JPH0776594A - Production of saccharide-carboxylic acid and novel saccharide-carboxylic acid - Google Patents

Abstract

PURPOSE:To produce a saccharide-carboxylic acid in which carbon atom having hydroxymethyl or hemiacetal hydroxyl group or both are specifically oxidized from wide saccharides in a high selectivity and yield. CONSTITUTION:The characteristics of this method for producing a saccharide- carboxylic acid or a salt thereof comprise reacting saccharides having carbon atom having hydroxymethyl and/or hemiacetal hydroxyl groups or a derivative thereof with a microorganism, having the ability to oxidize the carbon atom having the hydroxymethyl and/or hemiacetal hydroxyl groups into carboxyl group and belonging to the genus Pseudogluconobacter or a treated substance thereof, producing and accumulating the corresponding carboxylic acid and collecting the resultant carboxylic acid. Furthermore, this novel saccharide- carboxylic acid is obtained by the method for production The prepared carboxylic acid has excellent characteristics such as excellence in stability to enzymes and improvement in solubility in water.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel sugar carboxylic acid or a salt thereof and a method for producing a new sugar carboxylic acid. Specifically, a method for efficiently producing a corresponding carboxylic acid from a saccharide having a primary hydroxyl group (hydroxymethyl group) using a bacterium belonging to the genus Pseudogluconobacter or a treated product thereof, hydroxymethyl contained in the saccharide A novel sugar carboxylic acid whose group is oxidized to a carboxyl group,
A method for producing a corresponding carboxylic acid by oxidizing a carbon atom having a hemiacetal hydroxyl group of a saccharide having a hemiacetal hydroxyl group into a carboxyl group by using a bacterium belonging to the genus Pseudogluconobacter or a treated product thereof, and a hemiacetal hydroxyl group The present invention relates to a novel sugar carboxylic acid in a form in which a carbon atom having is oxidized to a carboxyl group.

[0002]

2. Description of the Related Art Microorganisms which catalyze the oxidation of hydroxymethyl groups to carboxyl groups are described in, for example, bacteria of the genus Acetobacter [Agric. Biol. Chem., Vol. 42, 233].
Item 1 (1978)], Gluconobacter bacteria [Agric. Biological Chemistry (Agric. B
iol. Chem.), Vol. 42, Item 2045 (1978)] or alcohol dehydrogenase of Pseudomonas bacteria [eg, Biochemical Journal (Biochem.J.
), 223, Item 921 (1984)], methanol dehydrogenase of a methanol bacterium [Agric. Biol. Chem.],
54, 3123 (1990)], etc., but their substrate specificity is limited, and there is no report that they act on other than alcohols such as methanol and ethanol.

Glucono is a microorganism which acts on sugars such as D-sorbitol and L-sorbose (hereinafter, may be simply referred to as sorbose) to catalyze the oxidation of the hydroxymethyl group to a carboxyl group. D-sorbitol dehydrogenase of Bacterium bacteria [Agric. Biol.
Chem.), Vol. 46, Item 135 (1982)], sorbose dehydrogenase [agricultural biological.
Chemistry (Agric. Biol. Chem.), Volume 55, 36
Section 3 (1991)], glucose dehydrogenase [Agric. Biological Chemistry (Agric. Bio
Chem.), Vol. 44, Item 1505 (1980)], etc., but all of them are insufficient in terms of versatility because of their limited substrate specificity. . In addition, a soil-separating bacterium already named Pseudogluconobacter saccharoketgenes produced a significant amount of 2-keto-L-gulonic acid from L-sorbose (JP-A-62-22828).
No. 8, JP-A-64-85088), or 2-keto-D-glucaric acid from D-glucose or the like (JP-A-62-2).
No. 28287) found to be generated.

On the other hand, of the polysaccharides, for example, dextran is a leuconostoc mesenteroid (Le
uconostoc mesenteroides) is a general term for high molecular weight glucans mainly composed of α1 → 6 bond produced from sucrose, and various chemical modification methods have been tried in the past, but in general chemical reactions, they are regioselective. With a very high reaction rate, it is difficult to obtain a product, and many side reaction products are given. As described above, many polysaccharides such as dextran are composed of homologues of various macromolecules with different molecular weights. In many cases, the substance of can not be clarified. ["Biotransformations in Preparative Organic Chemistry"]
ative Organic Chemistry HGDavis et al, Academic
(See Press)] In particular, it has been attempted to chemically oxidize dextran with sodium hypozincate, sodium bromate, chlorine, bromine, iodine, etc., but the structure of the product is not always clear. It has not been certified or is not fully supported. For example, regarding the chemical oxidation of dextran, Japanese application by Tosuni, Coelho and Patel (Patent Publication No. 61-23).
3001) and the like.

[0005]

As described above, the conventional
Oxidation of compounds such as sugars having hydroxymethyl groups to carboxylic acids using microorganisms was very limited to monosaccharides due to their substrate specificity. Further, the chemical oxidation of hemiacetal hydroxyl groups and hydroxymethyl groups of saccharides, especially oligosaccharides and polysaccharides, involves a number of side reactions as described above, which complicates the purification process and makes it possible to achieve a highly selective and efficient oxidation reaction. Technology was required. Thus, the object of the present invention is to obtain a hydroxymethyl group and / or a hemiacetal hydroxyl group in a high yield and with high selectivity by using a microorganism having a broad substrate specificity for producing industrially and industrially useful substances. It is to provide a method for producing a sugar carboxylic acid by oxidizing a carbon atom.

[0006]

[Means for Solving the Problems] As a result of intensive studies on various microorganisms in order to achieve the above object, the present inventors have found that Pseudogluconobacter saccharoketgenes has a hydroxymethyl group (--CH ₂ OH). It has been found that these groups of a wide range of sugars and / or their derivatives having carbon atoms with OH and / or hemiacetal OH are specifically oxidized to easily oxidize to sugar carboxylic acids with high yield and high selectivity. In addition, surprisingly, they found that the bacteria have extremely wide substrate specificity and extremely versatility, and further studies were conducted to complete the present invention. That is, the present invention provides 1) a monosaccharide derivative, an oligosaccharide or a derivative thereof, a polysaccharide or a derivative thereof having a hydroxymethyl group and / or a hemiacetal hydroxyl group, a carbon atom having the hydroxymethyl group and / or a hemiacetal hydroxyl group. A sugar carboxylic acid or a salt thereof characterized in that a microorganism belonging to the genus Pseudogluconobacter having the ability to oxidize to a carboxyl group or a processed product thereof is allowed to act to produce and accumulate a corresponding carboxylic acid, and the carboxylic acid is collected. The present invention relates to a production method, and 2) a novel sugar carboxylic acid in which at least one carbon atom having a hydroxymethyl group and / or a hemiacetal hydroxyl group of a saccharide or a derivative thereof is oxidized to a carboxyl group.

The microorganisms belonging to the genus Pseudogluconobacter which can be used in the present invention belong to the genus Pseudogluconobacter having an ability to oxidize carbon atoms having a hydroxymethyl group and / or a hemiacetal hydroxyl group of sugars to a carboxyl group. Any microorganism may be used,
Mutant strains obtained by a normal mutagenesis operation, for example, treatment with a mutagen such as nitrosoguanidine, ultraviolet irradiation treatment, or gene recombination are also included. Especially, Pseudogluconobacter saccharoketgenes is preferable.
More specifically, for example, European Patent Publication No. 22
The following strains described in No. 1,707 are listed as typical examples. Pseudo-Gluconobacter Saccharoketgenes K59
1s strain: FERM BP-1130, IFO 1446
4 Pseudogluconobacter Saccharoketgenes 12-
5 strains: FERM BP-1129, IFO 14465 Pseudogluconobacter saccharoketgenes TH
Strain 14-86: FERM BP-1128, IFO 1
4466 Pseudogluconobacter Saccharoketgenes 12-
15 strains: FERM BP-1132, IFO 1448
2 Pseudogluconobacter Saccharoketgenes 12-
4 strains: FERM BP-1131, IFO 14483 Pseudogluconobacter saccharoketgenes 22-
3 strains: FERM BP-1133, IFO 1148
4.

In the method of the present invention, the bacterial cells of the microorganism belonging to the genus Pseudogluconobacter may be allowed to act,
Alternatively, the processed product may be used. As the treated product, for example, a culture solution of these microorganisms can be used. Further, enzymes produced by these microorganisms may be used. However, in the case of the microorganism of the genus Pseudogluconobacter of the present invention, the enzyme is usually accumulated in the cells. Usually, it is convenient to use the bacterial cells themselves and bring them into contact with and act on the starting saccharides to generate carboxylic acids. Especially, it is preferable to use resting cells. The cells or the culture solution thereof can be produced, for example, according to the method described in JP-A No. 64-85088. That is, a seed culture is performed from a slant, then a main culture is performed to obtain a fermentation broth, and, if necessary, this fermentation broth is centrifuged, the precipitate is collected, and then washed with a saline solution several times to obtain The obtained precipitate can be subjected to a cell reaction. Further, the microorganism is a nutrient source that can be utilized by the strain under aerobic conditions, that is, carbon source (carbohydrate such as glucose, sucrose, starch or organic matter such as peptone, yeast extract), nitrogen source (ammonium salts, urea or Corn steep liquor,
Inorganic and organic nitrogen compounds such as peptone, inorganic salts (potassium, sodium, calcium, magnesium, iron,
Salts such as manganese, cobalt, copper phosphate, thiosulfate),
And a liquid medium containing vitamins / coenzymes such as CoA, pantothenic acid, biotin, thiamine, riboflavin, FMN (flavin mononucleoside) or amino acids such as L-cysteine and L-glutamic acid or natural products containing them as micronutrients. It can be cultured, and the culture solution thus obtained may be used in the method of the present invention.
Culturing can be performed at pH 4 to 9, preferably pH 6 to 8.

