JP2001252090A - Method for producing diglyceride - Google Patents

Abstract

PROBLEM TO BE SOLVED: To produce an oil and fat composition abundantly containing diglycerides by the method which permits the repeated reaction with an enzyme showing excellent stability, forms diglycerides in an excellent formation proportion and gives an abundant accumulation amount of diglycerides in a short reaction time. SOLUTION: This method for producing diglycerides is carried out by using a hydrophilic membrane holding fixed position specific lipase.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing an oil or fat composition having an excellent diglyceride production rate.

[0002]

2. Description of the Related Art It has been revealed that diglyceride has effects such as prevention of obesity and suppression of weight gain (Japanese Patent Application Laid-Open No. Hei 4-30).
No. 0828), and attempts have been made to blend it into various foods. If a glyceride mixture (oil composition) containing diglyceride at a high concentration is used for the oil phase, an edible oil-in-water emulsion composition having a rich taste even when the amount of fat is reduced and having a good flavor can be obtained. Is reported to be obtained (Japanese Patent No. 2848849).

[0003] As a method for producing diglycerides, specifically hydrolyzing the ester bond of S _n -2 position of oil using 2-lipase (Patent No. 2671349 discloses),
1,3- having transesterification activity on a substrate containing glycerin and a fatty acid or a fatty acid lower alcohol ester
Method for ester synthesis using lipase (Japanese Patent No. 213)
No. 5885) is known.

[0004] All of these reactants coexist with water or a lower alcohol, and must be separated from the oil or fat composition in a separate step. A method using a membrane reactor capable of simultaneously performing a diglyceride production reaction and removing water or a lower alcohol from the produced oil / fat composition has been proposed. For example, a method of contacting and reacting a fatty acid ester such as oil and fat with water containing lipase via a thin film (Japanese Patent No. 1844)
No. 457), a method in which lipase is sandwiched between a hydrophilic porous membrane and a hydrophobic porous membrane, and oil and fat are present on both sides of the lipase to cause a reaction in a central lipase layer (Japanese Patent No. 167).
No. 4165).

[0005]

However, these methods have a low diglyceride reaction generation rate and have poor accumulation of diglyceride even when substances involved in the reaction are repeatedly passed through the membrane reactor.

An object of the present invention is to provide a method for producing a diglyceride in which a reactant involved in the production of diglyceride is repeatedly passed through a membrane reactor to produce a diglyceride, and the accumulation of diglyceride is excellent.

[0007]

Means for Solving the Problems The present inventors have found that the use of a membrane reactor having a position-specific lipase immobilized on a hydrophilic membrane results in an increase in the rate of diglyceride reaction production.

[0008] That is, the present invention provides a diglyceride which reacts a substance selected from fats and oils, fatty acids and esters thereof, and a substance selected from water and glycerin with a hydrophilic membrane on which a regiospecific lipase is immobilized. It provides a manufacturing method.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The process for producing diglyceride of the present invention is carried out by reacting an oil or fat with water or glycerin, or by reacting a fatty acid or an ester thereof with glycerin.

The position-specific lipase used in the present invention is 2
Regioselective lipase (2-lipase), 3-selective lipase (3-lipase) and 1,3-selective lipase (1,
3-lipase). Regiospecific lipase are widely distributed microorganisms, animals, plants or the like in nature, e.g., Rhizopus (Rhizopus) genus Aspergillus (Aspergillus)
Sp., Mucor (Mucor) genus Penicillium (Penicillium)
Sp., Achromobacter (Achromobacter) genus Alcaligenes (Alcaligenes) genus, etc. derived from microorganisms of the lipase, animal pancreatic lipase, tongue glands lipase, lipoprotein lipase, and the like.

[0011] Among the regiospecific lipase, particularly preferably 1,3-lipase, Rhizopus (Rhizopus) genus Aspergillus (Aspergillus) genus Mucor (Mucor) microorganism-derived lipases, pancreatic lipase, and more specifically Is Rhizopus delemar : Rhizopus japonicus , Rhizopus nibeus
zopus niveus), Aspergillus niger (Aspergillus
niger ), Mucor javanicus ,
Lipases derived from Mucor miehei or the like are preferred.

