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Amino sugar-containing glucan, method for producing same and use of same

A technology of dextran and amino, applied in the field of dextran, can solve fatal problems

Active Publication Date: 2013-09-11
EZAKI GLICO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Glucosamine-containing dextran containing unremoved glucosamine-1-phosphate has a fatal disadvantage that it cannot be used as a medicine because the medicinal material does not allow impurities to be mixed

Method used

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  • Amino sugar-containing glucan, method for producing same and use of same
  • Amino sugar-containing glucan, method for producing same and use of same
  • Amino sugar-containing glucan, method for producing same and use of same

Examples

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Effect test

preparation example 1

[0395] (Preparation Example 1: Preparation of α-glucan phosphorylase derived from hyperthermophilus VF5)

[0396] The α-glucan phosphorylase derived from hyperthermophilus VF5 was recombinantly produced by the following method.

[0397] (A) Preparation of α-glucan phosphorylase gene derived from hyperthermophilus VF5

[0398] Chemically synthesize the amino acid sequence (the amino acid sequence described in SEQ ID NO: 2 of the Sequence Listing) that encodes the α-glucan phosphorylase gene derived from Hyperthermophilus VF5 by methods known to those skilled in the art; The amino acid sequence obtained from the base sequence No. 491380 to No. 493458 of AE000657 of ACCESSION No. AE000657 of the GenBank base sequence database of the National Center for Biotechnology Information (NCBI) (Accession No. Nucleic acid (also referred to as "α-glucan phosphorylase gene") of nucleotide sequence No. 491380 to No. 493458 of AE000657). An Ndel site is created upstream of the translation in...

preparation example 2

[0404] (Preparation Example 2: Preparation of Branched Glucan (B))

[0405] 50 g of waxy cornstarch (manufactured by SANWA CORNSTARCH Co., LTD) was suspended in 1000 ml of 10 mM sodium phosphate buffer (pH 7.0), and the suspension was heated to about 100°C to gel the waxy cornstarch . 200,000 units of the highly thermostable branched enzyme prepared according to the method described in Example 1 of Japanese Laid-Open Publication No. 2000-216581 were added to the starch slurry cooled to 70°C to prepare a reaction solution, and then allowed to react at 70°C for 16 Hour. The reaction solution was heated at 100° C. for 20 minutes, centrifuged at 6500 rpm for 10 minutes, and the supernatant was filtered through a membrane with a pore size of 0.8 μm. Then, the filtrate was subjected to gel filtration chromatography (AKTA purifier) ​​system (column: HiPrep produced by GE Healthcare TM 26 / 10 Desalting) to remove low molecular weight polysaccharides. 1000 ml of the filtrate was di...

preparation example 3

[0407] (Preparation Example 3: Preparation of Branched Glucan (P))

[0408] An aqueous solution of sucrose was prepared by dissolving 150 g of sucrose in 1000 ml of distilled water, and the solution was filtered through a membrane with a pore size of 0.2 μm. Mix sucrose aqueous solution (800 ml), 20 ml of 5% branched-chain dextran (B) aqueous solution (the 5% aqueous solution of branched-chain glucan (B) prepared in Preparation Example 2 of the above-mentioned branched-chain glucan 0.2 μm membrane filtration), 4 ml of 1 M sodium phosphate buffer (pH 7.0), 1800 units of recombinant Streptococcus mutans Sucrose phosphorylase, 1200 U of the α-glucan phosphorylase prepared in Preparation Example 1 of the present application, and 6000 U of the α-glucan phosphorylase prepared in Preparation Example 2 according to Example 1 of Japanese Laid-Open Publication No. 2000-316581 The high thermostable branched enzyme prepared by the method described above was used to adjust the liquid vol...

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Abstract

The purpose of the present invention is to provide a glucosamine-containing glucan, a modified product of the glucosamine-containing glucan and a conjugate of the glucosamine-containing glucan. This glucosamine-containing glucan is a glucosamine-containing glucan wherein at least one glucosamine residue is bound, by an alpha-1,4 bond, to each of two or more non-reducing terminals among a plurality of non-reducing terminals of a branched alpha-1,4 glucan but no glucosamine residue is present at any position other than non-reducing terminals of the branched alpha-1,4 glucan, and the glucan has a polymerization degree from 15 to 4*10<5> (inclusive). This glucosamine-containing glucan can be provided by having alpha-glucan phosphorylase act on an aqueous solution that contains a branched alpha-1,4 glucan and glucosamine-1-phosphate.

Description

technical field [0001] The present invention relates to dextran having at least 1 (preferably 2 or more) amino monosaccharide residues on each of at least 1 non-reducing end, its modified products and conjugates, as well as its preparation method and its use. More preferably, the present invention relates to glucans having at least 1 (preferably 2 or more) glucosamine residues on each of at least 1 non-reducing end, modified products and conjugates thereof, and their Preparation method and its use. The dextran, modified products and conjugates thereof of the present invention may have the function of non-covalently binding to nucleic acid to increase the apparent molecular weight of nucleic acid. Background technique [0002] The active ingredients of drugs are rapidly changing from chemically synthesized stable low-molecular-weight compounds to unstable substances that are easily degraded in the blood, such as proteins, antibodies, and nucleic acids. Therefore, it is nec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/26A61K47/36C08B37/00
CPCC12P19/26C08B37/0009C08B37/0063C08B31/00C08B35/00C08B37/0012A61K49/0054C12N15/87A61K9/5161A61K47/59A61K47/6455A61K47/36A61P43/00
Inventor 鹰羽武史久保亚希子柳濑美千代
Owner EZAKI GLICO CO LTD
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