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A step forward to improve recombinant protein production in Pichia pastoris : from specific growth rate effect on protein secretion to carbon-starving conditions as advanced strategy.
Garcia Ortega, Xavier
Adelantado Vallvé, Núria
Ferrer i Alegre, Pau (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Montesinos Seguí, José Luis (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Valero Barranco, Francisco (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)

Data: 2016
Resum: The recombinant protein production platform based on the GAP promoter and Pichia pastoris as a host has become a very promising system from an industrial point of view. The need for highly productive bioprocesses gives grounds for the optimization of fermentation strategies maximizing yields and/or productivities, which are often associated with cell growth. Coherent with previous studies, a positive effect of high specific growth rate (μ) on the productivity was observed in carbon-limited chemostat cultivations secreting an antibody fragment. Notably, no significant impact of this factor could be observed in the balance intra- and extracellular of the product. Accordingly, fed-batch cultures operating at a constant high μ were conducted. Furthermore, short carbon-starving periods were introduced along the exponential substrate feeding phase. Strikingly, it was observed an important increase of specific production rate (qP) during such short carbon-starving periods in relation to the exponential substrate feeding intervals. Therefore, the application of carbon-starving periods as an innovative operational strategy was proposed, resulting into increments up to 50% of both yields and total production. The implementation of the proposed substrate feeding profiles should be complementary to cell engineering strategies to improve the relation qP vs μ, thereby enhancing the overall bioprocess efficiency.
Nota: Número d'acord de subvenció MINECO/CTQ2013-42391-R
Nota: Número d'acord de subvenció AGAUR/2014/SGR-452
Drets: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial 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. Creative Commons
Llengua: Anglès
Document: article ; recerca ; acceptedVersion
Matèria: Antibody fragment ; Carbon starvation ; Fed-batch ; GAP promoter ; Heterologous protein production ; Optimized feeding strategy ; Pichia pastoris ; Protein secretion
Publicat a: Process biochemistry, Vol. 51, Issue 3 (March 2016) , p., ISSN 1359-5113

DOI: 10.1016/j.procbio.2016.02.018

Disponible a partir de: 2017-03-31

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