JP2009195158A - Method for producing monogalacturonic acid - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for simply producing monogalacturonic acid at a low cost, which has been difficult to obtain in a high yield until now. <P>SOLUTION: The method for producing monogalacturonic acid comprises, in treating a pectin-containing natural product, preferably beet, beet pulp, apple, apple fiber etc., with an enzyme to have activity acting on the pectin-containing natural product and liberating monogalacturonic acid so as to obtain monogalacturonic acid, directly treating the pectin-containing natural product with the enzyme without separating and extracting pectin from the natural product. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a method for producing monogalacturonic acid, an enzyme-treated product containing monogalacturonic acid at a high concentration, and a food or drink containing the same.

  Galacturonic acid is an acidic monosaccharide that constitutes the polysaccharide pectin contained in the cell walls and mesenchymal tissues of most land plants, and is partly methylesterified in pectin. Pectin generally contains arabinose, galactose, mannose, and rhamnose in addition to galacturonic acid, and the ratio is known to vary depending on the type and maturity of the plant.

  So far, with regard to the production method of monogalacturonic acid and / or oligogalacturonic acid, a method of adding an acid and heating and a method of decomposing with an enzyme have been reported. As a method of decomposing by adding an acid, for example, a method of decomposing by heating pectin with a nitric acid aqueous solution as described in Non-Patent Document 1, and a method of decomposing pectin with sulfuric acid as described in Non-Patent Document 2. A method of heating at 100 ° C. by acidification and a method of thermally decomposing pectin in a formic acid aqueous solution as disclosed in Patent Document 1 are disclosed.

As a method for decomposing with an enzyme, for example, Patent Document 2 discloses a method in which pectin or pectic acid is brought into contact with an immobilized enzyme. Patent Document 3 discloses a method in which pectin is heated to 80 ° C. to 100 ° C. A method for producing oligogalacturonic acid by reducing the molecular weight and then reacting with pectinase is disclosed. Patent Document 4 is an application for a mineral absorption promoter containing oligogalacturonic acid having a degree of polymerization of 1 to 9. In the examples, commercially available pectinase is used in a 5% aqueous solution of apple pectin as a method for producing oligogalacturonic acid. It is said that an oligogalacturonic acid having a degree of polymerization of 6 has been produced. Furthermore, in Example 1 of Patent Document 5, monogalacturonic acid, digalacturonic acid, trigalacturonic acid, tetragalacturonic acid, pentagalacturonic acid, by dissolving polygalacturonic acid in an acetic acid buffer and reacting by adding pectinase, It is disclosed that hexagalacturonic acid and heptagalacturonic acid were obtained in proportions of 12%, 16%, 18%, 14%, 10% and 2%, respectively.
Biochem. J. et al. 94,617 (1965) Carbohydrate Res. 20, 351-356 (1971) JP 2003-327602 A JP-A-6-205687 JP-A-10-226701 JP-A-5-316997 JP-A-8-245399

  However, the method of decomposing with an acid is decomposed to oligogalacturonic acid despite the use of a strong acid and heating at a high temperature, but the yield of monogalacturonic acid was very low. For example, Non-Patent Document 2 describes a method in which sulfuric acid is added to a solution of sodium salt of pectic acid obtained by repeating alkali deesterification treatment from apple pectin, and heat treatment is performed at 100 ° C. for 17 hours. It can only be decomposed into oligogalacturonic acid having a degree of polymerization. The decomposition product obtained in Patent Document 1 is oligogalacturonic acid having a polymerization degree of 40 or less. Furthermore, in the method of decomposing with an acid, the reaction solution has a low pH, and in addition, there is a limit to the reactors that can be used, such as corrosion resistance, for the reaction at high temperatures. Also, great effort and facilities are required for neutralizing and removing the acid.

  Enzymatic degradation can be achieved by using exo-type polygalacturonase, but until now there has been extensive research on enzymes that produce oligogalacturonic acid, but the degree of esterification and molecular weight of pectin have been studied. However, there are few reports on enzymes that efficiently release monogalacturonic acid, and even if reported at the laboratory level, the enzymes are very expensive. Yes, it was not industrially usable.

  In addition, the monogalacturonic acid and / or oligogalacturonic acid production methods disclosed so far all use pectin and / or polygalacturonic acid extracted and purified from plant tissues as raw materials. What was obtained by this method contained a large amount of oligogalacturonic acid and esterified product, and there was no method for obtaining monogalacturonic acid with high purity. The reason that pectin and / or polygalacturonic acid must be extracted and purified to use as a raw material is that plant tissues contain cellulose, hemicellulose, lignin, protein, etc. in addition to pectin, and can directly degrade polysaccharides. This is because a large amount of monosaccharides and oligosaccharides other than galacturonic acid is released, which makes purification difficult.

