JPH02501622A - Method for producing cis-9-octadecenoic acid composition - Google Patents

Abstract

A process for producing a high purity oleic acid composition utilizing enzymatic hydrolysis is disclosed. The process involves obtaining high oleic sunflower seed oil which oil contains triglycerides having fatty acid moieties of oleic acid in an amount of 60 % or more, preferably 80 % or more and further wherein the ratio of linoleic moiety to oleic moiety is less than about 0.25, preferably less than about 0.09. The oil obtained from the high oleic sunflower seed oils is subjected to enzymatic hydrolysis by contacting the oil with hydrolase enzymes and/or various combinations of hydrolase enzymes within an aqueous medium at a temperature in the range of 20-60 DEG C and a pH in the range of about 4.5 to about 10. The oil, hydrolase enzyme and water are agitated so that hydrolysis occurs at the oil water interface and the acid moieties of the triglycerides are separated away. An oleic acid layer is allowed to form and separate away from the aqueous medium and the aqueous medium is then separated away to provide a highly pure oleic acid composition.

Description

[Detailed description of the invention] name of invention Method for manufacturing cis-9-octadecenoic acid compositions Field of the Invention This invention generally relates to the production of cis-9-octadecenoic acid (i.e., oleic acid). Concerning the field of manufacturing methods. More specifically, the invention provides high oleic castor Wari seed oil (hjgh olefcsunflower 5eed oil) Concerning the production of oleic acid in very pure form by enzymatic hydrolysis, and As a result, the highly pure oleic acid composition derived from such hydrolysis has been Approximately 10 billion pounds of fatty acids are produced in the United States.

Approximately 80% of the fatty acids produced come from the industrial hydrolysis of tallow (tal low). It is derived from (M, W, Forgo, Baile 's Industrial Oil and Raw Materials, 4th edition, 2 volumes, edited by D. Stern, See J. Riley, Niy York, 1982, p. 379. Yo). Most fatty acids are produced by industrial fat crushing methods. However However, recently the use of enzymes has been promoted in connection with the hydrolysis of fats to produce fatty acids. There is growing interest in its use. Traditional method of using high pressure steam to crush fat The advantages of using enzymes compared to the above method are mainly the following (1): (2) and (3): (1) It is a more specific reaction, so it is cleaner. and a pure product can be obtained; (2) the energy required is lower and more and (3) the resulting sweet water is purer, i.e., this addition of water The glycerin-water mixture obtained from decomposition is purer. Oleic acid sandawa Cis-9-octadecenoic acid is present in natural fats and oils or biological lipids. It is a monounsaturated fatty acid. Oleic acid is CH3(CH2)7(Cl): CH) (CH2)7cooH, and is known as red oil (X Ru. Oleic acid is a very important substrate in both industrial and biological fields. Ru. Cleaner and purer products can be used in combination with products such as pharmaceuticals. is inherently safer; it also allows for the use of purer starting materials. Thus, purer refined chemical derivatives can be produced.

Oleic acid is most commonly produced from high-pressure steam fat using tallow as the starting material. Obtained from the crushing process. When producing oleic acid from such a fat crushing process , oleic acid is generally not available in pure form. A very purified me Inric acid is colorless and odorless, and has excellent stability against oxidative deterioration. These characteristics Oleic acid is used in combination with many food and pharmaceutical products to Always useful.

Pure oleic acid has excellent physical, chemical and physiological properties. can be used safely.

Because of these properties, oleic acid is often used in the field of fine or specialty chemistry. Effective and widely used. For example, oleic acid is used in pharmaceuticals, cosmetics and foods. It is widely used in the biochemical field related to biosensors and biosurfactants. Application examples have been found in the area. Oleic acid also has biological effects. exciting electronics field, and many other rapidly developing industries. Relevant applications are found in the field of technology.

Many uses of oleic acid require it to be very pure.

Commercially available oleic acids generally have different numbers of carbon atoms and different numbers of double bonds. Contains fatty acid homologs. Furthermore, commercially available oleic acid is often available in a variety of small amounts. Contains a large amount of impurities. Impure oleic acid compositions have poor color, odor, stability and safety have undesirable properties or characteristics. Due to these characteristics, many Such compositions cannot be fully applied in the field of technology.

Chemical and physical processes are used to produce highly purified oleic acid. The process was published in U.S. Patent No. 344.601.856 (July 22, 1986). published in ). However, many of the more common oleins The acid purification method is described in Ai Sen'ichi's 'Yogu Ichi Shushuou Wataru' (4th edition, 2 volumes, John Lyre). Lee and Sons (John Wiley & 5ons), two knee yoke, 19 82, (p, 379-387)). Disclosed in Bayesian literature The method described is currently considered to be the most commonly used method commercially. .

that enzymes can be used to produce fatty acids from triglycerides; The vitality of the hydrolysis reaction resulting from applying enzymes such as It has been indicated above that applications may be found in the field of technology. for example Quantitative determination of mono-, di- and especially triglycerides in human body fluids is used in the clinical diagnosis of many diseases and disorders such as: arteriosclerosis, Various metabolic disorders due to diabetes mellitus, nephrosis, bile duct obstruction, and endocrine disturbances.

For clinical analysis, glycerol esters are generally first hydrolyzed. , it is necessary to liberate glycerol and the corresponding fatty acids. this Related methods have been found to be useful for glycerol ester analysis. Enzyme compositions are disclosed in US Pat. No. 4,056,442.

This patent discloses compositions useful for hydrolysis in aqueous media: This aqueous medium contains a 15 to 95 units of Arrhizus arrhizus lipase, and LZ zL! Cylindracea (combined with Candida 11ndr+) lipase 5~ Contains a mixture of 85 units.

Certain methods and compositions for the hydrolysis of triglycerides are described in U.S. Pat. No. 4,259,440 (this is a Miles Laboratories, Inc. published on March 31, 1981) Ru. This method converts S/f-ase and cholesterol esterase into glycerol. In combination with a triglyceride analysis system, the step of adding triglycerides and the glycerin determining the amount of triglycerides present based on the amount of roll; . Other patents representing enzymatic hydrolysis of triglycerides generally include patent no. There is No. 59.440.