The culture time varies depending on the microorganisms used and the composition of the medium, etc., but is preferably 10-100.
It's time. A suitable temperature range for performing the culture is 10 to
40 degreeC, Preferably it is 25-35 degreeC. By adding a rare earth element to the medium at the time of culturing, the target substance can be produced more efficiently. The rare earth elements added to the medium include scandium (Sc), yttrium (Y), lanthanum (La), and cerium (C).
e), praseodymium (Pr), neodymium (Nd), samarium (Sm), europium (Eu), gadolinium (Gd), terbium (Tb), dysprosium (D)
y), holmium (Ho), erbium (Er), thulium (Tm), ytterbium (Yb) and lutetium (Lu). These rare earth elements may be added as metal powder or metal pieces, and they are also used as compounds such as chlorides, carbonates, sulfates, nitrates, oxides or oxalates. They may be used alone or two or more kinds of rare earth elements such as cerium carbonate and lanthanum chloride may be used at the same time. Further, a crude product obtained in the process of separating and refining various elements can also be used. The amount of rare earth element added to the medium may be selected within a range that does not inhibit the growth of the microorganism used,
Generally, the range of 0.000001 to 0.1% (W / V), preferably 0.0001 to 0.05% (W / V) is effective. As a method of addition to the medium, it may be added to the medium in advance, but it may be added intermittently during the culture or continuously.

In carrying out the present invention, the saccharide to be subjected to the reaction may be brought into contact with the microorganism using water or a solvent miscible with water, for example, dissolved or suspended in a solvent such as methanol, acetone or polyethylene glycol. . The amount of solvent used may be selected within a range that does not delay the reaction, and the substrate concentration is usually 0.1 to 20% (W / V),
Preferably, the range of 1 to 5% is effective. The temperature range suitable for carrying out the oxidation reaction by the microorganism of the present invention is 1
The temperature is 0 to 40 ° C, preferably 25 to 35 ° C. The reaction is preferably carried out under aerobic conditions. For example, while aerating air at 0.1 to 5 liters / minute, the reaction can be stirred at 50 to 2000 rpm if necessary. The reaction time is the primary hydroxyl group and // that substitutes the saccharide used for the reaction.
Or, depending on the properties of the hemiacetal hydroxyl group, 5 minutes ~
It is usually from 1 hour to 24 hours for 3 days. This reaction is p
It is preferable to adjust H. It is generally effective to carry out the treatment in the range of pH 4 to 9, preferably pH 6 to 8. The base used for pH adjustment can be used as long as it does not inhibit the reaction. For example, sodium hydroxide, potassium hydroxide,
Inorganic salts such as calcium carbonate, magnesium hydroxide, ferrous hydroxide, sodium morpholinoethane sulfonate,
Organic salts such as morpholinoethane sulfonate calcium can also be used. Moreover, by adding an anion exchange resin as desired, it is not necessary to add the above-mentioned neutralizing agent such as an alkali metal salt for adjusting the pH, and the reaction can be selectively controlled. The method of adding the anion exchange resin is particularly preferable in the case of selectively reacting to obtain one equivalent of the oxidant. The anion exchange resin used here may be any anion exchange resin as long as it adsorbs the produced carboxylic acid. Among them, styrene type and acrylic type anion exchange resins are preferable. Specifically, for example, Amberlite (trade name, Organo Co.) IRA-400, IRA-40
1, IRA-402, IRA-410, IRA-90
0, IRA-910, IRA-35, IRA-68, I
RA-94S, etc. Diaion (trade name, Mitsubishi Kasei)
SA-10A, SA-20A, PA-306, PA-3
08, PA-406, WA-10, WA-11, WA-
20, WA-30 and the like. These anion exchange resins are saccharides added as substrates (monosaccharide derivatives having a hydroxymethyl group and / or a hemiacetal hydroxyl group,
When the oligosaccharide or its derivative, the polysaccharide or its derivative) disappears in the reaction solution, the stirring is stopped, the reaction solution and the anion exchange resin are separated, and the anion exchange resin is added with an appropriate eluent, The product is obtained by elution. Examples of the eluent include an aqueous solution of sodium chloride, an alkali metal salt, or an acidic aqueous solution of hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, or the like. The sugar carboxylic acid thus eluted and accumulated can be collected and purified by a known means or a method similar thereto.

According to the method of the present invention, the primary hydroxyl group (hydroxymethyl group) is specifically oxidized to give the corresponding sugar carboxylic acid as sugars such as D-glucose, D-fractol, D-galactose, D-ribose, Derivatives of monosaccharides such as D-mannose and L-sorbose [eg, D-
Glucosamine, N-acetyl-D-glucosamine, N-
Amino sugars such as acetyl-chitobiose and tri-N-acetyl-chitotriose, ascorbic acid-related compounds such as glucosyl-L-ascorbic acid and L-ascorbic acid, inosine, adenosine, uridine, guanosine, cytidine, thymidine and 2-deoxy. Inosine, 2-
Nucleic acid related compounds such as deoxyadenosine, 2-deoxyuridine, 2-deoxyguanosine, 2-deoxycytidine, and 2-deoxythymidine, and streptozotocin (streptozocin)], sucrose, lactose, palatinose, raffinose, lactosucrose, glucosyl Oligosaccharides such as sucrose, galactosyl sucrose, xylobiose, starch-based sugars such as maltotriose, maltotetraose, isomaltotriose, panose, maltitol, amino sugars such as validamycin A, cellobiose, cellotriose, cellohexaose. Cellooligosaccharides, such as
Stevioside, rebaudioside-A, rebaudioside-C, rebaudioside-D, rebaudioside-
E, zulcoside-A, rubusoside [compounds of R ₁ = β-Glc, R ₂ = -β-Glc in the following formula (II)], steviol glycosides, oligosaccharides such as mogroside or derivatives thereof, and cyclodextrin, Examples include soluble starch, dextrin, dextran, polysaccharides such as β-1,3-glucan, and derivatives thereof. The oligosaccharide and polysaccharide derivatives also include sugar carboxylic acids in which the carbon atom having a hemiacetal hydroxyl group of the oligosaccharide or polysaccharide is oxidized to a carboxyl group by the method of the present invention described below.

According to the method of the present invention, hemiacetal hydroxyl groups are specifically oxidized to give sugar carboxylic acids as saccharides such as dextran, cellulose, chitin, amylose, amylopectin, maltotriose, panose, isomaltose, cellobiose, lactose, Maltose etc. are mentioned. The oligosaccharide and polysaccharide derivatives are, for example, sugar carboxylic acid such as dextranyl glucuronic acid in which the hydroxymethyl group of the primary hydroxyl group of the oligosaccharide or polysaccharide is oxidized to a carboxyl group by the above-described method of the present invention. Is also included. Among sugar carboxylic acids obtained from these sugars, for example, at least one hydroxymethyl group of palatinose is oxidized to a carboxyl group, and at least one hydroxymethyl group of sucrose is oxidized to a carboxyl group. Sugar carboxylic acid, sugar carboxylic acid in which at least one hydroxymethyl group of D-trehalose is oxidized to a carboxyl group, sugar carboxylic acid in which at least one of hydroxymethyl group of maltosyl-β-cyclodextrin is oxidized to a carboxyl group , 2-O-α-D-glucopyranosyl-L-ascorbic acid has at least one hydroxymethyl group oxidized to a carboxyl group, a sugar carboxylic acid, and streptozotocin has at least one hydroxymethyl group having a carboxyl group. Sugar carboxylic acid oxidized to silyl group, sugar carboxylic acid in which hydroxymethyl group of heptulose is oxidized to carboxyl group, formula (I)

[Chemical 4] A sugar carboxylic acid having at least one hydroxymethyl group of maltodextrin represented by the formula (II)

[0013]

[Chemical 5] A sugar carboxylic acid in which at least one hydroxymethyl group of the steviol glycoside represented by formula (III) is oxidized to a carboxyl group.

[Chemical 6] Of at least one hydroxymethyl group of the validamycin A represented by the formula (1), a sugar carboxylic acid in which at least one hydroxymethyl group of mogroside is oxidized to a carboxyl group, and salts thereof, and dextran Formula (VIII) in which the hydroxymethyl group possessed is oxidized to a carboxyl group

[Chemical 7] (However, in the formula, n = 1 to 50, preferably 10 to 15)
The sugar carboxylic acid represented by [that is, dextranyl glucuronic acid (also referred to as glucuronyl dextrin, dextran-glucuronic acid)], a salt thereof, or a complex or complex of the metal salt with them (hereinafter simply referred to as a complex) And dextranyl glucuronic acid (sometimes referred to as dextran-glucuronic acid) have the hemiacetal hydroxyl group-containing carbon atom oxidized to a carboxyl group (IX)

[Chemical 8] (In the formula, n = 1 to 50, preferably 10 to 15) The sugar carboxylic acid (that is, glucuronyl dextranyl gluconic acid), a salt thereof, or a complex of them and a metal salt is industrially used. It is a useful novel compound.