  Because of this situation, when obtaining high purity monogalacturonic acid, extract pectin from plant tissue or purchase commercially available apple pectin, beet pectin, etc. After degradation or enzymatic degradation, it was necessary to remove oligogalacturonic acid, other monosaccharides, and esterified products, which made the process complicated and expensive.

  An object of this invention is to provide the manufacturing method of monogalacturonic acid which can obtain monogalacturonic acid which was difficult to obtain with high purity until now at low cost.

  As a result of intensive studies to solve the above problems, the present inventors have found that a composition containing monogalacturonic acid at a high concentration by allowing an enzyme to act directly on a natural product containing pectin, particularly beet and apple. It was found that higher purity monogalacturonic acid can be obtained by adsorbing to an anion exchange resin and eluting with an alkaline aqueous solution.

  That is, according to the first aspect of the present invention, an enzyme having an activity of liberating monogalacturonic acid by acting on a natural product containing pectin is allowed to act on the natural product containing pectin to obtain monogalacturonic acid. The present invention is directed to a method for producing monogalacturonic acid characterized in that monogalacturonic acid is obtained by directly acting the enzyme without separating and extracting pectin from the product, The method for producing monogalacturonic acid as described above, wherein the product is at least one selected from the group consisting of beet, beet pulp, apple and apple fiber, and preferably acts on a natural product containing pectin to produce monogalacturonic acid. Enzymes with a liberating activity are Aspergillus niger and / or Is a process for producing the monogalacturonic acid, which is an enzyme derived from Aspergillus acculeatus.

  In the second aspect of the present invention, an enzyme having an activity of acting on a natural product containing pectin to release monogalacturonic acid is directly added to the natural product without separating and extracting the pectin from the natural product containing pectin. A monogalacturonic acid-containing enzyme-treated product obtained by acting is preferable, preferably monogalacturonic acid, oligogalacturonic acid, polygalacturonic acid, and the content of monogalacturonic acid in the total content of these methyl esters 70% by mass or more of the monogalacturonic acid-containing enzyme-treated product, more preferably, the monogalacturonic acid-containing enzyme-treated product is passed through an anion exchange resin to adsorb monogalacturonic acid, and then an alkaline aqueous solution. 65% purity by eluting the monogalacturonate by passing through A Monogarakutsuron acid containing an enzyme-treated product which is purified over amount%.

  In the third aspect of the present invention, an enzyme having an activity of acting on a natural product containing pectin to release monogalacturonic acid is directly acted on a natural product containing pectin to release monogalacturonic acid, and the above-mentioned. The gist of the present invention is a method for producing a monogalacturonic acid-containing enzyme-treated product, which is characterized in that the second monogalacturonic acid-containing enzyme-treated product is obtained.

  The fourth aspect of the present invention is summarized as a food or drink characterized by containing the above-described second monogalacturonic acid-containing enzyme-treated product of the present invention.

  According to the present invention, monogalacturonic acid, which has heretofore been difficult to obtain with high purity, can be easily obtained at low cost.

  The natural product containing pectin used in the present invention is not particularly limited as long as it contains pectin. For example, citrus fruits such as apple, pear, peach, cherry, strawberry, mandarin, lemon, grapefruit, ume, avocado, oyster , Fruits such as nectarine, pineapple, grape, melon, watermelon, vegetables such as cucumber, radish, carrot, celery, potato, sweet potato, olive, beet, tomato, soybean, azuki bean, lentil, cowpea, sweet bean, sweet pea It is done. Among these, cherries, citrus fruits, Japanese radish, beets, and apples are preferable, and beets and apples are particularly preferable because of the structure and properties of pectin.

  The form of the natural product containing pectin used in the present invention is not particularly limited, and a fresh product may be used as it is, or a squeezed juice or extract may be used. Considering the fact that fresh products have a high monosaccharide content, stable availability of raw materials, and cost, it is preferable to use squeezed juice and extract, and the dried ones are more stable in terms of storage stability. preferable. Preferable examples of these include squeezed rice cakes of various juices, commercially available products such as apple fiber and orange fiber sold by drying them, beet pulp, beet fiber, among them beet pulp, apple fiber However, it is most preferable because water-soluble monosaccharides are almost removed, it is inexpensive and easily available, and the properties of pectin are suitable.

