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Method of producing glycogen

A technology of glycogen and glucan, applied in the field of producing high molecular weight highly branched α-glucan

Active Publication Date: 2008-06-11
EZAKI GLICO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0054] The object of the present invention is, in order to solve the aforementioned problems, to provide a process for the production of highly branched and high molecular weight alpha-glucans, especially glycogen

Method used

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  • Method of producing glycogen
  • Method of producing glycogen
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Examples

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Embodiment 1

[0247] (Production Example 1: Recombinant production of BE derived from the wind-producing liquid bacteria VF5)

[0248] (A) Preparation of VF5 BE gene of P. aeruginosa

[0249] The gene encoding the amino acid sequence of SEQ ID NO:2 (SEQ ID NO:1) was chemically synthesized. The SD sequence was added upstream of the translation initiation codon of the gene, and a BamHI site was provided upstream of the SD sequence. An EcoRI site is provided downstream of the translation stop codon. This synthetic gene was cleaved with BamHI and EcoRI to prepare a gene fragment, which was then ligated into pUC19 (manufactured by Takara Shuzo Co., Ltd.) previously cleaved with BamHI and EcoRI using T4-DNA ligase to obtain plasmid pAQBE1.

[0250] (B) Expression of the BE gene of A. rhizogenes in Escherichia coli

[0251] Escherichia coli TG-1 was transformed with this plasmid, and the transformants were diluted and extracted on LB agar medium (100 μg / ml ampicillin, 1% tryptone manufactured b...

Embodiment 2

[0253] (Production Example 2: Recombinant production of BE derived from Bacillus stearothermophilus TRBE 14)

[0254] BE derived from Bacillus stearothermophilus TRBE 14 was recombinantly produced by the method shown in Non-Patent Document 12 from the Escherichia coli TG-1 strain carrying the plasmid pUBE821 shown in this document.

Embodiment 3

[0255] (Production Example 3: Recombinant production of MalQ derived from Thermus aquaticus (hereinafter referred to as TaqMalQ))

[0256] TaqMalQ was recombinantly produced by the method shown in Terada et al. (Applied and Environmental Microbillogy, vol. 65, pp. 910-915 (1999)) from the Escherichia coli MC1061 strain carrying the plasmid pFGQ8 shown in this document.

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Abstract

A method of producing glycogen comprising a step of: allowing a branching enzyme having the ability to synthesize glycogen to act on a substrate in a solution to produce a glycogen, wherein the substrate is an ±-glucan being linked mainly with ±-1,4-glucosidic bonds and having a degree of polymerization of 4 or more, and the number-average molecular weight of saccharides in the solution before initiation of the reaction is more than 180 but not more than 150,000. (The branching enzyme activity of the branching enzyme) / (the molecular-weight-decreasing activity of the branching enzyme) can be 500 or less. The branching enzyme can be a thermostable branching enzyme.

Description

technical field [0001] The present invention relates to a method of producing high molecular weight, highly branched alpha-glucans, especially glycogen. Background technique [0002] Alpha-glucans are polymers of alpha-D-glucose. Various forms of alpha-glucans exist in nature. Among alpha-glucans, typical examples are glycogen and starch. However, glycogen and starch differ significantly from each other in structure and physical properties. [0003] Glycogen is the main storage polysaccharide of animals, fungi, yeast and bacteria. Glycogen is water soluble and forms a milky white solution. The molecular structure of glycogen in animals has been well studied. Natural glycogen is homoglucan in which sugar chains of glucose (glucose) linearly bonded by α-1,4-glucosidic bonds are branched by α-1,6-glucosidic bonds, and the resulting Branches branch further to form a network structure. Natural glycogen consists of alpha-1,4-glucoside linked chains with an average degree of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/18C12N15/09C08B37/18C12R1/01C12N9/26C12R1/19C12N9/10
CPCC12P19/18C12N9/2411C12P19/04C12N9/107
Inventor 梶浦英树高田洋树鹰羽武史栗木隆
Owner EZAKI GLICO CO LTD
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