Recombinant hydrogen-producing cyanobacterium and uses thereof
A recombinant cyanobacterium comprising an oxygen-tolerant, hydrogen-evolving hydrogenase, kit, and methods of use.
1. A recombinant cyanobacterium comprising:
an expressible recombinant nucleic acid which encodes a functional microbial oxygen-tolerant, hydrogen-evolving [NiFe]-hydrogenase that comprises a sequence at least 90% identical to the nucleic acid sequence from nucleotides 5091 to 6110 of SEQ ID NO: 5;
wherein the hydrogenase is present in an amount effective to produce a measurable amount of hydrogen when the recombinant cyanobacterium is incubated aerobically, in the presence of a suitable light source, with water as the feed stock.
2. The recombinant cyanobacterium of claim 1 , wherein the suitable light source is solar energy or fluorescent light.
3. The recombinant cyanobacterium of one of claim 1 , wherein the cyanobacterium is from the group of unicellular cyanobacteria.
4. The recombinant cyanobacterium of claim 1 , wherein the recombinant cyanobacterium is Synochocystis or Synechococcus.
5. The recombinant cyanobacterium of claim 1 , wherein the oxygen-tolerant, hydrogen-evolving [NiFe]-hydrogenase is derived from Thiocapsa roseopersica.
6. The recombinant cyanobacterium of claim 1 , wherein the oxygen-tolerant, hydrogen-evolving [NiFe]-hydrogenase is derived from Ralstonia eutropha or Alteromonas macleodii.
7. The recombinant cyanobacterium of claim 1 , wherein the oxygen-tolerant, hydrogen-evolving [NiFe]-hydrogenase is derived from a marine bacterium.
8. The recombinant cyanobacterium of claim 1 wherein the measurable amount of hydrogen is detected using gas chromatography.
9. The recombinant cyanobacterium of claim 1 wherein the measurable amount of hydrogen is detected using a Clark Electrode System.
10. An expression vector comprising a nucleic acid encoding a functional microbial oxygen-tolerant hydrogen-evolving [NiFe]-hydrogenase that comprises a sequence at least 90% identical to the nucleic acid sequence from nucleotides 5091 to 6110 of SEQ ID NO: 5.
11. A cyanobacterium which comprises the expression vector of claim 10 .
12. A method for generating hydrogen from water, comprising culturing the recombinant cyanobacterium of claim 1 aerobically, under conditions effective to produce a measurable amount of hydrogen.