Production and secretion dynamics of prokaryotic Penicillin G acylase in Pichia pastoris
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F20%3A00534894" target="_blank" >RIV/61388971:_____/20:00534894 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11310/20:10411686 RIV/60461373:22330/20:43920754
Result on the web
<a href="https://link.springer.com/article/10.1007/s00253-020-10669-x" target="_blank" >https://link.springer.com/article/10.1007/s00253-020-10669-x</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1007/s00253-020-10669-x" target="_blank" >10.1007/s00253-020-10669-x</a>
Alternative languages
Result language
angličtina
Original language name
Production and secretion dynamics of prokaryotic Penicillin G acylase in Pichia pastoris
Original language description
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 q(p)), includes productivity of secreted (q(p,extra)) plus intracellularly accumulated (q(p,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 (q(p,extra)) and product retention (q(p,intra)) as functions of time, as well as the kinetics, i.e. productivity in relation to specific growth rate (mu). Within the time course, we distinguished (I) an initial phase with constant productivities, where the majority of product accumulated inside the cells, and q(p,extra), which depended on mu 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, q(p,extra) was linearly related to mu and was up to three times higher than in initial phase (I), while q(p,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.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10606 - Microbiology
Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2020
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Applied Microbiology and Biotechnology
ISSN
0175-7598
e-ISSN
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Volume of the periodical
104
Issue of the periodical within the volume
13
Country of publishing house
DE - GERMANY
Number of pages
14
Pages from-to
5787-5800
UT code for WoS article
000533813200001
EID of the result in the Scopus database
2-s2.0-85084981824