KR20030041615A - Method of recovering pinitol or chiro-inositol in high yield from soy fractions - Google Patents

Abstract

PURPOSE: A method for isolating chiro-inositol components from the soybean-processing by-products in higher yield is provided, thereby other sugar components except pinitol or chiro-inositol can be removed from the soybean-processing by-products, so that the chiro-inositol component can be isolated in higher yield. CONSTITUTION: The method for isolating chiro-inositol components from the soybean-processing by-products in higher yield comprises the steps of: (a) culturing microorganisms, yeast and fungi in the soybean-processing by-products to increase the content of pinitol or chiro-inositol as the chiro-inositol components; (b) centrifuging or filtering the cultured medium to remove a pellet of cells, water-insoluble materials and high molecular substances; and (c) passing the pre-treated solution through active carbon to adsorb or dissolve the pinitol or chiro-inositol, wherein the yeast is Saccharomyces sp. or Aspergillus sp.

Description

Method of recovering chiroinositol from soybean by-products in high yield {Method of recovering pinitol or chiro-inositol in high yield from soy fractions}

The present invention relates to a method for separating chiroinositol components from soybean processing by-products in high yield, and more specifically, soybean molasses produced in the process of producing tofu products or soy protein that are discarded after manufacturing tofu using soybeans. In addition, the present invention relates to a process for economically separating high yield of finitol or chiroinositol using a microbial treatment and activated carbon column from a solution obtained by hydrothermal extraction of defatted soybean meal.

Recently, as interest in functional foods increases, attempts are being made to systematically evaluate the effects of health foods on the basis of health functionalities and to discover new health functional foods. Soybean can be said to be a food material having a representative health function that is highlighted in this trend.

Soybean is known to have various physiological activities such as anti-cancer, anti-arteriosclerosis, antioxidant, hypoglycemic effect and antimicrobial effect as well as its nutritional aspects. Soybean ingredients known to have such physiological activity include isoflavones, saponins, lecithins, trypsin inhibitors and the like. Isoflavones have been shown to have anti-cancer effects and prevent osteoporosis, and are already sold as health foods in Korea as well as in developed countries such as the US and Japan. In addition, soybean oligosaccharides isolated from sugar components in soybeans, such as raffinose and stachyose, have been proved to be effective for intestinal useful bacteria and have been commercialized in Japan.

Soybeans are expected to contain other biologically active ingredients, and many studies have been conducted. Among them, a particularly interesting component is a hypoglycemic effect when administered to patients with type 2 (insulin-independent) diabetes. It is known that there is a Cairo inositol and its methyl ether type of finitol.

Chiro-inositol and pinitol are well known substances, and chiroinositol is an isomer of myo-inositol as its structural isomer. In addition, finitol has a structure in which a methyl group is linked to an ether bond to carbon 3 of Cairo inositol.

The hypoglycemic effect of Cairoinositol has been reported since the early 1990s (Ortmeyer et al., Endocrinol . 132: 646-651, 1993; Huang et al., Endocrinol . 132: 652-657, 1993; Farese et al., Proc. Natl. Acad. Sci. USA , 91: 11040-11044, 1994; Fonteles et al., Diabetologia 39: 731-734, 1996). In addition, Cairoinositol has no side effects such as gastrointestinal disorders, liver damage, and hypoglycemia when overused by existing oral hypoglycemic agents, and it is included in natural foods, so it can be used safely when developing it as a health supplement or pharmaceutical material. Your chances of success are high. In addition, Cairoinositol is known to be effective in the treatment of obesity, polycystic ovary, etc. (Nestler JE et al., New Eng. J. Med ., 340: 1314-1320, 1999). In addition, pinitol, which is a predominant chiroinositol component of soybean, has been found to have an equivalent hypoglycemic effect by itself (US Pat. No. 5,827,896; Narayanan et al, Current Science , 56 (3): 139-141, 1987).

Until now, in the method for preparing chiroinositol, a method of hydrolyzing pinitol (methyl ether of di-chiroinositol) extracted from plant leaves (Anderson et al., Ind. Eng. Chem ., 45: 593-596, 1953), A method of converting myo-inositol to chiroinositol by an organic chemical method (Shen et al., Tetrahedron Letters , 131: 1105-1108, 1990) has been disclosed in the literature, but such known manufacturing processes are time consuming. It takes a long time and is not economical because the yield is low, and the method of synthesizing from kasugamycin (U.S. Patent No. 5,091,596) also has a low yield and is considered to be a considerably high manufacturing price. In order to develop a method for producing lysine, edible resources, especially soybeans, processed soybeans and pine needles, containing chiroinositol, The method and a method for separating and purifying chiroinositol components by acid hydrolysis from this edible resource (Korean Patent Application No. 10-2000-12881, Korean Patent Application No. 10-2000-12882) As a further improved process, a method using an activated carbon column was developed to use a conventional zeolite (US Pat. No. 4,482,761), a method using a cation exchange resin (US Pat. No. 5,096,594), and a method using an anion exchange resin. (US Pat. No. 5,482,631) has been patented for a more economical separation method of finitol and chiroinositol (US Patent Application No. 10-2001-001611), and also prior art techniques for glycine glycoside from soybean processing by-products. Alternatively, glycosides or phosphorus can be solved by using microorganisms such as bacteria, yeast, and mold to solve the disadvantage of failing to recover phosphorus compounds. Compound such as to develop a method for more effectively to recover pinitol and Cairo inositol by giving the switching elements present in different forms in pinitol and Cairo inositol was bar filed patent (Republic of Korea Patent Application No. 10-2001-44677).

