@article{080435d1e7934e19b8d5dc4a6a5e69e0,
title = "Structure of a Minimal α-Carboxysome-Derived Shell and Its Utility in Enzyme Stabilization",
abstract = "Bacterial microcompartments are proteinaceous shells that encase specialized metabolic processes in bacteria. Recent advances in simplification of these intricate shells have encouraged bioengineering efforts. Here, we construct minimal shells derived from the Halothiobacillus neapolitanus α-carboxysome, which we term Cso-shell. Using cryogenic electron microscopy, the atomic-level structures of two shell forms were obtained, reinforcing notions of evolutionarily conserved features in bacterial microcompartment shell architecture. Encapsulation peptide sequences that facilitate loading of heterologous protein cargo within the shells were identified. We further provide a first demonstration in utilizing minimal bacterial microcompartment-derived shells for hosting heterologous enzymes. Cso-shells were found to stabilize enzymatic activities against heat shock, presence of methanol co-solvent, consecutive freeze-thawing, and alkaline environments. This study yields insights into α-carboxysome assembly and advances the utility of synthetic bacterial microcompartments as nanoreactors capable of stabilizing enzymes with varied properties and reaction chemistries. ",
author = "Tan, {Yong Quan} and Samson Ali and Bo Xue and Teo, {Wei Zhe} and Ling, {Lay Hiang} and Go, {Maybelle Kho} and Hong Lv and Robinson, {Robert C.} and Akihiro Narita and Yew, {Wen Shan}",
note = "Funding Information: This work was supported by grants from the National Research Foundation Synthetic Biology Research and Development Program (SBP-P3). A part of this work was supported by JSPS KAKENHI (18H02410) to A.N. Y.Q.T. and L.H.L. are supported by the NUS Graduate School for Integrative Sciences and Engineering scholarship. Funding Information: BL21(DE3)-Gold Δ lac strain was a kind gift from Jeff Hasty. The authors acknowledge Dr. Lu Ting Liow for providing advice on bacterial promoters. They also thank Professors Kaoru Mitsuoka, Research Center for Ultra-High Voltage Electron Microscopy (supported by Nanotechnology Platform Program, MEXT, Japan, A-19-OS-0052), and Kenji Iwasaki, Institute for Protein Research (supported by the Cooperative Research Program of Institute for Protein Research, Osaka University), both in Osaka University for access to the cryo-electron microscopy microscopes for data optimization and collection. This research was undertaken on the MX1 beamline at the Australian Synchrotron, part of ANSTO. Publisher Copyright: {\textcopyright} 2021 The Authors. Published by American Chemical Society.",
year = "2021",
month = oct,
day = "11",
doi = "10.1021/acs.biomac.1c00533",
language = "English",
volume = "22",
pages = "4095--4109",
journal = "Biomacromolecules",
issn = "1525-7797",
publisher = "American Chemical Society",
number = "10",
}