JP2007204717A - Method for obtaining mushroom-derived polysaccharide - Google Patents

Method for obtaining mushroom-derived polysaccharide Download PDF

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JP2007204717A
JP2007204717A JP2006028737A JP2006028737A JP2007204717A JP 2007204717 A JP2007204717 A JP 2007204717A JP 2006028737 A JP2006028737 A JP 2006028737A JP 2006028737 A JP2006028737 A JP 2006028737A JP 2007204717 A JP2007204717 A JP 2007204717A
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hot water
extraction
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mushroom
water
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Shinnosuke Miyauchi
信之助 宮内
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Nagaoka Univ Of Technology
国立大学法人長岡技術科学大学
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Abstract

<P>PROBLEM TO BE SOLVED: To provide a new method for obtaining mushroom-derived polysaccharides, wherein the polysaccharides having physiological activities such as antitumor activity can be effectively extracted and fractionated from a polypore mushroom without using various reagents. <P>SOLUTION: The method for obtaining mushroom-derived polysaccharides comprises a method of fractionation to obtain polysaccharides from a polypore mushroom by a pressurized hot water method, wherein the temperature condition is within a range of 140-250°C and the pressure condition is within a range of 0.1-10 MPa. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an acquisition method for extracting and fractionating schizophyllan, which is attracting attention as a polysaccharide having physiological activity such as antitumor activity, from mushrooms of porous fungi.

  The mushroom cell wall is an aggregate of natural macromolecules composed of polysaccharides, proteins, lipids and the like. Among them, polysaccharides are the most important components and occupy about 40 to 70% of the cell wall. Although this polysaccharide gives a unique shape to the cell wall and is thought to play a major role as a skeletal material that forms fruiting bodies, details of its composition, hierarchical structure, and chemical and physical properties are unknown. . For example, the structure and composition of the major parts of the fungi are not known at all, especially for the fungi of the basidiomycetous fungi such as the maitake mushrooms and garlic mushrooms of the hard fruit bodies, and for the porous fungi such as the oyster mushrooms, kawatake mushrooms and kofukisarno mushrooms that have a harder skin. It was.

  On the other hand, it has been said that mushrooms have long been effective as a Japanese and Chinese medicine, as a folk medicine, in addition to numerous medicinal effects such as tonicity, sedation, and blood pressure lowering, as well as intractable diseases such as cancer. In recent years, it has been demonstrated that the basis of the antitumor activity and physiological activity is a β- (1 → 3) glucan, a polysaccharide structure composed of β-glucan, that is, a schizophyllan type glucan. Various extraction methods such as hot water extraction method, extraction method using acid and alkali repeatedly, and enzymatic decomposition method have been shown so far. However, a general method for isolating the glucan from the cell wall of a mushroom having a complex multi-composition has not been shown. In particular, as described above, there is no method for efficiently extracting and fractionating mushrooms having a hard appearance such as cypress mushrooms or porous fungi having a harder skin.

  In such a situation, the present inventor has made various investigations. As a result, for mushrooms such as maitake mushrooms, the lipids are sufficiently removed in advance with acetone, and then hydrogen bonds are preliminarily reserved with sodium hypochlorite (NaClO). When it is destroyed, α-glucan is removed, so-called sticky substance is removed, and when hydrogen bond strength is weakened with dimethyl sulfoxide (DMSO) or the like, it hardly contains protein or chitin, It has been found that α-type and β-type complex carbohydrates can be obtained almost selectively (Non-patent Document 1).

And this inventor has proposed the method of acquiring a polysaccharide selectively from the mushroom of a basidiomycete Hidanastake based on the knowledge by the said nonpatent literature 1 (patent document 1).
JP 2005-133069 A Proceedings of the 50th Hokuriku branch meeting of the Society of Polymer Science, Japan, page 61 (2001)

  However, in the studies so far by the inventors, for example, it is necessary to use various reagents such as NaClO and DMSO in both Non-Patent Document 1 and Patent Document 1, and the skin is still hard. For this reason, it has not been possible to selectively obtain polysaccharides from porous fungi mushrooms whose polysaccharide composition is unknown.

  Therefore, the invention of this application is based on such a background, and a new mushroom that can efficiently extract and fractionate a polysaccharide having physiological activity such as antitumor activity from a porous fungus mushroom without using various reagents. It aims at providing the polysaccharide acquisition method of origin.

  The present invention is a method for obtaining a polysaccharide derived from a mushroom that solves the above-mentioned problem. First, a method for fractionating and obtaining a polysaccharide from a porous fungus mushroom by a pressurized hot water method, wherein the temperature condition is 140. And the pressure condition is in the range of 0.1 MPa to 10 MPa. Secondly, in the above method, secondly, the porous fungus mushroom is a fungus, a bamboo shoot, or a kofuki. It is characterized by the fact that it is a Sarno-Shokusake.

  Third, the polysaccharide is a schizophyllan-type glucan, and the present invention is fourthly characterized in that the polysaccharide is β- (1 → 4) glucan, β- (1 → 3 ) One or more of glucan and β- (1 → 6) glucan structures.

  According to the first to fourth aspects of the present invention as described above, schizophyllan-type glucan attracting attention as a polysaccharide having physiological activity such as antitumor activity, specifically, β- (1 → 3) glucan and β -Β-glucan having a structure such as (1 → 6) glucan can be efficiently extracted from mushrooms of porous fungi without using various reagents.

  The invention of this application has the features as described above, and the best mode for carrying out the invention will be described below.

  The present invention is a method for fractionating and obtaining a polysaccharide from a porous fungus mushroom by a pressurized hot water method. Specifically, the temperature condition of the pressurized hot water is in the range of 140 ° C. to 250 ° C., and the pressure condition is in the range of 0.1 MPa to 10 MPa.

