N-carbobenzyloxy (N-CBZ)-deprotecting enzyme and uses therefor
AssigneeBristol-Myers Squibb Company
InventorsVenkata B. Nanduri, Ramesh N. Patel, Steven L. Goldberg, Robert M. Johnston
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Abstract
This invention relates to isolated or recombinant N-carbobenzyloxy-deprotecting enzyme polypeptides, and variants and fragments thereof, that catalyze the removal of carbobenzyloxy from carbobenzyloxy-protected amino acids and alcohols. Also related are isolated nucleic acids encoding N-carbobenzyloxy-deprotecting enzyme polypeptides, and variants, and fragments thereof, as well as vectors and host cells comprising these nucleic acids. The invention also relates to methods of obtaining isolated nucleic acids, polypeptides, and antibodies, and methods of using the polypeptides in various reactions for industrial or pharmaceutical applications.
Claims15
An isolated nucleic acid comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:2, or the complement thereof.
An isolated nucleic acid comprising the nucleotide sequence, encoding a N-carbobenzyloxy-deprotecting enzyme polypeptide, deposited as ATCC Accession Number PTA-5051.
An isolated nucleic acid which hybridizes under stringent conditions to any one of:
- (a) the nucleotide sequence SEQ ID NO:1;
- (b) nucleotides 1–1278 of SEQ ID NO:1;
- (c) the nucleotide sequence of SEQ ID NO:3;
- (d) the complement of (a), (b) or (c);
- or (e) a nucleotide sequence that differs from (a), (b, (c) or (d) due to degeneracy of the genetic code, wherein said isolated nucleic acid encodes a polypeptide having catalytic activity, wherein said catalytic activity comprises deprotection of a N-carbobenzyloxy-protected amino acid, and wherein said stringent hybridization conditions are hybridization in 50% formamide, 5×Denhardt's solution, 5×SSPE, and 0.2% SDS at 42° C., followed by washing in 0.1×SSPE and 0.1% SDS at 65° C.
An isolated nucleic acid molecule comprising a nucleotide sequence sharing at least 95% sequence identity with SEQ ID NO:
- 1, wherein said nucleotide sequence encodes a polypeptide having catalytic activity, and wherein said catalytic activity comprises deprotection of a N-carbobenzyloxy-protected amino acid.
Dependencies are inferred from the claim text.