Modulation of sulfate permease for photosynthetic hydrogen production
First Claim
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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.
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Abstract
Sustained hydrogen production is obtained by the culturing of a genetically-modified algae, where the ability of the chloroplasts to intake sulfate is reduced or eliminated compared to wild-type algae. The alga is cultured in a sealed environment in a liquid or solid medium that contains sulfur, and hydrogen is generated continuously. Alternatively, the algae may be cultured in the presence of bacteria that also produce hydrogen gas. The hydrogen produced can be collected and used as a clean energy source.
32 Citations
31 Claims
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1. A method of hydrogen gas generation, comprising the steps of:
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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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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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.
- 2;
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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.
- 1 and a sequence with 90% or more sequence homology to SEQ ID NO;
- 12. A genetically-modified algae wherein the sulfate uptake pathway is downregulated to 50% or less relative to a native, wild-type, unmodified algae.
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17. A composition, comprising:
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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. - View Dependent Claims (18)
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19. An assay for detecting low levels of sulfur uptake in a sample of genetically-modified green algae comprising the steps of:
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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.
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20. An isolated antisense oligonucleotide consisting of a nucleotide sequence that is complementary to SEQ ID NO:
- 2.
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21. An isolated antisense oligonucleotide comprising a sequence complementary to codons 118to 412of SEQ ID NO 2.
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22. An expression vector comprising an antisense sequence complementary to codons 118 to 412 of SEQ ID NO:
- 2.
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23. A composition, comprising:
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a sulP1 strain of Chlamydomonas reinhardtii;
anda Rhodobacter sphaeroides bacterium that is anaerobic and photosynthetic. - View Dependent Claims (24)
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25. A process for producing hydrogen comprising culturing a combination of sulP1 strain of Chlamydomonas reinhardtii and Rhodobacter sphaeroides with Clostridruim sp.
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26. A method of generating hydrogen gas, comprising the steps of:
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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. - View Dependent Claims (27)
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28. A method for generating hydrogen gas, comprising the steps of:
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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. - View Dependent Claims (29)
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30. A method of generating hydrogen gas, comprising the steps of:
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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. - View Dependent Claims (31)
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Specification