Nanolipoprotein particles comprising a natural rubber biosynthetic enzyme complex and related products, methods and systems

Patent metadata

Field	Value
Publication number	US-9303273-B2
Application number	US-201414199973-A
Country	US
Kind code	B2
Filing date	Mar 6, 2014
Priority date	May 9, 2008
Publication date	Apr 5, 2016
Grant date	Apr 5, 2016

Abstract

Provided herein are nanolipoprotein particles that comprise a biosynthetic enzyme more particularly an enzyme capable of catalyzing rubber or other rubbers polymerization, and related assemblies, devices, methods and systems.

First claim

What is claimed is:

1. A nanolipoprotein particle comprising
a target protein,
a membrane forming lipid, and
a scaffold protein,
wherein the target protein is a natural rubber transferase complex (RuT), wherein the natural rubber transferase complex is provided within a discoidal membrane lipid bilayer formed by the membrane forming lipid and stabilized by the scaffold protein, and wherein the membrane lipid bilayer is attached to the RuT through interaction of the natural rubber transferase complex hydrophobic region with the membrane lipid bilayer.

2. The nanolipoprotein particle of claim 1 , wherein the natural rubber transferase complex is a collection of enzymes capable of catalyzing formation of natural polyisoprene rubber.

3. The nanolipoprotein particle of claim 1 , wherein the natural rubber transferase complex, is an isolated or purified naturally occurring rubber transferase.

4. The nanolipoprotein particle of claim 1 , wherein the nanolipoprotein particle is immobilized to a support.

5. The nanolipoprotein particle of claim 4 , wherein the support is a solid support.

6. A method to perform rubber biosynthesis, the method comprising:
providing reagents for performing said rubber biosynthesis;
providing the nanolipoprotein particle of claim 1 ; and
contacting the reagents with the nanolipoprotein particle of claim 1 for time and under conditions to allow the reaction to occur, thus performing said rubber biosynthesis.

7. The method of claim 6 , wherein the natural rubber transferase complex, is an isolated or purified naturally occurring rubber transferase.

8. The method of claim 6 wherein the nanolipoprotein particle of claim 1 is immobilized on a solid support.

9. The method of claim 6 , wherein the rubber biosynthesis is directed to rubber production.

10. A system to perform a rubber biosynthesis, the system comprising:
one or more nanolipoprotein particles of claim 1 ; and
reagents for performing the rubber biosynthesis catalyzed by the natural rubber transferase complex,
wherein the one or more nanolipoprotein particles of claim 1 and the reagents can be contacted for a time and under condition to allow the rubber biosynthesis to occur.

11. The system of claim 10 , wherein the natural rubber transferase complex, is an isolated or purified naturally occurring rubber transferase.

12. A device for performing a rubber biosynthesis, the device comprising:
a substrate compartment, a catalyst compartment, and a product compartment, with the substrate compartment in communication with the product compartment through the catalyst compartment, wherein
the catalyst compartment comprises the nanolipoprotein particle according to claim 1 arranged in a biosynthetic assembly; and
the substrate compartment is configured to comprise monomers for the rubber biosynthesis catalyzed by the natural rubber transferase complex and to allow contact between said monomers and said natural rubber transferase complex.

13. The device of claim 12 , wherein the nanolipoprotein particle is comprised in the catalyst compartment immobilized to a solid support.

14. The device of claim 12 , further comprising a product compartment in communication with the catalyst compartment, the product compartment configured to comprise the product produced following the reaction of a substrate with the biocatalyst.

15. The device of claim 12 , wherein the natural rubber transferase complex is an isolated or purified naturally occurring RuT.

Assignees

• L Livermore Nat Security Llc
• Us Agriculture

Inventors

• Hoeprich Paul D
• Whalen Maureen

Classifications

• C12N9/1085

  transferring alkyl or aryl groups other than methyl groups (2.5) · CPC title

• C12N9/96

  Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates · CPC title

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title

• C12P5/007 Primary

  containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00) · CPC title

• C12Y205/0102

  Rubber cis-polyprenylcistransferase (2.5.1.20) · CPC title