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Engineering bacterial microcompartments with heterologous enzyme cargos.

ABSTRACT: Bacterial microcompartments (BMCs) are intracellular proteinaceous organelles devoid of a lipid membrane that encapsulates enzymes of metabolic pathways. Salmonella enterica synthesizes propanediol-utilization BMCs containing enzymes involved in the degradation of 1,2-propanediol. BMCs can be designed to enclose heterologous proteins, paving the way to engineered catalytic microreactors. Here, we investigate broader applicability of this design principle by directing three different enzymes to the BMC. We demonstrate that ?-galactosidase, esterase Est5, and cofactor-dependent glycerol dehydrogenase can be directed to the BMC and copurified with the microcompartment shell in a catalytically active form. We show that the BMC shell protects enzymes from pH-dependent but not from temperature stress. Moreover, we provide evidence that the heterologously expressed BMCs act as a moderately selective diffusion barrier for lipophilic small molecules.

SUBMITTER: Wagner HJ

PROVIDER: S-EPMC6999380 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature

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Engineering bacterial microcompartments with heterologous enzyme cargos.

Wagner Hanna J HJ Capitain Charlotte C CC Richter Karsten K Nessling Michelle M Mampel Jörg J

Engineering in life sciences 20160908 1

Bacterial microcompartments (BMCs) are intracellular proteinaceous organelles devoid of a lipid membrane that encapsulates enzymes of metabolic pathways. <i>Salmonella enterica</i> synthesizes propanediol-utilization BMCs containing enzymes involved in the degradation of 1,2-propanediol. BMCs can be designed to enclose heterologous proteins, paving the way to engineered catalytic microreactors. Here, we investigate broader applicability of this design principle by directing three different enzym ...[more]

PMID: 32624727

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