  The “pressurized hot water” in the present invention is subcritical water, and this subcritical water has a maximum hydrolyzing power around 250 ° C., and can decompose organic substances into low molecules that dissolve in water at high speed. In addition, by using subcritical water, the ability to extract oil is strong while being water, and the oil in the organic matter can be extracted almost 100% instantaneously. Usually, subcritical water is water having a temperature and pressure below the critical point of water. More specifically, the critical point of this water is that the temperature of the water is 374 ° C. and the pressure is 218 atm (647 K, 22.1 MPa), so that the density of water and water vapor becomes equal. (Gas) is indistinguishable. This is the critical point.

  In consideration of this point, in order to extract the active ingredient of the mushroom more efficiently, the temperature of the pressurized hot water (subcritical water) in the present invention is in the range of 140 ° C. to 250 ° C. as described above. Is preferred. Also, the pressure is preferably in the range of 0.1 MPa to 10 MPa as described above. The raw water used as pressurized hot water (subcritical water) can be used as ordinary tap water, but is sufficient as ion-exchanged water, distilled water, filtered filtered water (for example, ultrafiltered water), etc. It is preferable to use purified water.

  In the present invention, the target mushrooms may be various types of mushrooms of porous fungi whose structure of polysaccharide is unknown and extraction is not easy because the epidermis has been hard. For example, Hilotake, Kawaratake, Kofukisarunoshikake, and the like.

  The polysaccharide extracted from any mushroom is preferably a schizophyllan glucan having an anticancer function, and the polysaccharide is β- (1 → 4) glucan, β- (1 → 3) glucan and It may be one or more of β- (1 → 6) glucan structures.

  In the present invention, pretreatment with acetone or the like may be performed as necessary. For example, when the extracted component is applied to food or the like, it is preferable that no pigment is contained. For this purpose, decolorization treatment may be performed as a pretreatment using acetone.

Further, in more detail about the present invention, the porous fungus <A> Agaric (Pycnoporus)
Extraction of polysaccharides from (Coccineus), <B> Coralus versicolor and <C> Pressurized hot water extraction of polysaccharides from Elfvingia applanata will be described as examples. Both are extracted at 150 ° C. or lower and around 180 to 210 ° C., but the yield is particularly large in the latter range.

  The present invention is not limited to the following examples.

<Example 1: Extraction of polysaccharides from Pycnoporus coccineus>
In order to study various physiological functions, the temperature rising pattern is intended to make components that are not extracted by normal hot water extraction solubilize water by reducing the molecular weight using the excellent hydrolytic ability of pressurized hot water. The extraction experiment was carried out by two-step temperature increase divided into low temperature and high temperature, and each Brix (sugar concentration) was measured, and the yield was calculated. Brix estimates the solute concentration from the refractive index of the solution and was used to confirm the end point of extraction.

In the extraction process, as a bamboo shoot sample, a fruit body cut with scissors to a particle size of 2 to 5 mm was used as an experimental sample. In addition, the water | moisture content in a sample was 11.33 wt%. Further, various physical properties of the samples were examined by the following methods.
<1> Cold water soluble content About 2.0 g of a sample was precisely weighed into a 500 mL beaker, and 300 mL of distilled water was added. The contents were treated for 48 hours at room temperature (20 ° C.) with occasional mixing. Thereafter, the contents were subjected to suction filtration using a 1G3 glass filter of known weight, and further washed with distilled water at room temperature, and the extraction residue was collected on the filter. The glass filter containing the extraction residue was transferred to a dryer set at 105 ° C. and dried to a constant weight. The cold water soluble content (wt%) was calculated according to the following equation.

<2> Hot water-soluble component About 2.0 g of a sample was precisely weighed into a 200 mL Erlenmeyer flask, and 100 mL of distilled water was added. Subsequently, a reflux condenser was attached to the flask, and the flask was transferred into boiling aqueous solution and held there for 3 hours. Thereafter, the content was suction filtered using a 1G3 glass filter of known weight, and further washed with hot water, and the extraction residue was collected on the filter. The glass filter containing the extraction residue was transferred to a dryer set at 105 ° C. and dried to a constant weight. The hot water soluble content (wt%) was calculated according to the following formula.

<3> Acetone Extraction About 2.0 g of a sample was precisely weighed into a cylindrical filter paper having a known weight and transferred to the extraction section of a Soxhlet extractor. About 200 mL of acetone was added to a 300 mL extraction flask, the extractor was assembled, transferred to a water bath set at 70 ° C., and Soxhlet extraction was started. After performing extraction for 12 hours, the extract was transferred into a 300 mL round bottom flask with a known weight and subjected to vacuum distillation using a rotary evaporator to obtain an acetone extract in the flask. The flask was transferred to a dryer set at 105 ° C. and dried to a constant weight. The acetone extract (wt%) was calculated according to the following formula.

<4> Ash content About 1.0 g of a sample was precisely weighed on a magnetic dish having a known weight, then transferred to an electric furnace, and ashed at 700 ° C. for 1.5 hours. The solid content remaining on the magnetic dish was regarded as ash, and the concentration was calculated according to the following formula.

<5> Pressurized hot water treatment A schematic diagram of the pressurized hot water flow reactor used in FIG. 1 is shown.

The reactor is mainly a distilled water tank, a high-pressure pump for feeding distilled water to a high-pressure field, a preheating serpentine, a percolation reactor (SUS316, 24 mm id 63 mm length, 28 mL), for stopping the reaction. It consists of a cooler, a back pressure valve for adjusting the pressure, and a cracked liquid collection receiver. The serpentine tube should be immersed in an oil bath (up to 200 ° C.) or in a molten salt solution (for example, KCO 3 : NaNO 2 : NaNO 3 = 53: 40: 7, 200 ° C.) depending on the target temperature. By heating. The hot water temperature is measured with an X thermocouple thermometer (made by CHINO) inserted at both ends of the reactor, and the temperature of the heating medium in which the serpentine tube is immersed and the temperature in the reactor are kept constant, and the temperature is raised appropriately. Controlled by external heating with an infrared furnace to ensure speed.
(1) Experimental procedure 1) Pressurized hydrothermal decomposition About 3.0 g of a sample was charged in a percolator reactor, and both ends of the reactor were placed on a sintered filter (Swalock, SS-16-VCR-2-5M) having a pore size of 5 μm. After capping, the sample was connected to the reaction apparatus, and the sample was decomposed and extracted by circulating steam and pressurized hot water at the target temperature at a predetermined flow rate. This extraction experiment was performed twice as Run1 and Run2. The conditions of each hot water temperature and pressure at that time were as shown in Table 1 below.

