Improved recombinant expression and purification of functional plant Rubisco

FEBS Letters
R H WilsonM Hayer-Hartl

Abstract

Improving the performance of the key photosynthetic enzyme Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) by protein engineering is a critical strategy for increasing crop yields. The extensive chaperone requirement of plant Rubisco for folding and assembly has long been an impediment to this goal. Production of plant Rubisco in Escherichia coli requires the coexpression of the chloroplast chaperonin and four assembly factors. Here, we demonstrate that simultaneous expression of Rubisco and chaperones from a T7 promotor produces high levels of functional enzyme. Expressing the small subunit of Rubisco with a C-terminal hexahistidine-tag further improved assembly, resulting in a ~ 12-fold higher yield than the previously published procedure. The expression system described here provides a platform for the efficient production and engineering of plant Rubisco.

References

Oct 26, 2005·Photosynthesis Research·Archie R Portis
Nov 9, 2005·Methods in Enzymology·Rohan T BakerPhilip G Board
Jun 19, 2007·Cell·Sandra SaschenbreckerManajit Hayer-Hartl
Jan 16, 2010·Nature·Cuimin LiuManajit Hayer-Hartl
Oct 21, 2010·Methods in Molecular Biology·David S KubienHeather J Kane
Jul 19, 2011·Nature Structural & Molecular Biology·Andreas BracherManajit Hayer-Hartl
Nov 8, 2011·Nature Structural & Molecular Biology·Mathias StotzManajit Hayer-Hartl
Nov 20, 2012·Journal of Experimental Botany·Martin A J ParrySpencer M Whitney
Oct 4, 2014·The Plant Journal : for Cell and Molecular Biology·Leila FeizDavid B Stern
Jul 1, 2015·Proceedings of the National Academy of Sciences of the United States of America·Donald R OrtXin Guang Zhu
Aug 4, 2015·Nature Structural & Molecular Biology·Thomas HauserManajit Hayer-Hartl
Nov 17, 2015·Nature Communications·Yi-Chin Candace TsaiOliver Mueller-Cajar
Mar 2, 2016·Scientific Reports·Robert H WilsonSpencer M Whitney
May 5, 2016·Molecular Plant·Bettina Bölter, Jürgen Soll
Oct 4, 2016·Current Opinion in Chemical Biology·Martin Hagemann, Hermann Bauwe
Jan 27, 2017·Annual Review of Plant Biology·Andreas BracherManajit Hayer-Hartl
Apr 27, 2017·Frontiers in Molecular Biosciences·Javaid Y BhatManajit Hayer-Hartl
Jun 6, 2017·Frontiers in Molecular Biosciences·Oliver Mueller-Cajar
Aug 27, 2017·Current Opinion in Biotechnology·Tobias J Erb, Jan Zarzycki
Nov 16, 2017·Plant Molecular Biology·Ragupathi Nagarajan, Kulvinder S Gill
Feb 6, 2018·The Plant Journal : for Cell and Molecular Biology·Rikard FristedtRoberta Croce
Mar 29, 2018·Biochemistry·Robert H Wilson, Manajit Hayer-Hartl
Apr 7, 2018·Metabolic Engineering·Lennart Schada von BorzyskowskiTobias J Erb
Jan 11, 2019·Plant Physiology·Martin K M Engqvist, Kersten S Rabe
Feb 5, 2019·Journal of Experimental Botany·Armin Kubis, Arren Bar-Even

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Citations

Apr 20, 2019·Traffic·Tobias WunderOliver Mueller-Cajar
Feb 16, 2020·Journal of Experimental Botany·Xin-Guang ZhuSusanne von Caemmerer
Oct 30, 2019·International Journal of Molecular Sciences·Yu Zhou, Spencer Whitney
Sep 20, 2020·The Journal of Biological Chemistry·Jediael NgOliver Mueller-Cajar
Feb 13, 2021·The Plant Journal : for Cell and Molecular Biology·Myat T LinMaureen R Hanson
Jul 25, 2020·Journal of Plant Physiology·Susanne von Caemmerer
Oct 9, 2021·Biochemical Society Transactions·Concepción IñiguezJeroni Galmés

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Methods Mentioned

BETA
protein assay
enzymatic assay

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