Modified .alpha.-amylases having altered calcium binding properties
AssigneeGenencor International, Inc.
InventorsRichard R. Bott, Andrew Shaw
Life of this patent
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Abstract
Novel .alpha.-amylase enzymes are disclosed in which a new calcium binding site is modified by chemically or genetically altering residues associated with that calcium binding site. The novel .alpha.-amylases have altered performance characteristics, such as low pH starch hydrolysis performance, stability and activity profiles.
Claims28
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to one or more of, Q298, G299, G301, Y302, L307, N309, Q340, F343, F403, H405, H406, D407, , G410, L427, I428, D430, G433, K436, N473, G474 and G475 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to Q298 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to after the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to G299 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to G301 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to after the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to Y302 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to after the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to L307 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby after the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to N309 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to after the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to Q340 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to F343 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to F403 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to H405 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to H406 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to after the characteristics of said calcium binding site and thereby after the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to D407 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to G410 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to after the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to L427 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to I428 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to D430 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to G433 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to K436 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby after the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to N473 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to G474 in Bacillus licheniformis.
An .alpha.-amylase comprising an A domain, a C domain and a calcium binding site, wherein said calcium binding site is associated with said A domain and said C domain comprises ligand residues in said A domain and/or said C domain, wherein said .alpha.-amylase is modified to alter the characteristics of said calcium binding site and thereby alter the performance of said .alpha.-amylase by substituting an amino acid residue at a position corresponding to G475 in Bacillus licheniformis.
Dependencies are inferred from the claim text.