Discrimination of riboflavin producing Bacillus subtilis strains based on their fed-batch process performances on a millilitre scale

Vester, Andrea; Hans, Michael; Hohmann, Hans‐Peter; Weuster‐Botz, Dirk

doi:10.1007/s00253-009-1966-z

Cited by 31 publications

(10 citation statements)

References 13 publications

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“…Therefore, the strains selected in these high‐throughput platforms for scale‐up may not be robust enough at industrial scale. High‐throughput experimental facilities that perform fed‐batch experiments with frequent glucose perturbations have been reported (Barz, Sommer, Wilms, Neubauer, & Cruz Bournazou, ; Cruz Bournazou et al, ; Vester, Hans, Hohmann, & Weuster‐Botz, ). Nevertheless, the objective of glucose perturbations in these works was to increase process dynamics information in optimal experimental design, and no reference to bioreactor heterogeneities or scale‐down efforts was made.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, the strains selected in these high-throughput platforms for scale-up may not be robust enough at industrial scale. High-throughput experimental facilities that perform fed-batch experiments with frequent glucose perturbations have been reported (Barz, Sommer, Wilms, Neubauer, & Cruz Bournazou, 2018;Cruz Bournazou et al, 2017;Vester, Hans, Hohmann, & Weuster-Botz, 2009).…”

Section: Introductionmentioning

confidence: 99%

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A model‐based framework for parallel scale‐down fed‐batch cultivations in mini‐bioreactors for accelerated phenotyping

Anane

García

Haby

et al. 2019

Biotech & Bioengineering

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Concentration gradients that occur in large industrial‐scale bioreactors due to mass transfer limitations have significant effects on process efficiency. Hence, it is desirable to investigate the response of strains to such heterogeneities to reduce the risk of failure during process scale‐up. Although there are various scale‐down techniques to study these effects, scale‐down strategies are rarely applied in the early developmental phases of a bioprocess, as they have not yet been implemented on small‐scale parallel cultivation devices. In this study, we combine mechanistic growth models with a parallel mini‐bioreactor system to create a high‐throughput platform for studying the response of Escherichia coli strains to concentration gradients. As a scaled‐down approach, a model‐based glucose pulse feeding scheme is applied and compared with a continuous feed profile to study the influence of glucose and dissolved oxygen gradients on both cell physiology and incorporation of noncanonical amino acids into recombinant proinsulin. The results show a significant increase in the incorporation of the noncanonical amino acid norvaline in the soluble intracellular extract and in the recombinant product in cultures with glucose/oxygen oscillations. Interestingly, the amount of norvaline depends on the pulse frequency and is negligible with continuous feeding, confirming observations from large‐scale cultivations. Most importantly, the results also show that a larger number of the model parameters are significantly affected by the scale‐down scheme, compared with the reference cultivations. In this example, it was possible to describe the effects of oscillations in a single parallel experiment. The platform offers the opportunity to combine strain screening with scale‐down studies to select the most robust strains for bioprocess scale‐up.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A model‐based framework for parallel scale‐down fed‐batch cultivations in mini‐bioreactors for accelerated phenotyping

Anane

García

Haby

et al. 2019

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…To avoid repeated optimisation in different cultivation formats, the fed-batch technology should be applied already in the first multiwell scale cultivation platforms, which is not a trivial task. Recently, new innovations paved the way by applying substrate autodelivery solutions [10,11] or by the use of robot based feeding [12]. …”

Section: Introductionmentioning

confidence: 99%

Novel approach of high cell density recombinant bioprocess development: Optimisation and scale-up from microlitre to pilot scales while maintaining the fed-batch cultivation mode of E. coli cultures

