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oai:recercat.cat:20.500.12327/12402025-10-22T11:34:11Zcom_2072_4428com_2072_4427col_2072_487898

Selecting Subpopulations of High-Quality Protein Conformers among Conformational Mixtures of Recombinant Bovine MMP-9 Solubilized from Inclusion Bodies Carratalá, Jose Vicente Gifre-Renom, Laia Roca-Pinilla, Ramon Villaverde, Antonio Alomar, Oscar Garcia-Fruitos, Elena Sánchez, Julieta María Ferrer-Miralles, Neus Producció Animal Producció de Remugants A detailed workflow to analyze the physicochemical characteristics of mammalian matrix metalloproteinase (MMP-9) protein species obtained from protein aggregates (inclusion bodies—IBs) was followed. MMP-9 was recombinantly produced in the prokaryotic microbial cell factories Clearcoli (an engineered form of Escherichia coli) and Lactococcus lactis, mainly forming part of IBs and partially recovered under non-denaturing conditions. After the purification by affinity chromatography of solubilized MMP-9, four protein peaks were obtained. However, so far, the different conformational protein species forming part of IBs have not been isolated and characterized. Therefore, with the aim to link the physicochemical characteristics of the isolated peaks with their biological activity, we set up a methodological approach that included dynamic light scattering (DLS), circular dichroism (CD), and spectrofluorometric analysis confirming the separation of subpopulations of conformers with specific characteristics. In protein purification procedures, the detailed analysis of the individual physicochemical properties and the biological activity of protein peaks separated by chromatographic techniques is a reliable source of information to select the best-fitted protein populations. 2025-10-22T11:34:11Z 2025-10-22T11:34:11Z 2025-10-22T11:34:11Z 2021-03-16 info:eu-repo/semantics/article Carratalá, Jose Vicente, Laia Gifre-Renom, Ramon Roca-Pinilla, Antonio Villaverde, Anna Arís, Elena Garcia-Fruitós, Julieta María Sánchez, and Neus Ferrer-Miralles. 2021. "Selecting Subpopulations Of High-Quality Protein Conformers Among Conformational Mixtures Of Recombinant Bovine MMP-9 Solubilized From Inclusion Bodies". International Journal Of Molecular Sciences 22 (6): 3020. doi:10.3390/ijms22063020. 1661-6596 http://hdl.handle.net/20.500.12327/1240 https://doi.org/10.3390/ijms22063020 eng International Journal of Molecular Sciences MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/AGL2015-00064-C02-01/ES/Validación del uso de las proteínas M-SAA3 y MMP-9 en la mejora del secado de la vaca de leche y optimización de su dosis efectiva mediante su nanoestructuración/ MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/RTA2015-00064-C02-02/ES/Validación del uso de las proteínas M-SAA3 y MMP-9 en la mejora del secado de la vaca de leche y optimización de su dosis efectiva mediante su nanoestructuración/ http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess Attribution 4.0 International MDPI