coexist with a carrier such as diatomaceous earth, activated carbon, plaster, zeolite, or sehalose. It is preferable to use a carrier-immobilized lipase immobilized on a carrier consisting of a porous solid and chitosan or a derivative thereof, as the carrier. The porous solid constituting the carrier includes:
One or more selected from the group consisting of florisil, diatomaceous earth, celite, silica gel, clay, corn cob, and sawdust are preferably used. In addition, the above-mentioned chitosan or chitosan derivative constituting the above-mentioned carrier includes chitosan, N-acyl chitosan, N-mixed acyl chitosan, N,O-acyl chitosan, N-allylidene chitosan, N-alkylidene chitosan, chitosan salt, and One or more compounds selected from the group consisting of these partial reactants are preferably used. These partial reactants refer to compounds formed by the reaction of a part of the functional groups of chitosan, that is, amino groups or hydroxyl groups. Furthermore, chitosan derivatives that are deacetylated in a homogeneous reaction system of chitin and have a deacetylation rate of 40 to 60% can also be effectively used. Further, the carrier in the immobilized lipase used in the present invention can further contain a super absorbent resin, and the super absorbent resin includes a water absorbent polyurethane resin,
Polyhydroxyethyl methacrylate, polyacrylic acid resin, starch-acrylic acid graft polymer (graft polymerization of acrylic acid to starch, neutralization,
(crosslinked with a small amount of crosslinking agent), starch-acrylonitrile graft polymer (starch is graft-polymerized with acrylonitrile by ceric salt radiation, hydrolyzed, purified, and dried), starch is converted to carboxymethyl with monochloroacetic acid. cellulose-acrylonitrile graft polymer, cellulose carboxymethylated with monochloroacetic acid and cross-linked with formalin, vinyl alcohol and acrylic acid copolymer, or vinyl acetate and methyl methacrylate copolymer. Examples include self-crosslinked polymers obtained by hydrolysis, polyvinyl alcohol intermolecularly crosslinked with dialdehyde or radiation, and crosslinked polyethylene oxide. These super absorbent resins may be used alone or in combination of two or more. Among these superabsorbent resins, those obtained by hydrolyzing and self-crosslinking starch-acrylic acid graft polymers, vinyl alcohol and acrylic acid copolymers, or vinyl acetate and methyl methacrylate copolymers are preferably used. obtain.
As the former, Sunwet IM-300 manufactured by Sanyo Chemical Industries, Ltd., and as the latter, Sumikagel S-50 manufactured by Sumitomo Chemical Industries, Ltd., are commercially available. The proportion (weight ratio) of the porous solid and the chitosan derivative used is preferably 0.05 to 1 part of the chitosan derivative per 1 part of the porous solid, and more preferably,
0.1 to 0.5 parts. When using a super absorbent resin, 0.05% of the super absorbent resin is used for 1 part of the porous solid.
The amount is preferably from 1 part to 1 part, more preferably from 0.1 part to 0.5 part. The immobilized lipase used in the present invention forms a gel of a chitosan derivative, disperses a porous solid in this gel, and then dries this dispersion to obtain a carrier.
It is produced by the method for producing an immobilized enzyme of the present invention, which is characterized in that the carrier retains lipase. In order to retain the enzyme (lipase) on the carrier, the enzyme can be effectively immobilized by drying and pulverizing the above dispersion to obtain the carrier, and mixing the carrier with an aqueous enzyme solution or an enzyme buffer solution. Furthermore, after thoroughly mixing the dried and pulverized product with the enzyme powder, water or a buffer solution can be sufficiently added and mixed to effectively immobilize the enzyme powder. After dispersing the porous solid in a gel made of chitosan derivative,
Examples of the drying method include adding and stirring the dispersion in acetone, forming a thin film and air drying it, spray drying, freeze drying, and the like. Alternatively, the immobilized lipase used in the present invention can be obtained by adding and mixing a super absorbent resin to a dried dispersion consisting of a chitosan derivative and a porous solid, and then adsorbing the enzyme (lipase) in the same manner as above. can be manufactured. The structure of the immobilized lipase produced in this way is such that the porous solid surface is coated with a gel made of a chitosan derivative, and the enzyme (lipase) is further adsorbed onto the chitosan derivative gel by adsorption, entrapment, ionic bonding, etc. It is assumed that it is fixed. In addition, in systems containing super absorbent resin,
It seems to have a structure in which the surface of a porous solid is coated with a gel made of a chitosan derivative and a superabsorbent resin, and the enzyme is further immobilized on the gel by adsorption, encapsulation, ionic bonding, or the like. Because of this structure, the immobilized lipase used in the present invention has a large surface area and becomes a highly active immobilized lipase. Also,
By selecting the particle size of the porous solid, separation,
It can be made into an immobilized lipase that is easily recovered. Also,
The surface part and the inside of the pores of the immobilized lipase are made of chitosan derivative gel or chitosan derivative and super absorbent resin gel, and the water content on the surface and inside of the immobilized lipase, which is the reaction site, can be freely controlled. It has the characteristic of being able to In other words, the above-mentioned immobilized lipase has the characteristic that it can retain not only water necessary and sufficient for the appearance of enzyme activity, but also water necessary and sufficient for the reaction. In the case of hydrolysis reactions of fats and oils, the system is usually a heterogeneous substrate reaction system consisting of a substrate and water, but in such reaction systems, emulsions are often formed at the interface, making it difficult to separate the decomposition products. This not only makes workability difficult, but also lowers the product recovery rate and increases process costs.
