EP0252666A2
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Chimeric Enzymes
Assignee: Novo Nordisk AS
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Filing: 1987-06-30
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Granted: —
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Expires: 2007-06-30
Abstract
Abstract Chimeric alpha-amylases are disclosed of the formula Q-R-L in which Q is a N-terminal polypeptide residue of from 55 to 60 amino acid residues which is at least 75 percent homologous to the 55 N-terminal amino acid residues in the Bacillus amyloliquefaciens alpha-amylase as described in Takkinen et al., J. Biol. Chem. 258 (1983) 1007-1013, R is a polypeptide residue of the general formula in which X₈ comprises His or Gln, X₉ comprises Gly or Ser, and X₁₀ comprises Ser or Asp; and L comprises a C-terminal polypeptide of from 390 to 400 amino acid residues which is at least 75 percent homologous to the 395 C-terminal amino acid residues in the Bacillus licheniformis 584 (ATCC 27811) alpha-amylase.
Inventors
- 👤 Borge Krag Diderichsen
- 👤 Helle Outtrup
- 👤 Martin Schülein
- 👤 Barrie Edmund Norman
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-05 03:03
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/24; C12N9/2402; C12N9/2405; C12N9/2408; C12N9/2411; C12N9/2414; C12N9/2417; C12P; C12P19/00; C12P19/14
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
114 words
A chimeric alpha-amylase comprising the general formula I in which Q comprises a N-terminal polypeptide of from 55 to 60 amino acid residues which is at least 75% homologous to the 55 N-terminal amino acid residues in the Bacillus amyloliquefaciens alpha-amylase as described in Takkinen et al., J. Biol. Chem. 258 (1983) 1007-1013; R comprises a polypeptide of the general formula Ia in which X₈ comprises His or Gln, X₉ comprises Gly or Ser, X₁₀ comprises Ser or Asp; and L comprises a C-terminal polypeptide of from 390 to 400 amino acid residues which is at least 75% homologous to the 395 C-terminal amino acid residues in the Bacillus licheniformis 584 (ATCC 27811) alpha-amylase.
Claim 2
Dependent Claim
114 words
A chimeric alpha-amylase comprising the general formula I in which Q comprises a N-terminal polypeptide of from 55 to 60 amino acid residues which is at least 75% homologous to the 55 N-terminal amino acid residues in the Bacillus amyloliquefaciens alpha-amylase as described in Takkinen et al., J. Biol. Chem. 258 (1983) 1007-1013; R comprises a polypeptide of the general formula Ia in which X₈ comprises His or Gln, X₉ comprises Gly or Ser, X₁₀ comprises Ser or Asp; and L comprises a C-terminal polypeptide of from 390 to 400 amino acid residues which is at least 75% homologous to the 395 C-terminal amino acid residues in the Bacillus licheniformis 584 (ATCC 27811) alpha-amylase.
Claim 3
Dependent Claim
19 words
The chimeric alpha-amylase according to claim 1, in which X₈ comprises His, X₉ comprises Gly, and X₁₀ comprises Ser.
Claim 4
Dependent Claim
19 words
The chimeric alpha-amylase according to claim 1, in which X₈ comprises Gln, X₉ comprises Ser, and X₁₀ comprises Asp.
Claim 5
Dependent Claim
16 words
The chimeric alpha-amylase according to claim 1, in which the homologies are at least 80 percent.
Claim 6
Dependent Claim
16 words
The chimeric alpha-amylase according to claim 1, in which the homologies are at least 90 percent.
Claim 7
Dependent Claim
58 words
The chimeric alpha-amylase according to claim 1, in which Q comprises an N-terminal polypeptide residue of the general formula Ib in which X₁ comprises Ala-Asn-Leu or Val, X₂ comprises Met or Thr, X₃ comprises Ser-Ala-Tyr or Ala-Glu-His, X₄ comprises Ala-Gly-His or Ser-Asp-Ile, X₅ comprises Thr or Leu, X₆ comprises Ala or Ser, and X₇ comprises Val or Asn.
Claim 8
Dependent Claim
31 words
The chimeric alpha-amylase according to claim 6, in which X₁ comprises Val, X₂ comprises Thr, X₃ comprises Ala-Glu-His, X₄ comprises Ser-Asp-Ile, X₅ comprises Leu, X₆ comprises Ser, and X₇ comprises Asn.
Claim 9
Dependent Claim
23 words
The alpha-amylase according to claim 4, 5, 6, or 7, in which L comprises a C-terminal polypeptide residue of the general formula Ic
Claim 10
Dependent Claim
93 words
A process for the production of a chimeric alpha-amylase comprising: (a) recombining in vivo the N-terminal coding region of the alpha-amylase gene of B. amyloliquefaciens with the C-terminal coding region of the alpha-amylase gene of B. licheniformis to form recombinants; (b) selecting the recombinants that produce a chimeric alpha-amylase that is thermostable and exhibits a reduced negative effect on the use of A. niger glucoamylase and B. acidopullulyticus pullulanase for the saccharification of starch; (c) culturing the selected recombinants in an appropriate growth medium, and (d) recovering the chimeric alpha-amylase from the culture.
Claim 11
Dependent Claim
93 words
A process for the production of a chimeric alpha-amylase comprising: (a) recombining in vivo the N-terminal coding region of the alpha-amylase gene of B. amyloliquefaciens with the C-terminal coding region of the alpha-amylase gene of B. licheniformis to form recombinants; (b) selecting the recombinants that produce a chimeric alpha-amylase that is thermostable and exhibits a reduced negative effect on the use of A. niger glucoamylase and B. acidopullulyticus pullulanase for the saccharification of starch; (c) culturing the selected recombinants in an appropriate growth medium, and (d) recovering the chimeric alpha-amylase from the culture.
Claim 12
Dependent Claim
46 words
A process for converting starch into high dextrose syrup, comprising: (a) reacting the starch with the chimeric alpha-amylase of claim 1, 2, 3, 4, 5, 6, or 7 to form oligosaccharides; and (b) reacting the oligosaccharides formed in step (a) with a glucoamylase to form dextrose.
Claim 13
Dependent Claim
39 words
A process for converting starch into high dextrose syrup, comprising: (a) reacting the starch with the chimeric alpha-amylase of claim 8 to form oligosaccharides; and (b) reacting the oligosaccharides formed in step (a) with a glucoamylase to form dextrose.
References Cited (Backward Citations)
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