# US 9,752,163 B2 · Process for the production of isoprenol from mevalonate employing a diphosphomevalonate decarboxylase > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,752,163 B2 - Document ID: US9752163B2 - Status: Granted - Kind: B2 - Application number: 14/792,669 - Filed: 2015-07-07 - Published: 2015-12-31 - Granted: 2017-09-05 - Earliest claimed: 2015-07-07 - Assignees: Global Bioenergies; Scientist of Fortune, S.A. - Applicants: Global Bioenergies; Scientist of Fortune SA - Inventors: Philippe Marliere; Maria Anissimova; Romain Chayot; Marc Delcourt - Web page: https://patentgrep.com/patent/US9752163B2 ## CPC - C12P 7/04: acyclic - C12N 9/88: Lyases (4.) - C12P 5/007: {containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00)} - C12Y 401/01033: Diphosphomevalonate decarboxylase (4.1.1.33), i.e. mevalonate-pyrophosphate decarboxylase - 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 ## Abstract Described is a method for the enzymatic production of isoprenol using mevalonate as a substrate and enzymatically converting it by a decarboxylation step into isoprenol as well as the use of an enzyme which is capable of catalyzing the decarboxylation of mevalonate for the production of isoprenol from mevalonate. Furthermore described is the use of mevalonate as a starting material for the production of isoprenol in an enzymatically catalyzed reaction. Also disclosed is a method for the production of isoprene comprising the method for the production of isoprenol using mevalonate as a substrate and enzymatically converting it by a decarboxylation step into isoprenol and further comprising the step of converting the produced isoprenol into isoprene as well as a method for the production of isoamyl alcohol comprising the method for the production of isoprenol using mevalonate as a substrate and enzymatically converting it by a decarboxylation step into isoprenol and further comprising the step of converting the produced isoprenol into isoamyl alcohol. ## Claims ### Claim 1 A method of producing isoprenol comprising: converting mevalonate into isoprenol by the enzyme of SEQ ID NO:2. ### Claim 2 (depends on 1) The method of claim 1 which is carried out in vitro. ### Claim 3 (depends on 1) The method of claim 1 wherein a co-substrate is added. ### Claim 4 (depends on 1) The method of claim 1 wherein ATP, a rNTP, a dNTP, a polyphosphate or pyrophosphate, or a mixture of any of these compounds is added. ### Claim 5 (depends on 1) The method of claim 1 further comprising culturing a recombinant microorganism or a plant cell overexpressing the enzyme to convert mevalonate into isoprenol. ### Claim 6 (depends on 5) The method of claim 5, wherein the recombinant microorganism is a bacterium, a fungus, a yeast or a microalgae. ### Claim 7 (depends on 5) The method of claim 5, wherein the method is carried out in a plant cell. ### Claim 8 (depends on 5) The method of claim 5, wherein the recombinant microorganism or plant cell has been genetically modified with a heterologous polynucleotide encoding the enzyme. ### Claim 9 (depends on 5) The method of claim 5, wherein the recombinant microorganism or plant cell has been genetically modified with a promoter that overexpresses the enzyme. ### Claim 10 (depends on 1) The method of claim 1 further comprising recovering isoprenol. ### Claim 11 (depends on 1) The method of claim 1 further comprising converting the isoprenol into isoprene. ### Claim 12 (depends on 11) The method of claim 11 further comprising recovering isoprene. ### Claim 13 (depends on 1) The method of claim 1 further comprising converting the isoprenol into isoamyl alcohol. ### Claim 14 (depends on 13) The method of claim 13 further comprising recovering isoamyl alcohol. ### Claim 15 A method of producing isoprenol comprising converting mevalonate into isoprenol by an enzyme at least 95% identical to the amino acid sequence of SEQ ID NO: 2, wherein the enzyme accepts mevalonate as a substrate and catalyzes a decarboxylation reaction to produce isoprenol. ### Claim 16 (depends on 15) The method of claim 15 which is carried out in vitro. ### Claim 17 (depends on 15) The method of claim 15 wherein ATP, a rNTP, a dNTP, a polyphosphate or pyrophosphate, or a mixture of any of these compounds is added. ### Claim 18 (depends on 15) The method of claim 15 further comprising culturing a recombinant microorganism or a plant cell overexpressing the enzyme to convert mevalonate into isoprenol. ### Claim 19 (depends on 18) The method of claim 18, wherein the recombinant microorganism is a bacterium, a fungus, a yeast or a microalgae. ### Claim 20 (depends on 18) The method of claim 18, wherein the method is carried out in a plant cell. ### Claim 21 (depends on 18) The method of claim 18, wherein the recombinant microorganism or plant cell has been genetically modified with a heterologous polynucleotide encoding the enzyme. ### Claim 22 (depends on 18) The method of claim 18, wherein the recombinant microorganism or plant cell has been genetically modified with a promoter that overexpresses the enzyme. ### Claim 23 (depends on 15) The method of claim 15 further comprising recovering isoprenol. ### Claim 24 (depends on 15) The method of claim 15 further comprising converting the isoprenol into isoprene. ### Claim 25 (depends on 24) The method of claim 24 further comprising recovering isoprene. ### Claim 26 (depends on 15) The method of claim 15 further comprising converting the isoprenol into isoamyl alcohol. ### Claim 27 (depends on 26) The method of claim 26 further comprising recovering isoamyl alcohol.