PHOTOCATALYTIC HYDROGEN PRODUCTION AND POLYPEPTIDES CAPABLE OF SAME

• US 20100203609A1
• Filed: 07/23/2008
• Published: 08/12/2010
• Est. Priority Date: 07/23/2007
• Status: Active Grant

• Current Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Original Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Inventors: Yacoby, Iftach, Benhar, Itai, Nelson, Nathan, Gazit, Ehud
• Granted Patent: US 9,181,555 B2
• Time in Patent Office: Days
• US Class Current: 435/168
• CPC Class Codes: C12M 21/02 Photobioreactors culturing algae A01G33/00, A01H4/001, C12N1/12
• CPC Class Codes: C12M 21/18 Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
• CPC Class Codes: C12M 31/10 by light emitting elements located inside the reactor, e.g. LED or OLED
• CPC Class Codes: C12M 47/10 Separation or concentration of fermentation products bioreactors combined with means for distillation or extraction of liquid fuel C12M43/02
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 9/0067 acting on hydrogen as donor (1.12)
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12P 3/00 Preparation of elements or inorganic compounds except carbon dioxide recovery of carbon dioxides as by-products C12F3/02
• CPC Class Codes: C12Y 112/98002 5,10-Methenyltetrahydromethanopterin hydrogenase (1.12.98.2)

Abstract

An isolated polypeptide comprising a hydrogen generating enzyme attached to a heterologous ferredoxin is disclosed, as well as polynucleotides encoding same, nucleic acid constructs capable of expressing same and cells expressing same. A method for generating hydrogen using the isolated polypeptide is also disclosed.

Claims

1. An isolated polypeptide comprising an algal Fe-only hydrogenase attached to a plant ferredoxin.

6. The isolated polypeptide of claim 1, further comprising a linker, capable of linking said algal Fe-only hydrogenase to said plant ferredoxin.

9. The isolated polypeptide of claim 6, wherein said linker is a peptide bond.

10. The isolated polypeptide of claim 9, as set forth in SEQ ID NOs:

24 or 25.

11. An isolated polynucleotide encoding the polypeptide of claim 1.

12. The isolated polynucleotide of claim 11, comprising a nucleic acid sequence as set forth in SEQ ID NOs:

1-6.

17. A cell comprising the isolated polynucleotide of claim 11.

18. The cell of claim 17, being selected from the group consisting of a cyanobacterial cell, an algal cell and a higher plant cell.

23. A method of generating hydrogen, the method comprising combining the polypeptide of claim 1 with an electron donor so as to generate an electron transfer chain, wherein said electron transfer chain is configured such that said electron donor is capable of donating electrons to the polypeptide of claim 1, thereby generating hydrogen.

24. The method of claim 23, wherein the generating hydrogen is effected under anaerobic conditions.

25. The method of claim 23, wherein said electron donor is selected from the group consisting of a biomolecule, a chemical, water, an electrode and a combination of the above.

29. The method of claim 25, wherein said biomolecule comprises Photosystem I (PSI) or rhodopsin.

35. The method of claim 23, further comprising harvesting the hydrogen following the generating.

36. The method of claim 23, wherein said combining is effected in a cell-free system.

38. The method of claim 23, wherein said combining is effected in a cellular system.

39. The method of claim 38, wherein said cellular system is selected from the group consisting of a cyanobacteria, an algae and a higher plant.

40. The method of claim 38, further comprising down-regulating an expression of endogenous ferredoxin in said cellular system.

41. A system comprising the polypeptide of claim 1 and an electron donor.

42. The system of claim 41, wherein said electron donor comprises an agent selected from the group consisting of a biomolecule, a chemical, water, an electrode and any combination of the above.

43. The system of claim 41, wherein said electron donor comprises PS-1 or rhodopsin.

51. The system of claim 42, wherein said biomolecule is comprised in particles.

56. The system of claim 43, being expressed in cells.

2. (canceled)

3. The isolated polypeptide of claim 2, as set forth in SEQ ID NOs:

23, 26, 27 or 28.

4-5. -5. (canceled)

7-8. -8. (canceled)

13-16. -16. (canceled)

19-22. -22. (canceled)

26-28. -28. (canceled)

30-34. -34. (canceled)

37. (canceled)

44-50. -50. (canceled)

52-55. -55. (canceled)

57. (canceled)

58. A bioreactor for producing hydrogen, comprising:

a vessel 321, holding a hydrogen producing system, said system comprising a suspension of an algal Fe-only hydrogenase attached to a plant ferredoxin and PSI;

a light providing apparatus comprising an optic fiber, said light providing apparatus being configured to provide light of a selected spectrum to said system; and

a gas liquid separation membrane for separating gas leaving the suspension from said suspension.

59. The bioreactor of claim 58, wherein said system comprises a suspension of cells.

60-62. -62. (canceled)