  The natural product containing pectin used in the present invention may be subjected to pretreatments such as crushing, washing, sterilization, and swelling before use, as long as the effects of the present invention are not impaired. The crushing can be performed by a conventionally known method, and a mixer, a juicer, a pulper, a mill or the like can be used. It can be expected that the enzymatic reaction proceeds rapidly by crushing. The washing can be performed by a conventionally known method, and can be carried out by usual washing with water or washing with an organic solvent such as hexane or ethanol. By washing with water, monosaccharides and oligosaccharides can be removed, and fats and oils can be removed by washing with an organic solvent. In addition, a surfactant or the like can be used as necessary when washing. Sterilization can be performed by a conventionally known method such as heat sterilization, ultraviolet irradiation, or gamma irradiation. Swelling can be performed by a conventionally known method. For example, there is a method of adding water and allowing to stand. At this time, the swelling action can be promoted by heating to 10 ° C to 121 ° C, preferably 30 ° C to 105 ° C, more preferably 50 ° C to 100 ° C. In particular, when a dried raw material is used, the enzyme reaction can be rapidly advanced by swelling.

  In carrying out the enzyme reaction in the present invention, it is desirable that water is present in an amount of 3 to 20 times, preferably 3 to 15 times, more preferably 5 to 12 times the solid content. If the amount of water is small, not only does the enzyme reaction hardly proceed, but there is also a problem that industrial production becomes difficult due to lack of fluidity, and if the amount of water exceeds this range, the enzymatic reaction will no longer proceed. Does not increase and only requires a large reactor.

  Enzymes used in the present invention include pectinase having an action of degrading pectin, polygalacturonase and oligogalacturonase having an action of degrading polygalacturonic acid, and pectin methyl having an action of removing the methyl ester group of pectin Examples include esterase, pectin lyase and pectate lyase that cleave α-1,4 bond by β-elimination reaction, and any may be used as long as they do not impair the effect of the present invention, such as Aspergillus niger, Aspergillus Aspergillus genus such as Acreatus, Aspergillus oryzae, Aspergillus pulberrentas, Trichoderma viride, Trichoderma genus such as Trichoderma reesei, Rhizopus derema, Rhizopus oryzae genus Rhizopus genus, Bacillus subtilis genus Bacillus Preferably, it is an enzyme produced by culturing Aspergillus niger and / or Aspergillus acreatas and producing this microorganism outside or inside the cell. This enzyme can be used in a crude state simply by crushing microorganisms or concentrating the medium and adding ammonium sulfate or trichloroacetic acid to precipitate, but preferably ultrafiltration, dialysis, gel filtration, DEAE Sepharose Used after purification such as purification.

  The enzyme can be obtained by culturing a microorganism, but a commercially available enzyme can also be purchased and used. As such commercially available enzymes, Sumiteam PX (manufactured by Shinnippon Chemical Industry Co., Ltd.), Sumiteam AP-2 (manufactured by Shinnippon Chemical Industry Co., Ltd.), Sumiteam ARS (manufactured by Shinnippon Chemical Industry Co., Ltd.), Sumiteam SPC (Shin Nippon Chemical) Industrial Co., Ltd.), Sumiteam MC (New Nippon Chemical Industry Co., Ltd.), Pectinase PL "Amano" (Amano Enzyme Co., Ltd.), Pectinase G "Amano" (Amano Enzyme Co., Ltd.), Pectinase GL "Amano" ( Amano Enzyme Co., Ltd.), Cellulosin PC5 (manufactured by HI Corporation), Cellulosin PE60 (manufactured by HI Corporation), Cellulosin PEL (manufactured by HI Corporation), Cellulosin ME (manufactured by HI Corporation), Pectinase SS (Yakult Pharmaceutical Co., Ltd.) Company-made), pectinase 3S ( Yakult Yakuhin Kogyo Co., Ltd.), Pectinase HL (Yakult Yakuhin Kogyo Co., Ltd.), ROHAPECT D5L (Higuchi Shokai Co., Ltd.), ROHAPECT D5S (Higuchi Shokai Co., Ltd.), ROHAPECT MA PLUS (Higuchi Shokai Co., Ltd.), ROHAPECT MAX (manufactured by Higuchi Shokai), ROCAPECT PTE (manufactured by Higuchi Shokai), ROHAPECT PL (manufactured by Higuchi Shokai), ROHAPECT B1 (manufactured by Higuchi Shokai), ROHAPECT VR-C (manufactured by Higuchi Shokai) ), ROHAPECT 7104 (manufactured by Higuchi Shokai), ROHAPECT DA6L (manufactured by Higuchi Shokai), ROHAPECT 10L (manufactured by Higuchi Shokai), ROHAPECT AP1 (manufactured by Higuchi Shokai), sucrase (Manufactured by MFC Lifetech Co., Ltd.), sucrase S (manufactured by MFC Lifetech Co., Ltd.), Pectinex Ultra SP-L (manufactured by Novozymes Japan Co., Ltd.), Pectinex 3XL (manufactured by Novozymes Japan Ltd.), Ultrazyme (Novozymes Japan Co., Ltd.), Vinozyme (Novozymes Japan Co., Ltd.), Citrozyme (Novozymes Japan Co., Ltd.), Olivex (Novozymes Japan Co., Ltd.), Novo Farm 12 (Novozymes) Japan Co., Ltd.), Vinoflow (Novozymes Japan Co., Ltd.), Beerzyme (Novozymes Japan Co., Ltd.), Pectinase <Nagase> (Nagase Biochemical Co., Ltd.), and the like.

