Cloned pullulanase
AssigneeMiller Brewing Company
InventorPatricia A. Bower
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Abstract
A method is disclosed for the expression of an active pullulanase enzyme in a microorganism host. In one aspect, a DNA construct contains a sequence encoding the pullulanase enzyme, except for the nucleotides necessary to encode the first two amino acids in mature pullulanase, and regulating sequences allowing expression of the coding sequence in a microorganism host. An advantageous DNA construct contains regulatory sequences permitting expression of the pullulanase in a yeast cell.
Claims22
A DNA construct capable of expressing an active rice pullulanase enzyme comprising a coding sequence according to that of SEQ ID NO:1 wherein the termination codon occurs at nucleotide positions 2647-2649, and regulatory sequences allowing expression of the coding sequence in a microorganism host selected from the group consisting of Saccharomyces, Bacillus, Aspergillus, Pichia, Kluyveromyces, and Escherichia coli, wherein said regulatory sequences are not operatively linked in nature with the coding sequence.
A DNA construct capable of expressing an active rice pullulanase enzyme comprising a coding sequence according to that of SEQ ID NO:2 wherein the termination codon occurs at nucleotide positions 2641-2643, and regulatory sequences allowing expression of the coding sequence in a microorganism host selected from the group consisting of Saccharomyces, Bacillus, Aspergillus, Pichia, Kluyveromyces, and Escherichia coli, wherein said regulatory sequences are not operatively linked in nature with the coding sequence.
A yeast according to ATCC Designation No. 74281.
A DNA construct capable of expressing an active rice pullulanase enzyme comprising a coding sequence according to that of SEQ ID NO:7 wherein the codon specifying the carboxyl-terminal amino acid of the pullulanase occurs at nucleotide positions 2644-2646, and regulatory sequences allowing expression of the coding sequence in a microorganism host selected from the group consisting of Saccharomyces, Bacillus, Aspergillus, Pichia, Kluyveromyces, and Escherichia coli, wherein said regulatory sequences are not operatively linked in nature with the coding sequence.
Dependencies are inferred from the claim text.