Design and Operation of Microbioreactor Systems for Screening and Process Development

Lattermann, Clemens; Büchs, Jochen

doi:10.1002/9783527683369.ch2

Cited by 13 publications

(15 citation statements)

References 119 publications

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“…For the characterization of operating conditions and identification of their possible transition into the unfavorable OP range, specific power input measurements (Büchs et al, ) and mixing time evaluations have been employed to date (Lattermann & Büchs, , pp. 35–76; Tan et al, ).…”

Section: Resultsmentioning

confidence: 99%

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Reassessing the out‐of‐phase phenomenon in shake flasks by evaluating the angle‐dependent liquid distribution relative to the direction of the centrifugal acceleration

Azizan

Sieben

Wandrey

et al. 2019

Biotech & Bioengineering

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Shake flasks are still the most relevant experimental tool in the development of viscous fermentation processes. The phase number, which defines the onset of the unfavorable out‐of‐phase (OP) phenomenon in shake flasks, was previously defined via specific power input measurements. In the OP state, the bulk liquid no longer follows the orbital movement of the imposed centrifugal force, which is for example, detrimental to oxygen transfer. In this study, an optical fluorescence technique was used to measure the three‐dimensional liquid distribution in shake flasks. Four new optically derived evaluation criteria for the phase transition between the in‐phase and OP condition were established: (a) thickness of the liquid film left on the glass wall by the rotating bulk liquid, (b) relative slope of the leading edge of bulk liquid (LB) lines, (c) trend of the angular position of LB, and (d) very high angular position of the leading edge. In contrast to the previously applied power input measurements, the new optical evaluation criteria describe the phase transition in greater detailed. Instead of Ph = 1.26, a less conservative value of Ph = 0.91 is now suggested for the phase transfer, which implies a broader operating window for shake flask cultivations with higher viscosities.

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Section: Resultsmentioning

confidence: 99%

“…Later work by Tan, Eberhard, and Büchs () supported the concept of the Phase number. They investigated the mixing time at different operating conditions and found that the mixing time drastically increases at decreasing Phase numbers approaching the critical value of 1.26 (Lattermann & Büchs, , pp. 35–76).…”

Section: Introductionmentioning

confidence: 99%

Reassessing the out‐of‐phase phenomenon in shake flasks by evaluating the angle‐dependent liquid distribution relative to the direction of the centrifugal acceleration

Azizan

Sieben

Wandrey

et al. 2019

Biotech & Bioengineering

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“…Different microbioreactor (MBR) systems have been developed in the past, enabling the tight monitoring and controlling of microbial cultivation experiments at elevated throughput . Next to microtiter plate (MTP) based systems, microscale bubble‐column and stirred tank reactor‐based MBR technologies with reaction volumes in the mL‐range were developed .…”

Section: Introductionmentioning

confidence: 99%

Less Sacrifice, More Insight: Repeated Low‐Volume Sampling of Microbioreactor Cultivations Enables Accelerated Deep Phenotyping of Microbial Strain Libraries

Hemmerich

Tenhaef

Steffens

et al. 2018

Biotechnology Journal

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With modern genetic engineering tools, high number of potentially improved production strains can be created in a short time. This results in a bottleneck in the succeeding step of bioprocess development, which can be handled by accelerating quantitative microbial phenotyping. Miniaturization and automation are key technologies to achieve this goal. In this study, a novel workflow for repeated low‐volume sampling of BioLector‐based cultivation setups is presented. Six samples of 20 μL each can be taken automatically from shaken 48‐well microtiter plates without disturbing cell population growth. The volume is sufficient for quantification of substrate and product concentrations by spectrophotometric‐based enzyme assays. From transient concentration data and replicate cultures, valid performance indicators (titers, rates, yields) are determined through process modeling and random error propagation analysis. Practical relevance of the workflow is demonstrated with a set of five genome‐reduced Corynebacterium glutamicum strains that are engineered for Sec‐mediated heterologous cutinase secretion. Quantitative phenotyping of this strain library led to the identification of a strain with a 1.6‐fold increase in cutinase yield. The prophage‐free strain carries combinatorial deletions of three gene clusters (Δ3102‐3111, Δ3263‐3301, and Δ3324‐3345) of which the last two likely contain novel target genes to foster rational engineering of heterologous cutinase secretion in C. glutamicum.

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“…Instead, small-scale tools such as shake flasks and microtiter plates (MTPs) have long been the tools of choice for such tasks. A comprehensive discussion of these cultivation systems and systematic strategies for scale-up can be found elsewhere [12,20,21,27]. Due to technical restrictions, many of the available systems remain limited to batch cultivation; therefore, batch mode screening is a frequently used standard strategy.…”

Section: Introductionmentioning

confidence: 99%

Parallelized microscale fed-batch cultivation in online-monitored microtiter plates: implications of media composition and feed strategies for process design and performance

Morschett

Jansen

Neuendorf

et al. 2020

Journal of Industrial Microbiology and Biotechnology

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Limited throughput represents a substantial drawback during bioprocess development. In recent years, several commercial microbioreactor systems have emerged featuring parallelized experimentation with optical monitoring. However, many devices remain limited to batch mode and do not represent the fed-batch strategy typically applied on an industrial scale. A workflow for 32-fold parallelized microscale cultivation of protein secreting Corynebacterium glutamicum in microtiter plates incorporating online monitoring, pH control and feeding was developed and validated. Critical interference of the essential media component protocatechuic acid with pH measurement was revealed, but was effectively resolved by 80% concentration reduction without affecting biological performance. Microfluidic pH control and feeding (pulsed, constant and exponential) were successfully implemented: Whereas pH control improved performance only slightly, feeding revealed a much higher optimization potential. Exponential feeding with µ = 0.1 h −1 resulted in the highest product titers. In contrast, other performance indicators such as biomass-specific or volumetric productivity resulted in different optimal feeding regimes.

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Design and Operation of Microbioreactor Systems for Screening and Process Development

Cited by 13 publications

References 119 publications

Reassessing the out‐of‐phase phenomenon in shake flasks by evaluating the angle‐dependent liquid distribution relative to the direction of the centrifugal acceleration

Reassessing the out‐of‐phase phenomenon in shake flasks by evaluating the angle‐dependent liquid distribution relative to the direction of the centrifugal acceleration

Less Sacrifice, More Insight: Repeated Low‐Volume Sampling of Microbioreactor Cultivations Enables Accelerated Deep Phenotyping of Microbial Strain Libraries

Parallelized microscale fed-batch cultivation in online-monitored microtiter plates: implications of media composition and feed strategies for process design and performance

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