Claims
- 1. A method of hydrogen gas generation, comprising the steps of:
culturing algae under illuminated conditions in a media comprising sulfur wherein sulfate permease expression of the algae is reduced relative to normal wild-type algae; sealing the algae culture from atmospheric oxygen; and collecting hydrogen gas evolved.
- 2. The method of claim 1, wherein the algae is a green algae and the algae comprises a genome which is artificially engineered to reduce sulfate permease expression relative to a wild-type algae.
- 3. The method of claim 2, wherein the algae is a unicellular, photosynthetic, anoxygenic algae.
- 4. The method of claim 1, wherein the algae is chosen from Rhodobacter sphaeroide and genetically modified Chlamydomonas reinhardtii.
- 5. The method of claim 1, wherein the algae is Rhodobacter sphaeroide an anoxygenic photosynthesis bacterium having a lineage of Proteobacteria; alphaproteobacteria, Rhodobacterales; Rhodobacteraceae.
- 6. The method of claim 1, wherein the algae is an isolated strain with downregulated expression of sulfate permease with 50% or less expression of sulfate permease relative to normal wild-type algae.
- 7. The method of claim 2, wherein the algae is genetically modified by insertion of an antisense sequence to CrcpSulP.
- 8. The method of claim 2, wherein the genetically-modified algae is modified by a technique chosen from insertion of an antisense strand of CrcpSulP, insertion of a sense strand of CrcpSulP, ablation of CrcpSulP and targeted gene deletion of CrcpSulP.
- 9. The method of claim 7, wherein the antisense sequence hybridizes to a portion of SEQ ID NO:2.
- 10. An isolated nucleotide sequence, chosen from SEQ ID NO:2; SEQ ID NO:3 and a sequence which hybridizes to any one of SEQ ID NO:2 and SEQ ID NO:3.
- 11. An isolated amino acid sequence selected from the group consisting of SEQ ID NO: 1 and a sequence with 90% or more sequence homology to SEQ ID NO: 1.
- 12. A genetically-modified algae wherein the sulfate uptake pathway is downregulated to 50% or less relative to a native, wild-type, unmodified algae.
- 13. The algae of claim 12, wherein the alga is a green algae.
- 14. The algae of claim 13, wherein expression of an endogenous CrcpSulP gene is downregulated by insertion of an antisense CrcpSulP polynucleotide into the genome of the algae.
- 15. The algae of claim 14, wherein the algae is Chlamydomonas reinhardtii.
- 16. The algae of claim 12, wherein the expression of the CrcpSulP gene is downregulated by an antisense sequence that hybridizes to a portion of the CrcpSulP mRNA transcript.
- 17. A composition, comprising:
water; algae growth nutrients; algae genetically modified for sulfate permease expression reduced by 50% or more relative to an unmodified wild-type version of the algae.
- 18. The composition of claim 17, wherein the algae is unicellular, photosynthetic, anoxygenic algae.
- 19. An assay for detecting low levels of sulfur uptake in a sample of genetically-modified green algae comprising the steps of:
a. culturing a genetically-modified sample of green algae in TAP media in lighted, anaerobic conditions; b. transferring an aliquot of the sample into a media comprising sulfur; c. culturing the aliquot in lighted conditions; and d. detecting the level of ARS activity in the aliquot, wherein an elevated level of aryl-sulfatase (ARS) activity is a positive indicator that the genetically-modified green algae is deficient in sulfur uptake compared to a wild-type algae.
- 20. An isolated antisense oligonucleotide consisting of a nucleotide sequence that is complementary to SEQ ID NO:2.
- 21. An isolated antisense oligonucleotide comprising a sequence complementary to codons 118to 412of SEQ ID NO 2.
- 22. An expression vector comprising an antisense sequence complementary to codons 118 to 412 of SEQ ID NO:2.
- 23. A composition, comprising:
a sulP1 strain of Chlamydomonas reinhardtii; and a Rhodobacter sphaeroides bacterium that is anaerobic and photosynthetic.
- 24. The composition of claim 23, further comprising a Clostridium sp having the lineage Bacteria; Firmicutes; Clostridia; Clostridiales; Clostridiaceae.
- 25. A process for producing hydrogen comprising culturing a combination of sulP1 strain of Chlamydomonas reinhardtii and Rhodobacter sphaeroides with Clostridruim sp.
- 26. A method of generating hydrogen gas, comprising the steps of:
providing in an aqueous media a sulP1 strain of Chlamydomonas reinhardtii and Rhodobacter sphaeroides bacteria; exposing the aqueous media to sunlight for a period of time and under conditions to allow for the generation of hydrogen.
- 27. The method of claim 26, further comprising:
providing Clostridium in the media.
- 28. A method for generating hydrogen gas, comprising the steps of:
subjecting a biomass comprising an algae to sunlight in a sulfur-containing media comprising carbon dioxide and inorganic nutrients for a period of time and under conditions so as to cause the algae to undergo oxygenic photosynthesis and to generate hydrogen gas; and subjecting an anaerobic photosynthetic bacterium in the media to sunlight for a period of time and under conditions so as to generate hydrogen from a nitrogenase/hydrogenase enzymatic system in the media.
- 29. The process of claim 28, further comprising:
inducing fermentation of the biomass the media of Chlamydomonas/Rhodobacter via Clostridium sp.
- 30. A method of generating hydrogen gas, comprising the steps of:
providing in an aqueous media a genetically-modified strain of Chlamydomonas reinhardtii providing a strain of Rhodobacter sphaeroides photosynthetic bacteria; exposing the aqueous media to sunlight for a period of time and under conditions to allow for the generation of biomass and hydrogen; subjecting an anaerobic photosynthetic bacterium in the media to sunlight for a period of time and under conditions so as to generate hydrogen from a nitrogenase/hydrogenase enzymatic system in the media; providing a strain of Clostridium in the media; and inducing fermentation of the biomass in the media via Clostridium sp.
- 31. The method of claim 30, wherein the genetically-modified algae is modified to decrease activity of sulfate permease by a technique selected from the group consisting of insertion of an antisense strand of a sulfate permease gene, insertion of a sense strand of a sulfate permease gene, ablation of the sulfate permease gene and targeted gene deletion of the sulfate permease gene.
CROSS-REFERENCE
[0001] This application claims the benefit of priority to U.S. Provisional Application No. 60/354,760, filed Feb. 4, 2002, and U.S. Provisional Application No. 60/377,902, filed May 2, 2002, both of which applications are incorporated herein by reference.
GOVERNMENT RIGHTS
[0002] The United States Government may have certain rights in this application pursuant to Grant FD-2002-35100-12278-MELI-08/04 and 1998-35100-6148, awarded by the USDA-NRI Competitive Grant.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60354760 |
Feb 2002 |
US |
|
60377902 |
May 2002 |
US |