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Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System

BACKGROUND: Protein downstream processing remains a challenge in protein production, especially in low yields of products, in spite of ensuring effective disruption of cell and separation of target proteins. It is complicated, expensive and time-consuming. Here, we report a novel nano-bio-purificati...

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Autores principales: Wu, Sha, Tian, Jiesheng, Xue, Xianle, Tang, Zongwen, Huang, Zekai, Hammock, Bruce D., Morisseau, Christophe, Li, Qing X., Xu, Ting
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Journal Experts 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10055543/
https://www.ncbi.nlm.nih.gov/pubmed/36993437
http://dx.doi.org/10.21203/rs.3.rs-2630343/v1
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author Wu, Sha
Tian, Jiesheng
Xue, Xianle
Tang, Zongwen
Huang, Zekai
Hammock, Bruce D.
Morisseau, Christophe
Li, Qing X.
Xu, Ting
author_facet Wu, Sha
Tian, Jiesheng
Xue, Xianle
Tang, Zongwen
Huang, Zekai
Hammock, Bruce D.
Morisseau, Christophe
Li, Qing X.
Xu, Ting
author_sort Wu, Sha
collection PubMed
description BACKGROUND: Protein downstream processing remains a challenge in protein production, especially in low yields of products, in spite of ensuring effective disruption of cell and separation of target proteins. It is complicated, expensive and time-consuming. Here, we report a novel nano-bio-purification system for producing recombinant proteins of interest with automatic purification from engineered bacteria. RESULTS: This system employed a complete genetic engineering downstream processing platform for proteins at low expression levels, referred to as a genetically encoded magnetic platform (GEMP). GEMP consists of four elements as follows. (1) A truncated phage lambda lysis cassette (RRz/Rz1) is controllable for lysis of Magnetospirillum gryphiswaldense MSR-1 (host cell). (2) A surface-expressed nuclease (NucA) is to reduce viscosity of homogenate by hydrolyzing long chain nucleic acids. (3) A bacteriogenic magnetic nanoparticle, known as magnetosome, allows an easy separation system in a magnetic field. (4) An intein realizes abscission of products (nanobodies against tetrabromobisphenol A) from magnetosome. CONCLUSIONS: In this work, removal of most impurities greatly simplified the subsequent purification procedure. The system also facilitated the bioproduction of nanomaterials. The developed platform can substantially simplify industrial protein production and reduce its cost.
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spelling pubmed-100555432023-03-30 Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System Wu, Sha Tian, Jiesheng Xue, Xianle Tang, Zongwen Huang, Zekai Hammock, Bruce D. Morisseau, Christophe Li, Qing X. Xu, Ting Res Sq Article BACKGROUND: Protein downstream processing remains a challenge in protein production, especially in low yields of products, in spite of ensuring effective disruption of cell and separation of target proteins. It is complicated, expensive and time-consuming. Here, we report a novel nano-bio-purification system for producing recombinant proteins of interest with automatic purification from engineered bacteria. RESULTS: This system employed a complete genetic engineering downstream processing platform for proteins at low expression levels, referred to as a genetically encoded magnetic platform (GEMP). GEMP consists of four elements as follows. (1) A truncated phage lambda lysis cassette (RRz/Rz1) is controllable for lysis of Magnetospirillum gryphiswaldense MSR-1 (host cell). (2) A surface-expressed nuclease (NucA) is to reduce viscosity of homogenate by hydrolyzing long chain nucleic acids. (3) A bacteriogenic magnetic nanoparticle, known as magnetosome, allows an easy separation system in a magnetic field. (4) An intein realizes abscission of products (nanobodies against tetrabromobisphenol A) from magnetosome. CONCLUSIONS: In this work, removal of most impurities greatly simplified the subsequent purification procedure. The system also facilitated the bioproduction of nanomaterials. The developed platform can substantially simplify industrial protein production and reduce its cost. American Journal Experts 2023-03-13 /pmc/articles/PMC10055543/ /pubmed/36993437 http://dx.doi.org/10.21203/rs.3.rs-2630343/v1 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
Wu, Sha
Tian, Jiesheng
Xue, Xianle
Tang, Zongwen
Huang, Zekai
Hammock, Bruce D.
Morisseau, Christophe
Li, Qing X.
Xu, Ting
Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System
title Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System
title_full Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System
title_fullStr Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System
title_full_unstemmed Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System
title_short Development of a Genetically Encoded Magnetic Platform in Magnetospirillum gryphiswaldense MSR-1 for Downstream Processing of Protein Expression System
title_sort development of a genetically encoded magnetic platform in magnetospirillum gryphiswaldense msr-1 for downstream processing of protein expression system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10055543/
https://www.ncbi.nlm.nih.gov/pubmed/36993437
http://dx.doi.org/10.21203/rs.3.rs-2630343/v1
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