JP5205569B2 - Process for producing novel polysaccharides - Google Patents

Description

  The present invention relates to a technique for easily extracting and fractionating a polysaccharide, and a thickener produced from the polysaccharide obtained using this technique.

  Thickeners used in foods are polysaccharides contained in cereals (such as starch), seaweeds (such as carrageenan, agar, and alginic acid), resins (such as gum arabic), and fruits (such as galactomannan and pectin). Manufactured as. In many cases, the polysaccharide thickener is localized at a specific part of the raw material, and can be extracted relatively easily by collecting the specific part (see, for example, Non-Patent Document 1).

On the other hand, plants such as Morohaya and Tsurumura Saki contain abundant polysaccharides having unique physical properties (see, for example, Non-Patent Document 2). In these plants, polysaccharides are distributed throughout the plant, and the specific site is difficult to recognize. For this reason, in order to obtain a polysaccharide, a considerable device is required. Using conventional technology, after extracting solvents such as acids and alkalis according to the properties of the thickening polysaccharide desired, extraction and purification are combined with column chromatography etc., so the process is complicated and between each process Extraction efficiency will be low. In addition, in alkaline extraction, only acidic polysaccharides are extracted, and extraction of other polysaccharides is difficult, so that the types of thickening polysaccharides obtained are limited by the means used (for example, non-existing polysaccharides). (See Patent Document 3).
Furthermore, the raw material containing the polysaccharide thickener increases in viscosity at the time of solvent extraction, requires time for filtration, and has poor workability. For this reason, it was necessary to reduce the viscosity by diluting with a large amount of solvent.

Naomichi Okazaki, Norio Sano, “Food Polysaccharides”, Yuki Shobo, November 25, 2001, p.43-204 Ohtani K, Okai K, Yamashita U, Yuasa I, and Misaki A. (1995). Biosci. Biotech. Biochem., 59, 378-381. Kazuo Matsuda, “Method for Separation and Purification of Polysaccharides”, Academic Publishing Center, February 28, 1987, p.19-113. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., and Smith F. (1956). Analytical Chemistry, 28, 350-356.

In the case where Morohaya or Tsurumura Saki, whose polysaccharide is distributed throughout the tissue, is used as a raw material, it is necessary to separate it from other components in order to obtain the desired polysaccharide.
The present invention has been made in view of the above circumstances, and the object of the present invention is to achieve polysaccharide extraction as easily as possible with fewer steps, and to improve workability by lowering the viscosity. Etc. is to provide.

As a result of intensive studies, the inventors have found that the above-mentioned problems can be solved by treating a raw material containing polysaccharides with an aqueous ammonium sulfate solution, and have basically completed the present invention. That is, by treating the raw material with an aqueous solution of ammonium sulfate, an increase in the aqueous solution viscosity due to the polysaccharide was suppressed, and at the same time, various polysaccharides were insolubilized and were easily separated from other components.
Thus, the polysaccharide extraction method according to the invention for solving the above-mentioned problems is characterized by using a 40% W / V to 80% W / V ammonium sulfate aqueous solution when extracting a polysaccharide from a raw material.

The polysaccharide extracted by the above invention is dispersed / dissolved in water, precipitated again, then dispersed / dissolved in water, and the supernatant is desalted to fractionate an appropriate polysaccharide. it can. Here, the dispersion / dissolution in water includes not only the case where the polysaccharide is completely dissolved in water, but also the case where a part of the polysaccharide is suspended in water in a dispersed state. The desalting treatment means a treatment for separating a salt (mainly, a salt derived from ammonium sulfate, a salt derived from a raw material, etc.) contained in the fractionated polysaccharide. As such desalting treatment, for example, methods such as gel filtration, ultrafiltration, and dialysis can be used.
The polysaccharide obtained in the present invention includes polysaccharides having various properties such as various acidic polysaccharides and alkaline polysaccharides. For this reason, it can fractionate by column chromatography, solvent extraction, etc. as needed, and can use a suitable thing according to a use.
Moreover, a thickener can be manufactured using the polysaccharide obtained by the said invention.

