Applied Cellular Physiology and Metabolic Engineering

Study of stationary phase metabolism via isotopomer analysis of amino acids from an isolated protein

  1. Afshan S. Shaikh1,†,
  2. Yinjie J. Tang2,†,
  3. Aindrila Mukhopadhyay3,4,†,
  4. Héctor García Martín3,4,
  5. Jennifer Gin3,4,
  6. Peter I. Benke3,4,
  7. Jay D. Keasling1,3,4,5,‡,*

Article first published online: 6 NOV 2009

DOI: 10.1002/btpr.325

Issue

Biotechnology Progress

Biotechnology Progress

Volume 26, Issue 1, pages 52–56, January/February 2010

Additional Information

How to Cite

Shaikh, A. S., Tang, Y. J., Mukhopadhyay, A., Martín, H. G., Gin, J., Benke, P. I. and Keasling, J. D. (2010), Study of stationary phase metabolism via isotopomer analysis of amino acids from an isolated protein. Biotechnology Progress, 26: 52–56. doi: 10.1002/btpr.325

Author Information

  1. 1

    Dept. of Chemical Engineering, University of California, Berkeley, CA 94720

  2. 2

    Dept. of Energy, Environmental and Chemical Engineering, Washington University, St Louis, MO 63130

  3. 3

    Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720

  4. 4

    Joint BioEnergy Institute, Emeryville, CA 94608

  5. 5

    Dept. of Bioengineering, University of California, Berkeley, CA 94720

  1. †

    The first three authors contributed equally in this study.

  2. ‡

    Jay D. Keasling has a consulting relationship with and a financial interest in Amyris and a financial interest in LS9, two biofuels companies.

*Dept. of Chemical Engineering, University of California, Berkeley, CA 94720

Publication History

  1. Issue published online: 5 FEB 2010
  2. Article first published online: 6 NOV 2009
  3. Manuscript Revised: 8 JUL 2009
  4. Manuscript Received: 22 JAN 2009

SEARCH

SEARCH BY CITATION

ARTICLE TOOLS

View Full Article with Supporting Information (HTML) Get PDF (101K)

Keywords:

  • secondary metabolites;
  • metabolic flux;
  • 13C;
  • green fluorescent protein;
  • proteinogenic amino acids

Abstract

Microbial production of many commercially important secondary metabolites occurs during stationary phase, and methods to measure metabolic flux during this growth phase would be valuable. Metabolic flux analysis is often based on isotopomer information from proteinogenic amino acids. As such, flux analysis primarily reflects the metabolism pertinent to the growth phase during which most proteins are synthesized. To investigate central metabolism and amino acids synthesis activity during stationary phase, addition of fully 13C-labeled glucose followed by induction of green fluorescent protein (GFP) expression during stationary phase was used. Our results indicate that Escherichia coli was able to produce new proteins (i.e., GFP) in the stationary phase, and the amino acids in GFP were mostly from degraded proteins synthesized during the exponential growth phase. Among amino acid biosynthetic pathways, only those for serine, alanine, glutamate/glutamine, and aspartate/asparagine had significant activity during the stationary phase. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010

View Full Article with Supporting Information (HTML) Get PDF (101K)