As the hydrophilic membrane used in the membrane reactor, it is convenient to use a thin film used in microfiltration, ultrafiltration, etc., and particularly preferred is a thin film used in ultrafiltration. Examples of the material of the hydrophilic film include cellulose derivatives such as cellulose nitrate and cellulose acetate, acrylic acid copolymers such as polyacrylonitrile, polypropylene ethylene vinyl alcohol copolymer, and polyvinyl alcohol. Among these hydrophilic films, cellulose derivatives, polyacrylonitrile and the like are preferable, cellulose acetate and polyacrylonitrile are more preferable, and polyacrylonitrile is particularly preferable.

The thickness of the hydrophilic film is 10 to 100 μm, preferably 20 to 50 μm, and particularly preferably 30 to 50 μm. The hydrophilic film may be any as long as it can fractionate a substance having a molecular weight of 40,000 or less.
It is preferable to use a substance capable of fractionating a substance having a molecular weight of 800.
Those which can fractionate 0 to 15000 substances are particularly preferred. The average pore diameter is 0.05 to 0.5 μm, particularly 0.07
5 to 0.2 μm is preferred. The thickness and molecular weight cutoff of these hydrophilic membranes are determined by the membrane reactor used based on the reaction rate, strength of the thin film and the like.

The regioselective lipase is immobilized on the hydrophilic membrane by ultrafiltration of the lipase solution. That is, a regioselective lipase solution using a buffer as a solvent is supplied to the shell side of the hydrophilic membrane at a flow rate of 0.5 to 5.0 L / min and circulated, and the pressure is 20,000 to 8
Filtration is performed at 000 Pa, and then a pressure difference between the shell side and the lumen side is similarly set to 20,000 to 80,80 through a buffer solution.
Circulating at 000 Pa, water remaining on the shell side is completely permeated and removed to the lumen side.

The production of the diglyceride of the present invention is carried out by hydrolysis or glycerolysis of fats and oils, and furthermore, by esterification or transesterification of fatty acid (or ester) with glycerin.

When the diglyceride of the present invention is produced by hydrolysis or glycerolysis of fats and oils, fats and oils such as beef tallow, lard, fish oil, whale fat, rapeseed oil, olive oil, soybean oil, coconut oil, palm oil, It may be any of vegetable oils such as linseed oil and safflower oil, and preferably includes fish oil, soybean oil, rapeseed oil and linseed oil. Generally, fats and oils are mainly composed of triglycerides, but monoglycerides, diglycerides, complex lipids (eg, phospholipids such as lecithin and kephalin), free fatty acids, long-chain alcohols, sterols, hydrocarbons (eg, squalene, etc.) It may contain fat-soluble vitamins, pigments and the like. Further, the extraction method may be any of a pressing method, a roasting method, a boiling method and the like.

The fatty acid used when the diglyceride production of the present invention is carried out by an esterification reaction is a saturated or unsaturated fatty acid having 2 to 24 carbon atoms, for example, butyric acid, valeric acid,
Caproic acid, enanthic acid, caprylic acid, pelargonic acid,
Capric acid, undecanoic acid, lauric acid, myristic acid, palmitic acid, zomaric acid, stearic acid, oleic acid, elaidic acid, linoleic acid, linolenic acid, arachidonic acid, gadrenic acid, arachinic acid, behenic acid, erucic acid, eicosapentaene Acid, docosahexaenoic acid and the like can be used. In particular, palmitic acid, zomaric acid, stearic acid, oleic acid, elaidic acid, linoleic acid, linolenic acid, arachidonic acid, gadrenic acid, arachinic acid, behenic acid, erucic acid, eicosapentaenoic acid, and docosahexaenoic acid are preferred. Further, the fatty acid may form an ester with a lower alcohol having 1 to 3 carbon atoms. Examples of the lower alcohol having 1 to 3 carbon atoms include methanol, ethanol, propanol, and isopropanol. These fatty acids or fatty acid esters can be used alone or in a mixture of two or more.

In the production of diglyceride using the membrane reactor of the present invention, reaction materials such as oils and fats, fatty acids (esters) are placed on the shell side of a hydrophilic membrane on which position-specific lipase is immobilized, and water and glycerin are placed on the lumen side. And the like. At this time, the circulating fluid flow rate on the shell side and the lumen side is 0.1 to 5.0 L / min, particularly 0.5 to 3.0 L / min.
L / min, pressure difference between shell side and lumen side is 20,000
~ 80,000 Pa, especially 40,000 ~ 60,000
The pressure is preferably set to Pa, and the pressure on the shell side is preferably increased. The reaction temperature and pH are appropriately selected and determined according to the position-specific lipase to be used, and the circulation is continued until the desired diglyceride is obtained.

