Please use this identifier to cite or link to this item: https://hdl.handle.net/2445/222164
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dc.contributor.authorFerrando Núñez, Jordi-
dc.contributor.authorMiñana i Galbis, David-
dc.contributor.authorPicart, Pere-
dc.date.accessioned2025-07-11T09:53:12Z-
dc.date.available2025-07-11T09:53:12Z-
dc.date.issued2024-06-25-
dc.identifier.issn1661-6596-
dc.identifier.urihttps://hdl.handle.net/2445/222164-
dc.description.abstractAchieving 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.extent21 p.-
dc.format.mimetypeapplication/pdf-
dc.language.isoeng-
dc.publisherMDPI-
dc.relation.isformatofReproducció del document publicat a: https://doi.org/https://doi.org/10.3390/ijms25136957-
dc.relation.ispartofInternational Journal of Molecular Sciences, 2024, vol. 13, p. 6957-
dc.relation.urihttps://doi.org/https://doi.org/10.3390/ijms25136957-
dc.rightscc-by (c) Jordi Ferrando et al., 2024-
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/-
dc.sourceArticles publicats en revistes (Biologia, Sanitat i Medi Ambient)-
dc.subject.classificationGenètica bacteriana-
dc.subject.classificationBacteris-
dc.subject.classificationBiopolímers-
dc.subject.otherBacterial genetics-
dc.subject.otherBacteria-
dc.subject.otherBiopolymers-
dc.titleThe Construction of an Environmentally Friendly Super-Secreting Strain of Bacillus subtilis through Systematic Modulation of Its Secretory Pathway Using the CRISPR-Cas9 System-
dc.typeinfo:eu-repo/semantics/article-
dc.typeinfo:eu-repo/semantics/publishedVersion-
dc.identifier.idgrec749241-
dc.date.updated2025-07-11T09:53:12Z-
dc.rights.accessRightsinfo:eu-repo/semantics/openAccess-
Appears in Collections:Articles publicats en revistes (Biologia, Sanitat i Medi Ambient)

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