One potential disadvantage of using enzymes to perform hydrolysis is the price; Enzymatic techniques have been developed that involve the immobilization of the molecule onto a substrate. U.S. Patent No. 4, No. 275,011 discloses a process for transesterification of oils and fats. This person The method involves treating such oils and fats with water-soluble microbial enzymes. Ru. The microbial enzyme is absorbed onto an inert, powdered, water-insoluble dispersant. So After this, the enzyme absorbed onto this inert substrate is recovered from the reaction medium.

i Saraomi! And High purity oleic acid is produced by enzymatic hydrolysis of high oleic content sunflower seed oil. A method of manufacturing is disclosed. This method involves very high oleic content. Obtaining sunflower seed oil is included. Sunflower seed oil contains triglycerides Contains. High oleic sunflower seeds as used in connection with the present invention The oil has an oleic content of 60% or more, more preferably 80% or more. oleic component content of 88% or more, even more preferably 88% or more. or more, most preferably an oleic component content of about 95%. have These high-content oleic oils can convert triglycerides into hydrolytic enzymes. Contains a highly purified oleic acid composition that undergoes enzymatic hydrolysis upon contact with Provides a reactant acid. The reaction medium resulting from enzymatic hydrolysis contains oleic acid, glycan Contains cerol, and many impure acids.

By performing this reaction in an aqueous medium, glycerol and other water-soluble compounds can be can be easily separated from the water-insoluble oleic acid.

In connection with this invention, the terms "high oleic sunflower seed oil", 'oleic "High content sunflower oil" and "high oleic content injection" are used synonymously and Seeds of the plant (contains triglycerides with fatty acid moieties) It refers to the oil extracted from the seeds. where 60% of such fatty acid moieties or or more (preferably 80% or more, more preferably 88% or more) More preferably, about 95%) is oleic acid. Furthermore, here , below, that is, the oleic part: linoleic part is 1: (0,25 or less). or less), preferably 1:0.09 or less, most preferably is 1: (0,09-0,01).

Additionally, the term “high purity oleic acid composition” and its analogs refer to “oleic acid composition” and its analogs. "high content sunflower oil" as starting material and disclosed herein in connection with the present invention. and the oleic acid composition obtained by carrying out the method as described. shows. A typical high purity oleic acid composition of the present invention has approximately the following physical properties: Has the characteristics: Dantaka! Natsume Hitoshi Specific gravity (IS, IlioC) 0.899 color (ASTM) L2.0 Color (Gardner) 5-6 %R200,13 Acid value 201 ° Iodine value 87.8 Titer 18℃ These physical parameters are based on the oleic content of the starting oil. It changes to a certain degree.

The first object of the present invention is to produce a An object of the present invention is to provide a method for producing a highly pure oleic acid composition.

Another object of the invention is such enzymatic hydrolysis of high oleic sunflower seed oil. Among the solution methods, using different enzymes, all of the triglycerides in sunflower oil can be extracted. The object of the present invention is to provide a method for effectively separating oleic acid.

An advantage of the present invention is that the hydrolysis of sunflower oil triglycerides is energy efficient. - It can be done in an efficient manner.

A feature of the present invention is that the reaction product generated from the hydrolysis of high oleic sunflower seed oil The product is a highly purified oleic acid with various applications in the field of biotechnology. The reason is that it is a composition.

(1) Agricultural plants (2) biochemical enzymatic hydrolysis; and (3) chemical engineering purification methods.

These are achieved by combining advances in the following areas: (1) details; Plant production based on the male'5 sterility method Breeding improvement; (2) use of specific lipases to hydrolyze triglycerides in oils; and (3) chemical reactions and processes specifically adapted to purify oleic acid. and physical separation methods.

These and other objects, advantages and features of the invention are described more fully below. By reading the details of the materials, methods and uses as described in It will become clearer. The accompanying general formulas and flowcharts forming part of this specification Identical symbols refer to identical molecular parts or steps throughout the text.

In a beautiful state! A narrow day Cis-9-octadecenoic acid (hereinafter referred to as oleic acid and pure oleic acid) Before disclosing and describing the method of making the present composition (as shown), this The invention is not limited to the particular methods and compositions described, and the methods and compositions themselves are of course It should be understood that this can be changed. The technical terms used here are: If the purpose is to describe only a particular embodiment, it is not intended to be limiting. should also be understood. The scope of the invention is defined by the appended claims. This is because it is limited.

As used in this specification and the appended claims, the singular form "a", "an" "and the" means plural unless the context clearly dictates otherwise. It should be pointed out that the term includes references to . Therefore, for example, “at Reference to “rfglyeride” includes mixtures of triglycerides and includes “a Reference to "enzyme" includes reference to mixtures of enzymes, and the Reference to "hydrolysis" includes multiple hydrolysis prongs.

The present invention combines technological achievements in essentially unrelated scientific areas. This is a unique method that provides highly pure oleic acid compositions. current Seeds containing oil with a particularly high content of oleic components are produced using plant breeding technology. can be obtained.

Using highly efficient and selective enzymes, this highly oleic compound is Fatty acids can be effectively removed from glycerides. This method uses energy - Performs hydrolysis in an efficient manner, improving both yield and purity.

The hydrolyzed triglyceride product is then subjected to chemical and physical purification. The preparation and isolation methods were applied to obtain very pure oleic acid compositions in high yields. It will be done. In order to fully describe and disclose this invention, three major aspects of the invention ( shown in the chart below) are first shown individually.

↓ ↓ (Margin below) 1) Kagori rib cage Sunflowers (genus Helianthus) are a worldwide source of vegetable oil. As a food source, it is second only to soybeans in terms of temperature. In the United States alone, it is mainly Approximately 4 million sunflowers are grown in Nesota each year. rice The average sunflower yield in the country is about 1200 to about 1400 per hectare. It is in the kilogram range. The oil content obtained from harvested seeds is based on dry weight. on average, about 40-45%. Oil content (as a percentage of total plant weight) Increasing both the yield of these sunflower plants is possible through plant variety modification. This is the main purpose of a good project. The present invention uses this sunflower as a raw material. Use the plant body.