The present inventors studied and tested a process that can further increase the economic efficiency in recovering the chiroinositol component from the soybean processing by-product, and after increasing the concentration of pinitol by converting the pinitol derivative in the soybean byproduct into pinitol through microbial treatment, the treatment solution After ethanol was removed and ethanol was removed from the supernatant from which the precipitate was removed by filtration or centrifugation, the purity of finitol was very high by using activated carbon. Finally, crystallization was performed by adding a solvent such as ethanol. It is possible to obtain a finitol product with a purity of 90% or more to complete the present invention.

Accordingly, it is an object of the present invention to provide a method for effectively enhancing the soybean processing by-product endinitol or chiolinositol content using microbial bioconversion.

It is another object of the present invention to provide a method for recovering finitol or chiroinositol from tofu or soy molasses or hot water extract of skim soybean using an activated carbon column.

It is still another object of the present invention to provide a method for recovering chiroinositol component in high yield using soybean by-products and microbial treatment and activated carbon adsorption methods.

Hereinafter, the configuration of the present invention will be described in detail. Other objects and advantages of the invention will be more clearly understood by the following examples.

1 is a schematic process diagram illustrating a process of separating finitol and chiroinositol from soybean by-product using an activated carbon column.

Figure 2 is a diagram showing the elution results of the tofu impurities adsorbed on the activated carbon packed column: at this time, the volume of the packed column (BV) is 50ml, eluent ethanol, ethanol concentration 10% (v / v) 500mL, 50% (v / v) 500 mL, eluent rate 500 mL / min, PI = finitol (10 times), CI = chiroinositol (10 times), OS = soy oligosaccharide.

Figure 3 is a diagram showing the results of elution of the tofu impurities adsorbed on the activated carbon packed column (packet column volume (BV) 500 mL, eluent ethanol, ethanol concentration 10% (v / v) 500 mL, 50% (v / v) 500 mL, eluent rate 500 mL / min: PI = pinitol (10-fold), CI = Cairoinositol (10-fold), OS = soy oligosaccharide).

Figure 4 is a schematic process diagram showing the process of separating the finitol and the chiroinositol from soybean by-products using a microbial treatment and activated carbon column.

Figure 5 is a preferred embodiment of the present invention, a figure showing the manufacturing process of high purity finitol product.

In a broad sense, the present invention relates to natural resources containing finitol and / or chiroinositol (hereinafter, the two compounds are referred to as the 'chiroinositol component'), such as medicinal beans, soybeans, soybeans, soybean leaves, soybean germ, skim soybeans, The present invention relates to a method for recovering high yield of finitol or cairoinositol from tofu sprouts, soy oligosaccharides, soybean pods, bean sprouts, pine needles, pine needles, pine bark, and the like. Cultivating microorganisms such as yeast, mold, etc. to increase the content of the compounds, selective adsorption and elution using activated carbon column to fractionate the compounds in high concentrations, and as a whole finite or chiroinositol from the above edible resources It provides a method of economically producing in high yield. The method of the present invention, which is significantly different from the conventional method, effectively removes other sugars in the edible resources, thereby significantly reducing the organic load in the finally discharged waste, and highly containing only the chiroinositol component. The fraction can be recovered in a small volume, so that the chiroinositol component is separated from the soybean product in high yield.

Soy fractions, which is a preferred edible resource containing a large amount of pinitol or chiroinositol in the present specification, are generated in the process of producing tofu or soy protein that is discarded after manufacturing tofu using soybean or soybean. It is used in a broad sense including soybean molasses, hot water extracted solution of skim soybean meal, or a mixed solution thereof. Usually, 1 g of soybean contains 3.4 to 6.8 mg of the chiroinositol compound, 15 to 20% of which is in the form of chiroinositol, 25 to 35% in the form of finitol, and the remaining 50 to 60% of the glycosides of chiroinositol or finitol Present in the form of phosphorus compounds. In addition, 15-20% is in the form of chiroinositol, 30-40% is in the form of finitol, and the remaining 40-55% is in the form of glycosides or phosphorus compounds of Cairoinositol or finitol. Specific examples include pinitol glycosides, pinitol phosphates, pinitol phytates, pinitol phospholipids, pinitol esters, and lipid-bound pinitol. As such, soybean by-products contain various kinds of chiroinositol compounds, most of which are present in the form of pinitol and also in the form of glycitol glycoside or phosphorus compound. Unlike the conventional finitol recovery process there was a problem that is discarded.

However, the present inventors have found that although the total amount of the chiroinositol component in the soybean sprouts released after soybean processing does not change, the content of free chiroinositol and pinitol gradually increases over time after the tofu sprouts are released. The results of this study are due to the fact that the derivatives of chiroinositol and pinitol in soybean by-products are converted to free chiroinositol and pinitol, which are effective in strengthening blood sugar, through microbial action such as bacteria, yeast, and fungi that naturally inhabit tofu products. It turned out that Through the microorganism treatment, the present inventors can convert the finitol derivative in soybean by-product to finitol, thereby maximizing the recovery rate of finitol in soybean by-product as well as effectively removing other sugars contained in soybean by-products in the subsequent separation and purification process. The treatment burden was reduced and the organic content of fermentation waste was greatly reduced.

As microorganisms used in the present invention, various kinds of bacteria, yeasts, and molds may be used, and reaction conditions of the conversion process, recovery rate of finitol, and removal efficiency of other sugars may vary depending on the type of microorganisms used. In particular, Saccharomyces carlsbergensis ( Saccharomyces carlsbergensis) of the microorganism was evaluated as the most preferable because it has the best ability to produce pinitol and also the ability to remove sugar components. After the microbial treatment is completed, the microorganisms are recovered through various subsequent recovery processes after removing the cells. In a preferred embodiment of the present invention, the dominant species among the microorganisms naturally inhabiting tofu and acting to increase the content of pinitol are purely separated and their ability to produce pinitol is shown in Table 1 below. Table 1 shows the results of measuring the change in the content of pinitol and chiroinositol after 5 days of inoculation by inoculating two types of yeasts and three kinds of bacteria which were naturally isolated after treating the tofu impurities at 121 ° C. for 15 minutes.