When an oxidative reaction is carried out by contacting the saccharide of the above-mentioned starting material with the above-mentioned bacterium belonging to the genus Pseudogluconobacter or a treated product thereof, the saccharide is
A bacterium belonging to the genus Pseudogluconobacter is also capable of being regioselectively and stepwise oxidized to specifically give a corresponding sugar carboxylic acid, reflecting the number and properties of primary hydroxyl groups and hemiacetal hydroxyl groups. It is a characteristic of the oxidation reaction by the body or processed products. When the target product is easily separated, the above-mentioned microorganism of the genus Pseudogluconobacter may be cultured in the above saccharide-containing medium. The culture conditions in this case can be the same as the method for obtaining the above culture solution. The sugar carboxylic acid thus produced and accumulated can be collected and purified by a known means or a method similar thereto. For example, filtration, centrifugation, treatment with activated carbon or adsorbent, solvent extraction,
Means such as chromatography, precipitation, and salting-out can be applied alone or in combination, and the desired product can be isolated and produced. As described above, when the oxidation is performed in the presence of the anion exchange resin, the reaction solution and the anion exchange resin are separated by a static method, a centrifugal separation method or the like, and the anion exchange resin is eluted with an eluent. After elution treatment, elution fractions containing the desired product are collected, and subjected to the above-mentioned known means or a method analogous thereto to isolate and purify the desired sugar carboxylic acid.

The complex of dextran carboxylic acid and iron salt can be generally produced by a known method for producing a complex of dextran and iron salt or according to the known method. For example,
It can also be produced by reacting dextran carboxylic acid with a sol of an iron salt such as ferric hydroxide. More preferably, dextran carboxylic acid is added to desalted and purified iron hydroxide sol by dialysis, and then the pH is adjusted to 8.0 to 10.
Then, a colloidal solution or suspension is obtained by heat treatment for 30 minutes in a heated temperature condition such as an autoclave at 100 ° C to 120 ° C. By this method, iron forms a dextran carboxylic acid derivative iron salt complex by a process such as salt formation, chelate formation and hydration. As will be described later, the elemental iron content of the colloidal suspension iron salt complex in the aqueous phase is about 50 in solution.
It is 250 mg / ml, and if desired, a high iron content iron salt complex can be prepared from a low iron content product by operations such as evaporation and concentration. The high iron content iron salt complex thus obtained is itself extremely stable for a long period of time, and in particular, has the property that its colloidal state is stably maintained in the aqueous phase. The complex of dextran carboxylic acid thus obtained and an iron salt such as ferric hydroxide is diluted with distilled water for injection, physiological saline or the like and parenterally administered to animals as an injection. Alternatively, in accordance with known pharmaceutical manufacturing methods, optionally using a pharmaceutically acceptable diluent or excipient, tablets, powders, granules, capsules, emulsions, liquids,
It can be orally administered as a premix agent or a syrup agent. In addition, it can be used as a single agent directly or after being dispersed in a carrier and mixed with feed, drinking water and the like.
Furthermore, if desired, each substance is separately formulated with a diluent, excipient, etc. that is acceptable for the drug substance, formulated into a diluent using a diluent, etc., and then mixed in feed, drinking water, etc. for administration. You can also do it. Furthermore, as described above, each separately formulated product can be administered separately to the same subject at the same time or at different times by the same route or different routes. The carboxylic acid may be obtained as a free form or as a salt by the method of the present invention. When it is obtained as a salt, it is converted into a free form by a conventional method, and when it is obtained as a salt, it is converted into a free form. It goes without saying that you can do it. Further, by allowing an alkali metal such as iron, lithium, sodium, potassium, etc., or an alkaline earth metal such as magnesium, calcium, etc. to be present in the medium, the salt thereof can be produced while the sugar carboxylic acid is produced and accumulated. Further, the obtained product can be identified by a conventional means such as elemental analysis, melting point, specific rotation, infrared absorption spectrum, NMR spectrum and chromatography.

The sugar carboxylic acid of the present invention thus obtained has various uses and characteristics. Examples of the steviol glycoside include the following compounds, and glucuronic acid type steviol glycoside derivatives obtained by oxidizing these are all novel substances.

[Table 1] The sugar carboxylic acid derivatives of various steviol glycosides containing these compounds were found to have improved sweetness and strong sweetness and refreshing taste. As a low-calorie, anti-cariogenic, enzyme-stable, highly sweet food material, it can be used in confectioneries, soft drinks, pickles, frozen desserts, etc. in accordance with conventional sweeteners. Further, since it is also excellent in terms of easy solubility, low toxicity, disintegrating property, degradability, etc., it can improve taste and can be used as a corrigent. For example, it is added to a formulation such as a tablet or a powder according to a conventional method,
It can be used by mixing. Also β-maltosyl-β
-The sugar carboxylic acid derivative of cyclodextrin is the first carboxylic acid derivative of cyclodextrins and has significantly improved solubility in water (solubility> 200 g).
/ 100 ml, water, 25 ° C). Therefore, for example, inclusion of a poorly soluble drug such as prostaglandin, steroid, and barbitolic acid with the sugar carboxylic acid of cyclodextrin of the present invention improves the water solubility and can be applied as an injection. When the sugar carboxylic acid is used as the clathrate as described above, it can be used in the same manner as the conventionally known clathrates. Further, β-D-fructosyl- (2 → 1) -α-D-glucuronic acid and α-D-glucuronyl- (1 → 2) -β obtained by reacting sucrose.
It was found that -D-6-fracturonic acid is a new sugar derivative having no sweet taste and is not decomposed by glucosidase, pectinase, glucuronidase and invertase. It is a sugar derivative that is more difficult to enzymatically decompose than sucrose. Therefore, it is useful as an indigestible, low-calorie sugar derivative as a food material for confectionery, cakes, frozen desserts and the like. The calcium salt, magnesium salt, and iron salt are useful as absorption improvers for these metal ions. Therefore, it can be added to confectioneries such as biscuits and soft drinks to be used for obtaining foods and drinks for preventing osteoporosis.

Further, β-D-fructosyl- (6 → 1) -α-D-glucuronic acid and α-D-glucuronyl- (6 → 1) -α-D- obtained by reacting palatinose.
Fracturonic acid is used as an indigestible, low-calorie anti-carious agent. Therefore, it can be used in various foods such as chewing gum and other confectionery in the same manner as ordinary sweeteners and seasonings. Further, α-D-glucuronyl-obtained by reacting D-trehalose
(1 → 1) -α-D-Glucuronic acid is useful as an indigestible moisturizer and a stabilizer for antibody preparations. Further, β-amino-2-deoxy-D-glucuronic acid obtained by reacting D-glucosamine is useful as a base material for cosmetics having excellent moisturizing properties. On the other hand, among nucleic acid derivatives obtained by reacting nucleosides such as inosine, especially 5'-
Carboxy-inosine and 5'-carboxy-adenosine have excellent taste properties and are useful as enzymatically stable taste agents, and are particularly useful as ingredients of foods and seasonings, especially for seasoning preserved foods. Available. In addition, 1-carboxy-2,7-anhydro-β obtained by reacting 2,7-anhydro-β-D-altro-heptulose
-D-Altro-heptulose can be used as a solubilizing agent for iron, calcium and magnesium having a weak sweetness in confectioneries such as biscuits and soft drinks. The sugar carboxylic acid of streptozotocin is used as an enzyme-resistant antibacterial agent and anticancer agent. Further, by utilizing its antibacterial effect, it can be used as it is or diluted with a solvent such as water to disinfect and sterilize a patient's room or limbs. The sugar carboxylic acid derivative of mogroside is useful as a high-intensity sweetener and can be used in soft drinks, confectionery, and the like.

Sugar carboxylic acid obtained by oxidizing maltodextrin is used as an indigestible, low-calorie food material.
It is used by adding and blending it into confectionery, cakes, frozen desserts, etc. by a conventional method. In addition, dextranyl glucuronic acid obtained by oxidizing a hydroxymethyl group of dextran to a carboxyl group is not only useful as a dextran per se or more stable and more useful as a more functional pharmaceutical additive, but is also a hydroxylated primary compound. (2) It exhibits excellent properties as a stabilizer for iron sol. Further, glucuronyl dextranyl gluconic acid in which a hydroxymethyl group of dextranyl gluconic acid is oxidized to a carboxyl group or a carbon atom having a hemiacetal hydroxyl group of dextranyl glucuronic acid is oxidized to a carboxyl group Alternatively, the salt thereof is useful as a pharmaceutical preparation material, and can be used as a viscosity-retaining agent for foods in the same manner as dextran. Examples of the metal salt forming a complex of dextranyl glucuronic acid and glucuronyl dextranyl gluconic acid, or a salt thereof and a metal salt include, for example, salts of valent, divalent or trivalent metals. And examples thereof include salts with metals such as calcium, magnesium, iron, sodium, lithium and potassium. Particularly, a complex with an iron compound such as ferric hydroxide can be used as an iron supplement for animal iron deficiency anemia in the same manner as a complex with dextran and iron as an anti-anemia agent. Also validamycin A
Is a compound represented by the above formula (III) and having a bactericidal activity against rice blight and the like, and the oxidant of this bacterial cell reaction is also resistant to glucosidase of the bacterial cell, It is useful as an agricultural fungicide with expected and improved persistence. The sugar carboxylic acid derivative of the ascorbic acid derivative also has improved stability against enzymatic degradation and is therefore used as an enzyme-stable antioxidant. As the antioxidant, it can be added to iron foods or the like and blended in the same manner as ascorbic acid. Thus, the sugar carboxylic acid of the present invention obtained by oxidizing the carbon atom having a hydroxymethyl group and / or a hemiacetal hydroxyl group of various sugars has not been heretofore provided with the characteristics that the sugar as a raw material does not have. Has utility.