  Among these, preferable examples include Sumiteam PX (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Sumiteam AP-2 (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Sumiteam SPC (manufactured by Shin Nippon Chemical Industry Co., Ltd.), pectinase PL “ Amano "(Amano Enzyme Co., Ltd.), Cellulosin PC5 (manufactured by HIBI Co., Ltd.), Cellulosin PE60 (manufactured by HIBI AI Co., Ltd.), Cellulosin PEL (manufactured by HIBI Corporation), pectinase SS (manufactured by Yakult Pharmaceutical Co., Ltd.), pectinase 3S (Manufactured by Yakult Pharmaceutical Co., Ltd.), sucrase N (manufactured by MFC Lifetech Co., Ltd.), sucrase S (manufactured by MFC Lifetech Co., Ltd.), Pectinex Ultra SP-L (manufactured by Novozymes Japan Ltd.), Pectinex 3XL ( Novozymes Japan stock Company), Ultrazyme (Novozymes Japan Co., Ltd.), Vinozyme (Novozymes Japan Co., Ltd.), Citrozyme (Novozymes Japan Co., Ltd.), Beerzyme (Novozymes Japan Co., Ltd.), Pectinase <Nagase> (manufactured by Nagase Seikagaku Corporation) can be mentioned, and more preferable examples include Sumiteam PX (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Sumiteam AP-2 (manufactured by Shin Nippon Chemical Industry Co., Ltd.), pectinase PL “Amano” (manufactured by Amano Enzyme), Cellulosin PE60 (manufactured by HIBI), Cellulosin PEL (manufactured by HIBI), Pectinex Ultra SP-L (manufactured by Novozymes Japan), Pectinex 3XL (Novo Zymes Japan Co., Ltd.).

  These enzymes can be used alone or in combination of two or more. In addition, other enzymes can be used in combination as long as the effects of the present invention are not impaired. Examples of the enzyme used in combination include cellulase, hemicellulase, mannanase, protease, lipase and the like, and cellulase and hemicellulase are preferable. By using cellulase and hemicellulase together, the solid content of pectin-containing natural substances can be solubilized, increasing fluidity and / or reducing insoluble content, improving filtration efficiency, and yielding galacturonic acid. Increase and reduction of waste can be expected.

  The kind of the combined enzyme is not limited, and for example, enzymes originating from Aspergillus, Trichoderma, Bacillus, Rhizopus and the like can be used. These enzymes can be purified from microorganisms and used, or commercially available enzymes can be purchased and used.