TABLE 1

Finitol Formability of Microorganisms Isolated from Tofu Purity

It can be seen from Table 1 that the type of microorganism contributes to an increase in the content of pinitol and chiroinositol, regardless of yeast or bacteria. The same experiments were conducted on several different microorganisms, indicating that an increase in the content of pinitol in soybean meal was caused by all microorganisms rather than by specific microorganisms. However, the growth rate of finitol was different according to the type of microorganism, and also the availability of sugars contained in pure water was different. In order to improve the completeness of the present invention, the inventors of the present invention 1) the production rate of pinitol is fast, 2) to remove sugars contained in the pure water, and 3) to screen the microorganisms that have been certified as safe for food. The results are shown in Table 2 below.

TABLE 2

Finitol Formation Effect of Various Microorganisms during Tofu Purity Treatment

In the case of yeast in the production speed of the pinitol is saccharide as MY process Carlsbad Bergen sheath (Saccharomyces carlsbergensis), saccharose in my process serenity non-zero (Saccharomyces cerevisae), if the fungus is Aspergillus or this (Aspergilus niger), Trichoderma irregularities having ( Trichoderma viride ), E. coli was particularly excellent in bacteria, Saccharomyces Carlsbergensis of the yeast was excellent. Saccharomyces Carlsbergensis and Aspergillus niger were found to be superior in sugar availability. In contrast to yeast and bacteria, fungi, including Aspergillus niger, increased the content of pinitol in the first two or three days after incubation, but decreased again afterwards. At the end of the cultivation period that people lacked, it was determined that the generated finitol is used as a nutrient. All of the microorganisms in Table 2 are those that are permitted to be used for one or more purposes on the Japanese Food Additives Code. The microorganisms for which the safety of food processing is generally secured are selected. Based on the above phenomena, it was determined that Saccharomyces Carlsbergensis was the most preferable microorganism as the microorganism for generating finitol.

Powdered degreased soybean meal, another raw material for the production of finitol, was extracted with hot water to remove undissolved powder, and even when treated with Saccharomyces Carlsbergensis, the concentration of finitol was 1.5-2.0. The result was similar to that of tofu sprouts. At this time, when the microorganisms were inoculated and cultured without removing the defatted soybean meal, the concentration of pinitol was further increased by 2.0 to 2.5 times. This may be due to the extraction of more glycosides present in the powder during the incubation period for several days.

To determine whether this increase in the concentration of finitol is caused by the action of specific enzymes produced by microorganisms, two kinds of microorganisms, Saccharomyces Carlsbergensis and Aspergillus niger, were incubated in tofu and then cultured with 0.45 μ filter paper. After complete removal of the mixture, the mixture was sterile and mixed with other pure products for two days. As a result, the concentrations of finitol and chiroinositol did not increase and no change in composition occurred. This fact suggests that the increase in the concentration of pinitol by microorganisms is due to a complex mechanism rather than one or two enzymes. The microorganism treatment for tofu sprouts or skim soybean extract may be carried out as it is or may be carried out, for example, after concentration of 10 to 20 times.

The present invention also provides a method of using an activated carbon column for recovering the chiroinotol component from the hot water extract of tofu, soybean molasses, or skim soybean. Tofu or skim soybean water extract contains a large amount of oligosaccharides in addition to finitol and chiroinositol. The present inventors have developed a method using an activated carbon column as a method for separating finitol and chiroinositol from other sugars. In the activated carbon column, pinitol and chiroinositol are adsorbed together with other sugars, and eluting while increasing the concentration of an organic solvent such as ethanol separates and elutes finitols and chiroinositol and other sugars (FIG. 1).

The process of the present invention for separating the chiroinositol component using the activated carbon column from the soybean by-product will be described in more detail with reference to the process for separation and purification shown in FIG. 1.

First process : pretreatment process

The pretreatment process performs two functions to increase the efficiency of the activated carbon process, which is a key process. First, insoluble matters in the soybean byproducts are removed using a centrifuge or a general filter. Failure to remove insoluble matters will block the activated carbon column and impede normal operation. Second, ultrafiltration removes polymers such as proteins contained in soybean processing by-products. Since the protein component is hardly removed after being adsorbed on activated carbon, it plays a role of significantly reducing the lifetime of activated carbon.

Second Process : Activated Carbon Filling Tower Adsorption Process

Through the pretreatment process, the sample in which the insoluble component and the polymer material are removed is passed through a column filled with activated carbon to adsorb the chiroinositol component and soy oligosaccharide. First, to increase the adsorption capacity, the pH is adjusted to 6 to 8 neutral with caustic soda. Samples of about 5 to 10 times the volume of activated carbon packed column can be processed at a time based on tofu with a total solid concentration of 25 g / L. The proper flow rate is 1 to 2 packed volume per hour (Bed Volume). ).

