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US4248966A
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Synthesis of isopenicillin derivatives in the absence of living cells
Assignee: Massachusetts Institute of Technology
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Filing: 1979-05-17
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Granted: 1981-02-03
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Expires: 1999-05-17
Abstract
Abstract Isopenicillin derivatives having antimicrobial properties and a general formula of: ##STR1## where R, R 1 , and R 2 are hydrogen, methyl, ethyl, propyl, isopropyl, or halogenated methyl, ethyl, propyl or isopropyl radicals and R 3 is ##STR2## are produced in a cell-free system using an extract from Cephalosporium acremonium. The starting materials for the synthesis consist of tripeptides composed of unsubstituted or β-substituted D-valine, unsubstituted or substituted L cysteine, and L- α-aminoadipic acid or its analogues. Certain enzymes in the cell-free extract are inactivated so that conversion does not proceed past the isopenicillin stage.
Inventors
- 👤 Arnold L. Demain
- 👤 Toshio Konomi
- 👤 Jack E. Baldwin
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 17:41
CPC Classifications
C; C12; C12P; C12P37/00; C12N; C12N1/00; C12N1/14; C12N1/145; C12R; C12R2001/00; C12R2001/645; C12R2001/745; C12R2001/75; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/911; Y10S435/925; Y10S435/926
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
61 words
A process for producing an isopenicillin derivative which exhibits antimicrobial activity, said process comprising the steps of: 1. providing a starting material selected from the group consisting of: ##STR18## wherein R, R 1 , and R 2 are the same or different and are selected from the group consisting of hydrogen, methyl, ethyl, propyl, and isopropyl and R 3 is ##STR19##
Claim 2
Dependent Claim
61 words
A process for producing an isopenicillin derivative which exhibits antimicrobial activity, said process comprising the steps of: 1. providing a starting material selected from the group consisting of: ##STR18## wherein R, R 1 , and R 2 are the same or different and are selected from the group consisting of hydrogen, methyl, ethyl, propyl, and isopropyl and R 3 is ##STR19##
Claim 3
Dependent Claim
80 words
providing a cell-free extract or permeabilized cells of Cephalosporium acremonium containing a certain late enzyme(s) which has been inactivated so that the conversion cannot proceed beyond the isopenicillin stage; 3. contacting the extract and the starting material in a reaction vessel; 4. providing ATP as an energy source to said reaction; and 5. allowing a component of the extract to react with said starting material for a sufficient amount of time to produce an isopenicillin derivative of the formula: ##STR20##
Claim 4
Dependent Claim
80 words
providing a cell-free extract or permeabilized cells of Cephalosporium acremonium containing a certain late enzyme(s) which has been inactivated so that the conversion cannot proceed beyond the isopenicillin stage; 3. contacting the extract and the starting material in a reaction vessel; 4. providing ATP as an energy source to said reaction; and 5. allowing a component of the extract to react with said starting material for a sufficient amount of time to produce an isopenicillin derivative of the formula: ##STR20##
Claim 5
Dependent Claim
18 words
The process as set forth in claim 1 wherein R, R 1 , and R 2 are hydrogen.
Claim 6
Dependent Claim
42 words
The process as set forth in claim 1 wherein R is hydrogen, one of R 1 and R 2 is methyl, and the other of R 1 and R 2 is selected from the group consisting of hydrogen, ethyl, propyl, and isopropyl.
Claim 7
Dependent Claim
27 words
The process as set forth in claim 1 wherein the ATP utilized is regenerated by an ATP regenerating system comprising a phosphate donor and a phosphotransferease enzyme.
Claim 8
Dependent Claim
21 words
The process as set forth in claim 4 wherein the phosphate donor is phosphoenolpyruvate and the phosphotransferase enzyme is pyruvate kinase.
Claim 9
Dependent Claim
23 words
The process as set forth in claim 1 wherein said cell-free extract is made by treating Cephalosporium acremonium cells with a lysing enzyme.
Claim 10
Dependent Claim
28 words
The process as set forth in claim 6 wherein said cell-free extract is made by treating Cephalosporium acremonium cells with endo β(1→3) glucanase, endo β(1→4) glucanase, and zymolyase.
Claim 11
Dependent Claim
22 words
The process as set forth in claim 1 wherein oxygen transfer is promoted by shaking the reaction components in the reaction vessel.
Claim 12
Dependent Claim
32 words
The process as set forth in claim 1 wherein mannitol and trace concentrations of KCl and MgSO 4 are included in the reaction and the system is buffered to about pH 7.2.
Claim 13
Dependent Claim
17 words
The process as set forth in claim 1 wherein, prior to step 3, said extract is frozen.
References Cited (Backward Citations)
US4002530A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-02-03 | 1984-08-04 | 1985-02-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-02-03 | 1988-08-04 | 1989-02-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-02-03 | 1992-08-04 | 1993-02-03 | $7700.00 | Window Closed / Lapsed |