# US 2019/0169656 A1 · Recombinant microorganism for the production of useful metabolites > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0169656 A1 - Document ID: US20190169656A1 - Status: Published application - Kind: A1 - Application number: 16/280,304 - Filed: 2019-02-20 - Published: 2019-06-06 - Earliest claimed: 2011-07-12 - Assignees: Scientist of Fortune, S.A. - Applicants: Scientist of Fortune, S.A. - Inventors: Philippe Marlière - Web page: https://patentgrep.com/patent/US20190169656A1 ## CPC - C12P 7/28: Acetone-containing products - C12N 9/16: acting on ester bonds (3.1) - C12N 9/88: Lyases (4.) - C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression - C12P 5/026: {Unsaturated compounds, i.e. alkenes, alkynes or allenes} - C12P 17/182: {Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system (alloxazine or isoalloxazine, e.g. riboflavine C12P25/00)} - C12P 19/32: having a condensed ring system containing a six-membered ring having two N-atoms in the same ring, e.g. purine nucleotides, nicotineamide-adenine dinucleotide - C12Y 301/03011: Fructose-bisphosphatase (3.1.3.11) - C12Y 401/02009: Phosphoketolase (4.1.2.9) - C12Y 401/02022: Fructose-6-phosphate phosphoketolase (4.1.2.22) - Y02P 20/52: using catalysts, e.g. selective catalysts - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES - Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS ## Abstract Described are recombinant microorganisms characterized by having phosphoketolase activity, having a diminished or inactivated Embden-Meyerhof-Parnas pathway (EMPP) by inactivation of the gene(s) encoding phosphofructokinase or by reducing phosphofructokinase activity as compared to a non-modified microorganism and having a diminished or inactivated oxidative branch of the pentose phosphate pathway (PPP) by inactivation of the gene(s) encoding glucose-6-phosphate dehydrogenase or by reducing glucose-6-phosphate dehydrogenase activity as compared to a non-modified microorganism. These microorganisms can be used for the production of useful metabolites such as acetone, isobutene or propene. ## Claims ### Claim 1 A recombinant microorganism characterized by: a) having phosphoketolase activity; b) (i) having a diminished or inactivated Embden-Meyerhof-Parnas pathway (EMPP) by inactivation of the gene(s) encoding phosphofructokinase or by being genetically modified so as to reduce phosphofructokinase activity as compared to a non-genetically modified microorganism; or (ii) not possessing phosphofructokinase activity; and c) (i) having a diminished or inactivated oxidative branch of the pentose phosphate pathway (PPP) by inactivation of the gene(s) encoding glucose-6-phosphate dehydrogenase or by being genetically modified so as to reduce glucose-6-phosphate dehydrogenase activity as compared to a non-genetically modified microorganism; or (ii) not possessing glucose-6-phosphate dehydrogenase activity, and d) furthermore characterized in that the activity of one or more of the enzymes selected from the group consisting of: formate C-acetyltransferase, classified as EC 2.3.1.54; acetate kinase, classified as EC 2.7.2.1 L-lactate dehydrogenase, classified as EC 1.1.1.28 phosphoenolpyruvate carboxylase, classified as EC 4;1.1.31 phosphoenolpyruvate carboxykinase, classified as (ATP) EC 4.1,1.49 succinate dehydrogenase, classified as EC 1.3.99.1 pyruvate decarboxylase, classified as EC 4.1.1.1 acetaldehyde dehydrogenase (acetylating), classified as EC 1.2.1.10 glycerol-3-phosphate dehydrogenase [NAD(P)+], classified as EC 1.1.1.94 acetolactate synthase, classified as EC 2.2.1.6 methylglyoxal synthase, classified as EC 4.2.3.3 formate hydrogenlyase, classified as EC 1.2.1.2 together with EC 1.12.1.2 is eliminated or reduced. ### Claim 2 (depends on 1) The recombinant microorganism of claim 1 further characterized by: d) having fructose-1,6-bisphosphate phosphatase activity. ### Claim 3 (depends on 1) The recombinant microorganism of claim 1, wherein the EMPP is further diminished or inactivated by inactivation of the gene(s) encoding glyceraldehyde 3-phosphate dehydrogenase or by being genetically modified so as to reduce glyceraldehyde 3-phosphate dehydrogenase activity as compared to a non-genetically modified microorganism. ### Claim 4 (depends on 1) The recombinant microorganism of claim 1 which has been genetically modified to have an increased phosphoketolase activity. ### Claim 5 (depends on 1) The recombinant microorganism of claim 1, wherein said microorganism is a fungus. ### Claim 6 (depends on 1) The recombinant microorganism of claim 1, wherein said microorganism is a bacterium. ### Claim 7 (depends on 6) The recombinant microorganism of claim 6, wherein the gene(s) encoding the PEP-dependent PTS transporter has/have been inactivated. ### Claim 8 (depends on 1) The recombinant microorganism of claim 1 further characterized in that it: e) is capable of converting acetyl-CoA into acetone. ### Claim 9 (depends on 8) The recombinant microorganism of claim 8 further characterized in that it: f) is capable of converting acetone into isobutene. ### Claim 10 (depends on 8) The recombinant microorganism of claim 8 further characterized in that it: g) is capable of converting acetone into propene. ### Claim 11 (depends on 1) Use of the recombinant microorganism of claim 1 for the conversion of glucose into acetyl-CoA. ### Claim 12 (depends on 8) Use of the recombinant microorganism of claim 8 for the conversion of glucose into acetone. ### Claim 13 (depends on 9) Use of the recombinant microorganism of claim 9 for the conversion of glucose into isobutene. ### Claim 14 (depends on 10) Use of the recombinant microorganism of claim 10 for the conversion of glucose into propene. ### Claim 15 (depends on 8) A method for producing acetone from glucose comprising the steps of: culturing the microorganism of claim 8 in a suitable medium; and optionally recovering said acetone from the culture medium. ### Claim 16 (depends on 9) A method for producing isobutene from glucose comprising the steps of: culturing the microorganism of claim 9 in a suitable medium; and optionally recovering said isobutene. ### Claim 17 (depends on 10) A method for producing propene from glucose comprising the steps of: culturing the microorganism of claim 10 in a suitable medium; and optionally recovering said propene. ### Claim 18 (depends on 6) The recombinant microorganism of claim 6, which is E. coli. ### Claim 19 (depends on 8) The recombinant microorganism of claim 8, wherein the fnr gene and/or the rpoS gene are deleted.