Step 3 : Elution from Activated Carbon Column

First, to remove the non-adsorbed components remaining in the activated carbon packed column is washed with 1 ~ 2 BV of distilled water. Suitable washing rates are 1 to 2 BV per hour. Then, in order to elute only the chiroinositol component adsorbed on the activated carbon, an eluent containing 1 to 2 BV of an organic solvent having a concentration of 5 to 20% (v / v) is supplied. Methanol, ethanol, isopropanol, or acetone may be used as the organic solvent. However, ethanol has been found to be the most suitable solvent in consideration of efficiency, safety, and economic efficiency. The pH of the eluent is suitably 3 to 4, in which the equilibrium constant of the chiroinositol component is lowered, and an appropriate feed rate is 0.5 to 2 BV per hour. Subsequently, a regeneration solution 1 BV containing the same solvent at a concentration of 40 to 80% is passed through to remove components strongly adsorbed to activated carbon such as oligosaccharides. The appropriate feed rate for this step is also 0.5 to 2 BV per hour. The eluate is fractionated into appropriate volume units, analyzed using high performance liquid chromatography (HPLC), and the fractions containing the chiroinositol component are collected together. When experience with the elution is accumulated, only the concentration of the solvent containing the chiroinositol component can be collected without a separate fractionation by measuring only the concentration of the solvent in the eluate using a refractometer. In the order of weak adsorption to activated carbon under elution conditions, the finitol fraction comes out first from 0.6-2.2 BV after initiation of elution, the chiroinositol fraction from 2.0-3.0 BV, and the oligosaccharide fraction from 2.6-4.0 BV. The finitol fraction contains 50 to 60% of finitol and 2 to 10% of the chiroinositol, and the total content of the chiroinositol component is 55 to 70%. The chiroinositol fraction contains 5 to 10% of finitol and 10 to 15%. Contains chiroinositol and does not completely separate from the oligosaccharide peak, thus containing a significant amount of oligosaccharides. When these two fractions are combined, the yield is 75-85% relative to the total amount of the chiroinositol component contained in the soybean processing by-product. The oligosaccharide fraction collected from 3.0 to 4.0 BV fraction contains only 0.1 to 0.5% of the chiroinositol component relative to the total solids. After passing through the regeneration solution, the solvent component remaining in the activated carbon is sufficiently removed by washing with 3 to 6 BV of distilled water, and the activated carbon can be reused for the next adsorption operation.

4th step : concentration of eluate and solvent recovery

Each eluate fraction was distilled under reduced pressure at a temperature of 60 ° C. or lower to recover the solvent in the fraction, and then concentrated for the subsequent process. Concentration is carried out until the solids content is at least 10%. The recovered solvent can be reused for the next operation after adjusting the concentration.

5th process : drying

The concentrate obtained in the fourth step is lyophilized or spray dried to obtain a white or yellow powder product.

An example of applying the basic process is as follows.

Application Process 1 : Combination with Isoflavone Recovery Process

Prior to the second step, isoflavone recovery, which is another useful ingredient contained in the soybean processing by-product, can be carried out. Recovery of isoflavones isoflavones obtained when the sugars of the isoflavone glycoside components are separated by using an adsorption resin (HP resin, Samyang Corp.) or by treating alpha-galactosidase enzyme. There is a method for recovering the precipitate (Korean Patent Publication No. 1998-032766). In the treatment of the adsorption resin for isoflavone recovery, the chiroinositol component is hardly adsorbed, whereas the protein component is substantially removed, and thus there is a pretreatment effect.

Application Process 2 : Combined with Soybean Oligosaccharide Recovery Process

In the oligosaccharide fraction generated in the fourth process, most of the oligosaccharides contained in the soybean processing by-products are recovered and many impurities such as salts and proteins other than oligosaccharides are removed. Obtain soy oligosaccharide products.

Application Process 3 : Retreatment of Cairo Inositol Fraction

The chiroinositol content can be increased by reprocessing the chiroinositol fraction using a separate activated carbon column. When the concentrated chiroinositol fraction is adjusted to pH 3 to 4 and chromatographed on an activated carbon column, only the chiroinositol having weak adsorptive power is eluted at this pH. Collect this Cairoinositol fraction to obtain a Cairoinositol product with a purity of 50% or more. In addition, a method of reprocessing by a method using a conventional anion exchange resin (US Pat. No. 5,482,631) is possible.

Application Process 4 : Manufacturing of High Purity Finitol Products

The concentrated solution of the finitol fraction obtained through the fourth step is further concentrated until the solid content is 50% or more, cooled, and then acetone is added in a volume of 1: 1 and left at a low temperature of 10 ° C. or lower for at least 12 hours. The precipitate is recovered by centrifugation or vacuum filtration and dried under reduced pressure to obtain a Finitol product having a purity of 95% or more.

Advantages of the activated carbon process of the present invention compared to the process using known ion exchange resins are: 1) high concentrations of samples of low concentration, such as tofu, since desorption is not possible until the cairoinositol component is strongly adsorbed and subjected to separate elution conditions. 2) the salt components contained in the sample are not adsorbed and are simply passed through, so no separate desalting process is required in the pretreatment step. 3) If the proper elution conditions are given, only the chiroinositol component is high. Since this is recovered in a small volume, it is effective to reduce the concentration burden in the subsequent process.

The separation method using the activated carbon column may be used to recover the chiroinositol along with a process in which a microbial treatment process is additionally combined before the pretreatment process of removing impurities in soybean by-products.

Basically, if activated carbon column is used, it can be divided into pretreatment process to remove insoluble components and proteins such as protein in soy processing by-products, activated carbon adsorption process, elution process from activated carbon column, and post-treatment process to recover chiroinositol component in powder form. More specifically, the pretreatment process of removing the insoluble material and the polymer material in the soybean processing by-products by centrifugation or filtration; Activated carbon packed column adsorption step of adsorbing the chiroinositol component by passing the sample in which the insoluble component and the polymer material is removed through the pre-treatment process through a column filled with activated carbon; And removing the non-adsorbed components by washing the activated carbon packed column subjected to the adsorption process with distilled water and continuously or stepwise concentration of an organic solvent such as methanol, ethanol, isopropanol, or acetone in a concentration range of 5-20% (v / v). It may include the step of eluting the chiroinositol component adsorbed on the activated carbon by supplying to increase.

