Methods for the enzymatic production of isoprene from isoprenol
AssigneesGlobal Bioenergies, Scientist of Fortune, S.A.
InventorsPhilippe Marliere, Maria Anissimova
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Abstract
Described are methods for the enzymatic production of isoprene which allow to produce isoprene from isoprenol and microorganisms which have been genetically modified so as to be able to produce isoprene from isoprenol. Also described are enzyme combinations which allow to convert isoprenol into isoprene as well as (micro)organisms which express such enzyme combinations.
Claims30
A method of producing isoprene from isoprenol comprising enzymatically converting isoprenol into isoprenyl monophosphate and then enzymatically converting isoprenyl monophosphate into isoprene.
A method of producing isoprene from isoprenol comprising enzymatically converting isoprenol into prenol by an isomerisation reaction, and then enzymatically converting prenol into prenyl monophosphate by a phosphorylation reaction and then enzymatically converting prenyl monophosphate into isoprene (Pathway C).
A method of producing isoprene from isoprenol comprising enzymatically converting isoprenol into isoprenyl sulfate and then enzymatically converting isoprenyl sulfate into isoprene.
A method of producing isoprene from isoprenol comprising enzymatically converting isoprenol into prenol by an isomerisation reaction, then enzymatically converting prenol into prenyl sulfate by a sulfurylation reaction and then enzymatically converting prenyl sulfate into isoprene (Pathway F).
A microorganism or a plant which expresses
- A) (a) a hydroxyethylthiazole kinase (EC 2.7.1.50);
- and
- (b) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), an beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), or a pinene synthase (EC 4.2.3.14);
- or
- B) (a) a hydroxyethylthiazole kinase (EC 2.7.1.50);
- and
- (b) an isopentenyl-diphosphate DELTA isomerase (EC 5.3.3.2);
- and
- (c) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), a beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14);
- or
- D) (a) a sulfotransferase (EC 2.8.2);
- and
- (b) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), a beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14), or a retinol sulfotransferase/dehydratase;
- or
- E) (a) a sulfotransferase (EC 2.8.2);
- and
- (b) an isopentenyl-diphosphate DELTA isomerase (EC 5.3.3.2);
- and
- (c) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), a beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14), or a retinol sulfotransferase/dehydratase;
- and which is capable of converting isoprenol into isoprene.
A composition comprising
- (A) (a) a hydroxyethylthiazole kinase (EC 2.7.1.50);
- and
- (b) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), a beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14);
- or
- B) (a) a hydroxyethylthiazole kinase (EC 2.7.1.50);
- and
- (b) an isopentenyl-diphosphate DELTA isomerase (EC 5.3.3.2);
- and
- (c) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), a beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14);
- D) (a) a sulfotransferase (EC 2.8.2);
- and
- (b) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), a beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14), or a retinol sulfotransferase/dehydratase;
- or
- E) (a) a sulfotransferase (EC 2.8.2);
- and
- (b) an isopentenyl-diphosphate DELTA isomerase (EC 5.3.3.2);
- and
- (c) a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), a beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14), or a retinol sulfotransferase/dehydratase.
A method of producing isoprene from isoprenol, wherein the method comprises converting isoprenol into isoprene using a combination of enzymes selected from:
- A) (a) a hydroxyethylthiazole kinase (EC 2.7.1.50);
- and
- (b) a terpene synthase, an isoprene synthase (EC 4.2.3.27), a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), an beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14);
- or
- B) (a) a hydroxyethylthiazole kinase (EC 2.7.1.50);
- and
- (b) an isopentenyl-diphosphate DELTA isomerase (EC 5.3.3.2);
- and
- (c) a terpene synthase, an isoprene synthase (EC 4.2.3.27), a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), an beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14);
- or
- D) (a) a sulfotransferase (EC 2.8.2);
- and
- (b) a terpene synthase, an isoprene synthase (EC 4.2.3.27), a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), an beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14), or a retinol sulfotransferase/dehydratase;
- or
- E) (a) a sulfotransferase (EC 2.8.2);
- and
- (b) an isopentenyl-diphosphate DELTA isomerase (EC 5.3.3.2);
- and
- (c) a terpene synthase, an isoprene synthase (EC 4.2.3.27), a monoterpene synthase, an alpha-farnesene synthases (EC 4.2.3.46), an beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), a pinene synthase (EC 4.2.3.14), or a retinol sulfotransferase/dehydratase.
A method of producing isoprene from isoprenol,
- wherein the method comprises converting isoprenol into isoprene using an enzyme selected from:
- (i) a terpene synthase, an isoprene synthase (EC 4.2.3.27), an alpha-farnesene synthases (EC 4.2.3.46), an beta-farnesene synthase (EC 4.2.3.47), a myrcene/(E)-beta-ocimene synthase (EC 4.2.3.15), or a pinene synthase (EC 4.2.3.14);
- or
- (ii) a sulfotransferase (EC 2.8.2).
Dependencies are inferred from the claim text.