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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 |
| Page(s): | 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 Computational Life Sciences (ICLS) Institute of Chemistry and Biotechnology (ICBT) |
| Appears in collections: | Publikationen Life Sciences und Facility Management |
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| File | Description | Size | Format | |
|---|---|---|---|---|
| 2020_Borcinova-etal_Production-secretion-dynamics-Penicillin-G-acylase.pdf | 1.2 MB | Adobe PDF |  View/Open |
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Borčinová, M., Raschmanová, H., Zamora, I., Looser, V., Marešová, H., Hirsch, S., Kyslík, P., & Kovar, K. (2020). Production and secretion dynamics of prokaryotic Penicillin G acylase in Pichia pastoris. Applied Microbiology and Biotechnology, 104(13), 5787–5800. https://doi.org/10.1007/s00253-020-10669-x
Borčinová, M. et al. (2020) ‘Production and secretion dynamics of prokaryotic Penicillin G acylase in Pichia pastoris’, Applied Microbiology and Biotechnology, 104(13), pp. 5787–5800. Available at: https://doi.org/10.1007/s00253-020-10669-x.
M. Borčinová et al., “Production and secretion dynamics of prokaryotic Penicillin G acylase in Pichia pastoris,” Applied Microbiology and Biotechnology, vol. 104, no. 13, pp. 5787–5800, 2020, doi: 10.1007/s00253-020-10669-x.
BORČINOVÁ, Martina, Hana RASCHMANOVÁ, Iwo ZAMORA, Verena LOOSER, Helena MAREŠOVÁ, Sven HIRSCH, Pavel KYSLÍK und Karin KOVAR, 2020. Production and secretion dynamics of prokaryotic Penicillin G acylase in Pichia pastoris. Applied Microbiology and Biotechnology. 2020. Bd. 104, Nr. 13, S. 5787–5800. DOI 10.1007/s00253-020-10669-x
Borčinová, Martina, Hana Raschmanová, Iwo Zamora, Verena Looser, Helena Marešová, Sven Hirsch, Pavel Kyslík, and Karin Kovar. 2020. “Production and Secretion Dynamics of Prokaryotic Penicillin G Acylase in Pichia Pastoris.” Applied Microbiology and Biotechnology 104 (13): 5787–5800. https://doi.org/10.1007/s00253-020-10669-x.
Borčinová, Martina, et al. “Production and Secretion Dynamics of Prokaryotic Penicillin G Acylase in Pichia Pastoris.” Applied Microbiology and Biotechnology, vol. 104, no. 13, 2020, pp. 5787–800, https://doi.org/10.1007/s00253-020-10669-x.
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