In addition, a method of recovering chiroinositol component using an activated carbon column first prepares a reagent in which the concentration of the chiroinositol component is increased through microbial treatment, removes the cells through centrifugation or filtration, and then adsorbs the activated carbon or performs other suitable processes. It is also possible to recover the chiroinositol or finitol in high purity, and more specifically, the method comprises the steps of increasing the content of pinitol or chiroinositol in the soybean by-product through the microbial treatment of the soybean by-product containing the chiroinositol component to be separated; A pretreatment step of removing the insoluble components and proteins such as proteins in the microbial cells and soybean processing by-products generated by culturing the microorganism through filtration or centrifugation; And separating the chiroinositol component from the pretreated chiroinositol-containing aqueous solution by chromatography using cation or anion exchange resin or selective chuck adsorption and elution using activated carbon.

In a preferred embodiment of the present invention, the method for recovering the chiroinositol component from soybean processing by-products is concentrated by soybean processing by-products and then cultured microorganisms such as bacteria, yeast or fungus content of pinitol or chiroinositol in soybean by-products Increasing the process; A pretreatment step of removing the insoluble material and the polymer material in the cells and soybean processing by-products generated during the microbial culture by centrifugation or filtration; And adsorbing and eluting the pretreated chiroinositol component-containing aqueous solution using an activated carbon column (FIG. 4).

On the other hand, an additional treatment process is required to produce a high purity finitol product having a content of 90% or more based on the above-described manufacturing method of pinitol. The crystallization process is essential to achieve a content of more than 90%. To enable the crystallization operation, impurities that inhibit the crystallization must be removed in advance, and the purity of finitol in the solution before crystallization must be 70% or more. As a result of analyzing the composition of the solution after the microorganism treatment, only 5 to 7% of the total solid content was finitol, and the rest was composed of protein, fat, other carbohydrates, and salts. When the solution is adsorbed onto the activated carbon column as it is, the treatment capacity of the activated carbon is lowered by the components other than these finitols, and it is difficult to increase the purity of the recovered finitol by more than 70%. To solve this problem, in order to increase the purity of finitol to 70% or more, the solution after the microbial treatment was concentrated to 50 to 70% (w / w) or more of total solid content, and then 95% ethanol solution of 1 to 3 times the volume of the solution was When added, pinitol was completely dissolved in the supernatant, while more than 75% of the total solids contained in the solution could be removed as a precipitate. After removing the precipitates by filtration or centrifugation, the supernatant was recovered and analyzed for composition. The content of pinitol in the total solids increased to 20% or more. The ethanol in the supernatant liquid was recovered by evaporation, and then evaporation was continued and completely removed while adding water over several times. The ethanol-free pinitol solution was eluted with 10% ethanol solution after adsorption on activated carbon column, and the purity of pinitol increased to 70% or more. Prior removal of the impurities in the solution by solvent treatment prior to injection of the activated carbon column, as described above, can not only obtain a high purity finitol product, but also increase the processing capacity of the activated carbon column and also irreversible adsorption of proteins and the like. There are many advantages, such as to alleviate the shortened phenomenon of activated carbon life. The solution produced in this way was concentrated so that the concentration of finitol to 600 g / L or more, and then crystallized by adding a solvent such as ethanol, it was possible to obtain a product of 90% purity or more.

Therefore, in another preferred embodiment of the present invention, the method for recovering the chiroinositol component from the soybean processing by-product in a high yield is to concentrate the soybean processing by-products and then treat microorganisms, such as bacteria, yeast or mold, to obtain finitol in the soybean by-product. Increasing the content of kaironiositol; Concentrating the microbial treatment solution and adding ethanol to precipitate insoluble materials and polymer materials in cells and soybean by-products produced during microbial culture; A pretreatment step of removing the precipitated materials and insoluble materials by centrifugation or filtration; And adsorbing and eluting the chiroinositol component by passing the pretreated solution through an activated carbon column. In one preferred embodiment, a method of recovering finitol from a soybean by-product in high yield includes (a) soybean by-product. Concentrating and treating microorganisms such as bacteria, yeast or mold to increase the content of pinitol or chiroinositol in soybean by-products; Concentrating the solution after the microorganism treatment to a total solid content of 50 to 70% (w / w) or more and precipitating solids contained in the solution by adding a volume of 95% ethanol to a volume of 1 to 3 times the solution; A pretreatment step of removing the precipitated material by centrifugation or filtration; (c) removing ethanol from the separation solution in which pinitol is concentrated in a large amount by the pretreatment step by evaporation; (d) adsorbing and eluting finitol by passing the ethanol removal solution through an activated carbon column; And (e) concentrating and crystallizing the eluted solution to produce a high purity finitol (FIG. 5).

In addition, the above-described microbial treatment method or the activity detection column adsorption method or the separation method combined with them are combined with the process of recovering other components in soybean by-products such as isoflavones, soy oligosaccharides, etc. to separate the desired active ingredient from the soybean by-products. You may.

Finitol or chiroyinositol prepared by the method of the present invention can be used in pharmaceuticals or foods to prevent or treat its complications such as diabetes, obesity, cataracts, etc., which are used in the form of a general pharmaceutical composition or chiroinosicol or It can also be used in the form of functional drinks or foods that contain finitol as an active ingredient.

Hereinafter, the configuration and working effects of the present invention will be described in more detail with reference to Examples. However, the following examples are only for the purpose of illustrating the present invention, and the scope of the present invention is not limited by these examples according to the gist of the present invention.

Example 1 Pure Water Treatment by Saccharomyces Carlsbergensis

Saccharomyces carlsbergensis ( Saccharomyces carlsbergensis ) was inoculated in sterile tofu pure incubated for 5 days to obtain the results shown in Table 3. After five days of incubation, finitol increased 2.31 times from 0.371 g / L to 0.857 g / L and 91% of total sugar was removed.

