US5173418A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Production in Escherichia coli of extracellular Serratia spp. hydrolases
Assignee: BENZON PHARMA A DANISH Ltd Co
•
Filing: 1990-02-07
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Granted: 1992-12-22
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Expires: 2010-02-07
Abstract
Abstract Extracellular Serratia spp. enzymes have been found to be excreted by another gram-negative organism harbouring a plasmid carrying DNA from Serratia spp. encoding the enzymes. This organism, e.g. E. coli, is therefore employed to produce the enzymes, specific examples of Serratia spp. enzymes produced are a nuclease, a lipase and a phosopholipase. The nuclease may be employed to remove nucleic acids from a biological material.
Inventors
- 👤 Soren Molin
- 👤 Michael Givskov
- 👤 Erik Riise
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 09:57
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C12N15/69; C12N1/00; C12N1/08; C12N15/11; C12N15/62; C12N15/625; C12N15/70; C12N15/74; C12N9/00; C12N9/14; C12N9/16; C12N9/18; C12N9/20; C12N9/22; C07; C07K; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/036
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
95 words
A method of producing an extracellular hydrolytic Serratia spp. enzyme selected from the group consisting of exocellular nuclease, exocellular phospholipase and exocellular lipase, comprising cultivating, E. coli in a culture medium, said E. coli harboring a hybrid plasmid which comprises DNA encoding an extracellular hydrolytic Serratia spp. enzyme selected from the group consisting of exocellular nuclease, exocellular phospholipase and exocellular lipase, operably linked to a promotor functional in said E. coli, under conditions conductive to the expression of said enzyme and its secretion into the culture medium, and harvesting the enzyme from the culture medium.
Claim 2
Independent Claim
95 words
A method of producing an extracellular hydrolytic Serratia spp. enzyme selected from the group consisting of exocellular nuclease, exocellular phospholipase and exocellular lipase, comprising cultivating, E. coli in a culture medium, said E. coli harboring a hybrid plasmid which comprises DNA encoding an extracellular hydrolytic Serratia spp. enzyme selected from the group consisting of exocellular nuclease, exocellular phospholipase and exocellular lipase, operably linked to a promotor functional in said E. coli, under conditions conductive to the expression of said enzyme and its secretion into the culture medium, and harvesting the enzyme from the culture medium.
Claim 3
Dependent Claim
16 words
A method according to claim 1 wherein the Serratia spp. enzyme is a Serratia spp. nuclease.
Claim 4
Dependent Claim
16 words
A method according to claim 1 wherein the Serratia spp. enzyme is a Serratia spp. phospholipase.
Claim 5
Dependent Claim
18 words
The method of claim 1 wherein the hybrid plasmid is a plasmid with a conditionally uncontrolled replication behavior.
Claim 6
Dependent Claim
11 words
The method of claim 1 wherein the DNA sequence is ##STR3##
Claim 7
Dependent Claim
37 words
A method according to claim 1 of producing the enzyme substantially free from other bacterial protein, in which the enzyme is secreted from the E. coli into the culture medium and is harvested from the culture medium.
Claim 8
Dependent Claim
28 words
The method of claim 1 wherein the DNA encodes a Serratia spp. nuclease which, prior to removal of the amino-terminal signal peptide, has the amino acid sequence ##STR4##
Claim 9
Dependent Claim
19 words
The method of claim 1 wherein the DNA encodes a Serratia spp. phospholipase encoded by the DNA sequence ##STR5##
Claim 10
Dependent Claim
12 words
A DNA fragment comprising a DNA sequence encoding a Serratia spp. phospholipase.
Claim 11
Dependent Claim
12 words
A DNA fragment comprising a DNA sequence encoding a Serratia spp. phospholipase.
Claim 12
Dependent Claim
13 words
A DNA fragment according to claim 9 wherein the DNA sequence is ##STR6##
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-12-22 | 1996-06-22 | 1996-12-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-12-22 | 2000-06-22 | 2000-12-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-12-22 | 2004-06-22 | 2004-12-22 | $7700.00 | Window Closed / Lapsed |