Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis

Metabolic flux analysis based on 13 C-derived constraints has proved to be a powerful method for quantitative physiological characterisation of one of the most extensively used microbial cell factory platforms, Pichia pastoris (syn. Komagataella spp.). Nonetheless, the reduced number of carbon atoms...

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Autores: Tomàs Gamisans, Màrius|||0000-0002-6076-6716, Ødum, Anders Sebastian Rosenkrans, Workman, Mhairi, Ferrer, Pau|||0000-0002-5287-4127, Albiol i Sala, Joan|||0000-0001-5626-429X
Tipo de recurso: artículo
Fecha de publicación:2019
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:281356
Acceso en línea:https://ddd.uab.cat/record/281356
https://dx.doi.org/urn:doi:10.1016/j.nbt.2019.01.005
Access Level:acceso abierto
Palabra clave:C-13-based meabolic flux analysis
Pichia pastoris
Komagataella spp
Glycerol
Genome-scale metabolic model
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spelling Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysisTomàs Gamisans, Màrius|||0000-0002-6076-6716Ødum, Anders Sebastian RosenkransWorkman, MhairiFerrer, Pau|||0000-0002-5287-4127Albiol i Sala, Joan|||0000-0001-5626-429XC-13-based meabolic flux analysisPichia pastorisKomagataella sppGlycerolGenome-scale metabolic modelMetabolic flux analysis based on 13 C-derived constraints has proved to be a powerful method for quantitative physiological characterisation of one of the most extensively used microbial cell factory platforms, Pichia pastoris (syn. Komagataella spp.). Nonetheless, the reduced number of carbon atoms and the symmetry of the glycerol molecule has hampered the comprehensive determination of metabolic fluxes when used as the labelled C-source. Moreover, metabolic models typically used for 13 C-based flux balance analysis may be incomplete or misrepresent the actual metabolic network. To circumvent these limitations, we reduced the genome-scale metabolic model iMT1026-v3.0 into a core model and used it for the iterative fitting of metabolic fluxes to the measured mass isotope distribution of proteinogenic amino acids obtained after fractional 13 C labelling of cells with [1,3- 13 C]-glycerol. This workflow allows reliable estimates to be obtained for in vivo fluxes in P. pastoris cells growing on glycerol as sole carbon source, as well as revising previous assumptions concerning its metabolic operation, such as alternative metabolic branches, calculation of energetic parameters and proposed specific cofactor utilisation. 22019-01-0120192019-01-01Articlehttp://purl.org/coar/resource_type/c_6501AMhttp://purl.org/coar/version/c_ab4af688f83e57aainfo:eu-repo/semantics/articleapplication/pdfhttps://ddd.uab.cat/record/281356https://dx.doi.org/urn:doi:10.1016/j.nbt.2019.01.005reponame:Dipòsit Digital de Documents de la UABinstname:Universitat Autònoma de BarcelonaInglésengMinisterio de Economía y Competitividad https://doi.org/10.13039/501100003329 CTQ2013-42391-RAgencia Estatal de Investigación https://doi.org/10.13039/501100011033 CTQ2016-74959-RAgència de Gestió d'Ajuts Universitaris i de Recerca https://doi.org/10.13039/501100003030 2014/SGR-452Ministerio de Educación, Cultura y Deporte https://doi.org/10.13039/501100003176 FPU12/06185open accesshttp://purl.org/coar/access_right/c_abf2Aquest 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/info:eu-repo/semantics/openAccessoai:ddd.uab.cat:2813562026-06-06T12:50:31Z
dc.title.none.fl_str_mv Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
title Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
spellingShingle Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
Tomàs Gamisans, Màrius|||0000-0002-6076-6716
C-13-based meabolic flux analysis
Pichia pastoris
Komagataella spp
Glycerol
Genome-scale metabolic model
title_short Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
title_full Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
title_fullStr Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
title_full_unstemmed Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
title_sort Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis
dc.creator.none.fl_str_mv Tomàs Gamisans, Màrius|||0000-0002-6076-6716
Ødum, Anders Sebastian Rosenkrans
Workman, Mhairi
Ferrer, Pau|||0000-0002-5287-4127
Albiol i Sala, Joan|||0000-0001-5626-429X
author Tomàs Gamisans, Màrius|||0000-0002-6076-6716
author_facet Tomàs Gamisans, Màrius|||0000-0002-6076-6716
Ødum, Anders Sebastian Rosenkrans
Workman, Mhairi
Ferrer, Pau|||0000-0002-5287-4127
Albiol i Sala, Joan|||0000-0001-5626-429X
author_role author
author2 Ødum, Anders Sebastian Rosenkrans
Workman, Mhairi
Ferrer, Pau|||0000-0002-5287-4127
Albiol i Sala, Joan|||0000-0001-5626-429X
author2_role author
author
author
author
dc.subject.none.fl_str_mv C-13-based meabolic flux analysis
Pichia pastoris
Komagataella spp
Glycerol
Genome-scale metabolic model
topic C-13-based meabolic flux analysis
Pichia pastoris
Komagataella spp
Glycerol
Genome-scale metabolic model
description Metabolic flux analysis based on 13 C-derived constraints has proved to be a powerful method for quantitative physiological characterisation of one of the most extensively used microbial cell factory platforms, Pichia pastoris (syn. Komagataella spp.). Nonetheless, the reduced number of carbon atoms and the symmetry of the glycerol molecule has hampered the comprehensive determination of metabolic fluxes when used as the labelled C-source. Moreover, metabolic models typically used for 13 C-based flux balance analysis may be incomplete or misrepresent the actual metabolic network. To circumvent these limitations, we reduced the genome-scale metabolic model iMT1026-v3.0 into a core model and used it for the iterative fitting of metabolic fluxes to the measured mass isotope distribution of proteinogenic amino acids obtained after fractional 13 C labelling of cells with [1,3- 13 C]-glycerol. This workflow allows reliable estimates to be obtained for in vivo fluxes in P. pastoris cells growing on glycerol as sole carbon source, as well as revising previous assumptions concerning its metabolic operation, such as alternative metabolic branches, calculation of energetic parameters and proposed specific cofactor utilisation.
publishDate 2019
dc.date.none.fl_str_mv 2
2019-01-01
2019
2019-01-01
dc.type.none.fl_str_mv Article
http://purl.org/coar/resource_type/c_6501
AM
http://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://ddd.uab.cat/record/281356
https://dx.doi.org/urn:doi:10.1016/j.nbt.2019.01.005
url https://ddd.uab.cat/record/281356
https://dx.doi.org/urn:doi:10.1016/j.nbt.2019.01.005
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Economía y Competitividad https://doi.org/10.13039/501100003329 CTQ2013-42391-R
Agencia Estatal de Investigación https://doi.org/10.13039/501100011033 CTQ2016-74959-R
Agència de Gestió d'Ajuts Universitaris i de Recerca https://doi.org/10.13039/501100003030 2014/SGR-452
Ministerio de Educación, Cultura y Deporte https://doi.org/10.13039/501100003176 FPU12/06185
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
https://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
https://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Dipòsit Digital de Documents de la UAB
instname:Universitat Autònoma de Barcelona
instname_str Universitat Autònoma de Barcelona
reponame_str Dipòsit Digital de Documents de la UAB
collection Dipòsit Digital de Documents de la UAB
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