The decomposition solution flowing out from the reactor was measured for Brix (%) over time and used as a measure of decomposition. The decomposition effluent was transferred to a 500 mL eggplant-shaped flask with a known weight, freeze-dried, and further vacuum-dried at 60 ° C. for 3 hours, and the water-soluble yield on the basis of the charged dry weight was calculated. The reactor residue was recovered after cooling and dried at 105 ° C. until a constant weight was reached.

  The decomposed extract that was solubilized in water and flowed out of the reactor passed through a back pressure valve (holding valve) after cooling and was collected in a receiver. The decomposition extract was measured for Brix (%) over time, and was handled as follows as an approximate decomposition guide.

  That is, the recovered water-soluble component (WS: Water Soluble) was concentrated under reduced pressure, water was removed by a freeze-drying method, and further, vacuum-dried (60 ° C., 30 hours) was weighed.

The residue in the reactor was collected in a beaker and weighed dry at 60 ° C. For each (water-soluble matter, residue), the residue yield based on the charged dry weight was calculated.
2) Solubility in various solvents The 140 ° C. and 200 ° C. fractions obtained in Run 2 were scraped with a spatula and dissolved in 6 mL of water, methanol, acetone, 1N NaOHaq, and visually. The solubility was examined.
(2) Experimental results 1) Various physical properties of samples Table 2 shows the yields of cold water soluble components, hot water soluble components, acetone extracts and ash components.

2) Pressurized hot water treatment <Verification of color change>
Although not shown in the figure, the color of fractionated effluent when yellow bamboo is sequentially treated with steam (140 ° C., 0.05 MPa) and pressurized hot water (140 ° C., 1 MPa, 200 ° C., 2 MPa). The change was verified.

As a result of the verification, at first, in the early stage of the water vapor fraction, components that seemed to be orange pigments flowed out, and gradually clouded components were recovered. Subsequently, in the initial fraction of pressurized hot water (140 ° C., 1 MPa), it was confirmed that a dark brown effluent was obtained and gradually became thinner as the water flow time increased. Moreover, when it verified also about the extract at the time of processing with pressurized hot water (200 degreeC, 2 Mpa), the color change was similar to the change in 140 degreeC.
<Brix (%) and yield of effluent>
FIG. 2 and FIG. 3 show the time-dependent changes in Brix (%) of the effluents of Run1 and Run2 shown in Table 1, respectively. In either case, when the temperature was maintained at 140 ° C., Brix (%) increased with the start of water flow, showed a maximum value, then decreased, and again reached a maximum at 200 ° C.

  Table 3 shows the yield of each fraction in the case of Run 1 (Table 1), which shows the change over time of Brix (%) in FIG. 2. In the table, the change over time of Brix (%) is shown in FIG. The yield of fractions and the like in the case of Run 2 shown (Table 1) was shown. As shown in Table 3, the yield of Run1 was low with water vapor, but increased with pressurization. Moreover, although the cumulative yield exceeded 100%, a considerable amount could be extracted. In addition, a considerable amount of Run 2 shown in Table 4 could be extracted.

And, as shown in Table 5, it can be estimated that the extracted main components of Run1 and Run2 were precipitated without dissolving in methanol and acetone, and it was estimated that polysaccharides were extracted. It can be judged that it is a polysaccharide.

3) Summary From the above results, as for Run1 and Run2, the oyster mushroom fruit body was solubilized by about 50 wt% by hot water treatment at 200 ° C., and it is clear from the change in Brix (%) of the eluate. It is considered that the components decomposed and extracted at 140 ° C. or lower and the components decomposed and extracted at 200 ° C. are mixed. The temperature of hot water extraction normally attempted is 100 ° C., and at most 121 ° C. by autoclave or the like. Therefore, the components decomposed and extracted at 140 to 200 ° C. as in the present invention are not water-solubilized and are insoluble in water. Will be treated.

  As shown in Tables 3 and 4 above, the yield of the 140-200 fraction is as high as about 60 wt%, a substance not found in ordinary hot water extracts, specifically a polysaccharide having beneficial physiological activity. Can be obtained.

That is, as a result of detailed examination of the extraction behavior of the bamboo shoot, the extract is classified into a component extracted at 140 ° C. or lower, a component extracted at around 200 ° C., and a residue. With normal hot water, the extraction rate was about 19 wt%, but with 140 ° C pressurized hot water extraction, it was 41 wt% and at 200 ° C, 89 wt%. That is, in the pressurized hot water method, the yield of 22 wt% increased from the normal pressure of 100 ° C., and the yield of 48 wt% increased further at 200 ° C.
<Example 2: Extraction of polysaccharides from Coriolus versicolor>
Next, using Kawaratake as a sample, a pressurized hot water treatment was performed to extract a polysaccharide.

  The apparatus and procedure used for the pressurized hot water treatment are basically the same as in Example 1, except that the temperature condition is in the range of 150 ° C. to 250 ° C., the pressure condition is in the range of 0.1 MPa to 10 MPa, The sample was decomposed and extracted by flowing pressurized hot water at a flow rate of 10 mL / min.

The result was as shown in FIG. (A) shows a one-step extraction at a constant speed up to 250 ° C., (B) shows a two-step extraction at 140 ° C. and 200 ° C., and (C) shows a three-step extraction at 150 ° C., 170 ° C., and 190 ° C. . In any of FIGS. 4A, 4B, and 4C, it was confirmed that Brix (%) increased from around 200 ° C.
<Example 3: Extraction of polysaccharides from Elfvingia applanata>
In addition, we also examined Kofukisarokoshikake. Polysaccharides were extracted using Kofukisarokoshikatake as a sample and subjected to pressurized hot water treatment. The apparatus and procedure used for the pressurized hot water treatment are basically the same as in Example 1, except that the temperature condition is in the range of 180 ° C to 240 ° C, the pressure condition is in the range of 0.1 MPa to 10 MPa, The sample was decomposed and extracted by flowing pressurized hot water at a flow rate of 10 mL / min.

