Production of cyclic terpenoids

• US 6,818,424 B2
• Filed: 08/24/2001
• Issued: 11/16/2004
• Est. Priority Date: 09/01/2000
• Status: Expired due to Fees

• Current Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Wang, Siqun, DiCosimo, Deana J., Odom, James M., Koffas, Mattheos
• Primary Examiner(s): Nashed, Nashaat T.
• Application Number:
  US09/938,956
• Publication Number:
  US 20020142408A1
• Time in Patent Office: 1,180 Days
• Field of Search: 435/166, 435/252.3
• US Class Current: 435/166
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 23/00 Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes containing heterorings C12P17/00
• CPC Class Codes: C12P 7/02 containing a hydroxy group
• CPC Class Codes: Y02E 50/30 Fuel from waste, e.g. synthetic alcohol or diesel

Abstract

A methanotrophic bacterium has been genetically engineered to produce cyclic terpenoids from geranyl pyrophosphate.

Claims

1. A method for the production of a monoterpene comprising:

a) providing a transformed Methylomonas cell comprising;

(i) geranyl pyrophosphate; and

(ii) at least one isolated nucleic acid molecule encoding a cyclic terpene synthase selected from the group consisting of limonene synthase, inene synthase, bornyl synthase, phellandrene synthase, cineole synthase and sabinene synthase under the control of suitable regulatory sequences;

(b) contacting the Methylomonas cell of step (a) under suitable growth conditions with an effective amount of a C1 carbon substrate whereby a monoterpene compound is produced.

2. A method according to claim 1 wherein the C1 carbon substrate is methane.

3. A method according to claim 1 wherein the Methylomonas is a high growth methanotrophic strain which comprises a functional Embden-Meyerof carbon pathway, said pathway comprising a gene encoding a pyrophosphate dependent phosphofructokinase enzyme.

4. A method according to claim 3, wherein the high growth methanotrophic bacterial strain optionally contains a functional Entner-Douderoff carbon pathway.

5. A method according to claim 3, wherein the high growth methanotrophic bacterial strain is Methylomonas 16a having the ATCC designation ATCC PTA 2402.

6. A method according to claim 1 wherein the monoterpene is selected from the group consisting of limonene, pinene, barnyl diphosphate, β

-phellandrene, 1,8-cineole, and sabinene.

7. A method according to claim 1 wherein the limonene synthase has the amino sequence as set forth in SEQ ID NO:

6.