TABLE 3

Pure water treatment effect by Saccharomyces carlsbergensis (unit: g / L)

Example 2 Pure Water Treatment with Saccharomyces cerevises

Saccharomyces cerevisae ( Saccharomyces cerevisae ) was inoculated in sterilized tofu incubated for 5 days to obtain the results shown in Table 4. After five days of incubation, finitol increased 1.82 times from 0.371 g / L to 0.676 g / L and 54% of total sugar was removed.

TABLE 4

Pure water treatment effect by Saccharomyces cerevises (unit: g / L)

Example 3 Pure Water Treatment by Saccharomyces Pastorinus

Saccharomyces pastorianus ( Saccharomyces pastorianus ) was inoculated in sterile tofu incubated for 5 days to obtain the results shown in Table 5. After five days of incubation, finitol increased 1.59-fold from 0.371 g / L to 0.590 g / L and 60% of total sugar was removed.

TABLE 5

Pure water treatment effect by Saccharomyces pastorianus (unit: g / L)

Example 4 Pure Water Treatment by Candida Utilities

Candida utilitis was inoculated in sterile tofu and incubated for 5 days to obtain the results shown in Table 6. Finitol was peaked at 0.539 g / L after 3 days of incubation at the initial 0.371 g / L and then decreased again. Only 31% of total sugar was removed.

TABLE 6

Pure water treatment effect by Candida utilitis (unit: g / L)

Example 5 Pure Water Treatment by Aspergillus Niger

Aspergillus niger ( Aspergilus niger ) was inoculated in sterile tofu incubated for 5 days to obtain the results shown in Table 7. Finitol peaked at 0.566 g / L after 3 days of incubation at the initial 0.371 g / L and then decreased again afterwards, almost disappearing after 5 days, and 94% of total sugar was removed.

Table 7

Pure water treatment effect by Aspergilus niger (unit: g / L)

Example 6 Pure Water Treatment with Penicillium Funiculosum

Penicillium funiculosum ( Penicillim funiculosum ) was inoculated in sterile tofu incubated for 5 days to obtain the results shown in Table 8. Finitol peaked at 0.534 g / L after 3 days of incubation at the initial 0.371 g / L and then decreased again and 49% of total sugar was removed.

TABLE 8

Pure water treatment effect by Penicillim funiculosum (unit: g / L)

Example 7 Treatment of Pure Water by Trichoderma Biride

Trichoderma viride was inoculated in sterile tofu and incubated for 5 days to obtain the results shown in Table 9. Finitol peaked at 0.709 g / L after a one-day incubation at an initial 0.371 g / L and then decreased again and 38% of total sugar was removed.

TABLE 9

Pure water treatment effect by Trichoderma viride (unit: g / L)

Example 8 Treatment of Pure Water by Bacillus Stearothermaphilus

Bacillus stearothermophilus ( Bacillus stearothermophilus ) was inoculated in sterile tofu incubated for 5 days to obtain the results shown in Table 10. After five days of incubation, finitol increased 1.39-fold from 0.371 g / L to 0.514 g / L and 29% of total sugar was removed.

TABLE 10

Pure water treatment effect by Bacillus stearothermophilus (unit: g / L)

Example 9 Pure Water Treatment by Escherichia Coli

E. coli ( Esherichia coli ) was inoculated in sterile tofu and incubated for 5 days to obtain the results shown in Table 11. After five days of incubation, finitol increased 1.98-fold from 0.371 g / L to 0.733 g / L and 31% of total sugar was removed.

TABLE 11

Pure water treatment effect by Escherichia coli (unit: g / L)

Example 10 Pure Water Treatment by Pseudomonas Amylodermosa

Pseudomonas amylodermosa ( Pseudomonas amylodermosa ) was inoculated in sterile tofu incubated for 5 days to obtain the results shown in Table 12. Finitol was peaked at 0.604 g / L after 1 day incubation at an initial 0.371 g / L, and then decreased again. Only 24% of total sugar was removed.

TABLE 12

Pure water treatment effect by Pseudomonas amylodermosa (unit: g / L)

Example 11 Treatment of Skim Soy Hot Water Extract and Skim Soy Suspension by Saccharomyces Carlsbergensis

Saccharomyces carlsbergensis ( Saccharomyces carlsbergensis ) was inoculated in sterile skimmed hot water extract and skim soybean suspension and incubated for 5 days to obtain the results shown in Table 13. After 5 days of incubation, the defatted soybean hot water extract increased 2.02 times from 0.596 g / L to 1.209 g / L, and in the case of defatted soybean suspension, finitol increased 2.51 times from 0.618 g / L to 1.551 g / L. .

TABLE 13

Effect of Treatment of Skim Soy Hot Water Extract and Skim Soy Suspension by Saccharomyces carlsbergensis (Unit: g / L)

Example 12 Separation of Chiroinositol Components Using Activated Carbon Column

Experiment 1: Investigation of adsorption equilibrium constant for activated carbon by pH

In order to investigate the suitability of activated carbon for the purpose of separating the chiroinositol component from other sugar components in the sample, the adsorption equilibrium constants of chiroinositol, finitol and sugar were determined for each pH in the presence of activated carbon. Indicated. Here, the adsorption equilibrium constant refers to the ratio of the concentration of a component adsorbed to the adsorbent under a given condition and the concentration remaining in the solution without being adsorbed.

TABLE 14

Adsorption Equilibrium Constants in the Presence of Activated Carbon of Cairoinositol, Finitol and Sugars

According to the above results, since all of the chieroinositol, finitol and sugar showed high equilibrium constants for activated carbon near the neutral pH, after adsorption under this condition, the chiroinositol and pinitol were eluted first. Therefore, it can be seen that these two components can be easily separated from sugar.

