US4536476A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Stable epimerase reagent, cyclase reagent and ring expansion reagent for cell-free production of cephalosporins
Assignee: Queen's University
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Filing: 1983-06-27
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Granted: 1985-08-20
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Expires: 2003-06-27
Abstract
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.
Inventors
- 👤 Saul Wolfe
- 👤 Donald Westlake
- 👤 Susan Jensen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 07:09
CPC Classifications
C; C07; C07K; C07K5/00; C07K5/04; C07K5/06; C07K5/06139; C12; C12N; C12N11/00; C12N11/18; C12N9/00; C12N9/90; C12P; C12P35/00; C12P35/06; C12P37/00; Y; Y10; Y10S; Y10S435/00; Y10S435/814; Y10S435/815; Y10S435/816; Y10S435/8215; Y10S435/822; Y10S435/886
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
92 words
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.
Claim 2
Dependent Claim
92 words
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.
Claim 3
Dependent Claim
22 words
An epimerase reagent as claimed in claim 1 derived from a prokaryotic organism selected from S. clavuligerus, S. cattleya and S. lipmanii.
Claim 4
Dependent Claim
12 words
An epimerase reagent as claimed in claim 1 derived from S. clavuligerus.
Claim 5
Dependent Claim
93 words
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.
Claim 6
Dependent Claim
93 words
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.
Claim 7
Dependent Claim
22 words
A cyclase reagent as claimed in claim 4 derived from a prokaryotic organism selected from S. clavuligerus, S. cattleya and S. lipmanii.
Claim 8
Dependent Claim
12 words
A cyclase reagent as claimed in claim 4 derived from S. clavuligerus.
Claim 9
Dependent Claim
93 words
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.
Claim 10
Dependent Claim
93 words
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.
Claim 11
Dependent Claim
23 words
A ring expansion enzyme as claimed in claim 7 derived from a prokaryotic organism selected from S. clavuligerus, S. cattleya and S. lipmanii.
Claim 12
Dependent Claim
13 words
A ring expansion enzyme as claimed in claim 7 derived from S. clavuligerus.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-08-19 | 1989-02-18 | 1989-08-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-08-19 | 1993-02-18 | 1993-08-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-08-19 | 1997-02-18 | 1997-08-20 | $7700.00 | Window Closed / Lapsed |