US6689589B2
Expired / Lapsed
US
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Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids
Assignee: CJ CheilJedang Corp
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Filing: 2001-11-09
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Granted: 2004-02-10
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Expires: 2021-11-09
Abstract
Abstract The gene encoding a 4-hydroxybutyryl-Co A transferase has been isolated from bacteria and integrated into the genome of bacteria also expressing a polyhydroxyalkanoate synthase, to yield an improved production process for 4HB-containing polyhydroxyalkanoates using transgenic organisms, including both bacteria and plants. The new pathways provide means for producing 4HB containing PHAs from cheap carbon sources such as sugars and fatty acids, in high yields, which are stable. Useful strains are obtaining by screening strains having integrated into their genomes a gene encoding a 4HB-CoA transferase and/or PHA synthase, for polymer production. Processes for polymer production use recombinant systems that can utilize cheap substrates. Systems are provided which can utilize amino acid degradation pathways, α-ketoglutarate, or succinate as substrate.
Inventors
- 👤 Gjalt W. Huisman
- 👤 Frank Skraly
- 👤 David P. Martin
- 👤 Oliver P. Peoples
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 05:38
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0008; C08; C08G; C08G63/00; C08G63/02; C08G63/06; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/82; C12N15/8241; C12N15/8242; C12N15/8243; C12N9/10; C12N9/1025; C12N9/1029; C12N9/1096; C12P; C12P7/00; C12P7/62; C12P7/625
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
64 words
A recombinant host having stably incorporated into the genome a gene encoding a heterologous enzyme selected from the group consisting of a polyhydroxyalkanoate synthase and a 4-hydroxybutyrate-CoA (4HB-CoA2) transferase, wherein when the host is provided with feedstock selected from the group consisting of carbohydrates, succinate, 4-hydroxybutyrate, α-ketoglutarate, and amino acids, the enzyme is expressed in a sufficient amount to produce polyhydroxybutyrate-co-polyhudrohexanoate or poly 4-hydroxybutyrate.
Claim 2
Dependent Claim
64 words
A recombinant host having stably incorporated into the genome a gene encoding a heterologous enzyme selected from the group consisting of a polyhydroxyalkanoate synthase and a 4-hydroxybutyrate-CoA (4HB-CoA2) transferase, wherein when the host is provided with feedstock selected from the group consisting of carbohydrates, succinate, 4-hydroxybutyrate, α-ketoglutarate, and amino acids, the enzyme is expressed in a sufficient amount to produce polyhydroxybutyrate-co-polyhudrohexanoate or poly 4-hydroxybutyrate.
Claim 3
Dependent Claim
19 words
The host of claim 1 having stably incorporated into its genome both a polyhydroxyalkanoate synthase and a 4HB-CoA transferase.
Claim 4
Dependent Claim
11 words
The host of claim 1 wherein the host is E. coli.
Claim 5
Dependent Claim
24 words
The host of claim 3 wherein the heterologous enzyme is a polyhydroxyalkanoate synthase and the host expresses an endogenous enzyme with 4HB-CoA transferase activity.
Claim 6
Dependent Claim
21 words
The host of claim 1 further comprising genes expressing enzymes selected from the group consisting of β-ketothiolase and acetoacetyl CoA reductase.
Claim 7
Independent Claim
73 words
A method for enhancing production of polymers containing 4HB in a host comprising stably incorporating into the genome of the host a gene encoding a heterologous enzyme selected from the group consisting of a polyhydroxyalkanoate synthase and a 4HB-CoA transferase; providing the host with feedstock selected from the group consisting of carbohydrates, succinate, 4-hydroxybutyrate, α-ketoglutarate, and amino acids. wherein the enzyme is expressed in a sufficient amount to produce polyhydroxybutyrate-co-polybydroxyhexanoate or poly 4-hydroxybutyrate.
Claim 8
Independent Claim
73 words
A method for enhancing production of polymers containing 4HB in a host comprising stably incorporating into the genome of the host a gene encoding a heterologous enzyme selected from the group consisting of a polyhydroxyalkanoate synthase and a 4HB-CoA transferase; providing the host with feedstock selected from the group consisting of carbohydrates, succinate, 4-hydroxybutyrate, α-ketoglutarate, and amino acids. wherein the enzyme is expressed in a sufficient amount to produce polyhydroxybutyrate-co-polybydroxyhexanoate or poly 4-hydroxybutyrate.
Claim 9
Dependent Claim
22 words
The method of claim 6 wherein the host has stably incorporated into its genome both a polyhydroxyalkanoate synthase and a 4HB-CoA transferase.
Claim 10
Dependent Claim
13 words
The method of claim 6 further comprising enhancing expression of the heterologous enzyme.
Claim 11
Dependent Claim
29 words
The method of claim 8 wherein expression is enhanced by mutating the host followed by providing 4HB as a substrate and screening for polymer production by the mutated host.
Claim 12
Dependent Claim
33 words
The method of claim 6 further comprising providing a host expressing enzymes selected from the group consisting of α-ketoglutarate transaminase, glutamate-succinic semialdehyde transaminase, glutamate dehydrogenase, glutamate decarboxylase, 4-hydroxybutyrate dehydrogenase and 4-hydroxybutyryl CoA transferase.
Claim 13
Dependent Claim
23 words
The method of claim 6 further comprising providing a host expressing enzymes degrading arginine, glutamine or proline to produce gamma amino butyric acid.
Claim 14
Dependent Claim
33 words
A 4HB polymer produced by a recombinant host having stably incorporated into the genome a gene encoding a heterologous enzyme selected from the group consisting of a polyhydroxyalkanoate synthase and a 4HB-CoA transferase.
Claim 15
Dependent Claim
33 words
A 4HB polymer produced by a recombinant host having stably incorporated into the genome a gene encoding a heterologous enzyme selected from the group consisting of a polyhydroxyalkanoate synthase and a 4HB-CoA transferase.
Claim 16
Dependent Claim
31 words
A vector comprising, an isolated gene encoding a 4HB-CoA transferase under the control of a promoter for enhancing expression, wherein expression occurs after integration into the genome of a heterologous host.
Claim 17
Dependent Claim
31 words
A vector comprising, an isolated gene encoding a 4HB-CoA transferase under the control of a promoter for enhancing expression, wherein expression occurs after integration into the genome of a heterologous host.
References Cited (Backward Citations)
US4430430A
Cited
US5663063A
Cited
US5250430A
Cited
US5245023A
Cited
US5534432A
Cited
US4876331A
Cited
CA2006508A1
Cited
WO1991000917A1
Cited
US5502273A
Cited
WO1992019747A1
Cited
US5286842A
Cited
WO1993002194A1
Cited
WO1993002187A1
Cited
US5650555A
Cited
US5610041A
Cited
US5378616A
Cited
US5292860A
Cited
WO1993006225A1
Cited
WO1994011519A1
Cited
US5602321A
Cited
WO1994012014A1
Cited
US5516883A
Cited
US5461139A
Cited
WO1995020614A1
Cited
WO1995020615A1
Cited
US5563239A
Cited
WO1996020621A1
Cited
US6117658A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 2007-02-09 | 2007-08-11 | 2008-02-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2011-02-09 | 2011-08-11 | 2012-02-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2015-02-09 | 2015-08-11 | 2016-02-10 | $7700.00 | Window Closed / Lapsed |