Rational, combinatorial, and genomic approaches for engineering L-tyrosine production in Escherichia coli
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Santos-2012-Rational, combinatorial, and genomic.pdf
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Author(s)
Santos, Christine Nicole S.
Xiao, Wenhai
Stephanopoulos, Gregory
Date Issued
August 2012
Journal
Proceedings of the National Academy of Sciences of the United States of America
Publisher
National Academy of Sciences (U.S.)
Citation
Santos, C. N. S., W. Xiao, and G. Stephanopoulos. “Rational, Combinatorial, and Genomic Approaches for Engineering L-tyrosine Production in Escherichia Coli.” Proceedings of the National Academy of Sciences 109.34 (2012): 13538–13543. CrossRef. Web.
Version
Final published version
Abstract
Although microbial metabolic engineering has traditionally relied on rational and knowledge-driven techniques, significant improvements in strain performance can be further obtained through the use of combinatorial approaches exploiting phenotypic diversification and screening. Here, we demonstrate the combined use of global transcriptional machinery engineering and a high-throughput L-tyrosine screen towards improving L-tyrosine production in Escherichia coli. This methodology succeeded in generating three strains from two separate mutagenesis libraries (rpoA and rpoD) exhibiting up to a 114% increase in L-tyrosine titer over a rationally engineered parental strain with an already high capacity for production. Subsequent strain characterization through transcriptional analysis and whole genome sequencing allowed complete phenotype reconstruction from well-defined mutations and point to important roles for both the acid stress resistance pathway and the stringent response of E. coli in imparting this phenotype. As such, this study presents one of the first examples in which cell-wide measurements have helped to elucidate the genetic and biochemical underpinnings of an engineered cellular property, leading to the total restoration of metabolite overproduction from specific chromosomal mutations.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1073/pnas.1206346109