The present invention is particularly effective when a fat or oil is hydrolyzed to obtain a fat or oil composition having a high diglyceride content. In the present invention, the fat composition is a glycerin mono, di,
A mixture of tri-fatty acid esters, which comprises hydrolysis of fats and oils, esterification of glycerin with fatty acids (esters),
It is produced by performing a reaction such as glycerolysis between fat and oil and glycerin using a regiospecific lipase. In this case, after the hydrolysis step by the membrane reactor is completed, the oil and fat composition and free fatty acids, glycerin and the like are separated by means such as molecular distillation, low-temperature fractionation, and urea addition, and the content of monoglyceride and triglyceride and the like is also determined. By adjusting the composition, it is possible to produce an oil and fat composition having a high diglyceride content as expected.

[0020]

EXAMPLE Immobilization of regioselective lipase 25 dissolved in 300 mL of 50 mM phosphate buffer (pH 7)
10,000 units of 1,3-lipase (Nipase Sangyo Co., Ltd .: Lilipase A10) is circulated to the shell side of a pencil type module (Asahi Kasei Corp .: Microza ACP-0013, polyacrylonitrile membrane, effective area 0.017 m ² ). The solution was subjected to ultrafiltration at a pressure of 40,000 to 60,000 Pa to be immobilized. One unit of lipase activity is the amount of enzyme that releases 1 μmol of fatty acid per minute when olive oil is decomposed.

Example 1 A membrane reactor loaded with a pencil-type module immobilized with 250,000 units of 1,3-lipase A10 was loaded with 50 g of soybean oil on the shell side of the lipase-immobilized membrane and 50 mM phosphate buffer on the lumen side. 50 mL of the liquid (pH 7) was circulated for 2 hours at a flow rate of 500 mL / min, a pressure difference of 40,000 Pa (higher on the shell side) and a temperature of 37 ° C. Gas chromatographic analysis of the reaction product on the shell side revealed that the free fatty acid was 29% by weight, the monoglyceride was 14% by weight, the diglyceride was 41% by weight, and the triglyceride was 16% by weight. On the other hand, to 2 mL of 5 mM phosphate buffer (pH 7) in which 10,000 units of lipase A10 were dissolved, 2 g of soybean oil having the same ratio of lipase activity (unit) / fat weight (g) was added, and 37 ° C. After reacting for 24 hours while stirring at, the reaction product was analyzed by gas chromatography. As a result, 33% by weight of free fatty acid, 2% by weight of monoglyceride, and 19% by weight of diglyceride were obtained.
And triglyceride was 57% by weight. The diglyceride production method of the present invention was excellent in the amount of generated and accumulated diglyceride in a short time.

Example 2 The same reaction was carried out using fish oil instead of soybean oil of Example 1. In a membrane reactor using a 250,000 unit lipase-immobilized membrane, 35% by weight of diglyceride was formed after 2 hours. In contrast, in the stirring reaction method as a comparative example, the amount of diglyceride produced and accumulated was 21% by weight even after 48 hours.

[0023]

According to the present invention, there is provided an oil / fat composition having excellent enzyme stability, capable of repeating a reaction, having an excellent diglyceride production rate, producing a large amount of accumulated diglyceride in a short time, and having a large diglyceride content. Can be manufactured.

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 4B033 NA01 NA27 NB36 NB45 NB70 NC04 ND02 4B064 AD85 CA21 CA35 CA36 CA40 CB26 CC03 CD05 CD07 CD21 CD22 DA10 4H059 AA04 BA13 BA26 BA30 BA33 BB02 BB03 BB52 BC03 BC48 CA36 CA36 CA36

Claims (3)

[Claims] 1. A substance selected from fats and oils, fatty acids and esters thereof, and a substance selected from water and glycerin,
A method for producing diglyceride in which a hydrophilic membrane on which a position-specific lipase is immobilized is allowed to act to react. 2. The method for producing diglyceride according to claim 1, wherein the hydrophilic film is a polyacrylonitrile film or a cellulose acetate film. 3. The method for producing diglyceride according to claim 1, wherein the position-specific lipase is 1,3-lipase.