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]

---

1. Michigan State University, East Lansing, MI (United States)
2. 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.

• Research Organization: Michigan State Univ., East Lansing, MI (United States)
• Sponsoring Organization: USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health, National Institute of Allergy and Infectious Diseases (NIAID)
• Grant/Contract Number: FG02-91ER20021; 5 R01 AI114975-05; AC02-05CH11231
• OSTI ID: 1671303
• Alternate ID(s): OSTI ID: 1702961
• Journal Information: Metabolic Engineering, Vol. 54; ISSN 1096-7176
• Publisher: Elsevier
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Bacterial microcompartments
protein design
metabolic engineering
synthetic biology