Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses
Altres ajuts: acords transformatius de la UAB Altres ajuts: PIF scholarship from the Universitat Autònoma de Barcelona The yeast Komagataella phaffii (Pichia pastoris) is currently considered a versatile and highly efficient host for recombinant protein production (RPP). Interestingly, the regulate...
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ftuabarcelonapb:oai:ddd.uab.cat:289870 2024-09-15T17:41:03+00:00 Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses Sales-Vallverdú, Albert Gasset Franch, Arnau Requena-Moreno, Guillermo Valero Barranco, Francisco Montesinos SeguÃ, José Luis Garcia-Ortega, Xavier 2024 application/pdf https://ddd.uab.cat/record/289870 eng eng Agencia Estatal de Investigación PID2019-104666GB-I00 Agencia Estatal de Investigación PID2022-136936OB-I00 Agència de Gestió d'Ajuts Universitaris i de Recerca 2021/SGR-00143 Agència de Gestió d'Ajuts Universitaris i de Recerca FI-DGR2019 Microbial biotechnology Vol. 17, Issue 2 (February 2024), art. e14411 https://ddd.uab.cat/record/289870 urn:10.1111/1751-7915.14411 urn:oai:ddd.uab.cat:289870 urn:oai:egreta.uab.cat:publications/dd9e9628-ed90-412d-a906-5cf96454e77c urn:pure_id:412946141 urn:pmid:38376073 urn:pmcid:PMC10877992 urn:wos_id:001167092700001 urn:scopus_id:85185683651 urn:articleid:17517915v17n2e14411 urn:oai:pubmedcentral.nih.gov:10877992 urn:pmc-uid:10877992 open access Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades. https://creativecommons.org/licenses/by-nc-nd/4.0/ Lipase/genetics Oxygen/metabolism Pichia/metabolism Recombinant Proteins/metabolism Saccharomycetales Article 2024 ftuabarcelonapb 2024-08-06T14:30:51Z Altres ajuts: acords transformatius de la UAB Altres ajuts: PIF scholarship from the Universitat Autònoma de Barcelona The yeast Komagataella phaffii (Pichia pastoris) is currently considered a versatile and highly efficient host for recombinant protein production (RPP). Interestingly, the regulated application of specific stress factors as part of bioprocess engineering strategies has proven potential for increasing the production of recombinant products. This study aims to evaluate the impact of controlled oxygen-limiting conditions on the performance of K. phaffii bioprocesses for RPP in combination with the specific growth rate (μ) in fedbatch cultivations. In this work, Candida rugosa lipase 1 (Crl1) production, regulated by the constitutive GAP promoter, growing at different nominal μ (0.030, 0.065, 0.100 and 0.120h−1) under both normoxic and hypoxic conditions in carbon-limiting fed-batch cultures is analysed. Hypoxic fermentations were controlled at a target respiratory quotient (RQ) of 1.4, with excellent performance, using an innovative automated control based on the stirring rate as the manipulated variable developed during this study. The results conclude that oxygen limitation positively affects bioprocess efficiency under all growing conditions compared. The shift from respiratory to respiro-fermentative metabolism increases bioprocess productivity by up to twofold for the specific growth rates evaluated. Moreover, the specific product generation rate (qp) increases linearly with μ, regardless of oxygen availability. Furthermore, this hypoxic boosting effect was also observed in the production of Candida antarctica lipase B (CalB) and pro-Rhizopus oryzae lipase (proRol), thus proving the synergic effect of kinetic and physiological stress control. Finally, the Crl1production scale-up was conducted successfully, confirming the strategy's scalability and the robustness of the results obtained at the bench-scale level. Article in Journal/Newspaper Antarc* Antarctica Universitat Autònoma de Barcelona: Dipòsit Digital de Documents de la UAB |
institution |
Open Polar |
collection |
Universitat Autònoma de Barcelona: Dipòsit Digital de Documents de la UAB |
op_collection_id |
ftuabarcelonapb |
language |
English |
topic |
Lipase/genetics Oxygen/metabolism Pichia/metabolism Recombinant Proteins/metabolism Saccharomycetales |
spellingShingle |
