# US 8,771,424 B2 · Method for the producing alpha-1, 6-branched alpha-1, 4-glucans from sucrose

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- Number: US 8,771,424 B2
- Document ID: US8771424B2
- Status: Granted
- Kind: B2
- Application number: 13/194,719
- Filed: 2011-07-29
- Published: 2012-02-23
- Granted: 2014-07-08
- Earliest claimed: 1998-10-09
- Assignees: Bayer BioScience N.V.; Max-Planck-Gesellschaft zur Foderung der Wissenchaften E.V.
- Applicants: Martin Quanz
- Inventors: Martin Quanz
- Web page: https://patentgrep.com/patent/US8771424B2

## CPC

- C12N 9/1051: {Hexosyltransferases (2.4.1)}
- C07K 14/22: from Neisseriaceae (F)
- C12N 9/107: {1,4-Alpha-glucan branching enzyme (2.4.1.18)}
- C12P 19/04: Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
- C12P 19/16: produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin (non-biological hydrolysis of starch C08B30/00)
- C12P 19/20: produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}

## Abstract

Nucleic acid molecules which encode a branching enzyme from a bacterium of the genus Neisseria, vectors, host cell, plant cells and plants containing said nucleic acid molecules as well as starch obtainable from the plants described are described. Furthermore, an in-vitro method for producing α-1,6-branched α-1,4-glucans on the basis of sucrose and a combination of enzymes of an amylosucrase and a branching enzyme as well as the α-1,6-branched α-1,4-glucans obtainable by said method are described.

## Claims

### Claim 1

Starch prepared by the process that comprises extracting starch from transgenic plants or parts thereof that comprise transgenic plant cell containing a nucleic acid molecule linked to regulatory sequences guaranteeing the transcription in plant cells, wherein said nucleic acid molecule encodes a branching enzyme from a bacterium of the genus Neisseria selected from the group consisting of
(a) nucleic acid molecules encoding a protein which comprises the amino acid sequence depicted in SEQ ID NO. 2;
(b) nucleic acid molecules comprising the coding region depicted in SEQ ID NO. 1;
(c) nucleic acid molecules encoding a protein which comprises the amino acid sequence encoded by the insert in plasmid DSM 12425;
(d) nucleic acid molecules comprising the coding region for a branching enzyme, which is contained in the insert of the plasmid DSM 12425; and
(e) nucleic acid molecules encoding a protein the sequence of which has, in the first 100 amino acids, a homology of at least 65% to the amino acid sequence depicted in SEQ ID NO. 2;
wherein said starch has a phosphate content that is at last 20% reduced as compared to starch from a corresponding wild type plant.

### Claim 2 (depends on 1)

The starch according to claim 1, wherein the composition of the starch is modified in such a way that it has at least one of the properties selected from the group consisting of an increased gel texture, a reduced phosphate content, a reduced peak viscosity, a reduced pastification temperature, a reduced size of starch granules and a modified distribution of side-chains when compared to the starch from corresponding wild type plant.

### Claim 3 (depends on 1)

The starch according to claim 1, wherein said starch has a modified distribution of side-chains in that the proportion of side-chains with a DP of 6 to 8 is increased by at least 25% in comparison to the proportion of side-chains with the corresponding degree of polymerization of amylopectin from a corresponding wild-type plant.

### Claim 4 (depends on 1)

The starch according to claim 1, wherein said protein sequence has a homology of at least 95% to the amino acid sequence depicted in SEQ ID NO. 2.