IP Library › Granted Patent US 11,634,733
Granted Patent B2
US 11,634,733 · App. 16/022,878 · Granted Apr 25, 2023

Methods, materials, synthetic hosts and reagents for the biosynthesis of hydrocarbons and derivatives thereof

Inventors: Stephen Thomas Cartman (Redcar, GB); Jonathan Paul Combe (Redcar, GB); Leonard Keith Pattenden (Redcar, GB); Andrew Shaw (Redcar, GB); Massimiliano Zampini (Redcar, GB)
Assignee: INV NYLON CHEMICALS AMERICAS, LLC
C12P5/007C12N9/0006C12N9/1029C12N9/1205C12N9/1229C12N9/88C12N9/90C12N15/746C12Y207/04002C12Y401/01033C12Y401/03C12Y402/03027C12Y503/03002C12Y101/01034C12Y203/01009C12Y203/0301C12Y207/01036
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Quick Facts
Patent No.
US 11,634,733
App. No.
16/022,878
Granted
Apr 25, 2023
Kind
B2
Abstract

Genetically engineered hosts and methods for their production and use in synthesizing hydrocarbons are provided.

Claims (9)

1. A genetically engineered Cupriavidus host capable of producing hydrocarbons comprising one or more isoprene units, said genetically engineered Cupriavidus host comprising a genome-integrated synthetic operon comprising an exogenous nucleic acid sequence encoding a polypeptide having acetoacetyl-CoA C-acetyltransferase (AACT), an exogenous nucleic acid sequence encoding HMG-CoA reductase (HMGR) or hydroxymethylglutaryl-CoA synthase (HMGS) enzyme activity and an exogenous nucleic acid sequence encoding an isoprene synthase (IspS) enzyme

wherein the polypeptide having AACT enzyme activity that catalyzes the chemical reaction of 2 acetyl-CoA to CoA and acetoacetyl-CoA and has at least 90% sequence identity to SEQ ID NO:56 or is encoded by a nucleic acid sequence with at least 90% sequence identity to SEQ ID NO: 55, the polypeptide having HMGR enzyme activity that catalyzes the reaction of HMG-CoA (3-hydroxy-3-methylglutaryl-CoA) to mevalonic acid and has at least 90% sequence identity to SEQ ID NO:58, 60, 62, 64 or 66 or is encoded by a nucleic acid sequence with at least 90% sequence identity to SEQ ID NO:57, 59, 61, 63 or 65 or the polypeptide having HMGS enzyme activity that catalyzes the reaction of acetoacetyl-CoA to HMG-CoA and has at least 90% sequence identity to SEQ ID NO: 68, 70, 72 or 74 or is encoded by a nucleic acid sequence with at least 90% sequence identity to SEQ ID NO: 67, 69, 71 or 73, and wherein the IspS enzyme has at least 90% sequence identity to an amino acid sequence set forth in SEQ ID NOs: 7, 8, 29 or 30 and catalyzes the reaction dimethylallyl pyrophosphate to isoprene and diphosphate,

wherein said genetically engineered Cupriavidus host produces the hydrocarbon comprising one or more isoprene units from a gas stream comprising at least one of natural gas, methanol, ethanol, non-volatile residue, caustic wash from cyclohexane oxidation processes, or waste stream, or derivative thereof, in an amount at least 60 times greater than a Cupriavidus host cell harbouring a plasmid expressing only IspS.

2. The genetically engineered host of claim 1 , wherein said hydrocarbon comprises said one or more isoprene units as depicted in Formula I

or a salt or derivative thereof.

3. The genetically engineered host of claim 1 further comprising one or more enzymes selected from mevalonate kinase (MVK), phosphomevalonate kinase (MPK), mevalonate diphosphate decarboxylase (MDD) and isopentenyl diphosphate isomerase (IDI).

4. The genetically engineered host of claim 1 wherein the genome-integrated synthetic operon encodes one or more Enterococcus faecalis enzymes.

5. The genetically engineered host of claim 1 further comprising one or more plasmids encoding one or more enzymes of the lower and/or upper MVA pathways.

6. The genetically engineered host of claim 1 wherein the genome integrated synthetic operon comprises an exogenous nucleic acid sequence encoding a polypeptide having AACT enzyme activity, an exogenous nucleic acid sequence encoding a polypeptide having HMGR enzyme activity, an exogenous nucleic acid sequence encoding a polypeptide having HMGS enzyme activity and an exogenous nucleic acid sequence encoding a polypeptide having IspS enzyme activity.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 22, 2021
From: CARTMAN, STEPHEN THOMAS; COMBE, JONATHAN PAUL; PATTENDEN, LEONARD KEITH; SHAW, ANDREW; ZAMPINI, MASSIMILIANO
To: INVISTA NORTH AMERICA S.A.R.L.
Reel/Frame 055667/0673 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 7, 2021
From: INVISTA NORTH AMERICA, LLC
To: INV NYLON CHEMICALS AMERICAS, LLC
Reel/Frame 054914/0897 →
CHANGE OF NAME Recorded Dec 15, 2020
From: INVISTA NORTH AMERICA S.A.R.L.
To: INVISTA NORTH AMERICA, LLC
Reel/Frame 054765/0645 →
Continuity (2)
Provisional Application 62527595 · Jun 30, 2017
Related Publication 20190002926A1 · Jan 3, 2019