The result was as shown in FIG. (A) shows a one-step extraction at a constant temperature increase up to 250 ° C., and (B) shows a three-step extraction at 180 ° C., 210 ° C., and 240 ° C. In the case of Kofukisarokoshitake, it was confirmed that Brix (%) increased when the temperature exceeded 200 ° C. as shown in FIGS.
<Example 4: Characteristic analysis of polysaccharides extracted from Hilotake>
The comparative examination of the polysaccharide by pressure hot water extraction and the polysaccharide by chemical extraction was performed as follows.
(1) Polysaccharide gel chromatography by pressurized hot water extraction and chemical extraction 1) Apparatus The apparatus used was composed of a roller pump, a fraction collector, and a gel filtration column. As the gel filtration column, a Sepharose CL-43 gel was packed in a column having an inner diameter of 26 mm and a height of 100 cm.
2) Preparation of sample solution 15 mg each of Hilotake extracted polysaccharide by pressurized hot water extraction and Hilotake extracted polysaccharide by chemical extraction are added to 4 mL of 0.3 M NaOH, dissolved with stirring, and filtered through a 0.45 μm disposable filter. The filtrate was used as a sample solution.

In addition, here, the chemical extraction used a conventional extraction method (for example, JP-A-2005-133069) by alkali extraction using sodium hypochlorite in combination.
3) Sample addition and elution 1.5 mL of the sample was flowed at a flow rate of about 30 mL / h, and at the same time, the fraction collector (5.5 mL / tube) was started. The sample was eluted with degassed 0.3 M NaOH aqueous solution. Fractions were collected at 550 mL (= 100 tube min).
4) Analysis of each fraction Sugar analysis was carried out by measuring the absorbance at 490 nm by the phenol sulfuric acid method, and a chromatogram in which the absorbance was plotted against the amount of elution was obtained (FIGS. 6A and 6B).
5) Preparation of calibration curve It was prepared using blue dextran, pullulan and glucose as standard polysaccharides.

Sample preparation, addition and elution were carried out in the same manner as described above to obtain chromatograms. The blue dextran chromatogram was obtained from the absorbance at 265 nm. Then, the relationship between the elution amount and the molecular weight was obtained from a calibration curve (FIG. 6C).
(2) Molecular weight of extracted polysaccharide The molecular weight of the extracted polysaccharide was measured by Sepharose CL-43 gel filtration chromatography. The measurement results are shown in FIG. (A) shows pullulan and glucose, (B) shows a gel filtration chromatogram of blue dextran, and (C) shows a calibration curve obtained therefrom.

  FIG. 7 summarizes the results of gel filtration chromatogram of the extracted polysaccharide. (A) is a pressurized hot water extract at 140 ° C., (B) is a pressurized hot water extract at 200 ° C., (C) is a chemical extract with 1.25 M NaOH when 25 mL of NaClO is added. , (D) shows a chemical extract with 1.25M NaOH when 50 mL of NaClO was added. Although not shown in the figure, the 140 ° C. hot water extract of (A) is orange, the 200 ° C. hot water extract of (B) is brown, and the chemical extraction of (C) It was confirmed that both the product and the chemical extract of (D) were brown.

  As shown in FIG. 7, for the pressurized hot water extract, the molecular weight peak of the 140 ° C. extract (cellulose) is 1200 (degree of polymerization DP: 7.4), and the molecular weight peak of the 200 ° C. extract (schizophyllan + cellulose). Was 7000 (degree of polymerization DP: 43), and the molecular weight was higher at higher extraction temperatures.

  In addition, in the chemical extract using alkali, the molecular weight peak of the 1.25M NaOH extract at 25 mL of NaClO added was 79000 (degree of polymerization DP: 490), and 1.25M NaOH extracted at 50 mL of NaClO added. The molecular weight peak of the product was 27000 (degree of polymerization DP: 170). That is, the higher the amount of NaClO added, the smaller the molecular weight of the extract.

From the above results, it was found that polysaccharides having different molecular weights can be obtained by the extraction method. Moreover, in pressurized hot water, it was shown that schizophyllan was eluted on the high temperature side, and in the alkali extraction, schizophyllan was contained in the strong pretreatment with NaClO.
<Example 5: Examination of temperature condition and pressure condition affecting pressurized hydrothermal decomposition>
In Example 1, since it was speculated that different extracts were obtained at each temperature, the effects of hot water temperature and pressure were examined on the solubilizate yield.

The sample used was a bamboo shoot, and the fruit body was cut with scissors to a particle size of 2 to 5 mm. In addition, the water | moisture content in a sample was 11.33 wt%.
(1) Experimental procedure It was basically the same as the experimental procedure of Example 1, and the hot water temperature condition and pressure condition are shown in Table 6.

(2) Experimental results 1) About Run 3 In FIG. 8 (A), Brix (black square) of the effluent when treated with pressurized hot water under the experimental conditions shown in Run 3 in Table 6 above, treatment The change with time of temperature (-) was shown.

In Fraction 1, which was treated at 140 ° C. for 90 minutes, the color of the effluent became vermilion at the beginning of water flow and then became transparent. However, the Brix (%) value of the effluent hardly increased. Subsequently, when the temperature was raised to 250 ° C., the color of the effluent changed to blackish brown when the water flow temperature reached around 200 ° C., and at the same time, the Brix (%) value increased to 1.9. Thereafter, the Brix (%) value decreased, and Brix (%) showed zero at 140 minutes after the start of water flow. Regarding WS yield:
In Fraction 1, 41.1 wt%,
In Fraction 2, it was 43.3 wt%.

Fraction 1 had a low Brix (%) value despite the WS yield of 41.1 wt%.
2) About Run 4 In FIG. 8 (B), when the hot hydrothermal treatment was performed under the experimental conditions shown in Run 4 of Table 6 above, the Brix (black square) of the effluent and the treatment temperature (−) Change over time was shown.

