Recombinant cephalosporin C amidohydrolase in cephalosporin biosynthesis
AssigneeGlaxo Group Limited
InventorsMartin Ramsden, David Wilson Spence
Life of this patent
Estimating the term…
Abstract
The present invention relates to an enzyme process for the one-step conversion of cephalosporin C or a derivative thereof into 7-aminocephalosporanic acid or a corresponding derivative thereof. The one step conversion is effected using a cephalosphorin C amidohydrolase derived from Pseudomonas Vesicularis B965, or from any cephalosporin C amidolydrolase producing or potentially producing descendants thereof, or from expression of DNA derived from Pseudomonas Vesicularis B965 or any cephalosporin C amidohydrolase producing or potentially producing descendants thereof.
Claims14
A method of expressing cephalosporin C amidohydrolase in a eukaryotic or prokaryotic host cell comprising
- a) transferring the host cell with a recombinant DNA expression vector that comprises
- i) an expression control sequence that functions in the host cell, and
- ii) a recombinant DNA sequence encoding a cephalosporin C amidohydrolase operably linked to the expression control sequence, wherein said DNA sequence has the nucleotide sequence shown in SEQ ID NO:1;
- b) culturing the host cell transformed in step a) under conditions that allow for expression of said cephalosporin C amidohydrolase.
A method of expressing a polypetitde having cephalosporin C amidohydrolase activity, said polypeptide having the amino acid sequence set forth in SEQ ID NO:2, or an amino acid sequence having at least 90% homology thereto, in a eukaryotic or prokaryotic host cell comprising:
- a) transferring the host cell with a recombinant DNA expression vector that comprises
- i) an expression control sequence that functions in the host cell, and
- ii) a recombinant DNA molecule encoding said polypetide which is operably linked to the expression control sequence;
- b) culturing the host cell transformed in step a) under conditions that allow for expression of said polypetide having cephalosporin C amidohydrolase activity.
A recombinant DNA molecule comprising a DNA sequence which encodes a polypeptide which comprises the amino acid sequence set forth in SEQ ID NO:2, or a polypetide having at leat 90% homology thereto and which retains essentially the same cephalosporin C amidohydrolase activity of the polypeptide comprising the amino acid sequence of SEQ ID NO:2.
A recombinant DNA molecule comprising a DNA sequence that encodes the amino acid sequence set forth in SEQ ID NO:2.
An isolated DNA molecule that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:2 or a polypeptide having an amino acid sequence at least 90% homologous thereto and that has essentially the same cephalosporin C aminohydrolase activity as the polypeptide comprising the amino acid sequence of SEQ ID NO:2.
Dependencies are inferred from the claim text.