On average, over the past 10 years, the number of acres of sunflowers grown in the United States has been found to be rapidly expanding. This is partly due to the lack of free time. Some important points in the field of sunflower breeding and variety improvement of sunflower plants. By development. One important development was the discovery of cytoplasmic male sterility and the restoration of fertility. It was in the discovery of the gene for. This discovery supports the growth of hybrid sunflower plants. made possible. Hybrids produced using this method were introduced in the 1970s. was introduced in the early days of These sunflower plants have improved resistance to diseases. and increased uniformity in height and flowering, overcoming the diversity of open pollination. The yield increased by about 2S%. Furthermore, these hybrids are compatibility (this reduces dependence on insect pollinator population to obtain a good set of species) There was a high percentage of people who responded, “alleviation”. Sunflower plants using cytoplasmic male sterility method The development of the body is disclosed in U.S. Pat. No. 4,627°192, the contents of which are hereby incorporated by reference. It is shown.

The description of cytoplasmic male sterility (CMS) and genetic restoration of fertility in sunflowers is Fick et al., 1. Breeding and genetics of sunflowers. and @, p. 279-338 (J. F. Carter, M. 1978)). It's being served. The content is how to improve the cytoplasmic male sterility related to sunflowers. Laws and disclosures of genetic fertility restoration methods are presented herein. CMS The production of certain sunflower hybrids using It is described in the number. These contents apply to specific sunflower hybrids and this Reference is made herein for the disclosure of methods for producing such hybrids. cytoplasmic Male sterility is now a virtually non-functional mechanism for later use in producing hybrids. This is a method of selecting for the production of sunflower plants with pollen that is incapable of producing pollen. However, rice Other methods described in US Pat. No. 4,378,655 are also available. this Their methods include the presence of recessive genes and the application of chemical gameton eradicators. or the use of partial genetic sterility. Has a high level of self-incompatibility Plants that can also be used in the method of producing hybrids.

Sunflower oil mainly contains valmitic acid, stearic acid, oleic acid and linoleic acid. Consisting of oleic acid, linoleic acid and linoleic acid, which are found in traditional sunflower seed oil. It accounts for about 90% of the total fatty acid content. However, sunflower oil contains: It is known to contain 13 types of fatty acids: linoleic acid, oleic acid, Valmitic acid, stearic acid, lylunic acid, balmitoylic acid, arachidic acid, marga Phosphoric acid, Valmitic acid, and Behenic acid. (T, Cuprina et al. , a certain Jyokugyokudanmawa 2, and Himawari's vibe 1's LIE ( Agricultural and Horticultural Society, Yugoslavia, 1983). Its contents are Regarding the disclosure of surrounding oil content, it is indicated here). Unsaturated acids: 1, 2 or 3 free bonds. These unsaturated acids are, for example, These are leic acid, linoleic acid and lylunic acid. Other acids (e.g. stearic acid and and valmitic acid) are saturated.

It has been recognized that there is an inverse correlation between oleic acid and linoleic acid. There is. This is highly influenced by environmental factors such as temperature, especially during the growing season. Receive the sound. The total oleic acid content and linoleic content generally determine the acid content of the oil. It is about 90% of the amount. Therefore, as the linoleic content increases to about 10%, The rhein content is reduced by about 80%. This relationship is only a rule of thumb and cannot be solved strictly. It should not be interpreted.

Cooler northern climates generally produce sunflower seeds with higher lylunic acid content. It will be done. In contrast, high oleic acid values indicate seeds grown in warmer southern regions. It is more characteristic. Concentrates with high linoleic acid content are used in soft margarine and sugar. Preferred in sunflower oil used in Rada dressing.

High oleic acid content is more preferred for many other applications.

For example, high oleic wrinkled sunflower seed oil of the present invention (which has a high purity of oleic (including the production of phosphoric acid). The purity of oleic acid is linoleic acid of conventional crude sunflower oil derived from seeds grown in favorable climates. Oxidative stability is the oxidative stability of crude oil extracted from sunflower seeds grown in the north. It is almost twice as many as the sex.

In the context of the present invention, the terms "variety" and "varietal" refer to the species (sunflower (Helia) nthus Annuus)) (for example, “C/L/Bennett (P ervenets)) are used synonymously.

They share certain fixed characteristics that distinguish them from other varieties within that wing. have. A variant used in connection with the present invention refers to the total number of individuals within a variant. may show significant variation.

This is primarily based on Mendelian trait segregation of descendants in successive generations. Ru. “Strains” are distinguished from “variants” and are created through several generations of self-pollination. In general (though not exclusively), there are , represents a group of plants. Furthermore, a “line” is defined as a single parent plant using tissue culture methods. The present disclosure is intended to encompass a group of plants that propagate vegetatively from objects. It is defined broadly enough. Create new hives using such strains Developing the lid is disclosed in U.S. Pat. ．． It is described in No. 624. These contents include disclosure of such tissue culture methods. As shown here. A variety or strain indicates that it has a particular characteristic. If it is genetically inbred within the following range, it is considered “pure breeding” for this trait. considered: the extent to which this purely bred variety or line is self-pollinated to the extent that no significant amount of independent trait segregation among offspring is observed.