  Examples of such commercially available enzymes include Sumiteam AC (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Sumiteam C (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd.), and Sumiteam BGA (Shin Nippon Chemical Industry Co., Ltd.). Sumiteam BGT (manufactured by Shinnippon Chemical Industry Co., Ltd.), Sumiteam X (manufactured by Shinnippon Chemical Industry Co., Ltd.), Cellulosin T2 (manufactured by Hibiii Corporation), Cellulosin AL (manufactured by Hibiii Corporation), Cellulosin AC40 ( HB IAI Co., Ltd.), Cellulosin TP25 (H.I.B.I. Co., Ltd.), Cellulosin HC100 (H.I.B.I. Co., Ltd.), Cellulase A "Amano" (Amano Enzyme Co., Ltd.), Cellulase T "Amano" (Amano Enzyme Co., Ltd.), Hemicellulase "Amano" Zyme Co., Ltd.), Cellulase “Onoka” R-10 (Yakult Pharmaceutical Co., Ltd.), Cellulase “Onoka” 3S (Yakult Pharmaceutical Co., Ltd.), Cellulase Y-NC (Yakult Pharmaceutical Co., Ltd.), Pancerase BR (manufactured by Yakult Pharmaceutical Co., Ltd.), Macero Team A (manufactured by Yakult Pharmaceutical Co., Ltd.), Cellulase TP Kyowa (manufactured by Kyowa Kasei Co., Ltd.), etc. Company), Sumiteam BGT (manufactured by Shin Nippon Chemical Industry Co., Ltd.), Cellulosin T2 (manufactured by HIBI Co., Ltd.), Cellulase “Onozuka” 3S (manufactured by Yakult Pharmaceutical Co., Ltd.), Cellulase TP Kyowa (manufactured by Kyowa Kasei Co., Ltd.) Can be mentioned.

  The amount of the enzyme as described above is not limited as long as the effect of the present invention is not impaired, but is preferably 0.01% by mass to 5% by mass, more preferably 0%, based on the solid content of the natural product containing pectin. 0.05% by mass to 1% by mass. If it is less than this range, it may not be efficiently decomposed into monogalacturonic acid, and even if it is added more than this range, further increase in yield can no longer be expected.

  The conditions for the enzyme reaction are not particularly limited, and the reaction is preferably carried out at the optimum temperature and pH of the enzyme used. For example, an enzyme derived from Aspergillus niger or Aspergillus acreatas is reacted at a temperature in the range of 30 ° C to 60 ° C, preferably 40 ° C to 60 ° C, and 2.0 to 9.0, preferably 2.5 to 8. The reaction can be carried out in a pH range of 0.0.

  The enzyme reaction time is not particularly limited as long as the effects of the present invention are not impaired, but is preferably 1 hour to 120 hours, more preferably 3 hours to 72 hours, and further preferably 10 hours to 48 hours. If it is shorter than this range, pectin may not be sufficiently decomposed and the yield of monogalacturonic acid may be reduced. Even if the reaction is longer than this range, further increase in yield can no longer be expected.

  In the present invention, the monogalacturonic acid released by the enzymatic reaction has a content of 70% by mass or more, preferably 75% by mass in the total content of monogalacturonic acid, oligogalacturonic acid, polygalacturonic acid and / or their methyl esters. % Or more, more preferably 85% or more. Since the monogalacturonic acid-containing enzyme-treated product produced according to the present invention exhibits such a high monogalacturonic acid content, in producing a high-purity product of monogalacturonic acid important as a pharmaceutical raw material or functional food, The purification process can be simplified. Moreover, the effect resulting from galacturonic acid can be expressed more greatly.

  In the present invention, the monogalacturonic acid-containing enzyme-treated product obtained by the enzyme reaction can be used as it is, or can be used after being dried, powdered, etc., or further purified.

  As a further purification method, a purification method using an anion exchange resin can be used. In this method, since monogalacturonic acid is a carboxylic acid, it is adsorbed on an anion exchange resin and then monogalacturonic acid is desorbed. In order to pass through the anion exchange resin, the enzyme-treated product is subjected to solid-liquid separation by a conventionally known method, and the resulting sugar solution can be used.

  Examples of the anion exchange resin used include Diaion SA10A, Diaion SA11A, Diaion SA12A, Diaion NSA100, Diaion SA20A, Diaion SA21, Diaion AUBA120, Diaion PA306S, and Diaion PA306S from Mitsubishi Chemical Corporation. Diaion PA308, Diaion PA312, Diaion PA316, Diaion PA318L, Diaion HPA25, Diaion PA408, Diaion PA412, Diaion PA418, Diaion WA10, Diaion WA20, Diaion WA21J, Diaion WA30, Diaion SAF11AL Diaion SAF12A, Diaion PAF308L, Diaion WA30C, manufactured by Dow Chemical Co., Ltd. 2, 1 × 4, 1 × 8, 2 × 8, SBR-PC, Marathon A, Marathon A2, Monosphere 550A, 22, 66, Marathon WBA, Monosphere 77, Amberlite IRA400J manufactured by Organo Corporation Amberlite IRA402BL, Amberlite IRA404J, Amberlite IRA410J, Amberlite IRA411, Amberlite IRA458RF, Amberlite IRA478RF, Amberlite IRA900J, Amberlite IRA910CT, Amberlite IRA904, Amberlite IRA958, Amberlite IRA96, Amberlite IRS96 Light XT6050RF, Amber Jet 4002, Amber Jet 4010, Amber Jet 4400, Amber List A 1, and the like. Among these, Diaion WA10, Diaion WA20, Diaion WA21J, Diaion WA30 of Mitsubishi Chemical Corporation, which are weakly basic anion exchange resins, 66 manufactured by Dow Chemical Co., Marathon WBA, Monosphere 77 Amberlite IRA67, Amberlite IRA96SB, and Amberlite XT6050RF manufactured by Organo Corporation are more preferable.

