A designed bacterial microcompartment shell with tunable composition and precision cargo loading

Ferlez, Bryan; Sutter, Markus; Kerfeld, Cheryl A.

doi:10.1016/j.ymben.2019.04.011

A designed bacterial microcompartment shell with tunable composition and precision cargo loading

Journal Article · Tue May 07 00:00:00 EDT 2019 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2019.04.011· OSTI ID:1671303

Ferlez, Bryan ^[1]; Sutter, Markus ^[2]; Kerfeld, Cheryl A. ^[2]

Michigan State University, East Lansing, MI (United States); MSU Plant Research lab
Michigan State University, East Lansing, MI (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Microbes often augment their metabolism by conditionally constructing proteinaceous organelles, known as bacterial microcompartments (BMCs), that encapsulate enzymes to degrade organic compounds or assimilate CO₂. BMCs self-assemble and are spatially delimited by a semi-permeable shell made up of hexameric, trimeric, and pentameric shell proteins. Bioengineers aim to recapitulate the organization and efficiency of these complex biological architectures by redesigning the shell to incorporate non-native enzymes from biotechnologically relevant pathways. To meet this challenge, a diverse set of synthetic biology tools are required, including methods to manipulate the properties of the shell as well as target and organize cargo encapsulation. We designed and determined the crystal structure of a synthetic shell protein building block with an inverted sidedness of its N- and C-terminal residues relative to its natural counterpart; the inversion targets genetically fused protein cargo to the lumen of the shell. Moreover, the titer of fluorescent protein cargo encapsulated using this strategy is controllable using an inducible tetracycline promoter. Furthermore, these results expand the available set of building blocks for precision engineering of BMC-based nanoreactors and are compatible with orthogonal methods which will facilitate the installation and organization of multi-enzyme pathways.

View Accepted Manuscript (DOE)

Research Organization:: Michigan State University, East Lansing, MI (United States)

Sponsoring Organization:: National Institutes of Health, National Institute of Allergy and Infectious Diseases (NIAID); USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-05CH11231; FG02-91ER20021

OSTI ID:: 1671303

Alternate ID(s):: OSTI ID: 1702961

Journal Information:: Metabolic Engineering, Journal Name: Metabolic Engineering Vol. 54; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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