This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Santos, C. N. S.
Right arrow Articles by Stephanopoulos, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Santos, C. N. S.
Right arrow Articles by Stephanopoulos, G.
Agricola
Right arrow Articles by Santos, C. N. S.
Right arrow Articles by Stephanopoulos, G.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, February 2008, p. 1190-1197, Vol. 74, No. 4
0099-2240/08/$08.00+0     doi:10.1128/AEM.02448-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Melanin-Based High-Throughput Screen for L-Tyrosine Production in Escherichia coli{triangledown}

Christine Nicole S. Santos and Gregory Stephanopoulos*

Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Received 30 October 2007/ Accepted 12 December 2007

We present the development of a simple, high-throughput screen for identifying bacterial strains capable of L-tyrosine production. Through the introduction of a heterologous gene encoding a tyrosinase, we were able to link L-tyrosine production in Escherichia coli with the synthesis of the black and diffusible pigment melanin. Although melanin was initially produced only at low levels in morpholinepropanesulfonic acid (MOPS) minimal medium, phosphate supplementation was found to be sufficient for increasing both the rates of synthesis and the final titers of melanin. Furthermore, a strong linear correlation between extracellular L-tyrosine content and melanin formation was observed by use of this new medium formulation. A selection strategy that utilizes these findings has been developed and has been shown to be effective in screening large combinatorial libraries in the search for L-tyrosine-overproducing strains.

* Corresponding author. Mailing address: Department of Chemical Engineering, Room 56-469, Massachusetts Institute of Technology, Cambridge, MA 02139. Phone: (617) 253-4583. Fax: (617) 253-3122. E-mail: gregstep{at}mit.edu

{triangledown} Published ahead of print on 21 December 2007.

Applied and Environmental Microbiology, February 2008, p. 1190-1197, Vol. 74, No. 4
0099-2240/08/$08.00+0     doi:10.1128/AEM.02448-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Klein-Marcuschamer, D., Santos, C. N. S., Yu, H., Stephanopoulos, G. (2009). Mutagenesis of the Bacterial RNA Polymerase Alpha Subunit for Improvement of Complex Phenotypes. Appl. Environ. Microbiol. 75: 2705-2711 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»