Stable epimerase reagent, cyclase reagent and ring expansion reagent for cell-free production of cephalosporins
AssigneeQueen's University at Kingston
InventorsSaul Wolfe, Donald Westlake, Susan Jensen
Life of this patent
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Abstract
Cyclase, epimerase and a ring expansion enzyme are isolated separately from a cell free extract of a prokaryotic beta-lactam producing organism to provide three separate and stable enzyme reagents for commercial production of cephalosporins from peptide precursors. Isolation is carried out by using ammonium sulfate, gel filtration and ion exchange chromatography. The enzymes may be immobilized on a suitable support and the production of cephalosporins may be carried out continuously.
Claims9
A stable epimerase reagent containing epimerase having a molecular weight of about 60,000 capable of epimerizing isopenicillin N and analogs thereof to penicillin N and analogs thereof, derived from a cell free extract of a prokaryotic beta-lactam producing organism, said epimerase being separated from cyclase having a molecular weight of about 36,500 capable of cyclizing L-aminoadipyl-L-cysteinyl-D-valine and analogs thereof to isopenicillin and analogs thereof, and from a ring expansion enzyme having a molecular weight of about 29,000 and capable of ring expanding penicillin N and analogs thereof to desacetoxycephalosporin and analogs thereof.
A stable cyclase reagent containing cyclase having a molecular weight of about 36,500 capable of cyclizing L-aminoadipyl-L-cysteinyl-D-valine and analogs thereof to isopenicillin N and analogs thereof, derived from a cell free extract of a prokaryotic beta-lactam producing organism, said cyclase being separated from epimerase having a molecular weight of about 60,000 capable of epimerizing isopenicillin N and analogs thereof to penicillin N and analogs thereof, and from a ring expansion enzyme having a molecular weight of about 29,000 and capable of ring expanding penicillin N and analogs thereof to desacetoxycephalosporin and analogs thereof.
A stable ring expansion enzyme reagent containing a ring expansion enzyme having a molecular weight of about 29,000 and capable of ring expanding penicillin N and analogs thereof to desacetoxycephalosporin and analogs thereof, derived from a cell free extract of a prokaryotic beta-lactam producing organism, said epimerase being separated from cyclase having a molecular weight of about 36,500 capable of cyclizing L-aminoadipyl-L-cysteinyl-D-valine and analogs thereof to isopenicillin and analogs thereof and epimerase having a molecular weight of about 60,000 capable of epimerizing isopenicillin N and analogs thereof to penicillin N and analogs thereof.
Dependencies are inferred from the claim text.