Characterization of oxygen transfer in a 24-well microbioreactor system and potential respirometric applications

Ramirez-Vargas, Rocio; Vital‐Jácome, Miguel; Camacho-Pérez, E.; Hubbard, Loe; Thalasso, F.

doi:10.1016/j.jbiotec.2014.06.031

Cited by 13 publications

(7 citation statements)

References 51 publications

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“…For the fermentation, the stains were inoculated at 1 × 10 6 cells per ml and incubated at 28 • C for 24 h and 245 rpm. The inoculum was added (100 µL) to 24-well plates (Fisher Scientific, Monterrey, Nuevo León, México), which working volume was 1.9 mL, then was fixed in a support with orbital shaking (5 mm diameter shaking) [18]. Each well of the plate containing the inoculum was previously adjusted to the amount of 1 × 10 6 cells per mL and 100 µL precursor and salt whose concentrations for each treatment (defined in Table 1), the remaining volume 1700 µL was sweet whey pretreated (see 2.2) [19].…”

Section: Fermentationmentioning

confidence: 99%

Optimization of 2-Phenylethanol Production from Sweet Whey Fermentation Using Kluyveromyces marxianus

Alonso-Vargas

Téllez‐Jurado

Aldapa³

et al. 2022

Fermentation

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The growing demand for natural products benefits the development of bioprocesses to obtain value-added compounds using residues such as sweet whey, which is rich in lactose. The yeast Kluyveromyces marxianus can ferment sweet whey to obtain 2-phenylethanol (2-PhEtOH), which is a superior alcohol with a rose aroma. Such fermentation only requires the addition of L-phenylalanine (precursor) and (NH4)2SO4 (salt). Therefore, it was sought to improve the fermentation conditions to produce 2-PhEtOH, which, in turn, would achieve the maximum decrease in the Chemical Oxygen Demand (COD) of the fermentation medium. With the use of the Response Surface Methodology and the application of a Central Composite Design for optimization, two parameters were evaluated as a function of time: salt concentration and precursor. The experimental data were adjusted to a second order polynomial, identifying that the precursor concentration presents a statistically significant effect. The best conditions were: 4.50 g/L of precursor and 0.76 g/L of salt, with a maximum production of 1.2 g/L (2-PhEtOH) at 48 h and achieving a maximum percentage of COD removal of 76% at 96 h. Finally, the optimal conditions were experimentally validated, recommending the use of the model.

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Section: Fermentationmentioning

confidence: 99%

Optimization of 2-Phenylethanol Production from Sweet Whey Fermentation Using Kluyveromyces marxianus

Alonso-Vargas

Téllez‐Jurado

Aldapa³

et al. 2022

Fermentation

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“…Oxygen-transfer capacity in MTP relies mainly upon the specific surface area for oxygen transfer [ 21 ]. Ramirez-Vargas et al (2014) claimed that the oxygen transfer rate was inversely proportional to fill volume, particularly at higher shaking frequencies in microtiter plates, which affected the cell growth and protein production.…”

Section: Discussionmentioning

confidence: 99%

Microtiter miniature shaken bioreactor system as a scale-down model for process development of production of therapeutic alpha-interferon2b by recombinant Escherichia coli

et al. 2018

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BackgroundDemand for high-throughput bioprocessing has dramatically increased especially in the biopharmaceutical industry because the technologies are of vital importance to process optimization and media development. This can be efficiently boosted by using microtiter plate (MTP) cultivation setup embedded into an automated liquid-handling system. The objective of this study was to establish an automated microscale method for upstream and downstream bioprocessing of α-IFN2b production by recombinant Escherichia coli. The extraction performance of α-IFN2b by osmotic shock using two different systems, automated microscale platform and manual extraction in MTP was compared.ResultsThe amount of α-IFN2b extracted using automated microscale platform (49.2 μg/L) was comparable to manual osmotic shock method (48.8 μg/L), but the standard deviation was 2 times lower as compared to manual osmotic shock method. Fermentation parameters in MTP involving inoculum size, agitation speed, working volume and induction profiling revealed that the fermentation conditions for the highest production of α-IFN2b (85.5 μg/L) was attained at inoculum size of 8%, working volume of 40% and agitation speed of 1000 rpm with induction at 4 h after the inoculation.ConclusionAlthough the findings at MTP scale did not show perfect scalable results as compared to shake flask culture, but microscale technique development would serve as a convenient and low-cost solution in process optimization for recombinant protein.

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“…This has led to the development of specialized, complex micro-reactor systems for scale down studies, where process conditions can be better controlled. 8,[14][15][16] The efforts in developing these systems again stem from a lack of robustness in biological systems and a concerted effort to improve the throughput of process optimization.…”

Section: Introductionmentioning

confidence: 99%

Two-stage Dynamic Deregulation of Metabolism Improves Process Robustness & Scalability in EngineeredE. coli

Hennigan

et al. 2020

Preprint

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We report improved strain and bioprocess robustness as a result of the dynamic deregulation of central metabolism using two-stage dynamic control. Dynamic control is implemented using combinations of CRISPR interference and controlled proteolysis to reduce levels of central metabolic enzymes in the context of a standardized two-stage bioprocesses. Reducing the levels of key enzymes alters metabolite pools resulting in deregulation of the metabolic network. The deregulated network is more robust to environmental conditions improving process robustness, which in turn leads to predictable scalability from high throughput small scale screens to fully instrumented bioreactors as well as to pilot scale production. Additionally, as these two-stage bioprocesses are standardized, a need for traditional process optimization is minimized. Predictive high throughput approaches that translate to larger scales are critical for metabolic engineering programs to truly take advantage of the rapidly increasing throughput and decreasing costs of synthetic biology. In this work we demonstrate that the improved robustness of E. coli strains engineered for the improved scalability of the important industrial chemicals alanine, citramalate and xylitol, from microtiter plates to pilot reactors.

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Characterization of oxygen transfer in a 24-well microbioreactor system and potential respirometric applications

Cited by 13 publications

References 51 publications

Optimization of 2-Phenylethanol Production from Sweet Whey Fermentation Using Kluyveromyces marxianus

Optimization of 2-Phenylethanol Production from Sweet Whey Fermentation Using Kluyveromyces marxianus

Microtiter miniature shaken bioreactor system as a scale-down model for process development of production of therapeutic alpha-interferon2b by recombinant Escherichia coli

Two-stage Dynamic Deregulation of Metabolism Improves Process Robustness & Scalability in EngineeredE. coli

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