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Biotechnology and Bioengineering
Article . 2024 . Peer-reviewed
License: CC BY
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Adaptive laboratory evolution in a novel parallel shaken pH‐auxostat

Authors: Burak Sarikaya; Hendrik Bück; Gino Pohen; Filipe Rodrigues; Karsten Günster; Katrin Wefelmeier; Katharina Miebach; +2 Authors

Adaptive laboratory evolution in a novel parallel shaken pH‐auxostat

Abstract

AbstractAdaptive laboratory evolution (ALE) is a widely used microbial strain development and optimization method. ALE experiments, to select for faster‐growing strains, are commonly performed as serial batch cultivations in shake flasks, serum bottles, or microtiter plates or as continuous cultivations in bioreactors on a laboratory scale. To combine the advantages of higher throughput in parallel shaken cultures with continuous fermentations for conducting ALE experiments, a new Continuous parallel shaken pH‐auxostat (CPA) was developed. The CPA consists of six autonomous parallel shaken cylindrical reactors, equipped with real‐time pH control of the culture medium. The noninvasive pH measurement and control are realized by biocompatible pH sensor spots and a programmable pump module, to adjust the dilution rate of fresh medium for each reactor separately. Two different strains of the methylotrophic yeast Ogataea polymorpha were used as microbial model systems for parallel chemostat and pH‐auxostat cultivations. During cultivation, the medium is acidified by the microbial activity of the yeast. For pH‐auxostat cultivations, the growth‐dependent acidification triggers the addition of fresh feed medium into the reactors, leading to a pH increase and thereby to the control of the pH to a predetermined set value. By controlling the pH to a predetermined set value, the dilution rate of the continuous cultivation is adjusted to values close to the washout point, in the range of the maximum specific growth rate of the yeast. The pH control was optimized by conducting a step‐response experiment and obtaining tuned PI controller parameters by the Chien‐Hrones‐Reswick (CHR) PID tuning method. Two pH‐auxostat cultivations were performed with two different O. polymorpha strains at high dilution rates for up to 18 days. As a result, up to 4.8‐fold faster‐growing strains were selected. The increased specific maximum growth rates of the selected strains were confirmed in subsequent batch cultivations.

Country
Germany
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Keywords

info:eu-repo/classification/ddc/570, 570, continuous cultivation, pH sensor spot, Hydrogen-Ion Concentration, Culture Media, Bioreactors, PID tuning, PID control, Saccharomycetales, Fermentation, pH‐auxostat, Directed Molecular Evolution, adaptive laboratory evolution

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    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
3
Top 10%
Average
Average
Green
hybrid