Nanolipoprotein particles comprising hydrogenases and related products, methods and systems

Patent metadata

Field	Value
Publication number	US-9688718-B2
Application number	US-35247209-A
Country	US
Kind code	B2
Filing date	Jan 12, 2009
Priority date	Jan 11, 2008
Publication date	Jun 27, 2017
Grant date	Jun 27, 2017

Abstract

Provided herein are nanolipoprotein particles that comprise a membrane associated hydrogenase and related assemblies, devices, methods and systems.

First claim

What is claimed is:

1. A nanolipoprotein particle comprising
a membrane associated hydrogenase,
a membrane forming lipid, and
a scaffold protein,
wherein the membrane associated hydrogenase is heteromeric and multimeric; and
wherein the nanolipoprotein particle further comprises a cell membrane fragment, the cell membrane fragment comprising the membrane associated heteromeric and multimeric hydrogenase in a catalytically active form,
the membrane forming lipid and the scaffold protein are at a mass ratio between 1:1 and 20:1 and the scaffold protein and the membrane associated heteromeric and multimeric hydrogenase are at a mass ratio between 1:0.025 and 1:1.

2. The nanolipoprotein particle of claim 1 , wherein the membrane associated hydrogenase is a [Ni/Fe] hydrogenase or a [Fe/Fe] hydrogenase.

3. The nanolipoprotein particle of claim 1 , wherein the membrane associated hydrogenase is a [Ni/Fe] hydrogenase from Allochromatium vinosum, Methanosarcina barkeri, Escherichia coli , and Rhodospirillum rubrum Desulfomicrobium baculatum and Ralstonia species.

4. The nanolipoprotein particle of claim 1 , wherein the membrane associated hydrogenase is a [Ni/Fe] hydrogenase from Pyrococcus Furiosus.

5. A biocatalyst assembly comprising the nanolipoprotein particle of claim 1 , wherein the nanolipoprotein particle is immobilized to a support.

6. The biocatalyst assembly of claim 5 , wherein the support is a solid support.

7. The biocatalyst assembly of claim 5 , wherein the support is an electrically conductive support.

8. A method to perform a chemical reaction catalyzed by a membrane associated hydrogenase, the method comprising:
providing reagents for performing said chemical reaction,
providing the nanolipoprotein particle of claim 1 , wherein the membrane associated hydrogenase is the membrane associated hydrogenase catalyzing the chemical reaction; and
contacting the reagents with the nanolipoprotein particle for a time and under conditions to allow the reaction to occur, thus performing said chemical reaction.

9. The method of claim 8 wherein the nanolipoprotein particle is immobilized on a solid support.

10. The method of claim 8 , wherein the chemical reaction is hydrogen production.

11. The method of claim 10 wherein the hydrogen production is performed as a solution phase reaction.

12. The method of claim 10 , wherein the conditions include a temperature between from about 25 degrees C. and about 95 degrees C.

13. The method of claim 8 , wherein the membrane associated hydrogenase is a [Ni/Fe] hydrogenase.

14. The method of claim 8 , wherein the chemical reaction is oxidation of molecular hydrogen.

15. A system to perform a chemical reaction catalyzed by a membrane associated hydrogenase, the system comprising:
the nanolipoprotein particle of claim 1 , wherein the membrane associated hydrogenase is the membrane associated hydrogenase catalyzing the chemical reaction; and
reagents for performing the chemical reaction,
wherein the nanolipoprotein particle and the reagents can be contacted for a time and under conditions to allow the chemical reaction catalyzed by said membrane associated hydrogenase to occur.

16. The system of claim 15 , wherein the membrane associated hydrogenase is immobilized to a support.

17. The system of claim 15 , wherein the membrane associated hydrogenase is a [Ni/Fe] hydrogenase from Allochromatium vinosum, Methanosarcina barkeri, Escherichia coli, Rhodospirillum rubrum, Desulfomicrobium baculatum and Ralstonia species.

18. The system of claim 15 , wherein the membrane associated hydrogenase is a [Ni/Fe] hydrogenase from Pyrococcus Furiosus.

19. A device for performing a chemical reaction catalyzed by a membrane associated hydrogenase, the device comprising:
a substrate compartment, a hydrogenase compartment, and a product compartment, with the substrate compartment in communication with the product compartment through the hydrogenase compartment,
wherein
the hydrogenase compartment comprises the biocatalyst assembly of claim 5 ; and
the substrate compartment is configured to comprise a substrate for the chemical reaction catalyzed by the membrane associated hydrogenase and to allow contact between said substrate and said membrane associated hydrogenase.

20. The device of claim 19 , further comprising a product compartment in communication with the hydrogenase compartment, the product compartment configured to comprise hydrogen produced following the reaction of a substrate with the membrane associated hydrogenase.

21. A nanolipoprotein particle comprising
a membrane associated hydrogenase,
a membrane forming lipid, and
a scaffold protein,
wherein the membrane associated hydrogenase is heteromeric and multimeric in a catalytically active form; and
wherein the nanolipoprotein particle is obtained from a cell membrane fragment, the cell membrane fragment comprising the membrane associated heteromeric and multimeric hydrogenase,
the membrane forming lipid and the scaffold protein are at a mass ratio between 1:1 and 20:1 and the scaffold protein and the membrane associated heteromeric and multimeric hydrogenase are at a mass ratio between 1:0.025 and 1:1.

22. The nanolipoprotein particle of claim 1 , wherein the membrane forming lipid is dimyristoylphosphatidylcholine (DMPC), the scaffold protein is Apo E422K, and the DMPC, Apo E422K and the membrane associated heteromeric and multimeric hydrogenase have a mass ratio of about 4:1:0.05.

23. The nanolipoprotein particle of claim 21 , wherein the membrane forming lipid is dimyristoylphosphatidylcholine (DMPC), the scaffold protein is Apo E422K, and the DMPC, Apo E422K and the membrane associated heteromeric and multimeric hydrogenase have a mass ratio of about 4:1:0.05.

Assignees

• Baker Sarah E
• Chromy Brett A
• Henderson Paul
• Hoeprich Jr Paul D
• L Livermore Nat Security Llc

Inventors

• Baker Sarah E
• Chromy Brett A
• Henderson Paul
• Hoeprich Jr Paul D

Classifications

• C12P3/00

  Preparation of elements or inorganic compounds except carbon dioxide {(recovery of carbon dioxides as by-products C12F3/02)} · CPC title

• C07K1/145 Primary

  by extraction or solubilisation · CPC title

• Y10S977/799

  Containing biological material · CPC title

• C12N11/02 Primary

  Enzymes or microbial cells immobilised on or in an organic carrier · CPC title