Permeability of currently available microtiter plate sealing tapes fail to fulfil the requirements for aerobic microbial cultivation

Sieben, Michaela; Giese, H.; Grosch, Jan-Hendrik; Kauffmann, Kira; Büchs, Jochen

doi:10.1002/biot.201600054

Cited by 19 publications

(13 citation statements)

References 44 publications

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“…For all MTP glucose release experiments in 96-square-well and 48-round-well MTPs, “AeraSeal Film” (A9224, Sigma-Aldrich Chemie GmbH, Germany) sealings were used as a permeable sterile barrier [ 32 ]. “Polyolefin sealing foil” (900371-T, HJ-Bioanalytik GmbH, Erkelenz, Germany) was used for cultivations in 48-round-well MTPs.…”

Section: Methodsmentioning

confidence: 99%

Optimized polymer-based glucose release in microtiter plates for small-scale E. coli fed-batch cultivations

Keil

Dittrich

Lattermann³

et al. 2020

J Biol Eng

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Background Small-scale cultivation vessels, which allow fed-batch operation mode, become more and more important for fast and reliable early process development. Recently, the polymer-based feeding system was introduced to allow fed-batch conditions in microtiter plates. Maximum glucose release rates of 0.35 mg/h per well (48-well-plate) at 37 °C can be achieved with these plates, depending on the media properties. The fed-batch cultivation of fluorescent protein-expressing E. coli at oxygen transfer rate levels of 5 mmol/L/h proved to be superior compared to simple batch cultivations. However, literature suggests that higher glucose release rates than achieved with the currently available fed-batch microtiter plate are beneficial, especially for fast-growing microorganisms. During the fed-batch phase of the cultivation, a resulting oxygen transfer rate level of 28 mmol/L/h should be achieved. Results Customization of the polymer matrix enabled a considerable increase in the glucose release rate of more than 250% to up to 0.90 mg/h per well. Therefore, the molecular weight of the prepolymer and the addition of a hydrophilic PDMS-PEG copolymer allowed for the individual adjustment of a targeted glucose release rate. The newly developed polymer matrix was additionally invariant to medium properties like the osmotic concentration or the pH-value. The glucose release rate of the optimized matrix was constant in various synthetic and complex media. Fed-batch cultivations of E. coli in microtiter plates with the optimized matrix revealed elevated oxygen transfer rates during the fed-batch phase of approximately 28 mmol/L/h. However, these increased glucose release rates resulted in a prolonged initial batch phase and oxygen limitations. The newly developed polymer-based feeding system provides options to manufacture individual feed rates in a range from 0.24–0.90 mg/h per well. Conclusions The optimized polymer-based fed-batch microtiter plate allows higher reproducibility of fed-batch experiments since cultivation media properties have almost no influence on the release rate. The adjustment of individual feeding rates in a wide range supports the early process development for slow, average and fast-growing microorganisms in microtiter plates. The study underlines the importance of a detailed understanding of the metabolic behavior (through online monitoring techniques) to identify optimal feed rates.

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Section: Methodsmentioning

confidence: 99%

Optimized polymer-based glucose release in microtiter plates for small-scale E. coli fed-batch cultivations

Keil

Dittrich

Lattermann³

et al. 2020

J Biol Eng

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“…For all microtiter plate cultivations and glucose release experiments in 96-square-well plates, “AeraSeal Film” (A9224, Sigma-Aldrich Chemie GmbH, Germany) sealings were used as permeable sterile barrier [40]. For release experiments at 45 °C, an airtight, self-made silicone sealing was fixed on top of the plate.…”

Section: Methodsmentioning

confidence: 99%

Polymer-based controlled-release fed-batch microtiter plate – diminishing the gap between early process development and production conditions

