Claims
- 1. A carotenoid overproducing bacteria comprising the genes encoding a functional carotenoid enzymatic biosynthetic pathway wherein the dxs, idi and ygbBP genes are overexpressed and wherein the yjeR gene is down regulated.
- 2. A carotenoid overproducing bacteria comprising the genes encoding a functional carotenoid enzymatic biosynthetic pathway wherein the dxs, idi, ygbBP and ispB genes are overexpressed.
- 3. The carotenoid overproducing bacteria of claim 1 or 2 wherein the lytB and dxr gene is optionally overexpressed. ispB lytB and dxr yjeR
- 4. The carotenoid overproducing bacteria of claim 1 or 2 wherein the carotenoid enzymatic biosynthetic pathway consists of the genes dxs, dxr, ygpP, ychB, ygbB, lytB, idi, ispA, ispB crtE, crtB, crtI, and crtY.
- 5. The carotenoid overproducing bacteria of claim 4 wherein the carotenoid enzymatic biosynthetic pathway optionally additionally comprises the crtZ and crtW genes.
- 6. The carotenoid overproducing bacteria of any of claims 1-5 wherein the bacteria is selected from the group consisting Agrobacterium, Erythrobacter, Chlorobium, Chromatium, Flavobacterium, Cytophaga, Rhodobacter, Rhodococcus, Streptomyces, Brevibacterium, Corynebacteria, Mycobacterium, Deinococcus, Paracoccus, Escherichia, Bacillus, Myxococcus, Salmonella, Yersinia, Erwinia, Pantoea, Pseudomonas, Sphingomonas, Methylomonas, Methylobacter, Methylococcus, Methylosinus, Methylomicrobium, Methylocystis, Alcaligenes, Synechocystis, Synechococcus, Anabaena, Thiobacillus, Methanobacterium, Klebsiella, and Myxococcus.
- 7. The carotenoid overproducing bacteria of claim 6 wherein the bactera is E. coli.
- 8. The carotenoid overproducing bacteria of claims 1-3 wherein the dxs, dxr, ygpP, ychB, ygbB, lytB, idi, ispA, ispB are derived from a Methylomonas sp.
- 9. The carotenoid overproducing bacteria of any of claims 1-3 wherein the dxs, idi, ispB and ygbBP genes are under the control of a strong promoter.
- 10. The carotenoid overproducing bacteria of claim 9 wherein the strong promoter is selected from the group consisting of lac, ara, tet, trp, λPL, λPR, T7, tac, PT5, and trc.
- 11. The carotenoid overproducing bacteria of any of claims 1-3 wherein the dxs, idi, ispB and ygbBP genes are integrated in multicopy in the bacterial chromosome.
- 12. The carotenoid overproducing bacteria of any of claims 1-3 wherein the dxs, idi, ispB and ygbBP genes are present in multicopy in the bacteria on one or more plasmids.
- 13. The carotenoid overproducing bacteria of of claim 7 wherein the yjeR gene is down regulated by gene disruption.
- 14. The carotenoid overproducing bacteria of claim 13 wherein the disrupted yjeR gene has the nucleotide sequence as set forth in SEQ ID NO:63.
- 15. The carotenoid overproducing bacteria of either of any of claims 1-3 wherein the dxs, idi, ispB ygbBP and lytB genes are chromosomally integrated into the host cell genome.
- 16. A carotenoid overproducing bacteria selected from the group consisting of: a strain having the ATCC identification number PTA-4807 and a strain having the ATCC identification number PTA-4823.
- 17. A method for the production of a carotenoid comprising:
a) growing the carotenoid overproducing bacteria of any of claims 1-5, the bacteria overexpressing at least one gene selected from the group consisting of dxs, idi ygbBP, ispB, lytB, dxr, wherein yjeR is optionally downregulated, for a time sufficient to produce a carotenoid; and b) optionally recovering the carotenoid from the carotenoid overproducing bacteria of step (a).
- 18. A method according to claim 17 wherein the carotenoid is selected from the group consisting of antheraxanthin, adonixanthin, astaxanthin, canthaxanthin, capsorubrin, β-cryptoxanthin, didehydrolycopene, didehydrolycopene, β-carotene, ζ-carotene, δ-carotene, γ-carotene, keto-γ-carotene, ψ-carotene, ε-carotene, β,ψ-carotene, torulene, echinenone, gamma-carotene, zeta-carotene, alpha-cryptoxanthin, diatoxanthin, 7,8-didehydroastaxanthin, fucoxanthin, fucoxanthinol, isorenieratene, β-isorenieratene lactucaxanthin, lutein, lycopene, neoxanthin, neurosporene, hydroxyneurosporene, peridinin, phytoene, rhodopin, rhodopin glucoside, siphonaxanthin, spheroidene, spheroidenone, spirilloxanthin, uriolide, uriolide acetate, violaxanthin, zeaxanthin-β-diglucoside, zeaxanthin, and C30-carotenoids.
- 19. A method according to claim 18 wherein the carotenoid is produced at a level of at least about 6 mg per gram dry cell weight.
- 20. A method according to claim 18 wherein the bacteria is selected from the group consisting Agrobacterium, Erythrobacter, Chlorobium, Chromatium, Flavobacterium, Cytophaga, Rhodobacter, Rhodococcus, Streptomyces, Brevibacterium, Corynebacteria, Mycobacterium, Deinococcus, Paracoccus, Escherichia, Bacillus, Myxococcus, Salmonella, Yersinia, Erwinia, Pantoea, Pseudomonas, Sphingomonas, Methylomonas, Methylobacter, Methylococcus, Methylosinus, Methylomicrobium, Methylocystis, Alcaligenes, Synechocystis, Synechococcus, Anabaena, Thiobacillus, Methanobacterium, Klebsiella, and Myxococcus.
- 21. A method according to claim 20 wherein the bacteria is E. coli.
- 22. A method according to claim 17 wherein the dxs, idi, ygbBP, ispB and lytB genes are under the control of a promoter selected from the group consisting of lac, ara, tet, trp, λPL, λPR, T7, tac, PT5, and trc.
- 23. A method according to claim 17 wherein the dxs, idi, ispB, ygbBP and lytB genes are integrated in multicopy in the bacterial chromosome.
- 24. A method according to claim 17 wherein the dxs, idi, ispB, ygbBP and lytB genes are in multicopy in the bacteria on one or more plasmids.
- 25. A method according to claim 17 wherein the yjeR gene is down regulated by gene disruption.
- 26. A method according to claim 25 wherein the disrupted yjeR gene has the nucleotide sequence as set forth in SEQ ID NO:63.
- 27. A method according to claim 17 wherein the dxs, idi ispB, ygbBP and lytB genes are chromosomally integrated into the host cell genome.
Parent Case Info
[0001] This application claims the benefit of U.S. Provisional Application No. 60/434,618 filed Dec. 19, 2002.
Provisional Applications (1)
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Number |
Date |
Country |
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60434618 |
Dec 2002 |
US |