N-carbobenzyloxy (n-cbz)-deprotecting enzyme and uses therefor
AssigneeBristol-Myers Squibb Company
InventorsVenkata Nanduri, Ramesh Patel, Steven Goldberg, Robert Johnston
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Abstract
This invention relates to isolated or recombinant N-CBZ-deprotecting enzyme polypeptides that catalyze the removal of CBZ from CBZ-protected amino acids and alcohols, and variants, modifications, and fragments thereof. Also related are isolated nucleic acids encoding N-CBZ-deprotecting enzyme polypeptides, and variants, modifications, 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 polypeptides in various reactions for industrial or pharmaceutical applications.
Claims24
An isolated nucleic acid comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:2, or the complement thereof.
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A 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 polypeptide comprising amino acid sequence SEQ ID NO:2.
An isolated polypeptide comprising an amino acid sequence having at least 80% identity to the amino acid sequence of SEQ ID NO:
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A recombinant polypeptide comprising amino acid sequence SEQ ID NO:2.
An isolated nucleic acid comprising a nucleotide sequence that encodes an amino acid sequence sharing at least 95% sequence identity with the amino acid sequence of SEQ ID NO:2, and wherein said amino acid sequence encodes a polypeptide having catalytic activity, wherein said catalytic activity comprises deprotection of a N-carbobenzyloxy-protected amino acid.
Dependencies are inferred from the claim text.