Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-21535
Publication type: Article in scientific journal
Type of review: Peer review (publication)
Title: Production and secretion dynamics of prokaryotic Penicillin G acylase in Pichia pastoris
Authors: Borčinová, Martina
Raschmanová, Hana
Zamora, Iwo
Looser, Verena
Marešová, Helena
Hirsch, Sven
Kyslík, Pavel
Kovar, Karin
et. al: No
DOI: 10.1007/s00253-020-10669-x
10.21256/zhaw-21535
Published in: Applied Microbiology and Biotechnology
Volume(Issue): 104
Issue: 13
Pages: 5787
Pages to: 5800
Issue Date: 2020
Publisher / Ed. Institution: Springer
ISSN: 0175-7598
1432-0614
Language: English
Subjects: Pichia pastoris; Penicillin G acylase; Specific rate of product formation; Process optimisation; Secretion of a heterologous protein; Fedbatch bioreactor cultivation
Subject (DDC): 660.6: Biotechnology
Abstract: To take full advantage of recombinant Pichia pastoris (Komagataella phaffii) as a production system for heterologous proteins, the complex protein secretory process should be understood and optimised by circumventing bottlenecks. Typically, little or no attention has been paid to the fate of newly synthesised protein inside the cell, or its passage through the secretory pathway, and only the secreted product is measured. However, the system's productivity (i.e. specific production rate qp), includes productivity of secreted (qp,extra) plus intracellularly accumulated (qp,intra) protein. In bioreactor cultivations with P. pastoris producing penicillin G acylase, we studied the dynamics of product formation, i.e. both the specific product secretion (qp,extra) and product retention (qp,intra) as functions of time, as well as the kinetics, i.e. productivity in relation to specific growth rate (μ). Within the time course, we distinguished (I) an initial phase with constant productivities, where the majority of product accumulated inside the cells, and qp,extra, which depended on μ in a bell-shaped manner; (II) a transition phase, in which intracellular product accumulation reached a maximum and productivities (intracellular, extracellular, overall) were changing; (III) a new phase with constant productivities, where secretion prevailed over intracellular accumulation, qp,extra was linearly related to μ and was up to three times higher than in initial phase (I), while qp,intra decreased 4-6-fold. We show that stress caused by heterologous protein production induces cellular imbalance leading to a secretory bottleneck that ultimately reaches equilibrium. This understanding may help to develop cultivation strategies for improving protein secretion from P. pastoris.
Further description: Erworben im Rahmen der Schweizer Nationallizenzen (http://www.nationallizenzen.ch)
URI: https://digitalcollection.zhaw.ch/handle/11475/21535
Fulltext version: Published version
License (according to publishing contract): CC BY 4.0: Attribution 4.0 International
Departement: Life Sciences and Facility Management
Organisational Unit: Institute of Applied Simulation (IAS)
Institute of Chemistry and Biotechnology (ICBT)
Appears in collections:Publikationen Life Sciences und Facility Management

Files in This Item:
File Description SizeFormat 
2020_Borcinova-etal_Production-secretion-dynamics-Penicillin-G-acylase.pdf1.2 MBAdobe PDFThumbnail
View/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.