  In Fraction A, which was treated at 140 ° C. and 5 MPa for 30 minutes, vermilion liquid, which was thought to be derived from the pigment of the oyster mushroom, flowed out at the beginning of water flow, and a rapid increase in Brix (%) value was confirmed. The subsequent effluent changed to a light brown color, and the Brix (%) value also decreased to 0.1. Subsequently, in Fraction B obtained by lowering the treatment pressure to 1 MPa and performing the treatment, the color of the effluent started to become yellowish and the Brix (%) value increased to 0.8. Furthermore, although the water flow was continued, the color of the effluent became transparent, and even in Fraction C where the pressure was reduced to 0 MPa and the treatment was performed for 15 minutes, an increase in the Brix (%) value was not confirmed.

  And even if the processing temperature time at 140 ° C. was increased, the Brix (%) value was not increased, so that the processing temperature was increased to 200 ° C. and the pressure was increased to 5 MPa. The color of the effluent became black when it reached around 200 ° C., and at the same time, the Brix (%) value also increased to 0.6. The water flow continued further, but the color of the effluent became transparent and showed zero at 75 minutes after the start of water flow.

Regarding WS yield:
In Fraction A, 25.7 wt%,
In Fraction B, 12.3 wt%,
In Fraction C, 3.4 wt%,
In Fraction D, it was 46.5 wt%.
(3) Summary The 140 ° C., 5 MPa effluent fraction at Run 3, that is, the WS yield of Fraction 1 is 41.1 wt%, and this value is the 140 ° C. effluent fraction at which the pressure at Run 4 was changed, ie, Fraction A˜ Since it showed a value very close to the total 41.4 wt% of the WS yield of C, it is considered that the treatment pressure does not affect the WS yield.

  However, since the Brix (%) value once increased in Fraction B, the pressure may have some influence on the extract.

For the high temperature fraction (200, 250 ° C.), the WS yield in Run 3 and Fraction 2 treated at 250 ° C. was 43.3 wt%, and in Run 4 and Fraction D treated at 200 ° C., 45% It was found that even if the temperature was raised from 200 ° C. to 250 ° C., the WS yield was not affected.
<Example 6: Analysis of decomposition extract at each treatment temperature>
In the above-mentioned embodiment, when the bamboo shoot is treated with pressurized hot water, there may be a component that is decomposed and extracted at 100 ° C. or lower, a component that is decomposed and extracted at around 140 ° C., and a component that is decomposed and extracted at around 200 ° C. It could be confirmed.

  Therefore, after obtaining the decomposed extract at each temperature, the decomposed extract not mixed with the decomposed extract at other temperatures was obtained, and the structural analysis of the decomposed extract at each temperature was also performed.

An experimental sample was obtained by cutting the fruit body of Hilotake with scissors to a particle size of 2 to 6 mm. The moisture and ash concentrations in the sample were 11.33 wt% and 3.47 wt%, respectively.
(1) Experimental procedure 1) Extraction with hot water (100 ° C.) About 2.0 g of a sample was precisely weighed into a 500 mL round bottom flask, and 100 mL of distilled water was added. After attaching a reflux condenser, it was kept in a boiling water channel. After 3 hours, the flask was taken out from boiling water (100 ° C.), and the content was suction filtered through a 2G4 glass filter of known weight to separate the residue and the filtrate. The residue on the filter was dried at 105 ° C. together with the filter. The filtrate was transferred to a 500 mL eggplant flask, freeze-dried, and further vacuum-dried at 60 ° C. for 3 hours.
2) Pressurized hot water extraction (140 ° C)
0.4 g of 100 ° C. extraction residue was charged into a percolation reactor (internal volume 3.8 mL, SUS316), capped with a stainless sintered filter with a pore size of 5 μm, and pressurized hot water circulation as illustrated in FIG. The experiment was conducted in the same procedure as in Example 1 by connecting to a reactor.
3) FT-IR analysis of solubilized product About 1 mg of the solubilized product obtained at each temperature was mixed with about 200 mg of KBr in a mortar and ground, and then FT- The infrared absorption spectrum was measured with an IR spectrometer (Nexus-470, manufactured by Nicolet). Moreover, the spectrum was similarly measured about the crude refined polysaccharide obtained as a 50-% ethanol precipitation of a part of 100 degreeC extract for comparison.
4) Sugar analysis of lysate 1 mL of the collected extract of Run 4 at 100 ° C, 140 ° C, and 200 ° C was placed in an Eppendorf tube and centrifuged at 1200 rpm x 10 min. Filtration through a 45 μm membrane filter. The obtained filtrate was subjected to high-performance anion exchange chromatography (hereinafter, HPAE-PAD) using an electrochemical detector.

The analysis conditions are as follows.
Gradient pump: GP40, eluent: distilled water, 0.1 M NaOH aqueous solution, 1.0 M sodium acetate, 0.1 M NaOH aqueous solution,
Gradient conditions: Carbopac PA-1 as column, pulsed amperometric detector ED-40 as detector,
Column oven: LC-30 (30 ° C)
5) 13 C-NMR analysis of solubilized product Using DSX300 manufactured by BRUKER, the CP-MAS method was used. The CP contact time was 1 ms, the repetition time was 4 s, the MAS was 4000 Hz, and the number of integrations was 1024 times. Chemical shifts were expressed on a tetramethylsilane basis using glycine as a secondary standard.
6) HPLC analysis of lysate The lyophilized product of each lysate was dissolved in an eluent used for HPLC analysis at a concentration of about 20 mg / mL, and then filtered through a membrane filter having a pore size of 0.45 μm. Analysis was carried out. The HPLC analysis conditions were as follows.
Column: Excelpack SEC-12 + SEC-13,
Column oven: 70 ° C.
Eluent: acetonitrile: distilled water = 30: 70 (flow rate 0.8 mL / min),
Detector: UV (254 nm).