As indicated above, various fatty acids (e.g. stearic acid, oleic acid and Linoleic acid) is characteristic of the various seed oils obtained. Content of such acids can be expressed as a percentage of the total fatty acid content of the triglycerides that make up the oil. Ru. To describe the oil obtained from sunflower seeds used in connection with the present invention This method is used here. For example, for oleic acid content, described below. The unitless ratio of linoleic acid content is the linoleic acid content of the total fatty acid portion on triglycerides. Calculated by dividing the percent oleic acid by the similar percent of the oleic acid moiety. be done. Therefore, the smaller this number means that oleic acid has a lower value than lylunic acid. represents a greater percentage of

In the context of the present invention, an oil derived from sunflower seeds (herein, triglycerides of this oil) Cerides are the total fatty acid content of triglycerides present in this sunflower seed oil. A content of oleic acid moieties greater than about 60% (preferably 80%) relative to the amount It is desirable to use the For the amount of oleic acid in this seed, The ratio of amounts of linoleic acid is approximately less than 0.25. In other words, the oleic part and the linoleic moiety is 1:0.25, preferably 1:0.09. or less. More preferably, the amount of oleic acid moieties in the ridge is The ratio of amounts of nolic acid moieties ranges from about 0.01 to about 0.09. In other words, The ratio of leic acid:linoleic acid is 1:(0,09-0,01).

Selective removal of specific fatty acids from oil triglycerides, as shown below. It is possible to use enzymes such as However, this enzyme selection Often linoleic acid and oleic acid (which contain the same number of carbon atoms) , and where the unsaturation positions overlap).

Therefore, a high oleic component content (80% by weight or more, more preferably , 88% or more, most preferably about 95% by weight) It is particularly important to obtain oils with an unusually low linoleic acid content. present invention in connection with the sunflower seed oil being obtained from a substantially homogeneous collection of sunflower seeds. What you get is worth noting. Any particular sunflower in this collection The oil may contain a high or low content of oleic acid, and may have different ratios of linoleic acid.

However, triglycerides obtained from a substantially homogeneous collection of sunflower oil On average, a statistical mixture of 60% or more oleic material (preferably preferably 80% or more, more preferably 88% or more, most Preferably, an oil containing 95% or more oleic substances is provided. . The ratio of linoleic to oleic in this oil is preferably less than 0.25. preferably less than 0.09, more preferably about 1:(0,09-0,01 ) is within the range.

The oil extracted from the acid parts of this oil depends on the sunflower plant. It changes as the relative amount of minutes changes.

Typical oils used in connection with the present invention contain the following acid moieties in the percentage amounts indicated: Ni containing Olein (18 carbon atoms, 1 double bond) 80.0 linole (18 carbon atoms 8.1 Stearin (1B carbon atom, no double bonds) 5. 5 palmitine (16 carbon atoms, no double bonds) 4.2 behene (22 carbon atoms, two (no double bonds) 0.7 lin (18 carbon atoms, 3 double bonds) 0.2 Generally, ± Variations in IO% are within the scope of this invention.

As indicated above, breeding as described in U.S. Pat. No. 4.627.192 By this method, it is possible to obtain such high oleic content sunflower seed oil. Ru. The subject matter of this patent is hereby incorporated by reference with respect to the disclosure of such breeding methods. ing.

Additionally, U.S. Patent Application No. 769.502 (filed August 26, 1985) ) includes disclosure of sunflower breeding methods and methods for harvesting from such sunflowers. The disclosure of oils derived from seeds is provided herein.

The high oleic sunflower oil preferably used in connection with the present invention is the most commonly used Specifically, it can be obtained from the breeding method disclosed in US Pat. No. 4,627,192.

However, such oil is obtained from sunflower seeds grown in other ways. Maybe you can get it from your child. Therefore, the oleic component content as described above , and any sunflower seed oil having a ratio of linoleic acid to oleic acid , is useful for the present invention. produces seeds like this Plant matter may be obtained in many ways. Such methods include tissue culture Manipulation of plant material by using vectors to insert genetic material into target host plant cells. vectors and transcription systems (e.g., T1 plasmid vectors, microinjection of genetic material, and cauliflower mosaic virus vector) and gene isolation method and cow Yaraku talization methods are included.

2) J to Saramizu brother anti As shown above, there are many different ways to hydrolyze fats and oils. Ru. This method includes decomposition by saponification and acid hydrolysis. People like this The process involves the decomposition of triglycerides applying high temperatures and steam pressure, and Tvitchell's decomposition method is included. Generally, this kind of decomposition The fatty acid compositions obtained using the method are indicated by the fact that they have a darker color. It's not particularly pure.

The resulting sweet waters are totally (impure). Due to their impurity, the acids in their composition chemical instability and unsuitability for use in connection with many biotechnological applications. Sex happens. In order to purify the hydrolyzed products obtained from such methods, To achieve this, a distillation step that increases the amount of energy required to produce the final product is recommended. It is necessary to do so.

The necessary distillation steps will vary depending on the chain length of the fatty acid being isolated. chain length increases As the temperature increases, the temperature and degree of vacuum required to carry out the distillation also increases. This results in Additionally, additional energy is required, increasing costs. Furthermore, the distillation temperature , reactions can occur between the fatty acids themselves, resulting in polymerization and oxidative degradation. Jiru. This reduces the yield of fatty acids obtained from such processes. heavy In addition to the synthesis reactions that occur, some fatty acids are isomerized at their double bonds. As a result, a large amount of isomerized fatty acids are produced. This means that the yield of fatty acid products obtained decrease.

In view of the above problems, the present invention separates fatty acids from triglycerides in a pond. Therefore, no high temperature, high pressure methods were utilized. The present invention improves fat and oil content. Enzymatic hydrolysis is used to perform the solution. Enzyme addition carried out according to the invention Water splitting reactions are highly selective and require very low energy.

The selectivity of the reaction increases the amount of specific fatty acids removed from triglycerides; Therefore, the purity of the output obtained increases. Additionally, high temperatures are not required during hydrolysis. Since high temperatures are actually undesirable, fatty acids should be used to hydrolyze substances at high temperatures. However, the performance of enzymes derived from a particular microorganism is often specific to that substance. be. Accordingly, the enzymatic hydrolysis reactions used in connection with the present invention are as follows: This is only done for sunflower seed oils with high lenin content.

In order to obtain the present invention, such high oleic content sunflower seed oil is enzymatically added. The specific reaction conditions required to split water depend on the type and amount of enzyme, Type and amount of pH1 additive, temperature, and amount of water (these are carefully considered in relation to the inic acid composition and (necessary to obtain high yields) has been done. Adjustment of such parameters may vary depending on the hydrolysis rate and/or or increase selectivity.