  The anion exchange resin may be used as it is, but it is preferable to use it after converting it to OH type by a conventionally known method. Conversion to the OH type can be performed, for example, by passing sodium hydroxide or potassium hydroxide.

  The anion exchange resin must be used in an amount sufficient to adsorb monogalacturonic acid, and is preferably 0.5 L to 10 L with respect to 1 kg of the dry weight of the natural product containing pectin used for the enzyme reaction. More preferably, it is 1L-5L.

  As long as the rate of passing through the anion exchange resin does not impair the effects of the present invention, it can be passed at a general rate, preferably 0.5 to 5 times the amount of resin per hour, more preferably Is 1 to 3 times.

  After passing the sugar solution after the enzyme reaction through an anion exchange resin and adsorbing monogalacturonic acid, it is preferable to wash with water in order to remove excess sugar solution. Washing with water is preferably carried out until no sugar is contained in the effluent of the anion exchange resin, and this can be easily measured using, for example, a Brix saccharimeter, high performance liquid chromatography or the like. In general, the amount of water used for washing is 2 to 10 times, preferably 3 to 6 times the amount of resin.

  The monogalacturonic acid adsorbed on the anion exchange resin can be desorbed and eluted by passing a salt that can be exchanged with the monogalacturonic acid. The salt is not limited as long as the effect of the present invention is not impaired, but an alkaline aqueous solution is preferable because regeneration can be performed simultaneously. Examples thereof include sodium hydroxide, potassium hydroxide, magnesium hydroxide and the like. Sodium is most preferred.

  When elution of monogalacturonic acid is carried out with an alkaline aqueous solution, the effluent is acidic to neutral when a large amount of monogalacturonic acid is desorbed and eluted, but when the monogalacturonic acid is almost desorbed, the effluent is alkaline. Therefore, it is possible to collect only a fraction in which monogalacturonate can be collected at a high concentration using pH as an index.

  The monogalacturonic acid-containing enzyme-treated product recovered as described above has a monogalacturonic acid purity in a solid content of 65% by mass, preferably 80% by mass or more, and more preferably 90% by mass or more. This purity tends to increase if only the fraction from which a large amount of monogalacturonic acid flows is collected, and tends to decrease if the collected fraction is expanded, and can be adjusted according to the monogalacturonic acid-containing enzyme-treated product to be prepared. .

  The monogalacturonic acid-containing enzyme-treated product obtained by purification as described above can be concentrated, dried, powdered or the like by a conventionally known method. For the concentration, for example, an evaporator, a concentration can, an evaporator manufactured by Okawara Chemical Industries, a wall wetter manufactured by Kansai Scientific Machinery, or the like can be used. Drying can be performed by spray drying, freeze drying, drying under reduced pressure, or the like. Powdering may be performed as it is by spray drying, freeze drying, drying under reduced pressure, or the like. For example, excipients such as dextrin, lactose, and reduced starch degradation products may be added.

  The monogalacturonic acid-containing enzyme-treated product obtained in the present invention is made from a highly safe food and can be added to various foods and drinks. Such food or drink is not particularly limited, and the form of addition is not limited. Examples of such foods and beverages include processed noodles such as udon and pasta, processed meat products such as ham and sausage, processed fishery products such as kamaboko and chikuwa, processed milk products such as butter, milk powder and fermented milk, jelly and ice cream Processed foods such as desserts such as breads, confectionery and seasonings, and beverages such as soft drinks, alcohols, fruit juice drinks, vegetable juice drinks, milk drinks, carbonated drinks and coffee drinks. By adding the monogalacturonic acid-containing composition of the present invention to foods and drinks, for example, a pH adjusting action and a pH buffering action can be expected. As other effects, a diet action, a mineral absorption promoting action and the like can be expected.

  EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

In the present invention, the content of monogalacturonic acid is a value determined by the following method. That is, it measured on the following conditions by the high performance liquid chromatography method (HPLC method), and detected using the fluorescence detector.
HPLC: Waters Alliance
Column: Aminex HPX-87H manufactured by Bio-Rad
Mobile phase: 0.05 N sulfuric acid flow rate: 0.6 ml / min Temperature: 60 ° C
Detection: Differential refractive index

Example 1
1 L of water was added to 100 g of beet pulp (manufactured by Hokkaido Sugar Industry Co., Ltd.), 1.5 g of cellulosin PE60 manufactured by HI was added, and the reaction was shaken at 50 ° C. for 16 hours. After the reaction, filtration was performed with a Buchner funnel using Radiolite # 100 manufactured by Showa Chemical Industry Co., Ltd. as a filter aid, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid released in the sugar solution was as shown in Table 1, and a large amount of monogalacturonic acid was obtained.

Example 2
1 L of water was added to 100 g of beet fiber (manufactured by Nippon Sugar Sugar Co., Ltd.), 1.5 g of cellulosin PE60 manufactured by HI was added, and the reaction was shaken at 50 ° C. for 16 hours. After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid released in the sugar solution was as shown in Table 1, and a large amount of monogalacturonic acid was obtained.

Example 3
1 L of water was added to 100 g of apple fiber (manufactured by Nichiro Co., Ltd.), 1.5 g of cellulosin PE60 manufactured by HI was added, and the reaction was shaken at 50 ° C. for 16 hours. After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid released in the sugar solution was as shown in Table 1, and a large amount of monogalacturonic acid was obtained.

Example 4
1L of water is added to 100g of freeze-dried powder of Unshu orange juice (manufactured by Nippon Fruit Industry Co., Ltd.), which is a squeezed bowl of Unshu orange juice, and 1.5g of cellulosin PE60 made by Hibiai is added and shaken at 50 ° C for 16 hours Went. After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid released in the sugar solution was as shown in Table 1, and a large amount of monogalacturonic acid was obtained.

Example 5
1 L of water was added to 100 g of beet pulp (manufactured by Hokkaido Sugar Co., Ltd.), 1.5 g of Sumiteam PX manufactured by Shin Nippon Chemical Industry Co., Ltd. was added, and the reaction was shaken at 50 ° C. for 16 hours. After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid released in the sugar solution was as shown in Table 1, and a large amount of monogalacturonic acid was obtained.

Example 6
1 L of water was added to 100 g of beet pulp (manufactured by Hokkaido Sugar Co., Ltd.), 1.5 g of Sumiteam AP2-L manufactured by Shin Nippon Chemical Industry Co., Ltd. was added, and the reaction was shaken at 50 ° C. for 16 hours. After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid released in the sugar solution was as shown in Table 1.

Example 7
1 L of water was added to 100 g of beet pulp (Hokkaido Sugar Industry Co., Ltd.), 1.5 g of Pectinex Ultra SP-L manufactured by Novozymes Japan Co., Ltd. was added, and the reaction was shaken at 50 ° C. for 16 hours. . After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid released in the sugar solution was as shown in Table 1.

Example 8
Add 1 L of water to 100 g of beet pulp (Hokkaido Sugar Industry Co., Ltd.), add 1.5 g of Cellulosin PE60 manufactured by Hibiai Co., Ltd. and 0.1 g of Cellulosin T2 manufactured by Hibiai Co., Ltd., which is a cellulase, at 50 ° C. The reaction was shaken for 16 hours. After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The content of monogalacturonic acid liberated in the sugar solution was as shown in Table 1. The amount of recovered sugar solution increased and the amount of recovered galacturonic acid also increased.

Comparative Example 1
1 L of 0.4 mol / L hydrochloric acid was added to 100 g of beet pulp (Hokkaido Sugar Industry Co., Ltd.) and heated in a boiling water bath for 1 hour. After the reaction, filtration was performed in the same manner as in Example 1, and the resulting sugar solution was analyzed by HPLC. The sugar solution was somewhat viscous, and a large amount of pectin and oligogalacturonic acid were present in the sugar solution, but the monogalacturonic acid content could not be confirmed as shown in Table 1.

Comparative Example 2
The same operation was performed except that 0.4 mol / L sulfuric acid was used in place of the hydrochloric acid of Comparative Example 1. As in Comparative Example 1, monogalacturonic acid could not be confirmed in the sugar solution.

