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Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles
Bacterial microcompartments (BMCs) are proteinaceous organelles widespread among bacterial phyla and provide a means for compartmentalizing specific metabolic pathways. They sequester catalytic enzymes from the cytoplasm, using an icosahedral proteinaceous shell with selective permeability to metabo...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2018
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5996877/ https://www.ncbi.nlm.nih.gov/pubmed/29922315 http://dx.doi.org/10.3389/fpls.2018.00739 |
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author | Fang, Yi Huang, Fang Faulkner, Matthew Jiang, Qiuyao Dykes, Gregory F. Yang, Mengru Liu, Lu-Ning |
author_facet | Fang, Yi Huang, Fang Faulkner, Matthew Jiang, Qiuyao Dykes, Gregory F. Yang, Mengru Liu, Lu-Ning |
author_sort | Fang, Yi |
collection | PubMed |
description | Bacterial microcompartments (BMCs) are proteinaceous organelles widespread among bacterial phyla and provide a means for compartmentalizing specific metabolic pathways. They sequester catalytic enzymes from the cytoplasm, using an icosahedral proteinaceous shell with selective permeability to metabolic molecules and substrates, to enhance metabolic efficiency. Carboxysomes were the first BMCs discovered and their unprecedented capacity of CO(2) fixation allows cyanobacteria to make a significant contribution to global carbon fixation. There is an increasing interest in utilizing synthetic biology to construct synthetic carboxysomes in new hosts, i.e., higher plants, to enhance carbon fixation and productivity. Here, we report the construction of a synthetic operon of the β-carboxysome from the cyanobacterium Synechococcus elongatus PCC7942 to generate functional β-carboxysome-like structures in Escherichia coli. The protein expression, structure, assembly, and activity of synthetic β-carboxysomes were characterized in depth using confocal, electron and atomic force microscopy, proteomics, immunoblot analysis, and enzymatic assays. Furthermore, we examined the in vivo interchangeability of β-carboxysome building blocks with other BMC components. To our knowledge, this is the first production of functional β-carboxysome-like structures in heterologous organisms. It provides important information for the engineering of fully functional carboxysomes and CO(2)-fixing modules in higher plants. The study strengthens our synthetic biology toolbox for generating BMC-based organelles with tunable activities and new scaffolding biomaterials for metabolic improvement and molecule delivery. |
format | Online Article Text |
id | pubmed-5996877 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-59968772018-06-19 Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles Fang, Yi Huang, Fang Faulkner, Matthew Jiang, Qiuyao Dykes, Gregory F. Yang, Mengru Liu, Lu-Ning Front Plant Sci Plant Science Bacterial microcompartments (BMCs) are proteinaceous organelles widespread among bacterial phyla and provide a means for compartmentalizing specific metabolic pathways. They sequester catalytic enzymes from the cytoplasm, using an icosahedral proteinaceous shell with selective permeability to metabolic molecules and substrates, to enhance metabolic efficiency. Carboxysomes were the first BMCs discovered and their unprecedented capacity of CO(2) fixation allows cyanobacteria to make a significant contribution to global carbon fixation. There is an increasing interest in utilizing synthetic biology to construct synthetic carboxysomes in new hosts, i.e., higher plants, to enhance carbon fixation and productivity. Here, we report the construction of a synthetic operon of the β-carboxysome from the cyanobacterium Synechococcus elongatus PCC7942 to generate functional β-carboxysome-like structures in Escherichia coli. The protein expression, structure, assembly, and activity of synthetic β-carboxysomes were characterized in depth using confocal, electron and atomic force microscopy, proteomics, immunoblot analysis, and enzymatic assays. Furthermore, we examined the in vivo interchangeability of β-carboxysome building blocks with other BMC components. To our knowledge, this is the first production of functional β-carboxysome-like structures in heterologous organisms. It provides important information for the engineering of fully functional carboxysomes and CO(2)-fixing modules in higher plants. The study strengthens our synthetic biology toolbox for generating BMC-based organelles with tunable activities and new scaffolding biomaterials for metabolic improvement and molecule delivery. Frontiers Media S.A. 2018-06-05 /pmc/articles/PMC5996877/ /pubmed/29922315 http://dx.doi.org/10.3389/fpls.2018.00739 Text en Copyright © 2018 Fang, Huang, Faulkner, Jiang, Dykes, Yang and Liu. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Plant Science Fang, Yi Huang, Fang Faulkner, Matthew Jiang, Qiuyao Dykes, Gregory F. Yang, Mengru Liu, Lu-Ning Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles |
title | Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles |
title_full | Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles |
title_fullStr | Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles |
title_full_unstemmed | Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles |
title_short | Engineering and Modulating Functional Cyanobacterial CO(2)-Fixing Organelles |
title_sort | engineering and modulating functional cyanobacterial co(2)-fixing organelles |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5996877/ https://www.ncbi.nlm.nih.gov/pubmed/29922315 http://dx.doi.org/10.3389/fpls.2018.00739 |
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