However, the first method of the present invention using the above immobilized lipase
In the case of the hydrolysis reaction of fats and oils in the stage reaction, the immobilized lipase can retain sufficient moisture necessary for the hydrolysis reaction, so the reaction of the present invention can be carried out in a system that does not substantially contain free water. can be done,
Almost no emulsion is generated, the decomposition products are easily separated, and the product recovery rate is also high. In addition, in the case of esterification reactions and transesterification reactions using enzymes, they are often non-aqueous, but in the case of non-aqueous systems, the apparent reduction in enzyme activity is often caused by a temporary decrease in enzyme activity due to a decrease in water in the reaction site. cessation of activity does not necessarily correspond to inactivation of the essential enzyme. However, in the case of the esterification reaction of the second stage reaction of the present invention using the above-mentioned immobilized lipase, it is possible to easily replenish the inside of the immobilized lipase with water necessary and sufficient for the appearance of enzyme activity. It has the characteristic of being able to maintain the appearance of activity. In the case of the hydrolysis reaction of the first stage reaction in the present invention, the amount of the immobilized lipase used is preferably 3 to 40%, more preferably 6 to 20%, based on the substrate (raw oil and fat). be. The amount of enzyme is preferably 5 to 2000 U/g, more preferably 50 to 500 U/g, relative to the substrate. In the case of the esterification reaction in the second stage reaction, the amount of the immobilized lipase used is preferably 0.5 to 10%, more preferably 1.0 to 5.0%, based on the substrate. The amount of enzyme is preferably 20 to 10,000 U/g, more preferably 100 to 1,000 U/g relative to the substrate.
However, the activity unit (U) of the enzyme is determined by adding the enzyme to 5 ml of olive oil emulsion and 4 ml of 0.1M phosphate buffer.
When the reaction was carried out at 37°C for 30 minutes, one activity unit (U) was defined as each fatty acid produced corresponding to 0.06 ml of 0.05N aqueous sodium hydroxide solution. The same applies to the enzyme activity units in the Examples shown below. The substrate oils and fats used in the transesterification method of oils and fats using lipase of the present invention include general vegetable oils, animal oils, processed oils and fats, and mixtures thereof, such as soybean oil, cottonseed oil, rapeseed oil, olive oil, corn oil,
These include coconut oil, safflower oil, beef tallow, lard, and fish oil. In addition, glycerides with specific compositions, which are raw materials for cocoa butter substitutes, such as 1,3-distearo-2-oleoglyceride, 1-palmito-2
- When using oleo-3-stearoglyceride or 1,3-palmito-2-oleoglyceride as the target product of the transesterification reaction, use an oil or fat containing a large amount of oleic acid at the 2-position of the glyceride, such as olive oil or camellia oil. , sasanya oil, palm fat, monkey fat, iritupe butter, kokum butter, shea butter, maua butter, furwara butter, Borneo tallow butter, or fractionated fats and oils thereof. In the method of the present invention, it is preferable to carry out the reaction in a system to which a relatively large amount of water is added in the first stage reaction, that is, the reaction stage mainly consisting of the hydrolysis reaction. In other words, water should be added in an amount of 0.01 part by weight or more, preferably 0.02 part by weight or more, per 1 part by weight of fats and oils, and by adding an amount of water within this range, the temperature at which normal enzymatic reactions can proceed is 20. By mixing and stirring at ~50℃, the reaction reaches equilibrium in 1 to 4 hours.
A reaction product having a diglyceride content of 15 to 70% by weight, based on total glycerides, is obtained. The optimal amount of water is 0.02 to 0.10 parts by weight per 1 part by weight of fats and oils,
By using a system in which water is added in an amount within this range, the diglyceride content can be reduced to 20% of the total glyceride content.
