Method for producing α-1, 6-branched α-1, 4-glucans from sucrose

• US 7,638,311 B2
• Filed: 07/20/2007
• Issued: 12/29/2009
• Est. Priority Date: 10/09/1998
• Status: Expired due to Fees

• Current Assignee: Bayer Cropscience AG (Bayer AG), Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften e.V. (Max-Planck-Gesellschaft zur Frderung der Wissenschaften e)
• Original Assignee: Bayer Bioscience GmbH, Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften e.V. (Max-Planck-Gesellschaft zur Frderung der Wissenschaften e)
• Inventors: Quanz, Martin, Buttcher, Volker
• Primary Examiner(s): Fronda; Christian L
• Application Number:
  US11/781,228
• Publication Number:
  US 20080020432A1
• Time in Patent Office: 893 Days
• Field of Search: None
• US Class Current: 435/101
• CPC Class Codes: C07K 14/22 from Neisseriaceae (F)
• CPC Class Codes: C12N 9/1051 Hexosyltransferases (2.4.1)
• CPC Class Codes: C12N 9/107 1,4-Alpha-glucan branching enzyme (2.4.1.18)
• CPC Class Codes: C12P 19/04 Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
• CPC Class Codes: 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
• CPC Class Codes: C12P 19/20 produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose

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

1. A method for producing α

-1,6-branched α

-1,4-glucans in in-vitro systems comprising

(1) expressing in an in-vitro system a protein encoded by a nucleic acid molecule encoding a branching enzyme from a bacterium of the genus Neisseria selected from the group consisting of

(a) a nucleic acid molecule encoding a protein which comprises the amino acid sequence depicted in SEQ ID NO. 2;

(b) a nucleic acid molecule comprising the coding region depicted in SEQ ID NO. 1;

(c) a nucleic acid molecule encoding a protein which comprises the amino acid sequence encoded by the insert in plasmid DSM 12425; and

(d) a nucleic acid molecule comprising the coding region for a branching enzyme, which is contained in the insert of the plasmid DSM 12425;

(e) a nucleic acid molecule encoding a protein the sequence of which has, a homology of at least 95% to the amino acid sequence depicted in SEQ ID NO. 2, and wherein positions 1-7 are SEQ ID NO;

8, positions 52-57 are SEQ ID NO;

9, positions 88-94 are SEQ ID NO;

10, positions 100-104 are SEQ ID NO;

11, positions 109-113 are SEQ ID NO;

12and positions 115-119 are SEQ ID NO;

13;

(f) a nucleic acid molecule having more than 95% identity with SEQ ID NO;

1, wherein said nucleic acid encodes a protein having SEQ ID NO;

8 at positions 1-7, SEQ ID NO;

9 at positions 52-57, SEQ ID NO;

10 at positions 88-94, SEQ ID NO;

11 at positions 100-104, SEQ ID NO;

12 at positions 109-113, are SEQ ID NO;

13 at positions 115-119, SEQ ID NO;

14 at positions 134-138, SEQ ID NO;

15 at positions 147-151, SEQ ID NO;

16 at positions 162-166, SEQ ID NO;

17 at positions 168-172, SEQ ID NO;

18 at positions 199-203, SEQ ID NO;

19 at positions 217-221, SEQ ID NO;

20 at positions 225-230, SEQ ID NO;

21 at positions 244-248, SEQ ID NO;

22 at positions 282-287, SEQ ID NO;

23 at positions 299-304, SEQ ID NO;

24 at positions 312-317, SEQ ID NO;

25 at positions 356-360, SEQ ID NO;

26 at positions 379-383, SEQ ID NO;

27 at positions 412-416, SEQ ID NO;

28 at positions 439-444, SEQ ID NO;

29 at positions 467-472, SEQ ID NO;

30 at positions 505-509, SEQ ID NO;

31 at positions 553-558, SEQ ID NO;

32 at positions 686-690, SEQ ID NO;

33 at positions 739-746 and SEQ ID NO;

34 at positions 754-762 of SEQ ID NO;

2; and

(g) a nucleic acid molecule the sequence of which deviates from the sequence of the nucleic acid molecule of (f) due to the degeneracy of the genetic code;

(2) contacting said protein with sucrose and amylosucrase;

or

and isolating said α

-1,6-branched α

-1,4-glucans.