Regarding the type of enzyme, the enzymes used in connection with the present invention can be of different types, such as: can be divided into categories: (1) non-site-specific enzymes; Various enzymes, and specific combinations of enzymes, can be used to extract high oleic sunflower seed oil. In order to obtain high yield and high purity oleic acid based on the contribution of the starting material, the present invention It has been found to be particularly useful in this connection.

The high oleic sunflower seed oil used in connection with the present invention has the following general structure: Consisting of triglycerides having formula (1): Here, RSR' and R'' are the hydrocarbon moieties of the acid moiety, and 80% or is the oleic acid moiety. As also shown above, preferably 88 % or more of the acid moieties are oleic acid moieties, most preferably about 95% or more is an oleic acid moiety.

When a non-site specific enzyme is contacted with a triglyceride of general structure (I), This enzyme separates all fatty acid moieties at three positions and separates them into glycerol and Leaving a mixture with fatty acids. Site-specific enzymes are utilized in relation to triglycerides. When used, this site-specific enzyme generally targets two primary triglycerides. Separate the fatty acid part from the part. Therefore, this site-specific part 2 of this fatty acid moiety by 100% efficient reaction with triglycerides such as 73 are separated. When fatty acid specific enzymes are subjected to reaction with triglycerides , this enzyme reacts with fatty acid sites where specific fatty acids are located. (This acid is (generally recognized by specific unsaturated moieties). For example, enzymes are oleic Can only react with fatty acid moieties. This fatty acid moiety is unsaturated at the 9th δ carbon. have a part. However, such fatty acid-specific moieties are also subjected to reaction with the rhein moiety (which also has an unsaturation at the 9th carbon). It will be done.

In connection with the present invention, the yield and purity of the oleic acid product obtained is 0, oleic and high. The plant breeding method described above can be used to obtain high content sunflower seed oil. It was first discovered that the increase in Here, this triglyceride is It has a particularly high content of oleic moieties. Then, the phase of the various enzymes and reaction conditions is determined. The differences were studied in order to obtain the most possible use of the most possible starting materials. this is, Different enzymes can be used in different ways with different p■, additive amounts and water amounts. Made by mixing with. Therefore, this invention has certain fruit content sunflower seeds Involves obtaining high yields of relatively pure oleic acid using oil starting materials . Hydrolyzed products obtained by enzymatic hydrolysis at the water/oil interface The substance is composed of fatty acids and glycerol. Glycerol is dissolved in the aqueous phase. It is decomposable. This aqueous phase is then removed, leaving a relatively pure oleic acid composition. Remains in high yield.

Mix enzymes or use enzymes stepwise to obtain products in stepwise reactions. It is possible to use different types of enzymes in connection with the present invention by Ru. According to biochemical nomenclature, enzymes are divided into six groups. The present invention Including the use of hydrolytic enzymes (more specifically water-soluble lipases).

(Margin below) The following describes non-site-selective lipase-inducing methods used in connection with the present invention. Table of microorganisms: viseosum) The following are microorganisms that induce site-selective enzymes used in connection with the present invention. Here is the table: These microorganisms are selective enzyme raw materials for fatty acids with the 9th δ carbon atom. be.

Certain combinations of enzymes have been found to be particularly useful in connection with the present invention. ing. These combinations are derived from the following microbial combinations:

In addition to microbial enzyme sources, animal sources can be used in connection with this invention.

This animal raw material includes, for example, pig pancreatic lipase (Porcine pancr). eatic 1ipase), Bovine panc reatic 1ipase), and pig liver esterase (Porcine Enzymatic hydration performed in connection with the present invention For the decomposition reaction, the amount of enzyme utilized depends on the amount of triglycerides that can be hydrolyzed. Depends on. The amount of this enzyme is related to the hydrolysis of triglycerides. It is expressed in digits (U). The amount of enzyme utilized depends on the specific enzyme used. ,change. The present invention merely applies enzymes to high oleic content sunflower seed oil. Because there is not, 1. The amount and type of enzyme will vary substantially depending on the oil being hydrolyzed. No. However, other considerations (e.g. reaction time, temperature and reaction medium) pH of the enzymes used) will affect the enzyme plate utilized. In connection with the present invention, olein per gram of high-content sunflower seed oil, 10-s, ooo units, preferably is 10 to 100 or more units, more preferably 20 to 40 units of fermentation. It is useful to use the raw material.

To express how the amount of enzyme used varies with the enzyme raw material, We propose the following scope: 1) Candida Rugosa ): 7-10U/II1. eq, 23.8-34. O07g.

2) 19 Poreine Want Want Frog R: 100-1000U/ +i, eq, 340-3,40007g.

3) Geotrjchum candidum : 25υ/+m, eq, 85. O07g.

4) 9x 2 and 25! (Pseudo+nonas): 3407g.

5) Koyo Ox”-y! λノJi1 (Penictllium): I U/+, e q, 3.407g.

6) Asbergillus niger (Aser 1llus nier): Commerce When used in publicly recognized processes, generally smaller amounts of enzyme are used.

More preferred ranges for certain enzyme raw materials are as follows: 1) L Otsukokyu! (Candida): 23.8-34. O07g.

2) Bu9 (Porcine): 34007g.

3 daring silver 2) +) Mumu! (Geotrichum): 85. O07g, also 4) Pseudomonas (Pseudo+nonas): 3407 g.

5) Mu Blood Seal (German μ): 85. O07g or less6) 722 soilヱ15 ! (oral): 34υ/g or less fermentation carried out in connection with the present invention The elementary hydrolysis reaction is preferably carried out at about 0% of the amount of high oleic sunflower seed oil. ．． This is done in the presence of water in an amount ranging from 5 to 1.5 times. This water is hydrolyzed It must be present in sufficient quantity to be effective. however, Inclusion of too much water also reduces the efficiency of hydrolysis and reduces the aqueous phase of the reaction medium. are difficult to effectively remove and treat. The enzymatic hydrolysis reaction of the present invention involves oil/ This occurs at the water interface.

