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Less toxic metabolites, more chemical product
Date:10/29/2013

emicals continues to grow in importance. To date, the most productive microbial hosts have been those engineered with heterologous pathways for which they have little or no native regulation of the metabolites being expressed. However, such unregulated expression of heterologous enzymes can be toxic to the host, which can limit the production of the target chemical to well below levels that could be obtained.

"Although synthetic biology has made great strides in creating novel, dynamic genetic circuits, most control systems for heterologous metabolic pathways still rely on inducible or constitutive promoters," Keasling says. "Approaches developed to tailor expression strength by means of promoter libraries, mRNA stability or ribosome-binding are optimized for a particular growth phase or condition in the bioreactor, however, growth and environmental conditions change during the fermentation process."

Since the accumulation of intermediate metabolites to toxic levels in a microbe during a fermentation process can lead to a stress response, Keasling and his JBEI colleagues reasoned that it should be possible to tap a host microbe's native stress response system when metabolites accumulate. Transcript profiling of the E. coli genome allowed them to evaluate transcriptional response to a heterologous pathway and create a list of promoters that could be used to respond to intermediate toxicity.

"Using such promoters to regulate pathway expression in response to the toxic intermediate metabolites creates a link between the cell's metabolic state and the expression of the metabolic pathway," Keasling says. "This enables us to create biosensors that respond to and regulate pathway intermediates. In silico models have indicated, and we've demonstrated in this study that our approach can be used to improve production of a desired chemical over common inducible promoters and constitutive promoters of various strengths."

Keasling a
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Contact: Lynn Yarris
lcyarris@lbl.gov
510-486-5375
DOE/Lawrence Berkeley National Laboratory
Source:Eurekalert  

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