In the above-mentioned invention, the raw material is Morohaya, and a thickener containing a thickener produced from this polysaccharide and at least one of carrageenan, xanthan gum, locust bean gum, or guar gum is provided. Can do.
Usually, ammonium sulfate (ammonium sulfate) is used for purification at the final stage in an enzyme or protein extraction process (see, for example, Non-Patent Document 4). In the present invention, polysaccharides that were conventionally difficult to extract due to many extraction steps and high viscosity are treated with an aqueous solution of ammonium sulfate in the first step to reduce the fraction of the target polysaccharide and reduce the viscosity. It can be achieved at the same time.

Thus, the present invention is a technique relating to the production of polysaccharides characterized by an aqueous ammonium sulfate treatment.
Hereinafter, the present invention will be described in detail.
In the present invention, the target of the raw material is not limited, and the plant from which the thickening polysaccharide has been extracted by the conventional method (that is, the method of collecting and extracting the site where the thickening polysaccharide is localized) is extracted. Plants that have been distributed throughout and that are difficult to extract thickening polysaccharides by conventional methods (for example, those selected from the group consisting of Morohaya, Tsurumurasaki, Okra and Iseimo) ). Among these raw materials, particularly the latter plant can be preferably used. As the raw material, processed products such as pulverized or ground, heat-treated, and the like can be used. Furthermore, what defatted with organic solvents, such as hexane, ethyl acetate, acetone, and methanol, a squeezed rice cake, etc. can be used.

The polysaccharide referred to in the present invention refers to a polymer that is mainly composed of monosaccharides and emits a high viscosity when dispersed in a suitable solvent such as water.
The thickening agent as used in the field of this invention says what made polysaccharides the main component and adjusted the thickening effect according to the use to be used. For example, the polysaccharide is used as it is, a plurality of types of polysaccharides, or those obtained by adding salts to polysaccharides. Thickeners are used to increase the viscosity of foods, stabilize the emulsification of fats and oils, form gels to make jelly, etc., and give food a body feeling. Is not limited. Examples of thickeners include emulsion stabilizers, gelling agents, water retention agents and the like.

  In the present specification, the aqueous ammonium sulfate treatment (or ammonium sulfate treatment) means that a raw material is dispersed in water and ammonium sulfate is added. When ammonium sulfate is not added, the thickening effect due to the raw material is remarkable, but the viscosity of the aqueous solution decreases as the amount of ammonium sulfate added increases. When the decrease in viscosity is over, finish adding ammonium sulfate and leave it gently. Alternatively, when the amount of ammonium sulfate added in which the decrease in viscosity of the raw material is known is known, an aqueous ammonium sulfate solution having that concentration may be prepared in advance, and the raw material may be dispersed therein. Whichever method is used, the polysaccharide precipitates as an insoluble matter.

As a method for recovering the precipitated polysaccharide, for example, decantation, filtration or centrifugation is used. The recovered polysaccharide is dispersed and dissolved with a minimum amount of water.
By this ammonium sulfate treatment, polysaccharides and contaminants (for example, proteins and nucleic acids) can be separated. In addition, since the viscosity of the aqueous solution containing the polysaccharide can be temporarily reduced, the working efficiency in the manufacturing process can be remarkably improved.

  The raw material for obtaining the thickener derived from moroheiya in the present invention is a plant belonging to the genus Corcorus, and its scientific name is Corchorus olitorius. Morohaya leaves and stems can be used, but leaves are preferred. Various processed morohea products such as those obtained by grinding leaves or heat-treated by roasting can also be used. Furthermore, what was defatted with organic solvents, such as hexane, ethyl acetate, acetone, methanol, can also be used as a raw material.

The carrageenan referred to in the present invention is a polysaccharide extracted from red algae and containing anhydrogalactose and a sulfate ester of galactose as constituent sugars. Carrageenan includes κ (kappa), ι (iota), and λ (lambda) depending on the sulfate ester content. In the present invention, any carrageenan can be used, and among these, κ carrageenan is particularly preferable.
The xanthan gum as used in the field of this invention is a polysaccharide which Xanthomonas campestris produces outside a microbial cell.
The locust bean gum referred to in the present invention is also called a carob bean and is made from the endosperm portion of the seed of Ceratonia siliqua, a leguminous plant.
The guar gum referred to in the present invention is produced from the endosperm portion of the seed of the leguminous plant Cyamopsis tetragonolobus.
The moroheia-derived thickener of the present invention obtained by the above-described production method forms a gel when mixed with carrageenan and dissolved in water. In addition, mixing with xanthan gum raises the aqueous solution significantly. Such a moroheia-derived thickener has the property of thickening and gelling foods, and can be used for improving the texture, so it can be contained in, for example, foods, cosmetics, and pharmaceuticals.