As a comparison, 5-HMF (reagent: Tokyo Kasei) is also analyzed in the same manner, the retention time is measured, and the retention time of the chromatogram obtained by analyzing the Hilotake pressurized hot water solubilized product is compared with the retention time of the chromatogram. A comparison was made to confirm the presence or absence of generation.
(2) Experimental result 1) FT-IR analysis result of solubilized material Raw material subjected to analysis, 100 ° C hot water extract, 100 ° C hot water extract after ethanol precipitation, 140 ° C extract, 200 ° C extract, The result of each 200 ° C. extraction residue is shown in FIG. As shown in FIG. 9, it was confirmed that the infrared absorption spectra of the 140 ° C. fraction and the 200 ° C. fraction were very similar.

  And the comparison with the infrared absorption spectrum of the crude refined polysaccharide (crude glucan) prepared by carrying out ethanol precipitation of the hot water extract was also performed. The results are shown in FIG. (A) is 140 ° C. extraction, (B) is 200 ° C. extraction, and (C) is 100 ° C. extraction after ethanol precipitation.

As shown in FIG. 10, absorption of 6-membered glucose C—O—C was observed in the vicinity of 1048 cm −1 and β-glycoside bond was observed in the vicinity of 891 cm −1 , which is a characteristic of β-glucan. Carbohydrates that are thought to be derived from glucan were obtained at any temperature.

In addition, in the extract at 200 ° C., the 891 cm −1 absorption is slightly deviated, and this may be a possibility of α-type binding.

  In the 140 ° C. extraction of FIG. 9 and FIG. 10, the fraction that should have been extracted in the 140 ° C. fraction at 100 ° C. or less was recovered in the 140 ° C. fraction. About 140 degreeC pressurized hot water processing was performed. The results are shown in FIG. (A) shows 100 ° C. hot water extraction, and (B) shows a residue obtained by hot water extraction at 140 ° C. with respect to the residue extracted with 100 ° C. hot water.

  As shown in FIGS. 11 (A) and 11 (B), it was confirmed that there was a high possibility that the same structure appeared at both 100 ° C. and 140 ° C.

Further, the amides in the vicinity of 1555cm -1 to specific absorption of residue II (R-CO-NH- R '), chitin by that there are two absorption of beta-1-4-glycoside bond in the vicinity of 1153cm -1 Is presumed to be the center.

  Therefore, the results of comparing the infrared spectrum with the chitin reagent are shown in FIGS. 12 (A), 12 (B), and 12 (C). In the figure, (A) shows 100 ° C. extraction → ethanol precipitation, (B) shows the residue, and (C) shows chitin.

The residue and the reagent, as described above, and 1655 cm -1 around to the amide II (R-CO-NH- R '), there are two absorption of beta-1-4-glycoside bond in the vicinity of 1153cm -1, It is not present in extracts at 100 ° C. Since chitin was not decomposed in pressurized hot water around 200 ° C., it was suggested that the main component of the residue may be chitin.
2) Results of sugar analysis of solubilized product The results were as shown in FIG. (A) 100 ° C, (B) 140 ° C, (C) 200 ° C.

In any of the extracts, oligosaccharides were confirmed, and as the temperature increased, oligosaccharide absorption was significantly observed. The oligosaccharide observed at 200 ° C. had an increased number of peaks compared to 100 ° C. and 140 ° C., suggesting that complex decomposition occurred.
3) Results of 13 C-NMR analysis of solubilized product The solubilized product obtained by subjecting agaric mushrooms to pressurized hot water treatment at 100 ° C., 140 ° C. and 200 ° C. was subjected to 13 C-NMR analysis. As shown in FIG. 14, when these three spectra were compared, a peak appeared at substantially the same position. Although the position of glycosidic bond is different, it was almost the same as compared with the result of 13 C-NMR analysis of cellulose having monosaccharide glucose which is the source of glucan. I was able to confirm.

Furthermore, about a bamboo shoot, a 140 degreeC extract and a 200 degreeC extract are shown in FIG. 15 as a more detailed spectrum. As for the extract at 200 ° C., signals of 87.6 ppm corresponding to β1,3 glucan and signals of 62.2 ppm and 70.2 ppm corresponding to β1,6 glucan appear. 62.2 ppm is a signal related to C6 which is not glucosylated, and it can be seen from these that schizophyllan glucan is extracted. On the other hand, the signal around 79.0 ppm corresponds to β1,4 glucan (cellulose). This shows that cellulose-related sugars are extracted. This sugar does not exist in the periphyceae maitake mushrooms, or Hanabiratake mushrooms, and has the characteristic structure of a hump-like firmness including the following Kawaratake mushrooms and Kofukisarno mushrooms. This existence may be the reason why Hilotake and the like make it almost impossible to extract components with hot water. In the 140 ° C. extract, the components of the schizophyllan type are somewhat unclear.
4) HPLC analysis result of solubilized product As shown in FIG. 16, the result was obtained when agaric was treated with hot water under pressure at (a) 100 ° C., (b) 140 ° C. and (c) 200 ° C. The solubilized product was subjected to HPLC (HP1100) analysis, and 5-HMF production obtained as a secondary decomposition product of sugar was confirmed.

The analysis results of (d) 5-HMF (reagent) are also shown. FIG. 16 clearly shows that the 5-HMF peak is detected at about 38 minutes. As for the solubilized product, 5-HMF was detected only at 200 ° C. This was because the solubilized glucan was decomposed into 5-HMF via oligosaccharide and glucose at around 200 ° C. This coincided with the degradation reaction path of cellulose, which is a glucose polymer, in pressurized hot water.
<Example 7: Decomposition characteristics of acetone extraction residue>
In consideration of edible application, decolorization treatment using acetone was performed, followed by extraction with pressurized hot water.

The sample was cut from 2 to 5 mm in particle size using scissors. In addition, the water | moisture content in a sample was 11.33 wt%.
(1) Experimental procedure Basically, the hot water extraction was performed in the same experimental procedure as in Example 1. As a pretreatment for this pressurized hot water extraction, acetone extraction was performed. The temperature conditions and pressure conditions are as shown in Table 7.

