US5032512A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for producing compounds containing a terminal hydroxyl or epoxy group and microorganisms suitable therefor
Assignee: RIJKSUNIVERSITEIT TE GRONINGEN A STATE UNIVERSITY OF NETHERLANDS
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Filing: 1988-01-14
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Granted: 1991-07-16
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Expires: 2008-01-14
Abstract
Abstract This invention relates to a process for the microbiological production of compounds containing a terminal hydroxyl or epoxy group from an aliphatic substrate or a substrate with an aliphatic side chain, using microorganisms genetically engineered so that they have retained their capacity to perform the terminal oxidation of the substrate, but are no longer able to convert the resulting oxidation product further to any significant extent. Preferred substrates are n-alkanes, n-alkenes, and n-alkadienes containing 6-12 carbon atoms. Preferred micro-organisms are genetically engineered Pseudomonas oleovorans and Pseudomonas putida strains lacking an active plasmidic alkanol-dehydrogenase gene. The invention also relates to micro-organisms thus genetically engineered.
Inventors
- 👤 Bernard Witholt
- 👤 Roland G. Lageveen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 05:25
CPC Classifications
C; C12; C12P; C12P7/00; C12P7/02; C12P7/04; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/52; C12P17/00; C12P17/02
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
115 words
A process for producing oxidation products containing a terminal hydroxyl or epoxy group using a microorganism of the genus Pseudomonas having an alkane-hydroxylase enzyme system and genetically engineered to have an absence of an active plasmid encoded alkanol dehydrogenase, said microorganism performing a terminal oxidation of an aliphatic substrate or aliphatic side chain of a substrate of said enzyme system in an aqueous nutrient medium containing a bulk non-aqueous phase under aerobic conditions and avoiding further oxidation of said oxidation products by said alkanol dehydrogenase, said microorganism being resistant to the presence of said non-aqueous phase, said process comprising the steps of culturing said microorganism in said nutrient medium and recovering the oxidation products formed.
Claim 2
Dependent Claim
115 words
A process for producing oxidation products containing a terminal hydroxyl or epoxy group using a microorganism of the genus Pseudomonas having an alkane-hydroxylase enzyme system and genetically engineered to have an absence of an active plasmid encoded alkanol dehydrogenase, said microorganism performing a terminal oxidation of an aliphatic substrate or aliphatic side chain of a substrate of said enzyme system in an aqueous nutrient medium containing a bulk non-aqueous phase under aerobic conditions and avoiding further oxidation of said oxidation products by said alkanol dehydrogenase, said microorganism being resistant to the presence of said non-aqueous phase, said process comprising the steps of culturing said microorganism in said nutrient medium and recovering the oxidation products formed.
Claim 3
Dependent Claim
27 words
A process as claimed in claim 1 wherein said substrate is selected from the group consisting of n-alkanes, n-alkenes, n-alkadienes and mixtures thereof containing 6-12 carbon atoms.
Claim 4
Dependent Claim
22 words
A process as claimed in claim 1 wherein said microorganism is selected from the species Pseudomonas oleovorans or the species Pseudomonas putida.
Claim 5
Dependent Claim
29 words
A process as claimed in claim 1, wherein said microorganism is a Pseudomonas oleovorans strain in which at least the plasmid encoded alkanol-dehydrogenase gene has been removed or inactivated.
Claim 6
Dependent Claim
47 words
A process as claimed in claim 1, wherein said microorganism is a Pseudomonas oleovorans strain whose natural plasmid has been replaced by a plasmid which contains the alkBA/R genes coding for the alkane-hydroxylase system and which does not contain a gene coding for an active alkanol-dehydrogenase enzyme.
Claim 7
Dependent Claim
25 words
A process as claimed in claim 1, wherein said microorganism is a Pseudomonas oleovorans strain whose natural plasmid has been replaced by the plasmid pGEc41.
Claim 8
Dependent Claim
29 words
A process as claimed in claim 1, wherein said microorganism is cultured at a temperature of 20°-34° C. in a system comprising two liquid phases whose pH is 5-9.
Claim 9
Dependent Claim
20 words
A process as claimed in claim 1, wherein said reactor contents are stirred at a stirring rate of 500-1000 rpm.
Claim 10
Dependent Claim
25 words
A process as claimed in claim 1, wherein said nutrient medium contains a carbon source selected from the group consisting of pyruvate, citrate and glucose.
Claim 11
Dependent Claim
22 words
A process as claimed in claim 1 wherein said non-aqueous phase comprises an organic phase in which the oxidation products are soluble.
Claim 12
Dependent Claim
19 words
A process as claimed in claim 10 wherein said organic phase is dibutyl phthalate, cyclohexane, cyclohexanol or mixtures thereof.
Claim 13
Dependent Claim
31 words
A process as claimed in claim 1, wherein said microrganism is cultured at a temperature of 20°-34° C. in a system comprising two liquid phases whose pH is 6.8 to 7.0.
References Cited (Backward Citations)
US3813316A
Cited
CA1189807A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-07-15 | 1995-01-14 | 1995-07-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-07-15 | 1999-01-14 | 1999-07-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-07-15 | 2003-01-14 | 2003-07-16 | $7700.00 | Window Closed / Lapsed |