The relative amounts of oil and water can be varied for each group and must be adjusted to obtain optimal results. Must be. Generally, the water/oil conversion is most preferably carried out at a temperature of about 35°C ± 2°C. , (1-'/2): (1-1'/2), preferably 1: (1-1'/2) 2). Temperatures are typically around 38-4 with 1.3 site-specific enzymes. 0°C, and more generally this temperature may range from 20-60°C.

The pH of the enzymatic reaction mixture affects hydrolysis. Due to hydrolysis reaction, water is removed. Produces soluble carboxylic acids. However, l) B in the range of about 4.5 to about 10.0; (more preferably in the range of 5.5 to 9). It may be desirable to include. In addition, this reaction may be affected by other additives (which are commonly (although not useful in connection with the present invention).

The enzymatic hydrolysis of the present invention is preferably performed at a temperature range of about 20°C to 60°C, and more Preferably, it is carried out at a temperature range of about 30°C to 50°C. This temperature Temperatures above about 20°C are generally used to allow the reaction to proceed fast enough to be economical. must be kept clean. Also, this temperature is , must be kept below 60°C to avoid inactivating the enzyme. . The reaction mixture was continuously stirred to allow enzymatic hydrolysis of triglycerides to proceed. Stirring is also desirable.

The relative amounts of oil/water will vary with other factors (e.g., agitation volume, temperature, and enzyme raw materials). become The amount of oil/water interface can be controlled by stirring and to some extent by temperature. be affected by The ratio of oil to water is preferably 1:(11/2), etc. The ratio is preferably 1:1.2, and the temperature is preferably between 30°C and 50°C. be. Agitation is generally used to keep the oil and water phases in uniform dispersion. It is done at a fast speed.

The following examples illustrate how oleic acid compositions are made and the hydrated compositions of this invention. To provide those skilled in the art with a complete disclosure and description of how to carry out the decomposition reactions. , is provided.

This example is not intended to limit the scope of the invention. numbers used Efforts have been made to ensure accuracy with respect to values (e.g. amounts, temperatures, etc.). However, some experimental errors and deviations should be taken into account. instructions to others Otherwise, parts are parts by weight, temperature is in degrees Celsius, and pressure is atmospheric or is around that area.

Eketsu Goko Rugosa (Candida ■dazzle■,) (A+aano Int, Lipase (10U/+, eq, oil) obtained from Enzyme Co-) 1. Go.

Add 35 liters of distilled water to 1 unit. Next, high oleic content sunflower seeds Add 100 ta, eq. of oil. This mixture was heated at about 35°C to React at a temperature of 40°C for about 22-24 hours.

This mixture is allowed to settle, forming two layers. To obtain an oleic acid composition, water, Remove the lower layer containing glycerol and other soluble impurities. Hydrolysis is It ranges from about 88% to 94%.

Large Jlt color except that acetate buffer was added to the water to maintain the pH at about 5.5 during the reaction. , repeat the method of Example 1. Hydrolysis is comparable to Example 1. That is about 88% ~94%.

K standing plate standing 1000 units of hyegusu rugosa (Candida shin) and Unicilium Penicilliun+c cloium 100 unit Dissolve the solution in 35 ml of water. Next, high oleic content sunflower seed oil (which contains 80% or more oleic acid moieties) 100+m , eq, is added. This mixture is heated at a temperature of about 35°C to 40°C with stirring. Allow to react for about 6 hours. This mixture is allowed to settle, forming two layers. Oleic acid group The lower layer containing water, glycerol and other water-soluble impurities is removed to obtain the product. leave Hydrolysis increases to over 90% in 6 hours.

If continued, this hydrolysis increases to over 98% in 24 hours.

Tsunetane plate clay Mucor m1ehei 1000 units and Benishi! J ’7A 100 units of Shifu 0 Piu A (p6 Yuyu” difference 1 ふし匝岨), 35 meters of water Dissolve in l. Next, add 100 teq of high oleic sunflower seed oil. Add. The mixture is heated at a temperature of about 35°C to 40°C with stirring at about 22-2°C. Allow to react for 4 hours. This mixture is allowed to settle, forming two layers. Oleic acid composition Remove the bottom layer containing water, glycerol and other water-soluble impurities to obtain do. Hydrolysis is greater than 96%.

Real i column”- Penicillium c cooium 200 units of lipase obtained from Pseudomonas 2.000 units of lipase obtained from Nas) (101110+, eq, oil) Add about 70 ml of distilled water to each pot. Next, oleic-rich sunflower seeds 200% of child oil (which contains 80% or more oleic acid moieties) Add m, eq. This mixture is heated at a temperature of about 35°C to 40°C for about 22-24°C. Allow time to react. This mixture is allowed to settle, forming two layers. Oleic acid composition The lower layer containing water, glycerol and other water-soluble impurities is removed to obtain Ru. Hydrolysis is greater than 93%.

L plate medical school f Lipase obtained from rugosa (Candida) 1,000 Unit: MU! Lipase obtained from Mucoy m1ehei 1,000 units, and 2-year-old IJ '7-person cyclovilla A Approximately 1 unit of distilled water per 300 units of lipase obtained from Add 05a+1. Then 300 m of high oleic sunflower seed oil; Add eq. This mixture is heated at a temperature of about 35°C at a pH range of about 5.5. Allow to react for about 22-24 hours. This mixture is allowed to settle, forming two layers. me Contains water, glycerol and other water-soluble impurities to obtain an inic acid composition Remove the bottom layer. Hydrolysis ranges from about 80% to 95%.

K Tan Goko Left side! against lipase obtained from rugosa (Candida zosa) and add 10-100 ml of distilled water. Next, high oleic content sunflower seeds Add 10-100 m, eQ, of oil. This lipase is 10% per gram of oil. should be added to provide ~35 units of linkase. this mixture , for about 4-24 hours at a temperature of about 35°C. This mixture is allowed to settle to form two layers. Let it form. water, glycerol and other water-soluble compounds to obtain the oleic acid composition. Remove the underlying layer containing impurities.