Experiment 2: Component Composition of Tofu Purity

Tofu impurities used in the present invention were obtained and used at a tofu factory in Jinju, and the general and chiroinositol components in the tofu impurities are shown in Table 15.

TABLE 15

Ingredient Analysis Table of Tofu Purity

Experiment 3: Extraction of Hot Water from Defatted Soybean Meal

Degreasing soybean meal was pulverized and 800 g of water was added to 200 g of the powder passed through 20 meshes, followed by hot water extraction at 80 ° C. for 2 hours with stirring. The extract was centrifuged at 10,000 rpm to give 600 mL of solid soybean solution (8.3% solids, w / v). It was as Table 16 when the general component and the chiroinositol component in this solution were measured.

TABLE 16

Component Analysis Table of Defatted Soybean Meal Hot Water Extract

Experiment 4: Constituents of Soybean Molasses

Analysis of the general and chiroinositol components contained in soybean molasses obtained from various sources showed the results shown in Table 17.

TABLE 17

Soybean Molasses Analysis Table (Unit: g / 100g)

Experiment 5: Recovery of Chiroinositol Components in Tofu Purity

After filtering 4 L of tofu impurities composed of the same ingredients as in Example 2 to remove insoluble solids, the pH was adjusted to 8.0 and the sample was placed in a glass column (inner diameter 5 cm x length 30 cm) filled with 500 mL of activated carbon. Was injected at a flow rate of 500 mL per hour so that the chiroinositol component was adsorbed onto the activated carbon. Activated carbon was used granular activated carbon of 30 to 80 mesh size. After the adsorption of the sample was completed, 500 mL of distilled water was passed to wash out the unadsorbed impurities. Then, 500 mL of 10% (v / v) ethane was flowed into the solution and 50% (v / v) ethanol solution at a flow rate of 500 mL per hour to elute the components adsorbed on the activated carbon. Subsequently, it was washed with 2 L of distilled water to remove ethanol remaining in activated carbon and reused for the next adsorption operation. 100 mL of the eluted sample was collected from the start of elution with ethanol solution until the end of washing. Finitol and chiroinositol components and sugar components (sucrose, stachyose, raffinose, fructose and glucose) contained in each fraction were analyzed by HPLC. The analytical column was Dionex Carbopak MA-1 (Dionex U.S.A) and confirmed by a pulsed electrochemical detector. The buffer used was 60 mM NaOH and eluted for 90 minutes at a flow rate of 0.4 mL per minute. The concentration of ethanol in each fraction was analyzed using gas chromatography. 2 is a table showing the content of each component measured for each fraction. Finitol eluted in the 0.6-2.2 BV fraction and peaked at 1.6 BV. Cairoinositol was eluted in the 2.0-3.2 BV fraction and peaked at 2.6 BV. Oligosaccharides were eluted in the 2.4-4.2 BV fraction and peaked at 3.4 BV. The fractions of 0.8-2.2 BV containing the peaks of finitol were collected and analyzed.The 800 mL fraction of finitol containing 2.03 g / L finitol, 0.11 g / L of chiroinositol and 0.20 g / L oligosaccharides was analyzed. Obtained. The content of the chiroinositol component in the dry weight of the finitol fraction was 61.2%. The composition of 2.4 to 2.8 BV fractions containing chiroinositol peaks was analyzed to determine the composition of the chiroinositol fractions containing 0.1 g / L of finitol and 1.22 g / L of chiroinositol and 4.10 g / L of oligosaccharides. 300 mL was obtained. The content of the chiranoinositol component in the dry weight of the chiroinositol fraction was 16.0%.

Experiment 6: Recovery of Cairo Inositol Components from Defatted Soybean Meal Hot Water Extract

1.5 L of defatted soybean meal hot water extract composed of the same ingredients as in Experiment 3 was treated in the same manner as in Experiment 5 to obtain 900 ml of finitol fraction and 1.10 g / L, respectively, of the total solid content of the finitol fraction and the Cairo inositol fraction. 300 mL was obtained.

Experiment 7 Recovery of Chiroinositol Component from Soybean Molasses

200 g of US soybean molasses containing the ingredients shown in Table 17 of Experiment 4 was diluted with distilled water to a volume of 1.5 L. This solution was treated in the same manner as in Example 5 to obtain 800 mL of the finitol fraction and 300 mL of the chiranoinositol fraction having a concentration of 1.88 g / L and 1.40 g / L, respectively, in the total solids.

Experiment 8: Concentration and Drying of Eluate

800 mL of the finitol fraction obtained in Experiment 5 was concentrated under vacuum at 50 ° C or lower in the evaporation tube. Vacuum concentration was carried out until the volume of the concentrate was 40 mL, and the samples were collected and lyophilized to obtain 3.0 g of a pale yellow powder. The content of the chiroinositol component in this dry powder was 63.1%. 300 mL of the Cairoinositol fraction was also concentrated to 30 mL in the same manner and 2.4 g of pale yellow powder was obtained by lyophilization. The content of the cairoinositol component in this dry powder was 16.5%.

