Isoprene synthase variants with improved solubility for production of isoprene

• US 9,273,299 B2
• Filed: 04/11/2014
• Issued: 03/01/2016
• Est. Priority Date: 10/27/2011
• Status: Active Grant

• Current Assignee: Goodyear Tire & Rubber Company, Danisco USA Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Goodyear Tire & Rubber Company, Danisco USA Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Rife, Christopher L., Wells, Derek H.
• Primary Examiner(s): Chowdhury, Iqbal H
• Application Number:
  US14/251,311
• Publication Number:
  US 20140315275A1
• Time in Patent Office: 690 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C08L 7/00 Compositions of natural rubber
• CPC Class Codes: C12N 15/8243 involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 5/007 containing one or more isoprene units, i.e. terpenes carotenes C12P23/00
• CPC Class Codes: C12Y 402/03007 Pentalenene synthase (4.2.3.7)

Abstract

The present invention provides methods and compositions of variant polypeptides having isoprene synthase activity with improved solubility. In particular, the present invention provides isoprene synthase variant for increased isoprene production in recombinant host cells.

Claims

1. An isolated polypeptide variant having isoprene synthase activity, wherein the polypeptide variant comprises a substitution at residue X288 corresponding to SEQ ID NO:

1, and wherein the polypeptide has increased protein solubility compared a parent polypeptide which does not comprise the substitution at residue X288, wherein said polypeptide variant is 90% identical to isoprene synthase polypeptide of SEQ ID NO;

1.

2. The polypeptide variant of claim 1 wherein the substitution is at S288.

3. The polypeptide variant of claim 2 wherein the substitution is S288C.

4. The polypeptide variant of claim 1, where the polypeptide has at least 5% to at least 75% increased protein solubility compared to the parent polypeptide.

5. The polypeptide variant of claim 1 wherein the polypeptide is derived from a parent polypeptide isolated from a plant.

6. The polypeptide variant of claim 5 wherein the parent polypeptide is isolated from a plant species selected from poplar (Populus sp.), kudzu (Pueraria sp.), English oak (Quercus sp.) or willow (Salix sp.).

7. The polypeptide variant of claim 6 wherein the species is Populus sp.

8. The polypeptide variant of claim 7 wherein the species is P. alba, P. tremuloides, P. trichocharpa, or P. nigra.

9. The polypeptide variant of claim 6 wherein the species is Pueraria sp.

10. The polypeptide variant of claim 9 wherein the species is Pueraria montana.

11. The polypeptide variant of claim 6 wherein the species is Quercus sp.

12. The polypeptide variant of claim 11 wherein the species is Quercus rubur.

13. The polypeptide variant of claim 6 wherein the species is Salix sp.

14. The polypeptide variant of claim 13 wherein the species is S. alba or S. baylonica.