Patent US3640326A successfully refreshed from Google Patents.
Patent US4414332A successfully refreshed from Google Patents.
US4414332A
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US
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Endoproteinase-Lys-C and process for its preparation thereof
Assignee: Roche Diagnostics GmbH
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Filing: 1981-08-28
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Granted: 1983-11-08
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Expires: 2001-08-28
Abstract
Abstract The present invention provides endoproteinase-Lys-C from bacteria, consisting of a chain of molecular weight 35000 to 38000 Dalton, having a pH optimum at pH 7.7 and being inhibited by aprotinin but not being inhibited by alpha2-macroglobulin, alpha 1-antitrypsin and ethylenediamine-tetraacetic acid. The present invention also provides a process for obtaining this enzyme, wherein a culture broth of an appropriate bacterial strain which has been filtered or pre-purified by usual methods is chromatographed over carrier-fixed alpha2-macroglobulin-metal complex and the enzyme is obtained from the filtrate.
Inventors
- 👤 Jurgen Schrenk
- 👤 Peter Wunderwald
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 19:58
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/52; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
48 words
Endoproteinase-Lys-C from Lysobacterales which cleaves proteins at the carboxyl group of lysine and consists of a chain of molecular weight 35000 to 38000 Dalton, having a pH optimum at pH 7.7 and inhibited by aprotinin but not inhibited by α 2 -macroglobulin, α 1 -antitrypsin and ethylenediamine-tetraacetic acid.
Claim 2
Dependent Claim
48 words
Endoproteinase-Lys-C from Lysobacterales which cleaves proteins at the carboxyl group of lysine and consists of a chain of molecular weight 35000 to 38000 Dalton, having a pH optimum at pH 7.7 and inhibited by aprotinin but not inhibited by α 2 -macroglobulin, α 1 -antitrypsin and ethylenediamine-tetraacetic acid.
Claim 3
Dependent Claim
44 words
Process for obtaining endoproteinase-Lys-C as claimed in claim 1, which comprises chromatographing a culture broth of an endoproteinase-Lys-C producing strain of Lysobacterales which has been filtered or pre-purified by convention methods over carrier-fixed α 2 -microglobulin-metal complex, and obtaining said enzyme from the filtrate.
Claim 4
Dependent Claim
18 words
Process as claimed in claim 2, wherein the chromatographing is carried out in 0.01-0.1 M buffer, pH 6.5-9.
Claim 5
Dependent Claim
61 words
Process as claimed in claim 2, wherein the protein is precipitated from the filtered culture broth by means of ammonium sulphate, the precipitate is fractionated, after again dissolving, between 1.3 and 3 M ammonium sulphate and the fraction which is insoluble in this range is dissolved in water and, after dialysis, fractionated with acetone between 1.2 and 2.4 vol. of acetone.
Claim 6
Dependent Claim
40 words
Process as claimed in claim 4, wherein, after dissolving, the precipitate of the acetone fractionation is fractionated over a molecular sieve and the protein fractions eluted at the beginning of the chromatography subjected to the α 2 -macroglobulin-metal complex chromatography.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-11-07 | 1987-05-09 | 1987-11-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-11-07 | 1991-05-09 | 1991-11-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-11-07 | 1995-05-09 | 1995-11-08 | $7700.00 | Window Closed / Lapsed |