The sugar carboxylic acid derivative obtained by oxidizing a hydroxymethyl group to a carboxyl group in the present invention is generally enzymatically stable and has improved solubility in water, as compared with the hydroxymethyl derivative as a substrate. And, if desired, it can be made into a metal salt, which is expected to be a diet food material such as indigestion and less likely to be calorie in food applications, a metal absorption improving effect, and the like. In addition, the carboxylic acids of steviol glycosides (stevioside, rebauside, rubusoside, etc.) are surprisingly free from the tinge and bad aftertaste shown by stevioside and rubusoside, and have a refreshing sweetness and 100% sugar. It is useful as a sweetener having a sweetness of up to 250 times and substantially no calories. Further, the carboxylic acid of the cyclodextrin derivative in which the hydroxymethyl group of the present invention is oxidized to a carboxyl group has excellent characteristics in terms of easy solubility, low toxicity, disintegration, degradability, and the like. It is possible to include basic drugs. By such inclusion, solubilization, stabilization and taste improvement of the active ingredient can be achieved, and thus it is used as a corrigent or flavoring agent. It is also useful as a pharmaceutical base that utilizes its enzymatic degradability. More specifically, use as a more functional base for formulations such as tablets, capsules, pills, powders, granules, ointments, injections, syrups, suspensions and nasal drops. Is mentioned. Thus, the present invention provides various novel sugar carboxylic acids by enabling the oxidation reaction of hydroxymethyl groups of a wide range of sugars to carboxyl groups. Further, the sugar carboxylic acid obtained by oxidizing the hemiacetal hydroxyl group to a carboxyl group obtained in the present invention is enzymatically stable as compared with a saccharide having a hemiacetal hydroxyl group as a substrate, and the solubility in water is improved, and If desired, it can be made into a metal salt, which is expected to be a material for diet foods such as indigestion and less likely to become calorie in food applications, a metal absorption improving effect, and the like. Further, it is useful as a enteric agent utilizing the properties of stabilizing active ingredients and enzyme degradability in pharmaceutical preparation applications.

[0020]

INDUSTRIAL APPLICABILITY The present invention provides a sugar carboxylic acid in which a carbon atom having a hydroxymethyl group and / or a hemiacetal hydroxyl group is specifically oxidized to a carboxyl group from a wide range of sugars,
It can be produced with high selectivity and high yield. The obtained carboxylic acid has excellent properties such as excellent stability against enzymes and improved solubility in water.

[0021]

EXAMPLES Next, the present invention will be described more specifically by way of examples, but the present invention is not limited to these examples. Example 1 β-D-fructosyl- (2 → 1) -α-D
-Manufacture of sodium glucuronic acid Pseudogluconobacter saccharoketgenes (Pseu
dogluconobacter saccharoketogenes) TH14-86
Preparation of bacterial solution; Pseudogluconobacter saccharoketogenes T
H14-86 strain slant was added to 20 ml of the following medium.
Add platinum loops and add this to a 200 ml smooth flask for 30
Incubate for 1 day at 0 ° C. with stirring using a rotary shaker. Then, 1 ml of the above culture broth was taken per 20 ml of medium, and this was cultivated with shaking at 30 ° C. for 2 days to give a seed culture. On the other hand, a slant of Bacillus megaterium IFO 12108 strain is added to a 200 ml smooth flask of 1 white ear per 20 ml of a medium (described below), and shake culture is carried out at 30 ° C. for 2 days. Next, the following main culture was performed. TH14-86 above
100 ml of strain seed culture (Seed medium) and Bacillus me
1.5 ml of Seed medium of gaterium was added to 900 ml of the following main culture medium, and the mixture was shake-cultured at 30 ° C for about 20 hours to obtain Ps.
eudogluconobactersaccharoketogenes TH14-86
Was used as the bacterial solution. Cell reaction: A solution prepared by dissolving 30 g of sucrose in 200 ml of sterilized water was added to a 5 liter jar fermenter manufactured by Biot Co., and the bacterial solution 1 prepared as described above was added thereto.
1 liter was added, and then 800 ml of sterilized water was added to make the total volume 2 liters. Air is aerated at 1 liter / min while stirring at 32 ° C. and 800 rpm (rpm), and 10% Na
The pH of the OH solution was automatically adjusted to 6.3 with the progress of the reaction. Converted to monocarboxylic acid in 2 hours.

Separation and purification; 80 g of 2 liters of the above reaction solution
The cells were separated with a cooling centrifuge at 00 rpm, the bacterial cells were removed, and the supernatant was subjected to activated carbon (special Shirasagi chromatocarbon, 400 g) column chromatography, washed with water (1.2 liters), and then with 3 liters of water. After elution, the fractions of the target compound were collected and concentrated under reduced pressure to obtain 19.70 g of β-D-fructosyl- (2 → 1) -α-D-glucuronic acid sodium salt as a white powder. Na-salt; in D ₂ O (90 MHz) δppm 61.39, 62.84,
71.48, 72.42, 73.09, 73.65, 74.65, 76.88, 82.01, 9
2.70, 104.18, 176.49. Preparation of magnesium salt of β-D-fructosyl- (2 → 1) -α-D-glucuronic acid β-D-fructosyl- (2 → 1) -α-D-glucuronic acid sodium salt 1.89 g of water was dissolved in 10 ml of water, then, passed through a 1R-120 (H ⁺ type) column (50 ml), and 0.103 g of magnesium oxide was added to the passing liquid,
Stir for 0 minutes. Then Millipore filter (Type G
S 0.22 μm), the filtrate was concentrated, and the resulting viscous syrup was washed with ethanol / acetone to give a white powder β-
1.45 g of magnesium salt of D-fructosyl- (2 → 1) -α-D-glucuronic acid was obtained. Elemental analysis C ₂₄ H ₃₈ O ₂₄ Mg.6H ₂ O calculated value C, 34.20; H, 5.98 measured value C, 34.33; H, 5.82

Medium composition [for TH14-86 strain] Seed medium (common to the 1st and 2nd times) Lactose 1% Yeast extract (Prodibel) 1 (NH ₄ ) ₂ SO ₄ 0.3 Corn steep liquor 3 CaCO ₃ ( Super) 2 Before CaCO _{3 addition} pH = 7.0 For Bacillus megathorium strain [Seed medium] Sucrose 4% Proflo [N source, Traders Protein (T & P)] 4 K ₂ HPO ₄ 0.65 KH ₂ PO ₄ 0.55 NaCl 0.05 (NH ₄ ) ₂ SO ₄ 0.05 MgSO ₄ .7H ₂ O 0.005 Calcium pantothenate 0.025 Before sterilization pH = 7.0 [Main medium] Sucrose 0.05% sterile corn steep liquor 2 separately sterilized (NH _{4) 2} SO ₄ 0.3 separately sterilized FeSO ₄ · 7H ₂ O 0.1 separately sterilized V. B ₂ 1 mg / L pre-sterilization pH = 7.0 sorbose 10% by Bacteria LaCl ₃ 0.01% by sterile CaCO ₃ (Super) 4% by sterilization

Example 2 α-D-glucuronyl- (1 →
2) Preparation of -β-D-6-fracturonic acid In the same manner as in Example 1, α-D-glucuronyl- (1 →
2) Sodium salt, magnesium salt, and calcium salt of -β-D-fracturonic acid were obtained. Sodium salt; white powder ¹³ C-NMR (D ₂ O) ppm: 61.98, 71.96, 73.02, 73.
35; 73.69, 76.37, 77.03, 79.82, 93.97, 105.80, 17
5.03, 175.70. Magnesium salt; White powder Elemental analysis C ₁₂ H ₁₆ O ₁₃ Mg.5H ₂ O Calculated value C, 29.86; H, 5.43 Measured value C, 29.76; H, 5.51 Calcium salt ; white powder Elementary analysis _{C 12 H 16 O 13 Ca ·} 3H 2 O calculated C, 31.17; H, 4.80 Found C, 31.26; H, 4.99

Example 3 Pseudogluconobacter saccharoketogene described in Example 1
s) 30 g of palatinose and 1 liter of sterilized water were added to 1 liter of the bacterial solution, and the reaction was carried out at 32 ° C. and 800 rpm with aeration of air at 1.6 l / min for 1 hour, and then the reaction solution was rotated at 8000 rpm. Separate the cells with a cooling centrifuge to remove the bacterial cells, and then add the supernatant to activated carbon (special Shirasagi Chromatocarbon 40
0g) Subject to column chromatography and wash with water (2 liters)
Then, the fractions eluted with a 10% aqueous methanol solution (4 liters) and further with a 50% aqueous methanol solution (4 liters) were collected, concentrated and lyophilized to give β-D-fructosyl- (6 → 1)-. 35.8 g of sodium salt of α-D-glucuronic acid was obtained. IR1 of this sodium salt
20 (H type) column and concentrate the passing liquid to β-D.
-Fructosyl- (6 → 1) -α-D-glucuronic acid was obtained. ¹³ C-NMR (D ₂ O) ppm: 63.35, 68.60, 71.78, 72.5
2, 72.77, 73.45, 75.14, 75.99, 79.53, 98.81, 102.2
1, 176.93. Elemental analysis C ₁₂ H ₂₀ O ₁₂ · 4.5H ₂ O Calculated value C, 32.96; H, 6.68 Measured value C, 33.17; H, 5.84

Example 4 In the same manner as in Example 3, the following starting materials [described in ()] were used to obtain the corresponding sugar carboxylic acid.