(2) Experimental results FIG. 17 shows changes over time in Brix (black squares) and processing temperature (−) of the effluent when subjected to pressurized hot water treatment under the experimental conditions shown in Table 7 above. It was.

  The treatment was carried out at 140 ° C. for 45 minutes, and the Brix (%) value was increased to 0.1 within 5 minutes from the start. The effluent was light brown instead of vermilion because the bamboo shoot pigment was removed by acetone extraction. Although water flow was continued, since the Brix (%) value was 0.0, after that, the treatment temperature was raised to 200 ° C. and the treatment was performed. The color of the effluent became black when it reached around 200 ° C., and at the same time, the Brix (%) value also increased to 1.1. Furthermore, although water flow was continued, the color of the effluent became transparent and showed zero at 85 minutes after the start of water flow.

Regarding WS yield:
In Fraction 1, 25.5 wt%,
In Fraction 2, it was 64.2 wt%.
(3) Summary When the acetone extraction residue was used as a sample, the yield at 140 ° C. was 26 wt%, which was slightly lower than the conventional extraction. This is considered to have been affected by the extraction treatment with acetone. The reason why the extraction rate was high at 200 ° C. is thought to be due to the extraction of components that were not extracted at 140 ° C.
<Example 8: NMR analysis>
A 13 C-NMR analysis of 140 ° C. and 200 ° C. extract solubilized extracts of Kawaratake of Example 2 and Sarnosh mushroom of Example 3 was performed, and the spectra thereof are shown in FIGS. 18 and 19, respectively.
The same peak is observed for Kawaratake. That is, in the extract at 200 ° C., signals of 87.6 ppm corresponding to β1,3 glucan and signals of 62.2 ppm and 70.2 ppm corresponding to β1,6 glucan appear, and from these, schizophyllan type glucan is extracted. You can see that On the other hand, the signal around 80 ppm is a signal corresponding to β1,4 glucan (cellulose). Although the tendency is not changed even with Kofukisarokoshika, a peak corresponding to β1,4 glucan (cellulose) is seen at 140 ° C. In any case, these mushrooms make it difficult to extract polysaccharides in the usual way due to this cellulose structure. Only by the pressurized hot water method by destroying this structure can a polysaccharide with a schizophyllan type structure be extracted.

It is the schematic diagram which showed an example of the pressurized hot water flow type reaction apparatus used in this invention. In Example 1, it is the figure which showed the time-dependent change of Brix (%) of Run1. In Example 1, it is the figure which showed the time-dependent change of Brix (%) of Run2. It is the figure which showed the time-dependent change of Brix (%) of Kawaratake in Example 2, (A) is 1 step | paragraph extraction with constant temperature rising to 250 degreeC, (B) is 2 step | paragraph extraction at 140 degreeC and 200 degreeC. , (C) shows three-stage extraction at 150 ° C, 170 ° C and 190 ° C. It is the figure which showed the time-dependent change of the Brix (%) of Kofukisarokoshikake in Example 3, (A) is a one-step extraction at constant speed up to 250 ° C., (B) is 180 ° C., 210 ° C., 240 ° C. The three-stage extraction is shown. It is the figure which showed the result of having measured the molecular weight of the extraction polysaccharide in Example 4 by gel filtration chromatography, (A) is the gel filtration chromatogram of pullulan and glucose, (B) is the gel filtration chromatogram of blue dextran. (C) shows a calibration curve obtained from these chromatograms. It is the figure which illustrated molecular weight MW and polymerization degree DP based on the result of FIG. 6, (A) is a pressurized hot water extract at 140 degreeC, (B) is a pressurized hot water extract at 200 degreeC. , (C) shows a chemical extract with 1.25 M NaOH when 25 mL of NaClO is added, and (D) shows a chemical extract with 1.25 M NaOH when 50 mL of NaClO is added. It is the figure which showed the time-dependent change of Brix (%) of the effluent in Example 5, (A) shows Brix (%) of Run3, (B) shows Brix (%) of Run4. It is the figure which showed the FT-IR spectrum of the solubilizate in Example 6. It is the figure which showed the FT-IR spectrum in the comparative examination of the crude glucan in Example 6, and the solubilizate obtained by pressurized hot water extraction, (A) is 140 degreeC extraction, (B) is 200 degreeC. Extraction, (C) 100 ° C. extraction → ethanol precipitation (crude glucan) is shown. It is the figure which showed the FT-IR spectrum about the 140 degreeC extract in FIGS. 9 and 10, (A) is 100 degreeC hot water extraction, (B) is 140 degreeC pressurization about the residue which extracted 100 degreeC hot water. Shows hot water. It is the figure which showed the result of having compared and examined the FT-IR spectrum shown in FIG. 11, and the FT-IR spectrum of chitin. It is the figure in Example 6 which showed the saccharide | sugar analysis result of the solubilizate by HPAE-PAD. It is the figure which showed the 13 C-NMR analysis result of the solubilizate in Example 6. It is the figure which showed in detail the 13 C-NMR spectrum from about 140 degreeC and 200 degreeC extract of a garland. It is the figure which showed the analysis result of the solubilizate by HPLC in Example 6. It is the figure which showed the time-dependent change of Brix (%) of a bamboo shoot in Example 7. It is a 13 C-NMR spectrum about a 140 degreeC and 200 degreeC extract of Kawaratake. It is a 13 C-NMR spectrum about the 200 degreeC extract of Sarno-Shokushitake.

