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Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays
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Short-Circuiting the SAM-Cycle in Escherichia coli.

Zhong Li1,2, Xiaojin Wen1, Seseg B Bolotova1,3

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Researchers engineered a novel Escherichia coli strain that recycles S-adenosylhomocysteine (SAH) using a synthetic methyl donor, creating a Short-Circuited SAM-Cycle (SCSC). This breakthrough enables efficient methyltransferase biocatalysis and natural product production.

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Area of Science:

  • Biochemistry
  • Synthetic Biology
  • Microbial Engineering

Background:

  • Methylation is crucial for life, primarily using S-adenosylmethionine (SAM) as the methyl donor.
  • The natural SAM cycle involves complex one-carbon metabolism and homocysteine methylation.
  • Existing methods for SAM regeneration are often inefficient or require specific substrates.

Purpose of the Study:

  • To engineer a metabolically engineered Escherichia coli strain with a self-contained SAM cycle.
  • To establish a novel system for S-adenosylhomocysteine (SAH) methylation using a synthetic methyl donor.
  • To create a platform for methyltransferase optimization and isotope-labeled natural product synthesis.

Main Methods:

  • Construction of an Escherichia coli strain with a Short-Circuited SAM-Cycle (SCSC).
  • Identification of aryl sulfonate methyl esters as biocompatible methyl donors.
  • Utilizing methyltransferases that accept synthetic methyl donors for SAH methylation in vivo.
  • Employing in vivo selection for enzyme optimization.

Main Results:

  • Successfully created an E. coli strain maintaining its SAM pool exclusively via SAH methylation with a synthetic donor.
  • Identified a novel aryl sulfonate methyl ester as an effective and biocompatible methyl donor.
  • Demonstrated the utility of the SCSC strain for optimizing SAH-methylating enzymes.
  • Showcased the strain's capability for producing isotope-labeled natural products.

Conclusions:

  • The SCSC strain offers a novel and self-sufficient system for SAM regeneration.
  • This engineered microbe facilitates advancements in methyltransferase biocatalysis.
  • The SCSC platform holds potential for natural product discovery and bacterial metabolomics research.