Lipase/genetics Oxygen/metabolism Pichia/metabolism Recombinant Proteins/metabolism Saccharomycetales Sales-Vallverdú, Albert Gasset Franch, Arnau Requena-Moreno, Guillermo Valero Barranco, Francisco Montesinos SeguÃ, José Luis Garcia-Ortega, Xavier Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses |
topic_facet |
Lipase/genetics Oxygen/metabolism Pichia/metabolism Recombinant Proteins/metabolism Saccharomycetales |
description |
Altres ajuts: acords transformatius de la UAB Altres ajuts: PIF scholarship from the Universitat Autònoma de Barcelona The yeast Komagataella phaffii (Pichia pastoris) is currently considered a versatile and highly efficient host for recombinant protein production (RPP). Interestingly, the regulated application of specific stress factors as part of bioprocess engineering strategies has proven potential for increasing the production of recombinant products. This study aims to evaluate the impact of controlled oxygen-limiting conditions on the performance of K. phaffii bioprocesses for RPP in combination with the specific growth rate (μ) in fedbatch cultivations. In this work, Candida rugosa lipase 1 (Crl1) production, regulated by the constitutive GAP promoter, growing at different nominal μ (0.030, 0.065, 0.100 and 0.120h−1) under both normoxic and hypoxic conditions in carbon-limiting fed-batch cultures is analysed. Hypoxic fermentations were controlled at a target respiratory quotient (RQ) of 1.4, with excellent performance, using an innovative automated control based on the stirring rate as the manipulated variable developed during this study. The results conclude that oxygen limitation positively affects bioprocess efficiency under all growing conditions compared. The shift from respiratory to respiro-fermentative metabolism increases bioprocess productivity by up to twofold for the specific growth rates evaluated. Moreover, the specific product generation rate (qp) increases linearly with μ, regardless of oxygen availability. Furthermore, this hypoxic boosting effect was also observed in the production of Candida antarctica lipase B (CalB) and pro-Rhizopus oryzae lipase (proRol), thus proving the synergic effect of kinetic and physiological stress control. Finally, the Crl1production scale-up was conducted successfully, confirming the strategy's scalability and the robustness of the results obtained at the bench-scale level. |
format |
Article in Journal/Newspaper |
author |
Sales-Vallverdú, Albert Gasset Franch, Arnau Requena-Moreno, Guillermo Valero Barranco, Francisco Montesinos SeguÃ, José Luis Garcia-Ortega, Xavier |
author_facet |
Sales-Vallverdú, Albert Gasset Franch, Arnau Requena-Moreno, Guillermo Valero Barranco, Francisco Montesinos SeguÃ, José Luis Garcia-Ortega, Xavier |
author_sort |
Sales-Vallverdú, Albert |
title |
Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses |
title_short |
Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses |
title_full |
Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses |
title_fullStr |
Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses |
title_full_unstemmed |
Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses |
title_sort |
synergic kinetic and physiological control to improve the efficiency of komagataella phaffii recombinant protein production bioprocesses |
publishDate |
2024 |
url |
https://ddd.uab.cat/record/289870 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
Agencia Estatal de Investigación PID2019-104666GB-I00 Agencia Estatal de Investigación PID2022-136936OB-I00 Agència de Gestió d'Ajuts Universitaris i de Recerca 2021/SGR-00143 Agència de Gestió d'Ajuts Universitaris i de Recerca FI-DGR2019 Microbial biotechnology Vol. 17, Issue 2 (February 2024), art. e14411 https://ddd.uab.cat/record/289870 urn:10.1111/1751-7915.14411 urn:oai:ddd.uab.cat:289870 urn:oai:egreta.uab.cat:publications/dd9e9628-ed90-412d-a906-5cf96454e77c urn:pure_id:412946141 urn:pmid:38376073 urn:pmcid:PMC10877992 urn:wos_id:001167092700001 urn:scopus_id:85185683651 urn:articleid:17517915v17n2e14411 urn:oai:pubmedcentral.nih.gov:10877992 urn:pmc-uid:10877992 |
op_rights |
open access Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades. https://creativecommons.org/licenses/by-nc-nd/4.0/ |
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1810487124440383488 |