Keil

Dittrich

Lattermann³

et al. 2019

J Biol Eng

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Background Fed-batch conditions are advantageous for industrial cultivations as they avoid unfavorable phenomena appearing in batch cultivations. Those are for example the formation of overflow metabolites, catabolite repression, oxygen limitation or inhibition due to elevated osmotic concentrations. For both, the early bioprocess development and the optimization of existing bioprocesses, small-scale reaction vessels are applied to ensure high throughput, low costs and prompt results. However, most conventional small-scale procedures work in batch operation mode, which stands in contrast to fed-batch conditions in large-scale bioprocesses. Extensive expenditure for installations and operation accompany almost all cultivation systems in the market allowing fed-batch conditions in small-scale. An alternative, more cost efficient enzymatic glucose release system is strongly influenced by environmental conditions. To overcome these issues, this study investigates a polymer-based fed-batch system for controlled substrate release in microtiter plates. Results Immobilizing a solid silicone matrix with embedded glucose crystals at the bottom of each well of a microtiter plate is a suitable technique for implementing fed-batch conditions in microtiter plates. The results showed that the glucose release rate depends on the osmotic concentration, the pH and the temperature of the medium. Moreover, the applied nitrogen source proved to influence the glucose release rate. A new developed mathematical tool predicts the glucose release for various media conditions. The two model organisms E. coli and H. polymorpha were cultivated in the fed-batch microtiter plate to investigate the general applicability for microbial systems. Online monitoring of the oxygen transfer rate and offline analysis of substrate, product, biomass and pH confirmed that fed-batch conditions are comparable to large-scale cultivations. Furthermore, due to fed-batch conditions in microtiter plates, product formation could be enhanced by the factor 245 compared to batch cultivations. Conclusions The polymer-based fed-batch microtiter plate represents a sophisticated and cost efficient system to mimic typical industrial fed-batch conditions in small-scale. Thus, a more reliable strain screening and early process development can be performed. A systematical scale-down with low expenditure of work, time and money is possible. Electronic supplementary material The online version of this article (10.1186/s13036-019-0147-6) contains supplementary material, which is available to authorized users.

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“…To avoid cross‐contamination by the volatile solvent, each solvent concentration was applied separately. Nonreactive wells containing volatile substances were closed using nonpermeable sealing foils (EN77.1, Carl Roth GmbH, Karlsruhe, Germany) . Reactive wells were not sealed during measurements to avoid increasing experimental error because of impaired intrinsic optical properties of the sealing.…”

Section: Methodsmentioning

confidence: 99%

Thermodynamic activity‐based intrinsic enzyme kinetic sheds light on enzyme–solvent interactions

Grosch

Wagner

Nistelkas

et al. 2016

Biotechnology Progress

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The reaction medium has major impact on biocatalytic reaction systems and on their economic significance. To allow for tailored medium engineering, thermodynamic phenomena, intrinsic enzyme kinetics, and enzyme-solvent interactions have to be discriminated. To this end, enzyme reaction kinetic modeling was coupled with thermodynamic calculations based on investigations of the alcohol dehydrogenase from Lactobacillus brevis (LbADH) in monophasic water/methyl tert-butyl ether (MTBE) mixtures as a model solvent. Substrate concentrations and substrate thermodynamic activities were varied separately to identify the individual thermodynamic and kinetic effects on the enzyme activity. Microkinetic parameters based on concentration and thermodynamic activity were derived to successfully identify a positive effect of MTBE on the availability of the substrate to the enzyme, but a negative effect on the enzyme performance. In conclusion, thermodynamic activity-based kinetic modeling might be a suitable tool to initially curtail the type of enzyme-solvent interactions and thus, a powerful first step to potentially understand the phenomena that occur in nonconventional media in more detail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:96-103, 2017.

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Permeability of currently available microtiter plate sealing tapes fail to fulfil the requirements for aerobic microbial cultivation

Cited by 19 publications

References 44 publications

Optimized polymer-based glucose release in microtiter plates for small-scale E. coli fed-batch cultivations

Optimized polymer-based glucose release in microtiter plates for small-scale E. coli fed-batch cultivations

Polymer-based controlled-release fed-batch microtiter plate – diminishing the gap between early process development and production conditions

Thermodynamic activity‐based intrinsic enzyme kinetic sheds light on enzyme–solvent interactions

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