IP Library Granted Patent US 9,267,118
Granted Patent B2
US 9,267,118 · App. 12/673,587 · Granted Feb 23, 2016

Methods for identifying bacterial strains that produce L-tyrosine

Inventors: Gregory Stephanopoulos (Winchester, MA); Christine Nicole S. Santos (Cambridge, MA)
Assignee: Massachusetts Institute of Technology
C12N9/0071C12Q1/04C12Q1/26C12Y114/18001G01N2333/90245
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Quick Facts
Patent No.
US 9,267,118
App. No.
12/673,587
Granted
Feb 23, 2016
Kind
B2
Abstract

The invention relates to high-throughput screens for identifying bacterial strains capable of L-tyrosine production.

Claims (12)

1. A method for identifying bacterial strains that produce L-tyrosine, comprising:

culturing in a medium one or more bacterial strains capable of L-tyrosine production and which heterologously express a tyrosinase, wherein the tyrosinase comprises the amino acid sequence of SEQ ID NO: 4, and wherein the tyrosinase has the ability to convert L-tyrosine into melanin, and

determining the amount of melanin produced by the one or more bacterial strains, wherein production of melanin by a strain indicates that the strain produces L-tyrosine.

2. The method of claim 1 , wherein the amount of melanin produced by the one or more bacterial strains correlates positively with the amount of L-tyrosine produced by the one or more bacterial strains.

3. The method of claim 1 , wherein the amount of melanin produced by the one or more bacterial strains is detected by a visual method or a spectrophotometric method.

4. The method of claim 1 , wherein the pH of the culture medium is at least about 5.

5. A method for identifying bacterial strains that produce extracellular L-tyrosine, comprising:

culturing a reporter bacterial strain that heterologously expresses a tyrosinase in a sample of a culture medium in which one or more bacterial strains capable of producing extracellular L-tyrosine are being cultured or have been cultured, wherein the tyrosinase comprises the amino acid sequence of SEQ ID NO: 4, and wherein the tyrosinase has the ability to convert L-tyrosine into melanin, and

determining the amount of melanin produced by the reporter strain, wherein production of melanin by a reporter strain indicates that the one or more bacterial strains produces extracellular L-tyrosine.

6. The method of claim 5 , wherein the amount of melanin produced by the reporter strain correlates positively with the amount of L-tyrosine produced by the one or more bacterial strains.

7. The method of claim 5 , wherein the amount of melanin produced by the reporter strain is detected by a visual method or a spectrophotometric method.

8. The method of claim 5 , wherein the pH of the culture medium is at least about 5.

Assignments (2)
CONFIRMATORY LICENSE Recorded May 24, 2012
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: NATIONAL SCIENCE FOUNDATION
Reel/Frame 028274/0888 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 21, 2012
From: STEPHANOPOULOS, GREGORY; SANTOS, CHRISTINE
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 028243/0188 →
Continuity (2)
Provisional Application 60965149 · Aug 17, 2007
Related Publication 20110151496A1 · Jun 23, 2011