Example 13 High Recovery Process of Finitol

100 L of tofu with 0.387 g / L of finitol content was boiled for 20 minutes to remove various germs generated during the tofu manufacturing process, and then inoculated with 2 L of Saccharomyces calsbergensis spawn precooled to 30 ° C. After incubation for 72 hours while supplying sufficient air, the cells and insoluble solids were removed by centrifugation to obtain 95 L of supernatant. The concentration of finitol in the supernatant was increased to 0.793 g / L and the content of chiroinositol was almost unchanged. This solution was passed through a column filled with 10 L of activated carbon at a rate of 20 L per hour to adsorb finitol. After washing activated carbon with 10 L of distilled water, 10 L of 10% ethane was injected again, and 10 L of 10 L of ethanol was eluted. Thereafter, 10 L of 50% ethanol was injected at a rate of 10 L per hour to remove sugar components, washed with distilled water, and reused for the next adsorption. From the start of the injecting the eluent was fractionated in 2L units and the change in the pinitol and total sugar components in each fraction was measured to obtain the results as shown in FIG. Among them, 1.4 L of the fraction containing finitol was collected, concentrated and lyophilized to obtain 90.2 g of a pale yellow powder having a purity of 68.0% of finitol. The Cairoinositol fraction was not recovered due to its low concentration.

Example 14 Preparation of High Purity Finitol Products

1,000 L of tofu impurities containing finitol were concentrated 10-fold to a concentration of 100 L at a concentration of 0.35 g / L. The concentrate was cooled to 30 ° C., and then placed in a 150 L vessel, followed by inoculation with Saccharomyces Carlsbergensis in the same manner as in Example 13. After incubating for 48 hours while injecting a sufficient amount of air, the microbial cells and suspended solids were removed by centrifugation. Through this process, the content of pinitol increased to 7.78 g / L and the liquid volume after removal of the cells was 95.5 L. The solution was concentrated 5 times to 19.1 L and then 30 L of 95% ethanol solution was added. As a result, 76.5% of the solids contained in the concentrate were removed and the purity of finitol in the solids was increased 4 times to 25%. Ethanol contained in this solution was completely removed by repeated addition of water and evaporation. 20 L of the ethanol-free solution was adsorbed onto a 20 L volume of activated carbon column in the same manner as in Example 12. After the adsorption was completed, the activated carbon column was washed with 20 L of distilled water and eluted with 20 L of 10% ethanol. The solution was concentrated 20 times to 1 L, and then 1.5 L of 95% ethanol was added thereto and left at room temperature for 12 hours with gentle stirring. The precipitate generated at this time was recovered by vacuum filtration and vacuum dried at 40 ° C. for 12 hours to obtain 495 g of a pinitol white powder product having a content of 96.5%. Table 18 shows the material balance for each step of the process.

TABLE 18

Mass Balance of High Purity Finitol Products

As described above, the present invention not only maximizes the recovery rate of finitol by converting the derivative of finitol in soybean by-product through microbial treatment, but also effectively removes other sugars in the raw material, thereby reducing the burden on subsequent recovery process. Not only does it have an excellent effect of greatly reducing the organic load in the finally discharged waste, but also the use of activated carbon columns to provide a large volume of samples with low concentrations at the same capacity as compared to conventional ion exchange resins. It can be treated, and there is no need for a separate desalting step in the pretreatment step, and if the proper elution conditions are given, the highly concentrated fractions containing only the chiroinositol component are recovered in a small volume. And Inositol production method as a new archive is a microbial treatment step with activated carbon column separation process is combined from a soybean product in comparison to the prior art is very excellent effects that can be separated to yield a high Cairo inositol.

Claims (12)

(a) increasing the content of pinitol or chiroinositol, which is a chiroinositol component in soybean processing by-products, either alone or in a mixed culture of microorganisms such as bacteria, yeast and mold in soybean by-products; (b) a pretreatment step of removing the insoluble material and the polymer material in the cells and soybean processing by-products generated during the microbial culture by centrifugation or filtration; And (c) adsorbing and eluting pinitol or chiroinositol by passing through the activated carbon a sample from which the insoluble component and the polymer material are removed through the pretreatment process. . The method of claim 1, wherein the soybean processing by-products belong to any one of the following; (i) soybean or skim soybean meal itself or extract thereof (ii) Tofu products, which are by-products of the tofu manufacturing process. (iii) Soybean whey, a by-product of soy protein manufacturing (iv) The concentrate of (i) or (ii) above. The method of claim 1, wherein the microorganisms used belong to Saccharomyces genus. The method of claim 1 wherein the microorganisms used belong to the genus Aspergillus. 4. A method according to claim 3, characterized in that it is Saccharomyces Carlsbergensis. The method of claim 4, wherein Aspergillus niger. 2. The method according to claim 1, wherein the chiroinositol component is adsorbed onto the activated carbon adjusted to pH 6 to 8 for separation in the step (c). 8. The method according to claim 7, wherein the chiroinositol component adsorbed on the activated carbon is eluted using an organic solvent aqueous solution. The method of claim 8, wherein the organic solvent for eluting the chiroinositol component is ethanol, isopropanol, methanol or acetone. The method of claim 8, wherein the chiroinositol component is separated from other sugar components by continuously or stepwise increasing the concentration of the organic solvent for eluting the chiroinositol component. The method of claim 10, wherein the concentration of the organic solvent for eluting the chiroinositol component is 1 to 20% at the beginning and 20 to 100% at the end. (a ') concentrating soybean by-products and then treating microorganisms such as bacteria, yeast or mold to increase the content of pinitol or cairoinositol in soybean by-products; (b ') Concentrating the solution after the microbial treatment to a total solid content of 50 ~ 70% (w / w) or more, and then add a volume of 95% ethanol solution of 1 to 3 times the solution to precipitate the solid contained in the solution fair; (c ') a pretreatment step of removing the precipitated material by centrifugation or filtration; (d ') removing ethanol from the separation solution in which pinitol is concentrated in a large amount by the pretreatment step by evaporation; (e ') adsorbing and eluting pinitol or Cairo inositol by passing the ethanol removal solution through an activated carbon column; And (f ') separation of the chiroinositol component, comprising the step of concentrating the eluted solution and crystallizing the chiroinositol or finitol.