Patent US4116741A successfully refreshed from Google Patents.
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EP0066994A2
Expired / Lapsed
EP
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Production and use of genetically-modified microorganisms
Assignee: Imperial Chemical Industries Ltd
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Filing: 1982-05-21
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Granted: —
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Expires: 2002-05-21
Abstract
Abstract The genetic modification of microorganisms which can be grown upon methanol as a carbon source to produce single cell protein (SCP). By the modification the genes specifying the methanol oxidase (MO) and methanol dehydrogenase (MDH) enzymes, used in the normal metabolic pathways by which the microorganisms convert methanol to formaldehyde, are replaced by adh genes capable of expressing an alcohol dehydrogenase (ADH) enzyme catalyst for the oxidation of methanol to formaldehyde. The modified microorganism has increased energetic efficiency in utilizing carbon. Preferably the modified microorganism is derived from Methylophilus methylotrophus. The claims cover genetically modified microorganisms a method of making genetically modified microorganisms and a process for the production of single cell protein.
Inventors
- 👤 Andrew John Beardsmore
- 👤 Stephen Hugh Collins
- 👤 Keith Adrian Powell
- 👤 Peter James Senior
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-28 23:56
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0006; C12N1/00; C12N1/32; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/746; C12N15/78
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
86 words
A process for the production of single cell protein by. aerobically cultivating a genetically-modified, methanol-utilizing microorganism in an aqueous culture medium comprising methanol as a source of assimilable carbon and inorganic nutrients wherein the methanol-utilizing microorganism has been genetically modified to remove from it the gene or genes coding for the enzyme activity of methanol dehydrogenase (MDH) or methanol oxidase (MO) and to replace this by insertion of DNA material comprising an-adh gene capable of expressing an enzyme catalyst for the oxidation of methanol to formaldehyde.
Claim 2
Dependent Claim
86 words
A process for the production of single cell protein by. aerobically cultivating a genetically-modified, methanol-utilizing microorganism in an aqueous culture medium comprising methanol as a source of assimilable carbon and inorganic nutrients wherein the methanol-utilizing microorganism has been genetically modified to remove from it the gene or genes coding for the enzyme activity of methanol dehydrogenase (MDH) or methanol oxidase (MO) and to replace this by insertion of DNA material comprising an-adh gene capable of expressing an enzyme catalyst for the oxidation of methanol to formaldehyde.
Claim 3
Dependent Claim
23 words
A process according to claim 1 wherein the genetically modified, microorganism is derived from one or more strains of the species Methylophilus methylophus.
Claim 4
Dependent Claim
25 words
A process according to claim 2 wherein the genetically modified microorganism is derived from one or more of the strains NCIB 10508-10515 and 10592-10596 inclusive.
Claim 5
Dependent Claim
43 words
A process according to claim 1 wherein the genetically modified microorganism is derived from one or more of the strains FRI 2247, FRI 2248, ATCC 21704, ATCC. 21438, ATCC 21439, NRRL B-3458, FRI 1690, FRI 1691, FRI 1693, FRI 1694 and ATCC 31226.
Claim 6
Dependent Claim
68 words
Aerobically cultured microorganisms of one or more species which are capable of utilizing methanol as a source of carbon and energy which have been genetically-modified to remove the gene or genes coding for the enzyme activity of methanol dehydrogenase (MDH) or methanol oxidase (MO) and to replace this by DNA material comprising an adh gene capable of expressing an enzyme catalyst for the oxidation of methanol to formaldehyde.
Claim 7
Dependent Claim
68 words
Aerobically cultured microorganisms of one or more species which are capable of utilizing methanol as a source of carbon and energy which have been genetically-modified to remove the gene or genes coding for the enzyme activity of methanol dehydrogenase (MDH) or methanol oxidase (MO) and to replace this by DNA material comprising an adh gene capable of expressing an enzyme catalyst for the oxidation of methanol to formaldehyde.
Claim 8
Dependent Claim
22 words
Aerobically cultured microorganisms according to claim 5 produced by the genetic modification of one or more strains of the species Methylophilus methylotrophus.
Claim 9
Dependent Claim
24 words
Aerobically cultured microorganisms according to claim 6 produced by the genetic modification of one or more of the strains NCIB 10508-10515 and 10592-10596 inclusive.
Claim 10
Dependent Claim
42 words
Aerobically cultured microorganisms according to claim 5 produced by the genetic modification of one or more of the strains FRI 2247, FRI 2248, ATCC 21704, ATCC 21438, ATCC 21439, NRRL B-5458, FRI 1690, FRI 1691, FRI 1693, FRI 1694 and ATCC 31226.
Claim 11
Dependent Claim
121 words
. A method for making a genetically-modified, methanol-utilizing microorganism by replacing in a methanol-utilizing microorganism the gene or genes coding for the enzyme activity of methanol dehydrogenase (MBH) or methanol oxidase (MO) by DNA material comprising an adh gene capable of expressing an enzyme catalyst for the oxidation of methanol to formaldehyde which method comprises the steps of (1) obtaining from a first microorganism having the adh gene by in vitro or in vivo means a replicon containing said gene, (2) producing a mutant of a methanol-utilizing microorganism deficient in the gene or genes coding for the enzyme activity of methanol dehydrogenase (MDH) or methanol oxidase (MO), and (3) transferring said adh gene on a suitable vector into a second microorganism.
Claim 12
Dependent Claim
121 words
. A method for making a genetically-modified, methanol-utilizing microorganism by replacing in a methanol-utilizing microorganism the gene or genes coding for the enzyme activity of methanol dehydrogenase (MBH) or methanol oxidase (MO) by DNA material comprising an adh gene capable of expressing an enzyme catalyst for the oxidation of methanol to formaldehyde which method comprises the steps of (1) obtaining from a first microorganism having the adh gene by in vitro or in vivo means a replicon containing said gene, (2) producing a mutant of a methanol-utilizing microorganism deficient in the gene or genes coding for the enzyme activity of methanol dehydrogenase (MDH) or methanol oxidase (MO), and (3) transferring said adh gene on a suitable vector into a second microorganism.
Claim 13
Dependent Claim
18 words
A method according to claim 9 wherein the second microorganism is the ultimate recipient of the adh gene.
Claim 14
Dependent Claim
51 words
A method according to claim 9 wherein the second microorganism is an intermediate microorganism and DNA material comprising the adh gene is transferred via one or more intermediate microorganism(s) from the first microorganism to the motant deficient in the gene or genes coding for the enzyme activity of MDH or MO.
Claim 15
Independent Claim
25 words
A method according to any one of claims 9 to 11 wherein in step (1) the replicon containing the adh gene is a plasmid vector.
Claim 16
Independent Claim
29 words
A method according to any one of claims 9 to 12 wherein the first microorganism is a strain of the species Bacillus. stearpthemorphilus which exhibits adh activity on methanol.
Claim 17
Dependent Claim
25 words
A method according to claim 13 wherein the first microorganism is a strain of Bacillus stearothermorphilus var Non diastaticus which exhibits adh activity on methanol.
Claim 18
Dependent Claim
18 words
A method according to claim 14 wherein the first microorganism is Bacillus stearothermophilus strain D2 166 (NCIB 11739)).
Claim 19
Independent Claim
26 words
A method according to any one of claims 9 and 11 to 15 wherein an intermediate microorganism is used which is a strain of E. coli.
Claim 20
Independent Claim
17 words
A method for making a genetically modified microorganism substantially as described and as shown in the Example.
Claim 21
Independent Claim
17 words
A method for making a genetically modified microorganism substantially as described and as shown in the Example.
References Cited (Backward Citations)
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