[0027]

[Table 2]

[0028]

[Table 3]

[0029]

[Table 4]

[0030]

[Table 5]

[0031]

[Table 6]

Example 5 Pseudogluconobacter saccharoketogene described in Example 1
s) 30 g of stevia extract (steviol glycoside containing stevioside as a main component) was placed in 1 liter of the bacterial solution in a 5 liter jar fermenter manufactured by Biot Co., and then 1 liter of sterilized water was added to prepare a reaction solution. 32 this
While stirring at 800 ° C. at 800 ° C., air was aerated at 1.6 l / min, and 10% NaOH solution was automatically added dropwise as the reaction progressed so that pH = 6.3.
After 1.5 hours, the disappearance of the substrate was observed, so the reaction was stopped, and then 2 liters of the reaction solution was separated with a cooling centrifuge at 8000 rpm to remove the bacterial cells, and the supernatant was added to HP-20 (aromatic). Group synthetic adsorbent, Mitsubishi Kasei column (1.8 liters)
To 50% EtO after washing with H ₂ O (8 liters).
A glucuronic acid-type steviol glycoside mixture was obtained from the fraction eluted with H (5 liters). After concentration, freeze-drying gave 19.28 g of white powder. The product was found to have a strong sweetness and a refreshing taste quality.

This product was subjected to HPLC under the following conditions. HPLC: Column ODP developing _{solution: CH 3 CN: H 2 O} = 3: 7 added Flow rate 0.1% trifluoroacetic acid to the solution; 1.0 ml / min Temperature; 35 ° C. Detector: RI and UV HPLC The pattern (RI detection) is shown in FIG. TLC: Silica Gel Merck Kieselgel
60 F ₂₅₄ No 5715 Developing solvent: CHCl ₃ : MeOH: H ₂ O (6: 4: 1) Color development: Sulfuric acid: MeOH (1: 1) 110-120 ° C, 10 minutes, heating Rf = 2.6 It was ¹³ C-NMR (D ₂ O) ppm of this product: 177.87 (ester), 17
1.84 (COO ^-), the signal of the carbonyl carbon is observed,
A new carbon was observed based on the carboxy group. HPLC
From the above findings, it was found that the main component of this product is glucose bound to the C ₁₉ position of stevioside and a compound in which glucose bound to the C ₁₃ position is converted into the glucuronic acid type.

Example 6 In the same manner as in Example 5, 30 g of stevia extract (steviol glycoside containing stevioside as a main component), 30 g of pseudogluconobacter saccharoketgenes bacterium solution, 32 ° C., 800 rpm , 5% NaOH solution 5 is used for oxidation reaction under the condition that air is aerated at 1.6 l / min.
The reaction was stopped when 9 ml (corresponding to about 2 equivalents of oxidation) was added dropwise. The required oxidation reaction time was 21 hours. After completion of the reaction, the reaction solution was subjected to centrifugal filtration to remove bacterial cells, and the supernatant solution (1850 ml) was purified (the same method as in Example 5) to obtain a white powder of glucuronic acid-type steviol glycoside mixture Na. 13.0 g of salt was obtained. The product was found to have a strong sweetness and a refreshing taste quality. HPLC; Column ODP, temperature 35 ° C. Mobile phase; 0.1% trifluoroacetic acid was added to a solution of CH ₃ CN: H ₂ O = 3: 7 Flow rate: 1 ml / min HPCL pattern (RI detection) in [FIG. 2] Indicated.

Example 7 In the same manner as in Example 5, 30 g of stevioside was used as a substrate for oxidation with Pseudogluconobacter saccharoketgenes. 30 ml of 5% sodium hydroxide was consumed and the reaction time was 90 minutes. The reaction solution was purified to obtain 20 g of white powder. This product had a high sweetness and a refreshing sweetness. The HPCL pattern (RI detection) of this product is shown in FIG. 400 mg of this white powder was used as a reverse phase column ODP-50 column (φ21.5 × 300 mm).
Asahi Kasei) and an eluent system of water / acetonitrile (72:28) containing 0.1% trifluoroacetic acid were used to separate the main components, followed by freeze-drying to obtain 124 mg of a white powder.
This product was recrystallized from a mixed solvent of methanol: water = 9: 1 to give colorless needle crystals. Melting point 226-230 [deg.] C. (decomposition). IR (KBr) cm- ¹ : 3500-3250, 1715, 1605, 1080-1
010, 890.

[Table 7] Elemental analysis C ₃₈ H ₅₇ O ₁₉ Na · 1.5H ₂ O Calculated value C, 52.59; H, 6.97 Measured value C, 52.56; H, 7.15 From the above, it was obtained in this reaction. The main oxidation product is R ₁ = -β-Glc-2-β-Glc, R ₂ = in the above formula (II).
-Β-Glc UA sodium salt, that is, 13-O-β-sophorosyl represented by the following formula (IV), 19
It was concluded that this is -O-β-D-glucuronyl steviol sodium salt.

[Chemical 9]

Example 8 In the same manner as in Example 5, 30 g of stevioside was used as a substrate for oxidation with Pseudogluconobacter saccharoketgenes. 67 ml of 5% sodium hydroxide solution was consumed in 80 minutes, the reaction solution was purified and white powder 32
g was obtained. This product had a high sweetness and a refreshing sweetness. The HPCL pattern (RI detection) of this product is shown in FIG. From 1 g of this white powder, reverse phase column ODP-
0.22 g of the main oxidation product was fractionated using the HPCL technique using a gradient system of 50 (φ21.5 × 300 mm Asahi Kasei) and 2% acetic acid / acetonitrile.
Secondary ion mass spectrometry of this product
Rometry. SI-MS) m / z 869 (M + 2H ₂ 0 ・
H ⁺ ) peak could be observed. ¹³ C-NMR (d ₆ -pyridine) ppm: 15.12, 18.95, 2
0.21, 21.69, 27.78, 37.91, 37.96, 39.33, 39.41, 4
0.35, 42.12, 43.58, 43.91, 47.23, 53.55, 56.86, 6
2.57, 70.07, 71.54, 72.49, 72.75, 75.75, 76.16, 7
6.79, 76.83, 77.51, 77.72, 77.78, 86.00, 86.97, 95.
07, 97.60, 103.89, 103.94, 153.50, 171.26, 171.89,
176.23. From the above findings, this product is represented by the formula (II) in which R ₁ = −
β-Glc-2-β-Glc UA, R ₂ = -β-Glc UA
Salt of the compound of 13-O-β-D-glucuronyl-β-D-glucosyl, 19-O-β-D-glucuronyl steviol represented by the following formula (V):
It was concluded that it was a sodium salt.

[Chemical 10]

Example 9 According to the method of Example 5, 30 g of stevioside was placed in a 5 liter jar fermenter manufactured by Biot Co., and 1.6 liter of sterilized water was added to prepare a reaction solution. While stirring the mixture at 32 ° C. and 800 rpm, air was aerated at 1 liter / min, 20 g of calcium carbonate was added, and the mixture was reacted for 4 hours. Then, 2 liters of the reaction solution was separated at 8000 rpm in a cooling centrifuge to remove the cells, and the supernatant was added to HP-2.
0 (aromatic synthetic adsorbent, Mitsubishi Kasei) column (1.8 liters) was passed through, washed with H ₂ O (8 liters), and then 5
From the fraction eluted with 0% ethanol (5 liters),
A calcium salt of uronic acid type stevioside was obtained. After concentration, freeze-drying gave 12.98 g of white powder.
This was recrystallized from methanol to obtain colorless powdery crystals. A weak sweetness was observed in this product. ¹³ C-NMR (d ₆ -pyridine) ppm of this product; 15.93, 19.
74, 20.92, 21.00, 28.58, 38.66, 38.71, 40.06, 41.0
4, 41.86, 42.00, 42.91, 44.33, 48.19, 54.50, 57.6
3, 73.11, 73.30, 73.45, 73.97, 74.08, 76.53, 77.7
2, 77.86, 77.89, 78.34, 78.39, 84.79, 86.79, 95.84,
98.21, 105.46, 107.02, 153.70, 171.93, 172.18, 17
2.95, 176.78. From the above findings, this product has the following formula (II) with R ₁ =
-Β-Glc UA-2-β-Glc UA, R ₂ = -β-Gl
c UA compound calcium salt, that is, 13-O-β-D-glucuronyl-β-D-glucuronyl-19-O-β-D-glucuronyl steviol calcium salt represented by the following formula (VI): .

[Chemical 11]

Example 10 In the same manner as in Example 5, 10 g of rebaudioside A was produced.
Pseudogluconobacter saccharoketgenes cells (15.8 g) and sterilized water (2000 ml) were added as a substrate.
℃, 800 rotations, stirring under the conditions of 1.6 l / min of air, the reaction, 2% NaOH solution was added, pH
Controlled to 6.3. The reaction was stopped when 23 ml of 2% NaOH was consumed, and the mixture was centrifuged to separate the supernatant. The reaction time required 135 minutes. The supernatant was used in Example 5.
Treated by the same purification method as in 10.16 g of white powder
Got The HPLC pattern (RI detection) of this product is shown in FIG. IR (KBr) cm ^-1 : 3340, 1720, 1610, 1410, 1070 This product has a high sweetness and an excellent sweetness with a refreshing feeling.