  As a raw material for obtaining a thickener derived from Tsurumurasaki according to the present invention, Tsurumurasaki is a plant belonging to the genus Tsurumurasaki, and its scientific name is Basella rubra. Tsurmurasaki leaves and stems can be used. Various processed vineyards such as those obtained by grinding leaves and stems, heat-treated by roasting, etc. can be used, but it is preferable to use raw or lyophilized raw ones for extraction. Furthermore, what was defatted with organic solvents, such as hexane, ethyl acetate, acetone, methanol, can also be used as a raw material.

  The raw okra used in the present invention to obtain the okra-derived thickening agent is a plant belonging to the genus Trolloaoi, and the scientific name is Abelmoschus esculentus Moench. Okra fruits can be used. Various processed okra products such as those obtained by grinding fruit or heat-treated by roasting can also be used. Furthermore, what was defatted with organic solvents, such as hexane, ethyl acetate, acetone, methanol, can also be used as a raw material.

  The raw material for obtaining a thickener derived from Iseimo in the present invention is a plant belonging to the genus Yamanoimo, whose scientific name is Dioscorea batatas. Various processed products can be used, such as those obtained by polishing sesame potatoes, or heat-treated by roasting. Furthermore, what was defatted with organic solvents, such as hexane, ethyl acetate, acetone, methanol, can also be used as a raw material.

  In the present invention, the thickening polysaccharide can be easily fractionated from the raw material rich in the thickening polysaccharide. So far, it has become possible to produce thickening polysaccharides from raw materials that have been difficult to extract, and can provide new thickeners using thickening polysaccharides as raw materials, which can be used in foods, cosmetics, and pharmaceuticals. Furthermore, the moroheia thickener obtained using the present invention can improve the gel strength of carrageenan and clay of xanthan gum. These things can be expected to make a very important contribution to the food industry.

Next, embodiments of the present invention will be described with reference to the drawings. However, the technical scope of the present invention is not limited by these embodiments, and various forms can be made without changing the gist of the invention. Can be implemented. Further, the technical scope of the present invention extends to an equivalent range.
<Test Example 1> Method for producing Morohaya-derived thickening agent Morohaya dried powder (Agricultural Association Iga-cho Morohaya Production Association) 50 g was added to 1 liter of 50% w / v ammonium sulfate aqueous solution little by little with stirring. No increase in viscosity of the aqueous solution was observed. The mixture was stirred for 1 hour, and the precipitate was collected by centrifugation (6000 × g, 10 minutes). Dissolve the precipitate in 1 liter of water, add acetone until the final acetone concentration is 50% v / v to precipitate the polysaccharide, decant and centrifuge (6000 × g, 10 minutes) to collect the precipitate (Moloheiya precipitation 1).
Furthermore, acetone was added to the supernatant after centrifugation so that the acetone concentration was 80% v / v, and decantation and centrifugation (6000 × g, 10 minutes) were performed to recover the precipitate (Morohaya precipitation) 2).

  Morohaya Precipitation 1 and Morohaya Precipitation 2 were each washed with the concentration of acetone deposited, gradually increasing the acetone concentration, and finally washed with 100% acetone, and the precipitate was recovered by centrifugation (6000 × g, 10 minutes). . Further, the precipitate was dissolved in water and dialyzed (molecular weight: 14,000 separation). Then, it dried with the freeze dryer and what was obtained from Morohaya precipitation 1 and Morohaya precipitation 2 was used as Morohaya thickener 1 and Morohaya thickener 2, respectively.

According to the above method, the viscosity when 50 g of Morohaya dry powder was dispersed with 1 liter of ammonium sulfate aqueous solution (50% w / v) and when dispersed with 1 liter of water was measured with a Brookfield viscometer, and the viscosity was compared. . The conditions for measuring the viscosity were as follows.
Brookfield viscometer: B-type viscometer manufactured by Tokyo Keiki Co., Ltd. Jig: NO.1 (Ammonium sulfate treatment) and NO.3 (dispersed in water)
Rotation speed: 60rpm
Measurement temperature: 23 ℃
In addition, about the jig | tool, the number of the standard goods was shown, and the processed solution was shown in parenthesis after the number.