Claims (4)

  1.   A method for fractionating and obtaining polysaccharides from a porous fungus mushroom by a pressurized hot water method, wherein the temperature condition is in the range of 140 ° C. to 250 ° C. and the pressure condition is in the range of 0.1 MPa to 10 MPa. A method for obtaining a polysaccharide derived from a mushroom.
  2.   2. The method for obtaining a mushroom-derived polysaccharide according to claim 1, wherein the porous fungus mushroom is Hilotake, Kawaratake or Kofukisarokoshitake.
  3.   The method for obtaining a mushroom-derived polysaccharide according to claim 1 or 2, wherein the polysaccharide is a schizophyllan-type glucan.
  4.   2. The polysaccharide according to claim 1, wherein the polysaccharide is one or more of β- (1 → 4) glucan, β- (1 → 3) glucan and β- (1 → 6) glucan. 3. A method for obtaining a mushroom-derived polysaccharide according to 2.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010095579A (en) * 2008-10-15 2010-04-30 Kumamoto Univ Method for clarifying organic porous material
JP2011026525A (en) * 2009-07-29 2011-02-10 Wood One:Kk METHOD FOR EXTRACTING POLYSACCHARIDE COMPRISING beta-GLUCAN AS MAIN COMPONENT
JP2011103877A (en) * 2009-10-20 2011-06-02 Saga Univ Method for lowering viscosity of high-viscosity culture solution, and polysaccharide-containing culture solution produced by the method
CN102603906A (en) * 2011-12-30 2012-07-25 安徽丰原发酵技术工程研究有限公司 Preparation method of inonotus obliquus polysaccharide aqueous solution
JP2014221899A (en) * 2014-06-17 2014-11-27 株式会社ウッドワン METHOD FOR EXTRACTING POLYSACCHARIDE COMPRISING β-GLUCAN AS MAIN COMPONENT
CN104387486A (en) * 2014-11-11 2015-03-04 罗宇 High-yield preparation method of coriolus versicolor polysaccharide
CN104725521A (en) * 2015-03-31 2015-06-24 广东省微生物研究所 Amauroderma rude single-peak polysaccharide F212, and preparation method and application thereof
JP2016079135A (en) * 2014-10-17 2016-05-16 イビデン株式会社 Coprinus comatus extract

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002262820A (en) * 2001-03-07 2002-09-17 Isao Horiuchi Method for extracting active ingredient of mushrooms
WO2002087603A1 (en) * 2001-04-27 2002-11-07 Ajinomoto Co., Inc. Immunopotentiators
WO2002098440A1 (en) * 2001-06-01 2002-12-12 Ajinomoto Co., Inc. Drugs for diabetes
WO2002098433A1 (en) * 2001-06-01 2002-12-12 Ajinomoto Co., Inc. Drugs for intestinal diseases
WO2004033502A1 (en) * 2002-10-08 2004-04-22 Ricom Corporation Chitosan-containing polysaccharide, process for producing the same and use thereof
JP2005133069A (en) * 2003-10-09 2005-05-26 Ichimasa Kamaboko Co Ltd METHOD FOR ACQUISITION OF MUSHROOM-DERIVED beta-GLUCAN POLYSACCHARIDE AND MUSHROOM-DERIVED BETA-GLUCAN POLYSACCHARIDE
JP2005160326A (en) * 2003-11-28 2005-06-23 Kid:Kk beta-D GLUCAN-CONTAINING PICKLING LIQUID, METHOD FOR EXTRACTING beta-D GLUCAN, METHOD FOR MAKING VEGETABLE PICKLE CONTAINING beta-D GLUCAN, AND THE VEGETABLE PICKLE
JP2005272405A (en) * 2004-03-26 2005-10-06 Hokuto Corp Skin function improving agent and skin function improving food

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002262820A (en) * 2001-03-07 2002-09-17 Isao Horiuchi Method for extracting active ingredient of mushrooms
WO2002087603A1 (en) * 2001-04-27 2002-11-07 Ajinomoto Co., Inc. Immunopotentiators
WO2002098440A1 (en) * 2001-06-01 2002-12-12 Ajinomoto Co., Inc. Drugs for diabetes
WO2002098433A1 (en) * 2001-06-01 2002-12-12 Ajinomoto Co., Inc. Drugs for intestinal diseases
WO2004033502A1 (en) * 2002-10-08 2004-04-22 Ricom Corporation Chitosan-containing polysaccharide, process for producing the same and use thereof
JP2005133069A (en) * 2003-10-09 2005-05-26 Ichimasa Kamaboko Co Ltd METHOD FOR ACQUISITION OF MUSHROOM-DERIVED beta-GLUCAN POLYSACCHARIDE AND MUSHROOM-DERIVED BETA-GLUCAN POLYSACCHARIDE
JP2005160326A (en) * 2003-11-28 2005-06-23 Kid:Kk beta-D GLUCAN-CONTAINING PICKLING LIQUID, METHOD FOR EXTRACTING beta-D GLUCAN, METHOD FOR MAKING VEGETABLE PICKLE CONTAINING beta-D GLUCAN, AND THE VEGETABLE PICKLE
JP2005272405A (en) * 2004-03-26 2005-10-06 Hokuto Corp Skin function improving agent and skin function improving food

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010095579A (en) * 2008-10-15 2010-04-30 Kumamoto Univ Method for clarifying organic porous material
JP2011026525A (en) * 2009-07-29 2011-02-10 Wood One:Kk METHOD FOR EXTRACTING POLYSACCHARIDE COMPRISING beta-GLUCAN AS MAIN COMPONENT
JP2011103877A (en) * 2009-10-20 2011-06-02 Saga Univ Method for lowering viscosity of high-viscosity culture solution, and polysaccharide-containing culture solution produced by the method
CN102603906A (en) * 2011-12-30 2012-07-25 安徽丰原发酵技术工程研究有限公司 Preparation method of inonotus obliquus polysaccharide aqueous solution
JP2014221899A (en) * 2014-06-17 2014-11-27 株式会社ウッドワン METHOD FOR EXTRACTING POLYSACCHARIDE COMPRISING β-GLUCAN AS MAIN COMPONENT
JP2016079135A (en) * 2014-10-17 2016-05-16 イビデン株式会社 Coprinus comatus extract
CN104387486A (en) * 2014-11-11 2015-03-04 罗宇 High-yield preparation method of coriolus versicolor polysaccharide
CN104725521A (en) * 2015-03-31 2015-06-24 广东省微生物研究所 Amauroderma rude single-peak polysaccharide F212, and preparation method and application thereof

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