Example 11 Stevioside (30 g) was added to 1 liter of Pseudogluconobacter saccharoketgenes bacterium solution described in Example 1 in a 5 liter jar fermenter of Biot Co.,
Then, 1 liter of sterilized water was added, and Amberlite IRA-68 (hereinafter simply referred to as IRA-68) 20
0 ml was added to make a reaction solution. 32 ℃,
Air was aerated with 1.6 l / min while stirring at 800 rpm. After 2 hours, stevioside was no longer observed in the supernatant, so the reaction was stopped and the supernatant and IRA-68 were separated by centrifugation.
(4 × 24 cm), followed by elution with 2 L of 2N-saline solution, and the eluted fraction was passed through a HP-20 column (0.5 L). After washing with 1 liter of water, it was eluted with 0.8 liter of 50% methanol and concentrated to dryness to obtain 9.20 g of white powder. This product was recrystallized from a mixed solvent of methanol: water = 9: 1 to obtain colorless needle crystals. mp. 226-230 ° C (decomposition) This product was the same as that in Example 7, 13-O-β-sophorosyl-
It was certified as 19-O-β-glucuronyl steviol sodium salt (IV).

Example 12 According to the method described in Example 11, 30 g of Stevia extract (a mixture of at least 6 kinds of steviol glycosides) was added to 1 liter of Pseudogluconobacter saccharoketgenes bacterium solution, and 5 liter of Biot Co. 1 liter of sterilized water, and 200 ml of IRA-68 was further added to prepare a reaction solution. While stirring this at 32 ° C. and 800 rpm, air was aerated at 1.6 l / min. After 2 hours, stevioside was no longer observed in the supernatant, so the reaction was stopped, and the supernatant and IRA-68 were separated by centrifugation to separate IRA-68.
68 is packed in a column (φ4 x 25 cm) and then 2N-N
Elution was carried out with 1 liter of aCl, and the eluted fraction was passed through an HP-20 column (0.5 liter). After washing with 1 liter of water, elution with 1 liter of 50% methanol and concentration to dryness gave 8.9 g of a white powder. The HPLC pattern (RI detection) of this product is shown in FIG. IR (KBr) cm ^-1 : 3500-3200, 1720, 1705, 1610, 1
Showed 400.

Example 13 Glycotransfer Stevia Glycoside (SK Sweet, Sanyo Kokusaku Pulp)
(Manufactured by Co., Ltd.) 30 g was oxidized in the same manner as in Example 6,
Purification was performed to obtain 28.3 g of white powder. ¹³ C- of this product
Eight signals (179.50, 179.46, 179.40, 179.37, 179.32, 17) in the carbonyl carbon region by NMR (D ₂ O)
9.26, 177.10, 177.07 ppm), it was confirmed that at least one or more of the primary acid groups of the sugar was converted to a carboxylic acid. This product had a refreshing sweetness with a refreshing feeling.

Example 14 Mogroside V, a highly sweet glycoside contained in the fruits of Rakan fruit, 2
According to the method described in Example 11, 00 mg of Pseudogluconobacter saccharoketgenes 10 ml (0.32 g) was suspended in 3 ml of water, and then added to IRA.
-685 ml was added. This mixed solution was heated at 30 ° C. and 229
The mixture was stirred by rotation for 5 hours and a half. After leaving the reaction solution to stand and removing the supernatant, IRA-68 was packed in a column and washed with water (50 m
After l), elution was performed with 0.01N HCl, and the eluted fractions were collected and freeze-dried to obtain a white powder (32 mg). This product is IR (K
Since Br) cm ⁻¹ : 3300, 1600, 1410, 1060-1000, it was confirmed that the sugar chain was converted to the glucuronic acid type. This product had a sugar-like sweetness.

Example 15 1.0 g of rubusoside was dissolved in 60 ml of sterilized water.
IRA-68 (13 ml) was added, and then Pseudogluconobacter saccharoketogenes (30 ml) described in Example 1 was added.
Was added, and air was bubbled at 60 liters / minute while stirring at 32 ° C. and 600 rotations / minute, and the reaction was performed while stirring for 240 minutes. IRA-68 was taken out from the reaction solution, packed in a column, washed with 15 ml of water, and then washed with 2N-NaCl.
The solution was eluted with 200 ml of SV 0.5, the eluate was adsorbed on a HP-20 column (20 ml), washed with 150 ml of water and then with 50% MeOH solution (100 ml) SV0.
Elution with 5, the eluate was concentrated to dryness, and the precipitate was recrystallized from methanol to obtain 0.48 g of colorless granular crystals. mp 184-188 ° C (decomposition) elemental analysis C ₃₂ H ₄₇ O ₁₄ Na · 5H ₂ O theoretical value C, 49.99; H, 7.47 measured value C, 50.05; H, 7.54

[Table 8] From the above data, we conclude that the structure is represented by (VII).

[Chemical 12] In addition, this product (VII) was a good and highly sweet substance such as sugar, which had no tinge.

Example 16 Fursultiamine hydrochloride 10 mg Vitamin B ₂ 2 mg Taurine 1000 mg Benzoic acid 30 mg Butyl paraoxybenzoate 2.5 mg Glucuronic acid type stevioside sodium salt (IV) 50 mg Citric acid 100 mg Glycine 500 mg According to the above composition, drink according to the following method I got an agent. That is, butyl paraoxybenzoate and benzoic acid are dissolved in about 30 ml of water heated to about 70 ° C, and after cooling to about 25 ° C,
Fursultiamine hydrochloride, vitamin B _2, taurine, glucuronic acid type stevioside as a sweetener, citric acid, glycine were dissolved, 1N-Mars soda was added, after a 3.5 to pH, total volume by adding water Make up to 50 ml.

Example 17 30 g of precipitated calcium carbonate, 5 g of magnesium carbonate, 58.5 g of lactose and 6 g of hydroxypropyl cellulose were uniformly mixed, 300 ml of water was added, and the above mixture was granulated. The granules obtained are dried and then ground. The obtained powder and glucuronic acid type stevioside as a corrigent 0.05
g and 0.5 g of magnesium stearate were added and mixed, and the mixture was tableted according to a conventional method to give a diameter of 8.5.
One tablet of 200 mg in mm was obtained.

Example 18 Pseudogluconobacter saccharoketogene described in Example 1
s) Dextran-4 (molecular weight:
4000-6000; Extra Synthese Company (France)
(Manufactured by Biot Co., Ltd.) in a 5 liter jar fan mentor manufactured by Biot Co., Ltd. While stirring this at 32 ° C. and 600 rpm, air was aerated at 1.6 liter / minute to obtain 0.5% N.
The pH of the aOH solution was automatically dropped to 6.3 as the reaction proceeded. Stop the reaction in 3 hours,
The reaction solution (12.0 liters) was separated at 8000 rpm with a cooling centrifuge to remove bacterial cells and obtain a supernatant (1.98 liters).
The supernatant was filtered through a cellulose acetate membrane filter (φ0.2 to μm), and sterilized again, and the filtrate was amberlite IRA-68 (OH type) column (5
0 ml) and washed with a small amount of water (100 ml) to obtain 2 liters of passing liquid. This passing liquid was used as an HP-20 column (9
00 ml). Then elute with 2 liters of water,
3. The first 700 ml of the passing liquid is drained and then eluted.
3 liters were collected, and 13.8 ml of concentrated hydrochloric acid was added thereto to make it acidic, and this was filtered with a cellulose acetate membrane filter (φ0.2 μm), and the filtrate was added to 1000 ml of Sepha beads (trade name) SP205 (manufactured by Mitsubishi Kasei). Conducted. Then, it was washed with 700 ml of 0.05M hydrochloric acid and 2 liters of water, and then eluted with 2 liters of 20% MeOH solution,
The target fraction was obtained. This was concentrated under reduced pressure to obtain 14 g of dextranyl-glucuronic acid white powder. HP of this product
LC (Condition: Asahi Pack GS320 (φ7.6mm × 5
0 mm; manufactured by Asahi Kasei), mobile phase; water 1 ml / min, detection; R
I (Waters 410) and 200nm (Tosoh UV
-8020) A single peak was observed at Rt = 8.48 in a sample 0.5% aqueous solution (20 µl injection). (Rt of dextran 4 as a raw material was 10.83) In order to confirm the structure of this product, an enzyme digestion treatment experiment was performed. To 100 μl of a 1.5% aqueous solution of this product, add 100 μl of glucoamylase (Wako Pure Chemical Industries) solution (4 mg / ml), and add 30
The enzyme-treated solution was examined by HPLC by incubating at 0 ° C for one day. As a target experiment, dextran-4 was similarly treated with glucoamylase. As a result, it was found that this product did not undergo glucoamylase digestion. On the other hand, dextran-4 was digested with glucoamylase, the substrate disappeared, and glucose was newly formed. From this fact, this product has the above formula (VIII) (where n = 1
It was confirmed to be dextranyl glucuronic acid shown in 5).