Example 1
A product obtained by dispersing 50 g of a dried moroheiya powder in 1 liter of an aqueous ammonium sulfate solution (50% w / v) was used.
Comparative Example 1
A product obtained by dispersing 50 g of Morohaya dry powder in 1 liter of water was used.
The results of viscosity measurement for each solution are shown in Table 1.

Table 1
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Sample viscosity (mPa · s)
-------------------
Example 1 780
Comparative Example 1 50,000
-------------------

  As is apparent from the table, the viscosity of the solution (Comparative Example 1) in which the moroheiya dry powder was dispersed in water was 50000 mPa · s, which was very high. On the other hand, in a solution (Example 1) in which a moroheiya dry powder was dispersed in an aqueous ammonium sulfate solution, the viscosity was 780 mPa · s, which was about 1/64 of that of Comparative Example 2. From this, it was found that by using an aqueous ammonium sulfate solution, the viscosity of the aqueous solution containing the moroheiya dry powder remained low, and the workability was improved.

Next, the molecular weight distribution of Morohaya thickener 1 and Morohaya thickener 2 was measured to examine the degree of purification. The measurement conditions for the molecular weight distribution were as follows.
Column: Sepharose CL-4B manufactured by Amersham Biosciences, 47.5 x 1.1 cm
Eluent: pH 6.8 50 mM phosphate buffer (containing 0.1 M sodium chloride)
Flow velocity (linear velocity): 6.3cm / h
Sample: 1 ml of 1 mg / ml injection Temperature: 23 ° C
Detection: phenol sulfuric acid method (see Non-Patent Document 4)
Molecular weight reference materials: Dextran 2000, Dextran 500, Dextran 70, Dextran 40 manufactured by Amersham Biosciences

Example 2
1 mg of Morohaya thickener 1 was dissolved in 1 ml of the eluate and injected into the column. The eluate was fractionated by 1 ml and detected by the phenol sulfuric acid method described above.
Example 3
1 mg of Morohaya thickener 2 was dissolved in 1 ml of the eluate and injected into the column. The eluate was fractionated by 1 ml and detected by the phenol sulfuric acid method described above.
The results of molecular weight measurement are shown in FIG.

  As is clear from the figure, Morohaya thickener 1 has a molecular weight of about 2000 kDa and was purified to almost a single peak. Morohaya thickener 2 had a molecular weight distributed over a wide range from a molecular weight of 40 kDa or less to 2000 kDa or more. Non-Patent Document 2 reports that a thickening polysaccharide derived from moroheiya is obtained through water extraction, quaternary ammonium salt extraction, and purification by anion chromatography, and the molecular weight of the thickening polysaccharide. Was 500 kDa. In general, a polymer substance tends to have a lower molecular weight as the purification process becomes more complicated. In the method according to the present embodiment, compared to the polysaccharide obtained by the method reported in Non-Patent Document 2, the production of thickening polysaccharide from Morohaya was achieved with a very simple process. The molecular weight was about 4 times as large.

<Test Example 2> Thickener containing Morohaya thickener and carrageenan Next, the influence of Morohaya thickener on the gel strength of κ carrageenan was measured. The measurement conditions for the gel strength were as follows.
Measurement conditions for gel strength Device: Creep meter manufactured by Yamaden Co., Ltd. Measurement mode: Break strength test Jig: No.6
Measurement speed: 1mm / sec
Temperature: 23 ° C

Example 4
Morohaya thickener 1 was added to a carrageenan aqueous solution (0.9% w / v) to prepare a 0-0.1% w / v Morohaya thickener 1 + κ carrageenan aqueous solution. This was melted in a hot water bath, poured into a cylindrical shape (diameter 1 cm, height 2 cm), and left at 4 ° C. for 24 hours to obtain a gel.

Example 5
Morohaya thickener 2 was added to a carrageenan aqueous solution (0.9% w / v) to prepare a 0 to 0.1% w / v molohea thickener 2 + carrageenan aqueous solution. This was melted in a hot water bath, poured into a cylindrical shape (diameter 1 cm, height 2 cm), and left at 4 ° C. for 24 hours to obtain a gel.
A graph showing the relationship between the concentration of the moroheiya thickener and the breaking load of the gel is shown in FIG. As is apparent from the figure, the strength of the kappa carrageenan gel could be dramatically improved by adding Morohaya thickener 1 or 2 to the kappa carrageenan.