~60% by weight of reaction product is obtained. Next, in the second stage reaction, that is, the reaction stage mainly consisting of ester synthesis reaction, a fatty acid for the purpose of transesterification is added to the reaction product of the first stage reaction, and 20 to 50%
Mix and stir while maintaining the temperature at °C. By adding the fatty acid, the reaction of the system rapidly shifts from the hydrolysis reaction to the ester synthesis reaction, and the diglyceride produced in the first stage reaction is esterified by the ester synthesis reaction to obtain the desired triglyceride. As the lipase used in the method of the present invention, Rhizopus type, Aspergillus type, Candida type, Mucor type, pancreatic lipase, etc. can be used, and many of these are commercially available. Among these lipases, lipases having 1,3-position specificity for triglycerides are particularly preferred;
delemar), Rhizopus japonicus (Rhizopus
japonicus), Mucor japonicus (Mucor
japonicus), etc. In a more preferred method of the present invention, in order to achieve selective transesterification, the proportion of 1,2(2,3)-diglyceride in the diglyceride produced in the first stage reaction, that is, the hydrolysis reaction step, is reduced.
The hydrolysis reaction is carried out so that the amount is 70% by weight or more, more preferably 90% by weight or more. Diglycerides often have unstable structures where acyl group transfer reactions can occur, so the hydrolysis reaction temperature must be adjusted.
The temperature is preferably 40°C or less, and the reaction time is preferably within 10 hours when the reaction temperature is 40°C. A more preferred method of the present invention is characterized in that water in the reaction system is removed in the second stage reaction, that is, the ester synthesis reaction stage. In the ester synthesis reaction stage, adding fatty acids causes a shift in the reaction equilibrium, but further removing water from the reaction system accelerates the ester synthesis reaction rate and gradually slows down the hydrolysis reaction rate. . Moisture in the reaction system during the ester synthesis reaction stage is removed by effectively stirring and removing dry inert gas by passing dry inert gas into the reaction system and exhausting it outside the reaction system. be able to. The inert gas may be any gas such as nitrogen gas, argon gas, helium gas, etc., as long as it is non-explosive and non-reactive with oils and fats. Ventilation into the reaction system can be carried out by bubbling into the liquid phase in the reactor or by blowing into the gas phase. In the agitation removal of water by inert gas ventilation, the exhausted mixed gas is passed through a condenser cooled to below the freezing point of water by a refrigerant, and the water vapor contained in the mixed gas is trapped as ice, and the inert gas and water vapor are completely separated. The separated inert gas can be reused by further refluxing it into the reaction system. The amount of fatty acid added in the second reaction (ester synthesis reaction) step of the method of the present invention is preferably 0.4 to 2.0 parts by weight per 1 part by weight of fats and oils.
As the fatty acid, linear saturated or unsaturated fatty acids having 2 to 22 carbon atoms can be used, such as palmitic acid, stearic acid, oleic acid, etc. Furthermore, in addition to adding a predetermined amount of the fatty acid in its entirety at once, it is also possible to use a method in which it is added gradually as the reaction progresses. When using stearic acid, palmitic acid, etc., which have high melting points, as added fatty acids, the reaction temperature may become non-uniform. In such cases, dissolve the fatty acids in an organic solvent that is inactive to lipase. The reaction can be carried out in a homogeneous system. Examples of this type of organic solvent include n-hexane, industrial hexane, petroleum ether, etc., and can be used in an amount of 1 to 10 parts by weight per 1 part by weight of fatty acid. The reaction temperature in the present invention is 20 to 70°C in both the first stage reaction (hydrolysis reaction) and the second stage reaction (ester synthesis reaction), which is the same as in normal enzyme reactions.
It can be done with However, in the first reaction stage, since the acyl group transfer reaction of the produced diglyceride depends on the reaction temperature, it is not appropriate to carry out the reaction at a temperature of 50°C or higher, and it is desirable to carry out the reaction at a temperature of 40°C or lower. As mentioned above, in the method of the present invention, the transesterification reaction of oils and fats is configured as a two-step reaction of hydrolysis reaction and ester synthesis reaction, so the reaction is more efficient than a reaction system that uses a trace amount of water. Therefore, compared to the conventional one-stage reaction in which the reaction rate had to be sacrificed in order to keep the partial glyceride produced by hydrolysis low, the reaction rate is dramatically improved and the final product is improved. This is an innovative reaction operation method that can reduce the content of partial glycerides such as diglycerides and monoglycerides. Increasing the reaction rate not only reduces reactor operation time and enables an efficient and highly productive process, but also reduces the residence time of the enzyme or enzyme-containing composition in the reactor, thereby increasing the reaction rate. It also has the advantage that deactivation of the enzyme or change in shape of the enzyme-containing composition caused by stress or physical changes on the surface caused by stirring in the container can be reduced. Further, the method of the present invention eliminates the need for the complicated labor involved in drying operations in the preparation of conventional enzyme-containing compositions, and completely avoids deactivation of enzymes due to excessive drying.
