In vivo method for the production of tulipalin a via itaconic acid
AssigneesBase SE, Max-Planck-Gesellschaft zur Férderung der Wissenschaften e.V.
InventorsSrividhya SUNDARAM, Barbara NAVÉ, Tobias ERB, Joeline XIBERRAS, Birgit HOFF, Oskar ZELDER, Patrick KAISER, Ingeborg HEUSCHKEL, Robin MACK, Philipp WICHMANN, Patrick GERLINGER
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Abstract
The present invention is concerned with the in vivo production of tulipalin A (α-methylene-γ-butyrolactone). Provided are methods for the in vivo production of tulipalin A, recombinant cells or organisms for in vivo production of tulipalin A, enzymes needed for the in vivo production of tulipalin A, and nucleic acids for expression of those enzymes.
Claims20
-19. (canceled)
An in vivo method for producing tulipalin A (α-methylene-γ-butyrolactone) from itaconic acid, the method comprising culturing at least one recombinant cell or organism selected from bacteria and fungi in a fermentation medium, said recombinant cell or organism comprising:
- one or more recombinant nucleic acid molecules encoding for succinate-CoA ligase as a first enzyme;
- one or more recombinant nucleic acid molecules encoding for succinyl-CoA reductase as a second enzyme;
- and
- one or more recombinant nucleic acid molecules encoding for 3-sulfolactaldehyde reductase or one or more recombinant nucleic acid molecules encoding for succinate semialdehyde reductase as a third enzyme.
A recombinant cell or organism, said recombinant cell or organism being selected from bacteria and fungi and comprising:
- one or more recombinant nucleic acid molecules encoding for succinate-CoA ligase as a first enzyme;
- one or more recombinant nucleic acid molecules encoding for succinyl-CoA reductase as a second enzyme;
- and
- one or more recombinant nucleic acid molecules encoding for 3-sulfolactaldehyde reductase or one or more recombinant nucleic acid molecules encoding for succinate semialdehyde reductase as a third enzyme;
- wherein the recombinant cell or organism is
- (a) a natural producer of itaconic acid and/or
- (b) comprises one or more recombinant nucleic acid molecules encoding for cis-aconitate decarboxylase, preferably aconitate decarboxylase cadA, and/or one or more recombinant nucleic acid molecules encoding for citrate synthase gltA, preferably citrate synthase gltA.
A method for producing tulipalin A (α-methylene-γ-butyrolactone) from itaconic acid, the method comprising the steps of:
- (a) culturing at least one recombinant cell or organism selected from bacteria and fungi in a fermentation medium, said recombinant cell or organism comprising
- one or more recombinant nucleic acid molecules encoding for succinate-CoA ligase as a first enzyme;
- one or more recombinant nucleic acid molecules encoding for succinyl-CoA reductase as a second enzyme;
- and
- one or more recombinant nucleic acid molecules encoding for 3-sulfolactaldehyde reductase or one or more recombinant nucleic acid molecules encoding for succinate semialdehyde reductase as a third enzyme;
- (b) harvesting the at least one recombinant cell or organism;
- (c) lysing the at least one recombinant cell or organism to prepare a lysate;
- (d) incubating the lysate with itaconic acid under suitable conditions;
- and
- (e) isolating the tulipalin A.
A method for producing tulipalin A (α-methylene-γ-butyrolactone) from itaconic acid, the method comprising the steps of:
- (a) culturing in a fermentation medium:
- a first recombinant cell or organism selected from bacteria and fungi, said recombinant cell or organism comprising one or more recombinant nucleic acid molecules encoding for succinate-CoA ligase as a first enzyme;
- a second recombinant cell or organism selected from bacteria and fungi, said recombinant cell or organism comprising one or more recombinant nucleic acid molecules encoding for succinyl-CoA reductase as a second enzyme;
- and
- a third recombinant cell or organism selected from bacteria and fungi, said recombinant cell or organism comprising one or more recombinant nucleic acid molecules encoding for 3-sulfolactaldehyde reductase or one or more recombinant nucleic acid molecules encoding for succinate semialdehyde reductase as a third enzyme;
- (b) harvesting the first, second and third recombinant cell or organism;
- (c) lysing the first, second and third recombinant cell or organism to prepare a first, second and third lysate;
- (d) combining the first, second and third lysate in a suitable ratio, thereby producing a lysate mixture;
- (e) incubating the lysate mixture with itaconic acid under suitable conditions;
- and
- (f) isolating the tulipalin A.
Dependencies are inferred from the claim text.