Title
Rationale-based selection of optimal operating strategies and gene dosage impact on recombinant protein production in Komagataella phaffii (Pichia pastoris)
Date Issued
01 March 2020
Access level
open access
Resource Type
journal article
Author(s)
Garrigós-Martínez J.
Sánchez-Farrando M.
Valero F.
Garcia-Ortega X.
Montesinos-Seguí J.L.
Universitat Autònoma de Barcelona
Publisher(s)
John Wiley and Sons Ltd
Abstract
Its features as a microbial and eukaryotic organism have turned Komagataella phaffii (Pichia pastoris) into an emerging cell factory for recombinant protein production (RPP). As a key step of the bioprocess development, this work aimed to demonstrate the importance of tailor designing the cultivation strategy according to the production kinetics of the cell factory. For this purpose, K. phaffii clones constitutively expressing (PGAP) Candida rugosa lipase 1 (Crl1) with different gene dosage were used as models in continuous and fed-batch cultures. Production parameters were much greater with a multicopy clone (MCC) than with the single-copy clone (SCC). Regarding production kinetics, the specific product generation rate (qP) increased linearly with increasing specific growth rate (µ) in SCC; by contrast, qP exhibited saturation in MCC. A transcriptional analysis in chemostat cultures suggested the presence of eventual post-transcriptional bottlenecks in MCC. After the strain characterization, in order to fulfil overall development of the bioprocess, the performance of both clones was also evaluated in fed-batch mode. Strikingly, different optimal strategies were determined for both models due to the different production kinetic patterns observed as a trade-off for product titre, yields and productivity. The combined effect of gene dosage and adequate µ enables rational process development with a view to optimize K. phaffii RPP bioprocesses.
Start page
315
End page
327
Volume
13
Issue
2
Language
English
OCDE Knowledge area
Bioquímica, Biología molecular
Ingeniería de procesos
Scopus EID
2-s2.0-85074580586
PubMed ID
Source
Microbial Biotechnology
ISSN of the container
17517907
Sponsor(s)
This work was funded by MINECO and FEDER under Project CTQ2016‐74959‐R. The authors’ group is member of 2017‐SGR‐1462 and the Reference Network in Biotechnology (XRB) of Generalitat de Catalunya. M.A. Nieto‐Taype acknowledges award by the National Council of Science, Technology and Technological Innovation (CONCYTEC) through its executing unit, the National Fund for Scientific, Technological and Technological Innovation Development (FONDECYT) and J. Garrigós‐Martínez, a PIF scholarship from the Universitat Autònoma de Barcelona.
This work was funded by MINECO and FEDER under Project CTQ2016‐74959‐R.
Funding information This work was funded by MINECO and FEDER under Project CTQ2016-74959-R. This work was funded by MINECO and FEDER under Project CTQ2016-74959-R. The authors? group is member of?2017-SGR-1462 and the Reference Network in Biotechnology (XRB) of Generalitat de Catalunya. M.A. Nieto-Taype acknowledges award by the National Council of Science, Technology and Technological Innovation (CONCYTEC) through its executing unit, the National Fund for Scientific, Technological and Technological Innovation Development (FONDECYT) and J. Garrig?s-Mart?nez, a PIF scholarship from the Universitat Aut?noma de Barcelona.
Sources of information:
Directorio de Producción Científica
Scopus