Furthermore, in conventional methods that use extremely small amounts of water, it was essential to strictly control the initial water content in order to suppress hydrolysis, but in the method of the present invention, the initial water content is in the range of 3 to 10% by weight based on the fats and oils that serve as the substrate. If the water content is , the water can be easily removed by the water removal operation in the ester synthesis reaction stage of the second stage reaction, which is very effective in terms of operability. Furthermore, the method of the present invention does not require strict moisture control even when the enzyme (lipase) is repeatedly used, and the enzyme can be reactivated by adding water to the reaction system in the second and subsequent reactions. ,
Since high enzyme activity can be maintained at all times, stable reaction operations are possible. Moreover, according to the method of the present invention, the enzyme can be reused five or more times, and the economic efficiency of the process can be dramatically improved. Furthermore, in the method of the present invention, as mentioned above, free fatty acids are converted into alcohol esters by adding alcohol to the reaction system, and since the alcohol esters are hardly hydrolyzed, the hydrolysis of glycerides is greatly promoted. It also has the advantage that triglyceride can be easily recovered after the reaction. Furthermore, as described above, by using a carrier-immobilized lipase as the lipase, the above-mentioned effects can be further improved, and this will be made clear in the Examples shown below. 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 Using the apparatus shown in the flow sheet in FIG. 1, transesterification of oils and fats was carried out as follows. 38 g of palm soft part oil, immobilized lipase prepared by dissolving 103 mg of lipase derived from Rhizopus deremer (98000 U/g) in 2.0 g of water and adsorbing this to 2.0 g on a carrier, 120 g of n-hexane and 2.5 g of butyl alcohol, 40 Closed reactor A for 2 hours at °C.
The first stage reaction (hydrolysis reaction) was carried out by stirring with a stirrer 1a inside the reactor. After the reaction, a small amount of the reaction mixture was separated and analyzed as described below, and the results are shown in Table 1. The carrier used here was prepared as follows. 8 g of chitosan (manufactured by Kyowa Yushi Kogyo Co., Ltd., trade name Fronac N) was added and mixed in 60 g of a 10% acetic acid aqueous solution to form a chitosan acetate gel, and further, 440 g of water and 32 g of Celite were added to the gel. After making a homogeneous mixture, this was mixed dropwise into 2000 g of acetone, and the insoluble matter was collected by centrifugation, and the insoluble matter was further added and mixed in 1000 g of acetone, filtered, air-dried, and then mixed under vacuum. The mixture was dried to remove acetone and a carrier consisting of chitosan acetate and celite was obtained. Next, after the first stage reaction, stirring was temporarily stopped, 20 g of stearic acid (NAA-180, manufactured by NOF Corporation) was added, and stirring was resumed. Dry nitrogen is blown from the nitrogen reservoir D through the desiccant packed bed B using the pump C, and the entrained inert gas containing water vapor in the gas phase 2 where a vapor-liquid equilibrium relationship is established is removed from the reaction system. By evacuation, the water content in the reaction system was gradually lowered (ester synthesis reaction of the second stage reaction). The average residence time of the inert gas during this second stage reaction was about 3 seconds. The entrained inert gas containing n-hexane and water vapor passes through a surface condenser F cooled to around -20°C with dry ice, where the n-hexane liquefies and the water vapor becomes ice, forming a gas-liquid-solid state. The n-
Hexane was refluxed into reactor A. This second stage reaction was carried out for 12 hours. After the reaction of each stage is completed, a small amount of the product is collected and stored in the J.A. The method of J. Blum et al. [Lipid, 5,
601, (1970)] using hexamethyldisilazane (HMDS) and trimethylchlorosilane (TMOS) (manufactured by Wako Pure Chemical Industries, Ltd.), and analyzed by temperature-programmed gas chromatography. The results are shown in Table 1 below. As shown in Table 1, in the first stage reaction, diglycerides accounted for 43.3% by weight of the total glycerides, and monoglycerides accounted for 7.6% by weight of the total glycerides. On the other hand, about 70% of the fatty acids liberated by hydrolysis were converted into alcohol esters. After the fatty acid was added, diglyceride and monoglyceride gradually decreased in the second stage reaction due to the esterification reaction, and decreased to 11.3% by weight at the end of the second stage reaction, that is, 12 hours later. Next, n-butyl alcohol is added to esterify the free fatty acids remaining in the system.