<Test Example 3> Method for producing Tsurumurasaki-derived thickener Fresh Tsurumurasaki (cultivated at the Agriculture Research Department, Mie Prefectural Science and Technology Promotion Center) was treated with a freeze dryer to obtain a dry powder. 5 g of this dry powder was added little by little to 145 ml of 50% w / v aqueous ammonium sulfate with stirring. No increase in viscosity of the aqueous solution was observed. The mixture was stirred for 1 hour, and the precipitate was collected by centrifugation (6000 × g, 10 minutes). The precipitate was dissolved in 500 ml of water, acetone was added until the final acetone concentration reached 50% v / v to precipitate the polysaccharide, and the decanted and centrifuged (6000 × g, 10 minutes) precipitate was collected ( Tsurmurasaki Precipitation 1).
Further, acetone was added to the supernatant after centrifugation so that the acetone concentration was 80% v / v, and decantation and centrifugation (6000 × g, 10 minutes) were performed to collect the precipitate (Tsurumasaki Precipitation) 2).

  Tsurmurasaki Precipitation 1 and Tsurmurasaki Precipitation 2 were each washed with the concentration of acetone deposited, gradually increasing the acetone concentration, and finally washed with 100% acetone, and the precipitate was recovered by centrifugation (6000 × g, 10 minutes). . The recovered material was dried with a freeze dryer, and those obtained from Tsurmurasaki Precipitation 1 and Tsurmurasaki Precipitation 2 were designated as Tsurmurasaki Thickener 1 and Tsurmurasaki Thickener 2, respectively. The masses of the recovered Tsurumurasaki thickener were 0.165 g (thickener 1) and 0.073 g (thickener 2), respectively.

According to the above-mentioned method, the viscosity was measured with a Brookfield viscometer when 5 g of Tsurumura lyophilized powder was dispersed in 145 ml of an aqueous ammonium sulfate solution (50% w / v) and 145 ml of water, and the viscosities were compared. The conditions for measuring the viscosity were as follows.
Brookfield viscometer: Tokyo Keiki Co., Ltd. B-type viscometer Jig: BL adapter Rotation speed: 60rpm
Measurement temperature: 23 ℃

Example 6
A dispersion of 5 g of Tsurumasaki dry powder with 145 ml of an aqueous solution of ammonium sulfate (50% w / v) was used.
Comparative Example 2
A dispersion of 5 g of Tsurmurasaki dry powder with 145 ml of water was used.
The results are shown in Table 2.

Table 2
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Sample viscosity (mPa · s)
-------------------
Example 6 135
Comparative Example 2 750
-------------------

  As can be seen from the table, the viscosity of the solution (comparative example 2) in which the dry powder of Tsurmurasaki was dispersed in water increased to 750 mPa · s. On the other hand, the viscosity (135 mPa · s) of the solution (Example 6) in which the dry powder of Tsurmurasaki was dispersed in an aqueous solution of ammonium sulfate was about 1 / 5.5 that of Comparative Example 8. From this, it was found that by using an aqueous ammonium sulfate solution, the viscosity of the aqueous solution containing the dry powder of Tsurumura stays low, and the workability is improved.

The molecular weight distributions of Tsurmurasaki Thickener 1 and Tsurmurasaki Thickener 2 were measured to examine the degree of purification. The measurement conditions for the molecular weight distribution were as follows.
Column: Sepharose CL-4B manufactured by Amersham Biosciences, 47.5 x 1.1 cm
Eluent: pH 6.8 50 mM phosphate buffer (containing 0.1 M sodium chloride)
Flow velocity (linear velocity): 6.3cm / h
Sample: 1 ml of 1 mg / ml injection Temperature: 23 ° C
Detection: phenol sulfuric acid method (Non-patent Document 4)
Molecular weight reference materials: Dextran 2000, Dextran 500, Dextran 70, Dextran 40 manufactured by Amersham Biosciences.

Example 7
1 mg of Tsurmurasaki Thickener 1 was dissolved in 1 ml of the eluate and injected into the column. The eluate was fractionated by 1 ml and detected by the phenol sulfuric acid method described above.
Example 8
1 mg of Tsurumurasaki thickener 2 was dissolved in 1 ml of the eluate and injected into the column. The eluate was fractionated by 1 ml and detected by the phenol sulfuric acid method described above.
The results of molecular weight measurement are shown in FIG.

