Fractionation of yeast extract by nanofiltration process to assess key compounds involved in <scp>CHO</scp> cell culture improvement

Mosser, Mathilde; Kapel, Romain; Chevalot, Isabelle; Olmos, Éric; Marc, Ivan; Marc, Annie; Oriol, E.

doi:10.1002/btpr.2110

Cited by 16 publications

(13 citation statements)

References 31 publications

(87 reference statements)

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“…No oxidative stress response was observed in PYE medium, while it was highly evident in M2G medium. This could be due to the presence of readily available glutathione and other antioxidants derived from the yeast extract in PYE [ 108 ]. The proteomic reconfiguration, which was also only observed in M2G medium, would then be a consequence of the oxidative stress in this medium.…”

Section: Resultsmentioning

confidence: 99%

Environmental Conditions Modulate the Transcriptomic Response of Both Caulobacter crescentus Morphotypes to Cu Stress

et al. 2021

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Bacteria encounter elevated copper (Cu) concentrations in multiple environments, varying from mining wastes to antimicrobial applications of copper. As the role of the environment in the bacterial response to Cu ion exposure remains elusive, we used a tagRNA-seq approach to elucidate the disparate responses of two morphotypes of Caulobacter crescentus NA1000 to moderate Cu stress in a complex rich (PYE) medium and a defined poor (M2G) medium. The transcriptome was more responsive in M2G, where we observed an extensive oxidative stress response and reconfiguration of the proteome, as well as the induction of metal resistance clusters. In PYE, little evidence was found for an oxidative stress response, but several transport systems were differentially expressed, and an increased need for histidine was apparent. These results show that the Cu stress response is strongly dependent on the cellular environment. In addition, induction of the extracytoplasmic function sigma factor SigF and its regulon was shared by the Cu stress responses in both media, and its central role was confirmed by the phenotypic screening of a sigF::Tn5 mutant. In both media, stalked cells were more responsive to Cu stress than swarmer cells, and a stronger basal expression of several cell protection systems was noted, indicating that the swarmer cell is inherently more Cu resistant. Our approach also allowed for detecting several new transcription start sites, putatively indicating small regulatory RNAs, and additional levels of Cu-responsive regulation.

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Section: Resultsmentioning

confidence: 99%

Environmental Conditions Modulate the Transcriptomic Response of Both Caulobacter crescentus Morphotypes to Cu Stress

et al. 2021

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“…This could be due to yeastolate components binding to BSA, and thus changing the emission properties, identifying specific binding processes is not feasible due to the large number of potential interactions (e.g. BSA can interact with vitamin B12 and B6 (Zhang et al, 2008), both are present in yeastolate (Mosser et al, 2015). The fourth component (1%–2%) is weaker in the TSFS || data which might suggest that this originates from smaller yeastolate fluorophores.…”

Section: Resultsmentioning

confidence: 99%

Development of a rapid polarized total synchronous fluorescence spectroscopy (pTSFS) method for protein quantification in a model bioreactor broth

Boateng

Elcoroaristizabal

Ryder

2021

Biotech & Bioengineering

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Protein quantification during bioprocess monitoring is essential for biopharmaceutical manufacturing and is complicated by the complex chemical composition of the bioreactor broth. Here we present the early‐stage development and optimization of a polarized total synchronous fluorescence spectroscopy (pTSFS) method for protein quantification in a hydrolysate‐protein model (mimics clarified bioreactor broth samples) using a standard benchtop laboratory fluorometer. We used UV transmitting polarizers to provide wider range pTSFS spectra for screening of the four different TSFS spectra generated by the measurement: parallel (||), perpendicular (⊥), unpolarized (T) intensity spectra and anisotropy maps. TSFS|| (parallel polarized) measurements were the best for protein quantification compared to standard unpolarized measurements and the Bradford assay. This was because TSFS|| spectra had a better analyte signal to noise ratio (SNR), due to the anisotropy of protein emission. This meant that protein signals were better resolved from the background emission of small molecule fluorophores in the cell culture media. SNR of >5000 was achieved for concentrations of bovine serum albumin/yeastolate 1.2/10 g L–1 with TSFS||. Optimization using genetic algorithm and interval partial least squares based variable selection enabled reduction of spectral resolution and number of excitation wavelengths required without degrading performance. This enables fast (<3.5 min) online/at‐line measurements, and the method had an LOD of 0.18 g L–1 and high accuracy with a predictive error of <9%.

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“…Yeast extract plays an important role for growth of S. cerevisiae cells since it consists of the water-soluble portion of autolyzed yeast, which contains amino acids, peptides, carbohydrates, vitamins and minerals (Jiang et al 2010 ; Mosser et al 2015 ; Thomas et al 2002 ). To increase the ATP pool by reducing ATP demands for cell growth, the Kyokai 6 strain was cultivated in the medium containing various concentrations of yeast extract (0.0–5.0 g L −1 ).…”

Section: Resultsmentioning

confidence: 99%

Metabolome analysis of Saccharomyces cerevisiae and optimization of culture medium for S-adenosyl-l-methionine production

2016

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S-Adenosyl-l-methionine (SAM) is a fine chemical used as a nutritional supplement and a prescription drug. It is industrially produced using Saccharomyces cerevisiae owing to its high SAM content. To investigate the optimization of culture medium components for higher SAM production, metabolome analysis was conducted to compare the intracellular metabolite concentrations between Kyokai no. 6 (high SAM-producing) and laboratory yeast S288C (control) under different SAM production conditions. Metabolome analysis and the result of principal component analysis showed that the rate-limiting step for SAM production was ATP supply and the levels of degradation products of adenosine nucleotides were higher in Kyokai 6 strain than in the S288C strain under the l-methionine supplemented condition. Analysis of ATP accumulation showed that the levels of intracellular ATP in the Kyokai 6 strain were also higher compared to those in the S288C strain. Furthermore, as expected from metabolome analysis, the SAM content of Kyokai 6 strain cultivated in the medium without yeast extract increased by 2.5-fold compared to that in the additional condition, by increasing intracellular ATP level with inhibited cell growth. These results suggest that high SAM production is attributed to the enhanced ATP supply with l-methionine condition and high efficiency of intracellular ATP consumption.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0210-3) contains supplementary material, which is available to authorized users.

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Fractionation of yeast extract by nanofiltration process to assess key compounds involved in CHO cell culture improvement

Cited by 16 publications

References 31 publications

Environmental Conditions Modulate the Transcriptomic Response of Both Caulobacter crescentus Morphotypes to Cu Stress

Environmental Conditions Modulate the Transcriptomic Response of Both Caulobacter crescentus Morphotypes to Cu Stress

Development of a rapid polarized total synchronous fluorescence spectroscopy (pTSFS) method for protein quantification in a model bioreactor broth

Metabolome analysis of Saccharomyces cerevisiae and optimization of culture medium for S-adenosyl-l-methionine production

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