°・and, Oleic-rich sunflower seeds according to the above disclosure related to “plant breeding” Seed oil is obtained from a substantially homogeneous collection of seeds. orei as defined above High content sunflower seed oil contains the following triglycerides. this Triglycerides contain at least 60% of the oleic moiety on the triglyceride. It constitutes the fatty acid part. Furthermore, the resulting high oleic content sunflower seed oil is It has a particularly defined ratio of the amount of linoleic acid to oleic acid. “Enzyme hydrolysis In connection with the hydrolysis method described above under "Solution", high oleic content sunflower seeds By using oil, a hydrolysis reaction product with a very high oleic acid content can be obtained. It is possible to

The specific purity of oleic acid is the first oleic high content sunflower seed oil used It varies to some extent depending on. High oleic sunflower seeds as described above Utilizing seed oil and hydrolyzing this oil to obtain oleic acid Oleic acid compositions derived from ing. However, the oleic acid obtained from this high oleic sunflower seed oil The inic acid composition is prepared by physical and chemical methods such as those described in detail herein. Can be further purified.

First, after performing enzymatic hydrolysis, the reaction product is separated into two phases. The upper phase is Contains fatty acids removed from liglycerides. The lower aqueous phase is mainly and glycerol and certain impurities (which are derived from high oleic sunflower seed oil). It is made up of water in which water is dissolved. Therefore, this enzymatic hydrolysis product The first step in the purification of The goal is to separate the The remaining upper phase contains a very high concentration of oleic acid. Ru.

The properties of this oleic acid composition vary depending on the initial sunflower species used. . The following are characteristic of typical such oleic acid-rich compositions: Transmission Specific gravity (at 15.6℃) 0.899 Color (ASTM) L2.0 Color (Gardner) 5-6 %H2O0,13 Acid value 201 Iodine value: 87.8 Titer 18℃ 1” monthly 1 intense 80% oleic acid, 8.1% linoleic acid, 5.5% stearic acid, palmitic acid 4.2%, behenic acid 0.7%, and lylunic acid 0°2%.

This oil also contains some metals (e.g. Ca%BusZn and Fe) in small amounts. C (for example, 1-100 ppm). physical characteristics and chemical Each of the features varies to different extents, but typically varies by ±10%.

From the above, this upper phase contains impure fatty acids such as linoleic acid and oleic acid. other fatty acids with longer and/or shorter chains than phosphoric acids, and more unsaturated fats. fatty acids and fatty acids with low degree of unsaturation). These impurity fatty acids are by using one or more chemical or physical separation methods. , can be separated. For example, as described in U.S. Pat. No. 4,601.856. It is possible to use chemical separation methods. These contents are oleic acid Concerning the disclosure of chemical purification methods, there is provided herein.

The oleic acid-rich compositions of this invention result from the enzymatic hydrolysis process described above. It is obtained as the upper phase. This oleic acid-rich composition is available in several different types. contains fatty acids and other impurities listed above. This oleic acid composition For further purification, fatty acids with different chain lengths or different degrees of unsaturation can be used. It is possible to separate. more specifically, more than or more than 18 fatty acids having fewer carbon atoms, or more than one or less than one A fatty acid with high unsaturation is oleic acid (having 18 carbon atoms and a single (contains unsaturated bonds).

The high oleic fatty acid composition of the present invention can be obtained by subjecting this composition to low temperature treatment. Winter protection measures are taken. The temperature is gradually lowered until crystallization begins. Therefore, it is possible to separate fatty acids with a higher degree of saturation than oleic acid. Therefore , by gradually lowering the temperature, fats such as stearic acid and palmitic acid A point will be reached where the fatty acids will crystallize and precipitate within the composition. Then these The fatty acids are removed to provide a further purified high oleic acid composition.

Polar solvents (e.g. acetone and methanol) are (acid and palmitic acid) can be crystallized almost quantitatively. On the contrary , unsaturated acids, such as oleic acid, remain dissolved within the solvent. Therefore, the composition in an amount sufficient to crystallize impure palmitic acid and stearic acid in the substance, Incorporating acetone and/or methanol into the oleic-rich composition Separation can be achieved by this. After crystallization occurs, the crystallized impure pulp Filtration may be performed to separate mitic acid and stearic acid. Generally, a Setone methanol solvent is 3-4 liters of solvent per 1 liter of fatty acid. is added to this composition in a proportion of After adding the solvent, heat is added to perform crystallization. reduce the degree of Generally, this temperature is lowered to -10°C to -15°C, resulting in After crystallization occurs, filtration is performed using a vacuum rotary filter. Next The filter was then sprayed with chilled acetate to remove free oleic acid. It can be done. The solvent is removed from this oleic acid composition by flash distillation and vapor removal. is removed.

In addition to the methods indicated above, methods containing detergents (e.g. sodium decyl sulfate) By cooling this fatty acid composition in water, the oleic acid composition is purified. It is possible to do so. The crystals that form during the aqueous dispersion are covered with a film of detergent. Overturned. These crystals remain in the aqueous phase when this mixture is centrifuged. Ru. The oil phase is free of crystals and water.

It is also possible to purify oleic acid-rich compositions using fractional distillation. This way Such a method is described in U.S. Patent No. 2.054.096; 2, 224.984: 2. 322.056; and 2.674.570. All of these The contents of this document are set forth herein with respect to the disclosure of fatty acid fractionation and purification methods.

L direct five l First, the enzymatic hydrolysis method described in Example 1 above is carried out. After enzymatic hydrolysis , separate the lower aqueous phase, and separate the upper phase containing the oleic acid-rich composition of this invention. obtain. Approximately 1 liter of oleic acid composition is obtained and this 1 liter of oleic acid composition is obtained. Add 3 liters of methanol to the inic acid composition and heat between -10°C and -15°C. Reduce the temperature to a point. Maintain low temperature until crystals appear completely. crystallization A highly purified oleic acid composition is filtered to remove the get.