2.0g was added and the free fatty acid esterification reaction was carried out at 40°C for 5 hours. This treatment gave the final product an acid value of 20.6. In order to recover the triglyceride portion from this final product, deacidification and distillation processes may be used, which is very easy and has a high recovery rate.

【table】

[Table] Example 2 A reaction was carried out in the same manner as in Example 1 except that n-decyl alcohol was used as the alcohol. The amount of alcohol added in the first reaction stage is
A hydrolysis reaction was carried out using 5.3 g at 40°C for 4 hours.
In the second reaction stage, 20 g of stearic acid was added to the system, dehydration of the system was started, and the system was stirred for 8 hours to carry out the esterification reaction. When the diglyceride concentration in the total glyceride decreased to about 10%, an additional 5.0 g of n-decyl alcohol was added,
Free fatty acids were esterified. The results of the analysis conducted in the same manner as in Example 1 are shown in Table 2 below.

[Table] Example 3 N - 4.5 g of butyl alcohol was added, and the system was stirred for 4 hours while dehydrating the system using the system (apparatus) shown in FIG. 1 to esterify free fatty acids. The results of the analysis in this case are shown in Table 3 below.

【table】

【table】 [Brief explanation of drawings]

FIG. 1 is a flow sheet of an apparatus suitable for carrying out the present invention. A... Reactor, B... Desiccant packed bed, C... Pump, D... Nitrogen reservoir, E... Nitrogen cylinder, F... Condenser, 1a... Stirrer, 1... Liquid phase section, 2... Gas phase part.

Claims (1)

[Scope of Claims] 1. The production of fats and oils by lipase, which consists of two consecutive reactions: a first stage reaction mainly consisting of a hydrolysis reaction of fats and oils, and a second stage reaction mainly consisting of an ester synthesis reaction. A transesterification reaction method comprising:
1. A method for transesterification of oils and fats using lipase, which comprises adding alcohol in a step reaction step and/or a second step reaction step. 2. A method for transesterification of oils and fats using lipase according to claim 1, characterized in that the lipase is used fixed on a carrier consisting of a porous solid and a chitosan derivative. 3. A method for transesterifying fats and oils using a lipase according to claim 1 or 2, wherein the lipase is a lipase having 1,3-regiospecificity. 4. The method for transesterification of oils and fats using lipase according to any one of claims 1 to 3, wherein the first stage reaction is a reaction that yields 15 to 70% by weight of diglyceride based on the total glyceride. 5. The method for transesterification of oils and fats using lipase according to any one of claims 1 to 4, wherein the first stage reaction is a reaction that yields 20 to 60% by weight of diglyceride based on the total glyceride. 6 1,2 (2,3) in which the diglyceride obtained by the first stage reaction is 70% by weight or more of the total diglyceride
- is a diglyceride containing diglyceride,
A method for transesterifying fats and oils using a lipase according to any one of claims 1 to 5. 7 1,2 (2,3) in which the diglyceride obtained by the first stage reaction is 90% by weight or more of the total diglyceride
- is a diglyceride containing diglyceride,
A method for transesterifying fats and oils using a lipase according to any one of claims 1 to 6. 8. The method for transesterifying fats and oils using lipase according to any one of claims 1 to 7, wherein the second stage reaction is a reaction of adding a fatty acid to perform an ester synthesis reaction. 9 The amount of fatty acid added is 0.4 per part by weight of fats and oils.
9. A method for transesterifying fats and oils using the lipase according to any one of claims 1 to 8, wherein the amount is 2.0 parts by weight. 10. A method for transesterification of oils and fats using lipase according to any one of claims 1 to 9, wherein in the second stage reaction, water in the reaction system is removed. 11 Claims in which moisture is removed by continuously or intermittently passing dry inert gas into the reaction system and exhausting it outside the reaction system to simultaneously remove moisture within the reaction system. A method for transesterification of oils and fats using the lipase according to any one of Items 1 to 10. 12 Transesterification of oils and fats using the lipase according to any one of claims 1 to 11, in which the gas exhausted outside the reaction system is passed through a condenser to separate and remove moisture, and then refluxed into the reaction system. Reaction method.