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https://hdl.handle.net/2445/222164
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DC Field | Value | Language |
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dc.contributor.author | Ferrando Núñez, Jordi | - |
dc.contributor.author | Miñana i Galbis, David | - |
dc.contributor.author | Picart, Pere | - |
dc.date.accessioned | 2025-07-11T09:53:12Z | - |
dc.date.available | 2025-07-11T09:53:12Z | - |
dc.date.issued | 2024-06-25 | - |
dc.identifier.issn | 1661-6596 | - |
dc.identifier.uri | https://hdl.handle.net/2445/222164 | - |
dc.description.abstract | Achieving commercially significant yields of recombinant proteins in Bacillus subtilis requires</p><p>the optimization of its protein production pathway, including transcription, translation, folding,</p><p>and secretion. Therefore, in this study, our aim was to maximize the secretion of a reporter α-</p><p>amylase by overcoming potential bottlenecks within the secretion process one by one, using a clustered</p><p>regularly interspaced short palindromic repeat–Cas9 (CRISPR-Cas9) system. The strength of</p><p>single and tandem promoters was evaluated by measuring the relative α-amylase activity of AmyQ</p><p>integrated into the B. subtilis chromosome. Once a suitable promoter was selected, the expression</p><p>levels of amyQ were upregulated through the iterative integration of up to six gene copies, thus</p><p>boosting the α-amylase activity 20.9-fold in comparison with the strain harboring a single amyQ</p><p>gene copy. Next, α-amylase secretion was further improved to a 26.4-fold increase through the overexpression</p><p>of the extracellular chaperone PrsA and the signal peptide peptidase SppA. When the</p><p>final expression strain was cultivated in a 3 L fermentor for 90 h, the AmyQ production was enhanced</p><p>57.9-fold. The proposed strategy allows for the development of robust marker-free plasmidless</p><p>super-secreting B. subtilis strains with industrial relevance. | - |
dc.format.extent | 21 p. | - |
dc.format.mimetype | application/pdf | - |
dc.language.iso | eng | - |
dc.publisher | MDPI | - |
dc.relation.isformatof | Reproducció del document publicat a: https://doi.org/https://doi.org/10.3390/ijms25136957 | - |
dc.relation.ispartof | International Journal of Molecular Sciences, 2024, vol. 13, p. 6957 | - |
dc.relation.uri | https://doi.org/https://doi.org/10.3390/ijms25136957 | - |
dc.rights | cc-by (c) Jordi Ferrando et al., 2024 | - |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | - |
dc.source | Articles publicats en revistes (Biologia, Sanitat i Medi Ambient) | - |
dc.subject.classification | Genètica bacteriana | - |
dc.subject.classification | Bacteris | - |
dc.subject.classification | Biopolímers | - |
dc.subject.other | Bacterial genetics | - |
dc.subject.other | Bacteria | - |
dc.subject.other | Biopolymers | - |
dc.title | The Construction of an Environmentally Friendly Super-Secreting Strain of Bacillus subtilis through Systematic Modulation of Its Secretory Pathway Using the CRISPR-Cas9 System | - |
dc.type | info:eu-repo/semantics/article | - |
dc.type | info:eu-repo/semantics/publishedVersion | - |
dc.identifier.idgrec | 749241 | - |
dc.date.updated | 2025-07-11T09:53:12Z | - |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | - |
Appears in Collections: | Articles publicats en revistes (Biologia, Sanitat i Medi Ambient) |
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862709.pdf | 4.08 MB | Adobe PDF | View/Open |
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