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  Improved recombinant expression and purification of functional plant Rubisco

Wilson, R., Thieulin-Pardo, G., Hartl, F.-U., & Hayer-Hartl, M. (2019). Improved recombinant expression and purification of functional plant Rubisco. FEBS Letters, 593(6), 611-621. doi:10.1002/1873-3468.13352.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0003-E19E-D Version Permalink: http://hdl.handle.net/21.11116/0000-0003-E19F-C
Genre: Journal Article

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Wilson_et_al-2019-FEBS_Letters.pdf (Publisher version), 529KB
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© 2019 The Authors.

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 Creators:
Wilson, Robert1, Author              
Thieulin-Pardo, Gabriel1, Author              
Hartl, Franz-Ulrich2, Author              
Hayer-Hartl, Manajit1, Author              
Affiliations:
1Hayer-Hartl, Manajit / Chaperonin-assisted Protein Folding, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565153              
2Hartl, Franz-Ulrich / Cellular Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565152              

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Free keywords: ASSEMBLY CHAPERONE; PROTEIN; ACTIVASE; CARBOXYLASE/OXYGENASE; GENE; PHOTOSYNTHESIS; IDENTIFICATION; BIOGENESIS; ENZYMES; RBCXBiochemistry & Molecular Biology; Biophysics; Cell Biology; assembly; folding; molecular chaperones; protein expression; Rubisco; Rubisco activase;
 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 similar to 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.

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Language(s): eng - English
 Dates: 2019
 Publication Status: Published in print
 Pages: 11
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: ISI: 000462186700006
DOI: 10.1002/1873-3468.13352
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Project name : -
Grant ID : SFB1035
Funding program : -
Funding organization : Deutsche Forschungsgemeinschaft (DFG)

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Title: FEBS Letters
  Other : FEBS Lett.
Source Genre: Journal
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Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 593 (6) Sequence Number: - Start / End Page: 611 - 621 Identifier: ISSN: 0014-5793
CoNE: https://pure.mpg.de/cone/journals/resource/954925399501