Šiurkus¹,

Panula-Perälä

Horn

et al. 2010

Microb Cell Fact

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BackgroundBioprocess development of recombinant proteins is time consuming and laborious as many factors influence the accumulation of the product in the soluble and active form. Currently, in most cases the developmental line is characterised by a screening stage which is performed under batch conditions followed by the development of the fed-batch process. Performing the screening already under fed-batch conditions would limit the amount of work and guarantee that the selected favoured conditions also work in the production scale.ResultsHere, for the first time, high throughput multifactorial screening of a cloning library is combined with the fed-batch technique in 96-well plates, and a strategy is directly derived for scaling to bioreactor scale. At the example of a difficult to express protein, an RNase inhibitor, it is demonstrated that screening of various vector constructs and growth conditions can be performed in a coherent line by (i) applying a vector library with promoters and ribosome binding sites of different strength and various fusion partners together with (ii) an early stage use of the fed-batch technology. It is shown that the EnBase® technology provides an easy solution for controlled cultivation conditions in the microwell scale. Additionally the high cell densities obtained provide material for various analyses from the small culture volumes. Crucial factors for a high yield of the target protein in the actual case were (i) the fusion partner, (ii) the use of of a mineral salt medium together with the fed-batch technique, and (iii) the preinduction growth rate. Finally, it is shown that the favorable conditions selected in the microwell plate and shake flask scales also work in the bioreactor.ConclusionsCultivation media and culture conditions have a major impact on the success of a screening procedure. Therefore the application of controlled cultivation conditions is pivotal. The consequent use of fed-batch conditons from the first screening phase not only shortens the developmental line by guarantying that the selected conditions are relevant for the scale up, but in our case also standard batch cultures failed to select the right clone or conditions at all.

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“…Isett et al (2007) achieved feeding in the 24‐well µ‐24 system (Applikon Biotechnology Inc., Foster City, CA) by manually removing the reactor cassette and adding substrate aseptically in a biosafety cabinet. Other devices at both the microliter and milliliter scale have used robotics to intermittently add feed solution to the microreactor cultures (Legmann et al, 2009; Vester et al, 2009). Siurkus et al (2010) sought to achieve more continuous feeding using the Enbase controlled‐release technology (Panula‐Perala et al, 2008) to mimic a fed‐batch process in 96‐well plates used for high‐throughput library screening.…”

Section: Introductionmentioning

confidence: 99%

Fed‐batch microbioreactor platform for scale down and analysis of a plasmid DNA production process

Bower

Lee

Ram

et al. 2012

Biotech & Bioengineering

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The rising costs of bioprocess research and development emphasize the need for high‐throughput, low‐cost alternatives to bench‐scale bioreactors for process development. In particular, there is a need for platforms that can go beyond simple batch growth of the organism of interest to include more advanced monitoring, control, and operation schemes such as fed‐batch or continuous. We have developed a 1‐mL microbioreactor capable of monitoring and control of dissolved oxygen, pH, and temperature. Optical density can also be measured online for continuous monitoring of cell growth. To test our microbioreactor platform, we used production of a plasmid DNA vaccine vector (pVAX1‐GFP) in Escherichia coli via a fed‐batch temperature‐inducible process as a model system. We demonstrated that our platform can accurately predict growth, glycerol and acetate concentrations, as well as plasmid copy number and quality obtained in a bench‐scale bioreactor. The predictive abilities of the micro‐scale system were robust over a range of feed rates as long as key process parameters, such as dissolved oxygen, were kept constant across scales. We have highlighted plasmid DNA production as a potential application for our microbioreactor, but the device has broad utility for microbial process development in other industries as well. Biotechnol. Bioeng. 2012; 109:1976–1986. © 2012 Wiley Periodicals, Inc.

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Discrimination of riboflavin producing Bacillus subtilis strains based on their fed-batch process performances on a millilitre scale

Cited by 31 publications

References 13 publications

A model‐based framework for parallel scale‐down fed‐batch cultivations in mini‐bioreactors for accelerated phenotyping

A model‐based framework for parallel scale‐down fed‐batch cultivations in mini‐bioreactors for accelerated phenotyping

Novel approach of high cell density recombinant bioprocess development: Optimisation and scale-up from microlitre to pilot scales while maintaining the fed-batch cultivation mode of E. coli cultures

Fed‐batch microbioreactor platform for scale down and analysis of a plasmid DNA production process

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