Taxoid-14-beta-hydroxylases and methods for their use
AssigneeWashington State University Research Foundation
InventorsRodney B. Croteau, Anne Schoendorf, Stefan Jennewein
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Abstract
Oxygenase enzymes and the use of such enzymes to produce paclitaxel (Taxol®) related taxoids, as well as intermediates in the Taxol® biosynthetic pathway are disclosed. Also disclosed are nucleic acid sequences encoding the oxygenase enzymes. Particular oxygenase enzymes include the disclosed taxoid-14-beta hydroxylases.
Claims36
A purified polypeptide, comprising a polypeptide having an amino acid sequence that is at least 75% identical to SEQ ID NO:
- 67, or a fragment thereof, wherein the polypeptide has oxygenase activity.
An isolated nucleic acid molecule that:
- (a) hybridizes under low-stringency conditions with a nucleic acid probe, the probe comprising a nucleotide sequence according to SEQ ID 54, or a fragment thereof;
- and
- (b) encodes a protein having oxygenase activity.
An isolated nucleic acid molecule that:
- (a) has a nucleotide sequence at least 75% sequence identical to SEQ ID NO:
- 54;
- and
- (b) encodes a polypeptide having oxygenase activity.
A method for isolating a nucleic acid sequence, comprising:
- (a) hybridizing the nucleic acid sequence to at least 10 contiguous nucleotides of SEQ ID NO:
- 54;
- and
- (b) identifying the nucleic acid sequence as one that encodes an oxygenase.
A purified protein having oxygenase activity, comprising a polypeptide having an amino acid that:
- (a) comprises SEQ ID NO:
- 68;
- (b) differs from the amino acid sequence specified in (a) by one or more conservative amino acid substitutions;
- or
- (c) that is at least 70% identical to the sequence specified in either (a) or (b).
Dependencies are inferred from the claim text.