IP Library Granted Patent US 8,927,241
Granted Patent B2
US 8,927,241 · App. 13/249,388 · Granted Jan 6, 2015

Microbial engineering for the production of chemical and pharmaceutical products from the isoprenoid pathway

Inventors: Parayil K. Ajikumar (Cambridge, MA); Gregory Stephanopoulos (Winchester, MA); Too Heng Phon (Kent Vale, SG)
Assignees: Massachusetts Institute of Technology; National University of Singapore
C12P15/00C12P17/02C12P23/00C12P5/007
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Quick Facts
Patent No.
US 8,927,241
App. No.
13/249,388
Granted
Jan 6, 2015
Kind
B2
Abstract

The invention relates to the production of one or more terpenoids through microbial engineering, and relates to the manufacture of products comprising terpenoids.

Claims (15)

1. A method for making a product containing Myrcene or a derivative thereof, the method comprising:

providing an Escherichia coli ( E. coli ) that produces isopentyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) through an upstream methylerythritol pathway (MEP) and converts the IPP and DMAPP to Myrcene through a recombinantly expressed downstream synthesis pathway comprising Geranyl Diphosphate Synthase and Myrcene Synthase; and

culturing the E. coli to produce the Myrcene, wherein the accumulation of indole in the culture is controlled to below 100 mg/L to thereby increase Myrcene production; and optionally preparing a derivative of the Myrcene; and

incorporating the Myrcene or derivative into a fragrance product, a cosmetic product, oral care product, a pest control product, a cleaning product, a lubricant or surfactant, or a soap product.

2. The method of claim 1 , wherein accumulation of indole in the culture is controlled by balancing the upstream MEP pathway with the downstream synthesis pathway.

3. The method of claim 1 , further comprising, measuring the amount or concentration of indole continuously or intermittently.

4. The method of claim 1 , wherein accumulation of indole in the culture is maintained to below 50 mg/L.

5. The method of claim 1 , wherein accumulation of indole in the culture is maintained to below 10 mg/L.

6. The method of claim 1 , wherein the Myrcene is produced at 10 mg/L or more.

7. The method of claim 1 , wherein the Myrcene is produced at 100 mg/L or more.

8. The method of claim 1 , wherein the E. coli has additional copies of one or more of the dxs, idi, ispD and ispF genes of the MEP pathway.

9. A method for making a myrcene, the method comprising:

providing an Escherichia coli ( E. coli ) that produces isopentyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) through an upstream methylerythritol pathway (MEP) and converts the IPP and DMAPP to myrcene through a recombinantly expressed downstream synthesis pathway comprising Geranyl Diphosphate Synthase and Myrcene Synthase; and

culturing the E. coli to produce the myrcene, wherein the accumulation of indole in the culture is controlled to below 100 mg/L to thereby increase terpenoid production; and

recovering the myrcene.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 24, 2014
From: AJIKUMAR, PARAYIL K.; STEPHANOPOULOS, GREGORY
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 033163/0589 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 24, 2014
From: TOO, HENG-PHON
To: NATIONAL UNIVERSITY OF SINGAPORE
Reel/Frame 033163/0631 →
CONFIRMATORY LICENSE Recorded Sep 27, 2012
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 029045/0559 →
Continuity (4)
Continuation In Part 12943477 · Nov 10, 2010
Provisional Application 61280877 · Nov 10, 2009
Provisional Application 61388543 · Sep 30, 2010
Related Publication 20120107893A1 · May 3, 2012