Regarding that the present invention is considered to be the most practical and more preferred embodiment, Shown and described here. However, within the scope of this invention, its implementation Deviations from the embodiments may occur and will be apparent to those skilled in the art after reading this disclosure. It is recognized that variations may be made. This is considered a more preferred embodiment. It is further recognized that others may differ as to particular embodiments of the invention. Ru.

(Margin below) international search report 1ma-*IIay-h Touge Bifu<hmp*m, p:T/'JSEB1034BO international search report US 8803480 SA　25149

Claims (1)

[Claims] 1. A method of manufacturing an oleic acid composition, the method comprising the following steps: Obtaining high oleic content sunflower seed oil from an aggregate of sunflower seeds; The oil contains triglycerides, which contain 60% or more Contains a fatty acid moiety composed of oleic moieties; The ratio of linoleic moieties to The high oleic content sunflower seed oil is added to an aqueous medium under conditions that cause water decomposition. enzymatically hydrolyzing the oil by contacting it with a hydrolytic enzyme in the oil; forming an oleic acid layer and separating it from the aqueous glycerol-containing medium; and Beauty Removing the aqueous glycerol-containing medium to obtain a highly pure oleic acid composition. and. 2. 2. The method as claimed in claim 1, wherein the triglyceride comprises an oleic triglyceride. The ratio of linoleic moieties to moieties is less than 0.09. 3. 2. The method as claimed in claim 1, wherein the seeds are a substantially homogeneous set of seeds. and the oleic moiety is about 80% or more of the triglyceride. Present on Lyceride. 4. 4. The method as claimed in claim 3, wherein the oleic moiety is in an amount of about 88%. and is present on the triglycerides. 5. 3. The method claimed in claim 2, wherein the triglyceride contains an oleic triglyceride. The ratio of linoleic moieties to moieties ranges from about 0.09 to about 0.01. 6. 2. The method as claimed in claim 1, wherein the seeds are a substantially homogeneous set of seeds. the oleic moiety is present in an amount of about 95%, and the triglyceride moiety is present in an amount of about 95%; In cerides, the ratio of linoleic to oleic moieties ranges from about 0.09 to about 0. ．． The range is 01. 7. 2. The method as claimed in claim 1, wherein the enzymatic hydrolysis comprises about 4.5 to about 1 It is carried out at a pH in the range of 0.0 and at a temperature in the range of about 20<0>C to about 60<0>C. 8. The method as claimed in claim 7, wherein the temperature is in the range of 30°C to 50°C. and the pH ranges from about 5.5 to about 9.0. 9. 2. The method as claimed in claim 1, wherein the enzymatic hydrolysis carried out using a formulation of hydrolytic enzymes including . 10. 2. The method as claimed in claim 1, wherein the hydrolytic enzyme is derived from: selected from the group consisting of: Candida rugosa Candid; arugosa), Candida lipolytica a), Mucor pusilis, Geotrium cande Geotrichum candidum, Pseudomonas sp. eudomonassP. ), and Venicillium cyclobium (Pe nicillium cyclopium). 11. 2. The method as claimed in claim 1, wherein the hydrolase is a Candida spp. Candidarugosa and Benicillium cyclobium (P enicillium cyclopium). be. 12. 2. The method as claimed in claim 1, wherein the hydrolytic enzyme Aspergillus niger and Benicillium cyclobi Penicillium cyclopium It's Barze. 13. 2. The method as claimed in claim 1, wherein the hydrolytic enzyme is Mucormiehei, Candida rugosa osa) and Penicillium cyclobium pium). 14. 2. The method as claimed in claim 1, wherein the hydrolytic enzyme is Squirrel (Mucor Pusilis) and Benicillium cyclobium (Pen) icillium cyclopium). . 15. 2. The method as claimed in claim 1, wherein the hydrolase is a chromobacterium. Chromobactreium viscosum and Penicillium cyclopium Reverse derived from a combination of. 16. 2. The method as claimed in claim 1, wherein the hydrolytic enzyme is Mucormiehei and Peni It is a reverse derived from a combination of cillium cyclopium). 17. 2. The method as claimed in claim 1, wherein the hydrolase is a Pseudomonas Pseudomonas sp. and Pseudomonas sp. Reverse derived from a combination of Penicillium cyclopum) be. 18. The method as claimed in claim 1, wherein the aqueous medium is selected from the group consisting of: consisting essentially of water with a buffer selected from: 4.5 to about 9.5 Acetate and phosphate buffers such that the pH range can be maintained. 19. 19. The method as claimed in claim 18, wherein the water is added during enzymatic hydrolysis. Contains a pH buffer that can maintain a pH range of 5.5 to 9.0. 20. A method of manufacturing an oleic acid composition, the method comprising the steps of: : Oleic high content sunflower seed oil containing triglycerides is added to the triglycerides. temperature in the range of 30°C to 50°C and about 5.5 to about 9.0°C to hydrolyze the Site-selective and non-site-selective reverses in aqueous media at a pH in the range of enzymatically hydrolyzing the oil by contacting it with a formulation of , the triglycerides are fatty acids composed of 80% or more oleic acid. contains acid moieties, and the ratio of linoleic moieties to oleic moieties is approximately 0. ．． less than 25; separating the aqueous medium to obtain a highly pure oleic acid composition; and. 21. 21. The method as claimed in claim 20, further comprising: A solvent selected from the group consisting of acetone and methanol is added to the high purity olefin. adding to the inic acid composition; adjusting the temperature of the high purity oleic acid composition to about 0°C to - reducing to a point in the range of 20°C; maintaining the temperature in the range of 0°C to -20°C until crystallization of the fatty acids occurs; and Further purified oleic acid is filtered to remove the crystallized fatty acids. Obtaining a composition. 22. The method as claimed in claim 1 further comprising the steps of: The temperature of the high purity oleic acid composition is increased to a temperature in the range of -10°C to -15°C. to lower; Maintaining a temperature in the range of -10°C to -15°C until crystallization of the fatty acids occurs. ;and Filtering to remove the crystallized fatty acids.