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Engineering bacterial microcompartments with heterologous enzyme cargos.

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|July 7, 2020
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Bacterial microcompartments (BMCs) can enclose new enzymes, creating functional microreactors. These engineered BMCs protect enzymes from pH changes and act as diffusion barriers.

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Area of Science:

  • Microbiology
  • Biochemistry
  • Synthetic Biology

Background:

  • Bacterial microcompartments (BMCs) are protein-based organelles found in bacteria.
  • They compartmentalize metabolic enzymes, enhancing pathway efficiency.
  • Salmonella enterica uses BMCs for 1,2-propanediol degradation.

Purpose of the Study:

  • To explore the broader applicability of BMCs for enclosing heterologous enzymes.
  • To engineer BMCs as catalytic microreactors for diverse functions.
  • To assess the protective capabilities and diffusion properties of engineered BMCs.

Main Methods:

  • Directed expression of three distinct enzymes (β-galactosidase, esterase Est5, glycerol dehydrogenase) into BMCs.
  • Copurification of enzymes with the BMC shell.
  • Enzyme activity assays under varying pH and temperature conditions.
  • Investigation of BMCs as diffusion barriers for small lipophilic molecules.

Main Results:

  • Successfully targeted and co-purified active β-galactosidase, esterase Est5, and glycerol dehydrogenase with BMCs.
  • Demonstrated that BMC shells provide protection against pH-dependent inactivation.
  • Showed BMCs do not offer significant protection against thermal stress.
  • Provided evidence for BMCs acting as a selective diffusion barrier for lipophilic molecules.

Conclusions:

  • Bacterial microcompartments can be engineered to house and activate diverse heterologous enzymes.
  • Engineered BMCs offer partial protection to encapsulated enzymes, primarily against pH variations.
  • The BMC shell functions as a moderately selective diffusion barrier, influencing molecular transport.