As is apparent from the figure, Tsurmurasaki Thickener 1 has a molecular weight of about 1000 kDa and has been purified to almost a single peak. Tsurumurasaki thickening polysaccharide 2 had a molecular weight distributed over a wide range from a molecular weight of 40 kDa or less to 2000 kDa or more.
As described above, the production of the thickener from Tsurumura Saki was very easily achieved in the same manner as the production of Morohaya thickener.

<Test Example 4> Method for producing Morohaya-derived high-viscosity thickener 17 g of Morohaya dry powder described in Test Example 1 was added to 500 ml of a 50% w / v aqueous ammonium sulfate solution little by little with stirring. At this time, no increase in viscosity of the aqueous solution was observed. The mixture was stirred for 30 minutes, and the precipitate was collected by centrifugation (15000 × g, 10 minutes).
The precipitate was dissolved by adding 500 ml of water, and the electric conductivity was adjusted to about 2.0 S / m ² . After stirring for 30 minutes, the supernatant was removed by centrifugation (10000 × g, 10 minutes), and the precipitate was collected. Furthermore, 500 ml of water was added to the precipitate to adjust the electric conductivity to 0.96 S / m ² , and the same operation was repeated to collect the supernatant. The supernatant was desalted with a dialysis membrane (separated by a molecular weight of 14000 kDa), and Morohaya thickener 3 was obtained by lyophilization. The 1% w / w aqueous solution of Morohaya thickener 3 was slightly yellowish.

  Morohaya thickener 3 was dissolved in water to 1% w / w, and then twice the amount of ethanol was added to recover the precipitate. The precipitate was washed successively with high-concentration ethanol (final ethanol concentration 99.5% v / v), and then dried at 40 ° C. overnight (Morohaya thickener 4). The 1% w / w aqueous solution of Morohaya thickener 4 was almost colorless and transparent.

Morohaya thickeners 1, 2, 3, and 4 and commercially available high viscosity thickeners xanthan gum, locust bean gum, guar gum and Morohaya-derived thickeners produced by the method reported in Non-Patent Document 2 The viscosity of a 1% w / w aqueous solution was measured with a vibration viscometer and compared. The viscosity measurement conditions were as follows.
Vibrating viscometer: SV-10 made by A & D Co., Ltd.
Temperature: 25 ° C

Example 9
A product obtained by dispersing Morohaya thickener 1 in water (concentration 0.25 to 1.0% w / w) was used.
Example 10
A product obtained by dispersing Morohaya thickener 2 in water (concentration 0.25 to 1.0% w / w) was used.
Example 11
A product obtained by dispersing Morohaya thickener 3 in water (concentration 0.25 to 1.0% w / w) was used.
Example 12
A product obtained by dispersing Morohaya thickener 4 in water (concentration 0.25 to 1.0% w / w) was used.

Comparative Example 3
Xanthan gum dispersed with water (concentration 0.25 to 1.0% w / w) was used.
Comparative Example 4
Locust bean gum dispersed in water (concentration 0.25 to 1.0% w / w) was used.
Comparative Example 5
A guar gum dispersed in water (concentration 0.25 to 1.0% w / w) was used.
Comparative Example 6
A thickened polysaccharide derived from Morohaya produced by the method reported in Non-Patent Document 2 was used (concentration 0.25 to 1.0% w / w) dispersed in water.
The viscosity measurement results of each solution are shown in FIG.

As apparent from FIG. 4, the moroheia thickener 3 and the moroheia thickener 4 both exhibited high viscosity as the concentration increased. At 1% w / w, the viscosity was higher than that of xanthan gum, locust bean gum and guar gum, which are existing thickeners exhibiting high viscosity. In particular, Morohaya thickener 4 was 1% w / w, which was more than twice as high as locust bean gum (Comparative Example 5) which exhibited the highest viscosity among commercially available thickeners. On the other hand, Morohaya thickeners 1 and 2 and Comparative Example 3 had remarkably lower viscosities than Morohaya thickeners 3 and 4. Further, Comparative Example 6 exhibited a dark brown color as pointed out in non-patent literature, and was different from Morohaya thickeners 3 and 4 of the present invention.
In Test Example 4, the precipitate obtained with the aqueous ammonium sulfate solution was separated from the supernatant that was separated by the first hydration (500 ml) and the residue that was produced when the morohea thickener 3 was produced. Each of the supernatants was collected, and a thickener was produced by desalting and lyophilization with a dialysis membrane (molecular weight 14000 kDa separation). These 1% w / w aqueous solutions were not only low in viscosity, but were also clearly colored (green brown to brown).
From this, it was found that a transparent and high-viscosity thickener can be obtained by adjusting the mixing ratio of the Morohaya dry powder and the aqueous ammonium sulfate solution and devising extraction with water.