Example 9
30L jar fermenter was charged with 1.8kg of beet pulp (Hokkaido Sugar Industry Co., Ltd.), 18L of water was added, 27g of Cellulosin PE60 manufactured by HIBI Co., Ltd. and 1.8g of Cellulosin T2 manufactured by HI Co., Ltd. were added. The reaction was stirred for 24 hours at 50 ° C. The reaction product was subjected to solid-liquid separation using a pressure filter and a filter press precoated with Radiolite # 100. The obtained sugar solution had a Brix sugar content of 5.0% and was 16.03 L. As a result of HPLC analysis, 190 g of monogalacturonic acid was contained. This sugar solution was passed through a column filled with 2 L of anion exchange resin Diaion WA30 manufactured by Mitsubishi Chemical Co., Ltd., which had been regenerated to OH type with NaOH, with SV = 2, and the Brix sugar content of the column effluent was adjusted with ion exchange water. Washed to 0.0%. Next, a 0.4% sodium hydroxide aqueous solution was passed through with SV = 2, and the effluent was collected by 1 L. FIG. 1 shows the pH of each collected fraction, FIG. 2 shows the Brix sugar content, and FIG. 3 shows the amount of monogalacturonic acid. Monogalacturonic acid was discharged at a high concentration in fractions 1-14. Fractions 1 to 12 were combined and concentrated to obtain 690 ml of monogalacturonic acid-containing enzyme-treated product. This composition was a pale yellow transparent liquid having a Brix sugar content of 30.6%, a monogalacturonic acid content of 141 g, and a monogalacturonic acid purity of 80.6%.

Example 10
0.5 ml of the monogalacturonic acid-containing enzyme-treated product obtained in Example 9 was added to 350 ml of green tea. There was no change in the taste, color, and smell of green tea, and monogalacturonic acid-containing green tea was produced.

Example 11
20 ml of the monogalacturonic acid-containing enzyme-treated product obtained in Example 9 was added to 2 sets of white rice and 500 ml of water, and cooked with an electric rice cooker. 10g of cooked rice is soaked in 100g of water, crushed with a mixer, and centrifuged and analyzed by HPLC. As a result, 0.06g of monogalacturonic acid in 10g of rice contains monogalacturonic acid, even at high temperatures. It was thought that it was not decomposed or denatured.

It represents the pH of each fraction of the monogalacturonic acid eluate. It represents the Brix sugar content of each fraction of the monogalacturonic acid eluate. It represents the monogalacturonic acid content of each fraction of the monogalacturonic acid eluate.

Claims (8)

An enzyme having an activity of acting on a natural product containing pectin to release monogalacturonic acid is allowed to act on a natural product containing pectin to obtain monogalacturonic acid without separating and extracting pectin from the natural product. A method of producing monogalacturonic acid, wherein the enzyme is directly reacted to obtain monogalacturonic acid. The method for producing monogalacturonic acid according to claim 1, wherein the natural product containing pectin is one or more selected from the group consisting of beet, beet pulp, apple and apple fiber. 3. The enzyme having an activity of acting on a natural product containing pectin to release monogalacturonic acid is an enzyme derived from Aspergillus niger and / or Aspergillus aculeatus. Of producing monogalacturonic acid. Monogalacturon obtained by allowing an enzyme having an activity to act on a natural product containing pectin to release monogalacturonic acid directly on the natural product without separating and extracting the pectin from the natural product containing pectin Acid-containing enzyme-treated product. The monogalacturonic acid-containing enzyme-treated product according to claim 4, wherein the content of monogalacturonic acid in the total content of monogalacturonic acid, oligogalacturonic acid, polygalacturonic acid and their methyl esters is 70% by mass or more. A monogalacturonic acid-containing enzyme-treated product according to claim 5 is passed through an anion exchange resin to adsorb monogalacturonic acid, and then the aqueous solution is passed through an alkaline aqueous solution to elute the monogalacturonic acid salt to have a purity of 65% by mass or more. A purified monogalacturonic acid-containing enzyme-treated product. The enzyme having the activity of acting on a natural product containing pectin to release monogalacturonic acid is allowed to act directly on the natural product containing pectin to release monogalacturonic acid, so as to release monogalacturonic acid. A method for producing a monogalacturonic acid-containing enzyme-treated product according to claim 1, wherein the monogalacturonic acid-containing enzyme-treated product is obtained. Food / beverage products containing the monogalacturonic acid containing enzyme processed material in any one of Claims 4-6.