Example 19 Pseudogluconobacter Saccharoketogene was prepared according to Example 1.
s) K591S strain was used in a peptone yeast medium (PY) medium consisting of 1% Bactopeptone and 1% yeast extract at 30 ° C. and 230 rpm with shaking at pH 7.5.
Culture was carried out for a day. Then, transfer to the main culture, add 0.1% calcium chloride in peptone yeast medium (PY medium), shake at 30 ° C., 230 rpm,
It was cultured for 48 hours. Then, the culture was centrifuged at 10,000 rpm for 10 minutes to collect the cells, which was then washed with 500 ml of water to obtain 7.59 g of cells per liter of broth. Then, Dextran-4 (molecular weight 4000-6
000; 30 g of Extra Synthesis (French) was dissolved in 1 liter of water, then 56 g of wet bacterial cells was suspended in 1 liter of water, and the mixture was stirred at 32 ° C. and 800 rpm for 60 minutes with air for 1 minute. Aerated with 0.1% NaO
The reaction was carried out for 6.5 hours while controlling the pH to 6.3 with the H solution. The reaction solution was centrifuged, 2 liters of the supernatant was collected, filtered and sterilized with a membrane filter, passed through an HP-20 column (900 ml), and eluted with 2000 ml of water. Concentrated hydrochloric acid was added to the passing solution to a final concentration of 0.05M,
After acidifying and filtering with a membrane filter, the filtrate was passed through an SP-205 column (100 ml),
0.05M HCl (1000ml), then water (2000
Fractions eluted with 20% MeOH (2000 ml) were collected, concentrated and freeze-dried to obtain 14 g of white powder of dextranyl gluconic acid. HPLC of this product
[Conditions: Asahi Pack GS320 (φ7.6mm × 50m
m: manufactured by Asahi Kasei), mobile phase: water 1 ml / min, RI (Waters 410) and 200 nm (Tosoh UV-802).
0), 20 μl injection of 0.5 %% aqueous solution]
There was a single peak at = 8.70. When this product was digested with glucoamylase under the conditions described above, it was easily digested, and glucose and glucosylgluconic acid could be detected. Therefore, this compound was confirmed to be dextranyl gluconic acid.

Example 20 Pseudogluconobacter was prepared in the same manner as in Example 19.
Pseudogluconobacter Saccharoke
togenes), 50 g of the cells (wet cells) were added to a solution of 30 g of dextranyl glucuronic acid (prepared by the method described in Example 18) in 1 liter of water, and the temperature was 32 ° C. and 800 rpm. The mixture was agitated at 60 ° C. for 1 minute with stirring, and reacted for 21 hours while controlling the pH to 6.3 with a 0.1% NaOH solution. The reaction solution was centrifuged, 2 liters of the supernatant was collected, filtered and sterilized with a membrane filter, and passed through an HP-20 column (900 ml).
Elution with 1.5 liters of water, the desired fractions were collected, concentrated and freeze-dried to obtain 15 g of 12 g of a white powder of sodium salt of glucuronyl dextranyl gluconic acid represented by the above formula (IX). HPLC: Rt = 9.12 1 peak HPLC conditions; column: GS-320 Mobile phase: H ₂ O, flow rate 1 ml / min Detection: RI ¹³ C-NMR (D ₂ O) δppm: 179.554, 178.119, 17
Observed at 2.434 and its intensity is 1: 0.5: 0.5,
The attribute is that the carbonyl carbon of the glucuronic acid part is 179.55.
4 and the other was carbonyl carbon of gluconic acid part (assuming 1 part lactone type).

Example 21 50 ml of a 50% aqueous solution of ferric chloride hexahydrate was heated to 30 ° C., and then 50 ml of a 24% Na ₂ CO ₃ solution was stirred well at a rate of 0.4 ml for 1 minute. Add dropwise. Then,
3 ml of 50 ml of 10% dextranyl glucuronic acid solution using dextranyl glucuronic acid obtained in Example 18
0.4 ml of 16% Na ₂ CO ₃ solution was added dropwise at a rate of 1 / min.
The pH was adjusted to 4.3 by adding at a rate of / min. Next, add 200 ml of ethanol, stir vigorously to form a slurry, centrifuge (5000 rpm), collect the precipitate, then 40 m of water.
Dissolve in ethanol, mix well with 60 ml of ethanol, centrifuge to remove soluble matter, add 20 ml of water to the precipitate, disperse well, remove ethanol well with an evaporator under reduced pressure, and remove with a 10% NaOH solution to give a 0.1. Add 2 ml / min at a speed with good stirring to adjust pH to 5-6, 100 ° C, 2
After heating for 0 minutes, phenol was added to 4 mg / ml to obtain 22 ml of dextranyl glucuronic acid iron hydroxide sol solution. The intramuscular administration of 1 ml of this product to young pigs was found to have the effect of significantly improving the anemia condition. The dextranyl gluconate iron hydroxide sol can be produced by the same method, and the effect of remarkably improving anemia of young pigs was recognized.

Example 22 Method for producing iron hydroxide sol of dextranyl gluconic acid 100 g of ferric chloride (FeCl ₃ .6H ₂ O) was dissolved in 100 ml of water, and this solution was ultrasonicated with Branson B-220. The treatment is carried out for 1 hour to sufficiently dissolve it. Then add water,
Make up to 200 ml and transfer the solution to a 500 ml beaker. Then, while stirring 200 ml of 24% sodium carbonate solution well,
A light tan iron hydroxide sol was prepared by adding at a rate of 0.8 ml / min at 0 ° C. 100 ml of the iron hydroxide sol prepared in this way was sampled and transferred to a 300 ml beaker at 30 ° C.
Slowly add 50 ml of 10% dextranyl gluconic acid solution while stirring well. Then, a 16% NaCO ₃ solution, very slowly, added at 0.1-0.2 / min, the pH was adjusted to 4.3. Then ethanol 2
00 ml was added with stirring and the resulting precipitate was centrifuged and
Separated from the supernatant, 80 ml of water was added to the precipitate to suspend it well, 120 ml of ethanol was added to reprecipitate, and the precipitate was separated by centrifugation. This operation was repeated once more, and the resulting precipitate was suspended in 20 ml of water and stirred well to 10%.
Add NaOH solution at a rate of 0.1-0.2 ml / min, p
H was added to 6.0. This solution at 120 ℃ 10
After autoclaving for 1 minute, phenol (so as to contain 1%) was added as a preservative, followed by concentration with an evaporator to prepare an iron hydroxide sol solution of dextranyl gluconic acid. The product formed a stable iron hydroxide sol. The property was analyzed and it was as follows. Total iron salt concentration: 200 mg / ml, viscosity; 32 cP, conductivity 4
It was 7 mS / cm.

Example 23 50 ml of 50% aqueous solution of iron chloride ferric chloride hexahydrate by dialysis method of dextranyl glucuronic acid was heated to 30 ° C., and then 50 ml of 24% Na ₂ CO ₃ solution was added to 1 ml. 0 minutes.
The mixture was added dropwise at a speed of 30 ml with good stirring to obtain a slightly blackish ocher iron hydroxide sol (pH 1.54). Transfer this iron hydroxide sol to a dialysis membrane (dialyses tube),
It was dialyzed overnight in ultrapure water. The iron hydroxide sol after dialysis is about 17
0 ml, a pH of 4.47 was dark brown. Then, to this dialyzed iron hydroxide sol, 50 ml of a 10% aqueous solution of dextranyl glucuronic acid obtained in Example 18 was added, stirred well, and adjusted to pH 4.3 with a 16% Na ₂ CO ₃ solution. After autoclaving this at 100 ° C for 30 minutes, 1
The pH was adjusted to 12 by adding 0% NaOH solution and the pH was adjusted to 12 again.
Autoclave for 20 minutes at 1 ° C to completely dissolve it, and then hydrate the dark brown dextranyl glucuronic acid
An iron salt complex was obtained. Then, this was dialyzed with a dialysis membrane overnight and concentrated, and then phenol was added thereto at 4 mg / ml to obtain 22 ml of an injection of dextranyl glucuronic acid ferric hydroxide complex. This product was a stable sol solution, and its properties were analyzed as follows. Total iron salt concentration: 201 mg / ml, viscosity 23.9 cP, electric conductivity 3.7 mS / cm. The intramuscular administration of 1 ml of this product to young pigs was found to have the effect of significantly improving the anemia condition. The dextranyl glucuronic acid ferric hydroxide complex can also be produced by the same method, and the effect of remarkably improving anemia of young pigs was recognized.

Example 24 Pseudogluconobacter was prepared in the same manner as in Example 11.
30 g of rebaudioside-A was added to 1 liter of Saccharoketgenes bacterium solution in a 5 liter jar fermenter manufactured by Biot Co., and then 1.5 liter of sterilized water was added,
Further, 400 ml of IRA-68 was added to obtain a reaction solution. This was aerated with 1.6 l / min of air while stirring at 32 ° C. and 600 revolutions. Rebaudioside-A was no longer observed in the supernatant after 1 hour, so the reaction was stopped and the supernatant and IRA-68 were separated by centrifugation.
IRA-68 was packed in a column (φ4 × 40 cm), and then eluted with 2N-saline solution (8.1 liter), and the eluted fraction was passed through a HP-20 (1 liter) column for adsorption.
After washing with water (8 liters) and eluting with 10% to 50% ethanol, the desired sodium salt of glucuronic acid type rebaudioside-A was obtained. After concentration and freeze-drying, 19.6 g of colorless powder was obtained. This recrystallized from MeOH-H ₂ O, to give a colorless needles 10.6 g. mp 220 ℃ (decomposition) HP of this product
LC (ODP column, acetonitrile: water = 30: 7)
0) gave one peak at Rt = 9.70. Of this product
¹³ C-NMR (D ₂ O) ppm: 18.06, 21.52, 22.91, 24.
19, 30.89, 39.51, 40.15, 42.02, 43.01, 43.73, 44.5
8, 46.69, 46.79, 49.89, 56.17, 59.64, 63.51, 63.7
1, 64.37, 71.36, 72.43, 73.14, 74.40, 74.67, 76.2
9, 76.99, 78.12, 78.70, 78.72, 78.81, 78.93, 79.27,
79.38, 81.49, 87.99, 90.31, 96.64, 98.70, 104.87,
105.06, 107.32, 155.99, 177.79, 181.64 From the above findings, this product has the following formula (II):

[Chemical 13] It was concluded that the compound is a sodium salt of the compound, that is, represented by the following formula (X).