<Test Example 5> Thickener containing Morohaya thickener and xanthan gum Next, the influence of Morohaya thickener on the viscosity of xanthan gum was measured. Viscosity measurement conditions were as follows.
Viscosity measurement conditions Equipment: Vibrating viscometer SV-10 manufactured by A & D Corporation
Measurement temperature: 25 ℃

Example 13
Morohaya thickener 3 was prepared in 0.5% w / w aqueous solution and mixed with xanthan gum 0.5% w / w aqueous solution at a constant sugar concentration of 0.5% w / w.
The results are shown in FIG. It was found that Morohaya thickener 3 and xanthan gum exhibited the highest viscosity when approximately equal amounts were added.

Example 14
Next, the viscosity change of the solution when the thickener (sugar concentration 0.5% w / w) containing equal amounts of Morohaya thickener 3 and xanthan gum is boiled for 15 minutes and then cooled to 25 ° C. Examined.
The results are shown in FIG. By boiling treatment, the viscosity of the mixed solution increased slightly. From this, it was found that Morohaya thickener 3 has the ability to dramatically improve the viscosity of xanthan gum while maintaining heat resistance.

<Test Example 6> Gelation of moroheiya thickener Next, the reaction of moroheiya thickeners 1, 2 and 3 with divalent metal ions was observed.
A 1% w / w aqueous solution of Morohaya thickener was prepared and spread thinly on a glass petri dish having a diameter of 3 cm. Thereafter, a calcium chloride solution (100 mg / ml in terms of calcium ions) was sprayed and allowed to stand.

Example 15
1% w / w aqueous solution of Morohaya thickener 1 Example 16
Example 17 of 1% w / w aqueous solution of Morohaya Thickener 2
When 1% w / w aqueous solution of Morohaya thickener 3 was sprayed with calcium chloride on each thickener solution, only Morohaya thickener 3 (Example 17) gelled into a film. This gelation was found to proceed instantaneously upon contact with calcium ions.

  As described above, according to the present embodiment, the thickening polysaccharide can be easily fractionated from the raw material rich in the thickening polysaccharide (for example, Morohaya, Tsurumurazaki, etc.). So far, it has become possible to produce thickening polysaccharides from raw materials that have been difficult to extract, and can provide new thickeners using thickening polysaccharides as raw materials, which can be used in foods, cosmetics, and pharmaceuticals. Furthermore, the moroheiya thickener can improve the gel strength of kappa carrageenan and the viscosity of xanthan gum. These things can be expected to make a very important contribution to the food industry.

It is a graph which shows the molecular weight distribution of Morohaya thickener 1 and Morohaya thickener 2. It is a graph which shows the influence which Morohaya thickener 1 and Morohaya thickener 2 have on the gel strength of (kappa) carrageenan gel. It is a graph which shows the molecular weight distribution of Tsurumurasaki thickener 1 and Tsurumurasaki thickener 2. It is the graph which compared the viscosity of Morohaya thickener 1-4 and the various thickeners conventionally known. It is the graph which investigated the change of the viscosity when the Morohaya thickener 3 and xanthan gum were mixed. It is the graph which compared the viscosity before and behind the heating of the thickener (carbohydrate concentration of 0.5% w / w) containing equal amounts of Morohaya thickener 3 and xanthan gum.

Claims (4)

Jew, Malabar spinach, To extract the polysaccharide from raw materials selected from the group consisting of okra and Ise potatoes, by using ammonium sulfate aqueous solution of 40% W / V~80% W / V, Rukoto to precipitate the polysaccharide A method for extracting polysaccharides. The polysaccharide extracted by the method according to claim 1 is dispersed / dissolved in water, precipitated again, then dispersed / dissolved in water, and the supernatant is desalted. Fractionation method. A thickener produced by using the polysaccharide obtained by the method according to claim 1 or 2 . The said raw material is a moroheiya, and contains the thickener manufactured from this polysaccharide, and at least any one of a carrageenan, a xanthan gum, a locust bean gum, or a guar gum of Claim 3 characterized by the above-mentioned. Thickener.

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