[Chemical 14]

Test Example 6-O-α-D-glucuronyl (1 →
4) -α-D-glucosyl-β-cyclodextrin N
a Stability test of salt (1) against various enzymes was performed by the following method. As a comparative control, 6-O-α-maltosyl-β-cyclodextrin was used. Test method After adding a predetermined amount of each of the following enzymes to a test cyclodextrin aqueous solution of 10 mM, the mixture is allowed to stand in a warm bath at 37 ° C. Take aliquots of 500 μl each and store at 100 ℃ for 15
The enzyme is inactivated by heating for 1 minute and then centrifuged (15,000 rpm for 5 minutes). Further Millipore U
Filter with SY-1 (molecular weight cut off 10,000). 10
It was diluted twice and subjected to HPLC analysis under the following conditions. HPLC analysis conditions: Column; NH2P-50 (Asahipak) Mobile _{phase; CH 3 CN: H 2 O} = 48: 52 was added to PIC reagent 0.005M to. Flow rate: 0.8 ml / min Detection: RI From the HPIC of each sample obtained by the above operation, the residual ratio of cyclodextrin was calculated every 120 minutes.

Enzyme used and enzyme concentration;

[Table 9] Results 6-O-α-D-glucuronyl (1 → 4) -α-D-glucosyl-β-cyclodextrin Na salt (1) and 6-O-α-maltosyl-β-cyclodextrin remaining by each enzyme Fig. 11] to [Fig. 1]
4]. (1) was used as a control against the enzymatic treatment of α-amylase, glucoamylase and pullulanase.
It was found to be more stable than -O-α-maltosyl-β-cyclodextrin. On the other hand, in the study on the enzymatic treatment of pullulanase, 6-O-α-maltosyl-β-cyclodextrin as a control was decomposed by about 60%, whereas (1) was stable.

[Brief description of drawings]

FIG. 1 shows the HPLC pattern (RI detection) of the glucuronic acid type steviol glycoside mixture obtained in Example 5.

FIG. 2 shows the HPLC pattern (RI detection) of the glucuronic acid type steviol glycoside mixture obtained in Example 6.

FIG. 3 shows the HPLC pattern (RI detection) of the glucuronic acid type steviol glycoside sodium salt obtained in Example 7.

FIG. 4 shows the HPLC pattern (RI detection) of the glucuronic acid type steviol glycoside disodium salt obtained in Example 8.

FIG. 5 shows rebaudioside-A obtained in Example 10.
2 shows the HPLC pattern (RI detection) of 1 equivalent of the oxidant.

FIG. 6 shows the HPLC pattern (RI detection) of the glucuronic acid type steviol glycoside mixture obtained in Example 12.

FIG. 7 shows 6-O- (2-carboxyethyl) -β-cyclodextrin Na salt obtained in Example 4 and 6,6-
The chart of ¹³ C-NMR (270 MHz, D ₂ O) δ ppm of di-O- (2-carboxyethyl) -β-cyclodextrin is shown.

FIG. 8 is ¹³ C-NMR (270 MHz, D ₂ O) of 6-O-α-D-glucuronyl- (1 → 4) -α-D-glucosyl-α-cyclodextrin obtained in Example 4. δ
The chart of ppm is shown.

FIG. 9 is a ¹³ C-NMR (27 C-NMR of the 6-O-α-D-glucuronyl-β-cyclodextrin obtained in Example 4;
A chart of 0 MHz, D ₂ O) δ ppm is shown.

FIG. 10 shows 6-O- (2-carboxy2-hydroxyethyl) -β-cyclodextrin Na salt obtained in Example 4 and 6,6-di-O- (2-carboxy2-hydroxyethyl). ^{13 of} -β-cyclodextrin Na salt
_{C-NMR (270MHz, D 2} O) shows a chart of [delta] ppm.

FIG. 11 shows 6-O-α-D-glucuronyl (1 → 4) -α-D-glucosyl-β-cyclodextrin Na salt obtained in Test Example and control 6-O-α-maltosyl-
The result of the stability test with respect to (alpha) -amylase of (beta) -cyclodextrin is shown, and the time-dependent change of the residual rate when both compounds are (alpha) -amylase treatment is shown.

FIG. 12 shows 6-O-α-D-glucuronyl (1 → 4) -α-D-glucosyl-β-cyclodextrin Na salt obtained in Test Example and control 6-O-α-maltosyl-
The result of the stability test with respect to glucoamylase of (beta) -cyclodextrin is shown, and the time-dependent change of the residual rate when both compounds are treated with glucoamylase is shown.

FIG. 13 shows 6-O-α-D-glucuronyl (1 → 4) -α-D-glucosyl-β-cyclodextrin Na salt obtained in the test example and 6-O-α-maltosyl-control.
The results of the stability test of β-cyclodextrin against pullulanase are shown, and the time course of the residual ratio when both compounds are treated with pullulanase is shown.

FIG. 14 shows 6-O-α-D-glucuronyl (1 → 4) -α-D-glucosyl-β-cyclodextrin Na salt obtained in Test Example and control 6-O-α-maltosyl-
The result of the stability test with respect to (beta) -glucuronidase of (beta)-cyclodextrin is shown, and the time-dependent change of the residual rate at the time of treating both compounds with (beta) -glucuronidase is shown.

[Explanation of symbols]

In FIGS. 11 to 14, ● represents 6-O-α-maltosyl-cyclodextrin, and ▲ represents 6-O-α-D.
-Glucuronyl (1 → 4) -α-D-cyclodextrin Na salt is shown.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location C12P 19/56 7432-4B // (C12P 19/00 C12R 1:01) (C12P 19/56 C12R 1:01)

Claims (25)

[Claims] 1. A monosaccharide derivative, an oligosaccharide or a derivative thereof, a polysaccharide or a derivative thereof having a hydroxymethyl group and / or a hemiacetal hydroxyl group, and the carbon atom having the hydroxymethyl group and / or the hemiacetal hydroxyl group is a carboxyl group. A method for producing a sugar carboxylic acid or a salt thereof, which comprises reacting a microorganism belonging to the genus Pseudogluconobacter or a treated product thereof with the ability to oxidize to produce a corresponding carboxylic acid, accumulate the carboxylic acid, and collect the carboxylic acid. . 2. A monosaccharide derivative having a hydroxymethyl group, an oligosaccharide or a derivative thereof, a polysaccharide or a derivative thereof,
A microorganism belonging to the genus Pseudogluconobacter having the ability to oxidize the hydroxymethyl group to a carboxyl group or a treated product thereof is allowed to act to produce a corresponding carboxylic acid,
A method for producing a sugar carboxylic acid or a salt thereof, which comprises accumulating and collecting this. 3. The method according to claim 1 or 2, wherein the bacterial cells themselves of a microorganism belonging to the genus Pseudogluconobacter are allowed to act. 4. A sugar carboxylic acid in which at least one hydroxymethyl group of palatinose is oxidized to a carboxyl group or a salt thereof. 5. A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of D-trehalose is oxidized to a carboxyl group. 6. A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of maltosyl-β-cyclodextrin is oxidized to a carboxyl group. 7. 2-O-α-D-glucopyranosyl-L-
A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of ascorbic acid is oxidized to a carboxyl group. 8. A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of streptozotocin is oxidized to a carboxyl group. 9. A sugar carboxylic acid or a salt thereof in which the hydroxymethyl group of heptulose is oxidized to a carboxyl group. 10. A compound of formula (I): A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of maltodextrin represented by is oxidized to a carboxyl group. 11. Formula (II): A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of the steviol glycoside represented by is oxidized to a carboxyl group. 12. The sugar carboxylic acid according to claim 11, wherein the steviol glycoside is a stevioside in which R ₁ = -β-Glc-2-β-Glc and R ₂ = -β-Glc in the formula (II). Or its salt. 13. A steviol glycoside is represented by the formula (II): The sugar carboxylic acid or a salt thereof according to claim 11, which is rebaudioside-A which is 14. A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of validamycin A is oxidized to a carboxyl group. 15. A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of mogroside is oxidized to a carboxyl group. 16. A sugar carboxylic acid or a salt thereof in which at least one hydroxymethyl group of dextran is oxidized to a carboxyl group. 17. A complex of the sugar carboxylic acid or a salt thereof according to claim 16 and a metal salt. 18. A monosaccharide derivative, an oligosaccharide or a derivative thereof, a polysaccharide or a derivative thereof having a hemiacetal hydroxyl group, and a Pseudogluconobacter genus having an ability to oxidize a carbon atom having the hemiacetal hydroxyl group to a carboxyl group. A method for producing a sugar carboxylic acid or a salt thereof, which comprises reacting a microorganism to which the microorganism belongs or a treated product thereof to produce and accumulate a corresponding carboxylic acid and collecting the carboxylic acid. 19. The method according to claim 18, wherein the cells of a microorganism belonging to the genus Pseudogluconobacter are allowed to act. 20. The method according to claim 18, wherein the polysaccharide having a hemiacetal hydroxyl group is dextran. 21. The derivative of the polysaccharide having a hemiacetal hydroxyl group is dextranyl glucuronic acid.
The manufacturing method described. 22. A sugar carboxylic acid or a salt thereof in which a carbon atom having at least one hemiacetal hydroxyl group of dextranyl glucuronic acid is oxidized to a carboxyl group. 23. A complex of the sugar carboxylic acid or a salt thereof according to claim 22 and a metal salt. 24. A process for producing a complex of dextranyl glucuronic acid and ferric hydroxide, which comprises reacting dextranyl glucuronic acid with ferric hydroxide sol. 25. The method according to claim 1, wherein the oligosaccharide having a hydroxymethyl group is sucrose.