Media Composition, Microbial, Laboratory Scale

Cote, R.

doi:10.1002/9780470054581.eib613

Cited by 8 publications

(5 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Second, large quantities of additional complex medium components are fed into the fermenter during the fed-batch phase. This can result in osmotic problems and the presence of large quantities of undefined proteinaceous medium components can complicate downstream processing of product proteins [5]. Finally, use of glucose as a feed can present difficulties from acid formation due to overflow metabolism, especially when growth rates fluctuate.…”

Section: Resultsmentioning

confidence: 99%

Use of a stress-minimisation paradigm in high cell density fed-batch Escherichia coli fermentations to optimise recombinant protein production

Wyre

Overton

2014

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

Production of recombinant proteins is an industrially important technique in the biopharmaceutical sector. Many recombinant proteins are problematic to generate in a soluble form in bacteria as they readily form insoluble inclusion bodies. Recombinant protein solubility can be enhanced by minimising stress imposed on bacteria through decreasing growth temperature and the rate of recombinant protein production. In this study, we determined whether these stress-minimisation techniques can be successfully applied to industrially relevant high cell density Escherichia coli fermentations generating a recombinant protein prone to forming inclusion bodies, CheY-GFP. Flow cytometry was used as a routine technique to rapidly determine bacterial productivity and physiology at the single cell level, enabling determination of culture heterogeneity. We show that stress minimisation can be applied to high cell density fermentations (up to a dry cell weight of >70 g L(-1)) using semi-defined media and glucose or glycerol as carbon sources, and using early or late induction of recombinant protein production, to produce high yields (up to 6 g L(-1)) of aggregation-prone recombinant protein in a soluble form. These results clearly demonstrate that stress minimisation is a viable option for the optimisation of high cell density industrial fermentations for the production of high yields of difficult-to-produce recombinant proteins, and present a workflow for the application of stress-minimisation techniques in a variety of fermentation protocols.

show abstract

Section: Resultsmentioning

confidence: 99%

Use of a stress-minimisation paradigm in high cell density fed-batch Escherichia coli fermentations to optimise recombinant protein production

Wyre

Overton

2014

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

show abstract

“…BE (0.3%) was used to simulate foreign particles, which can also contribute to encasement. Beef extract is a mixture of peptides, amino acids, nucleotide fractions, organic acids, minerals, and some vitamins, and it is derived from infusion of beef (6). Saliva is a very dilute fluid, composed of more than 98% water, electrolytes, mucus, and enzyme.…”

Section: Methodsmentioning

confidence: 99%

Effects of Relative Humidity and Spraying Medium on UV Decontamination of Filters Loaded with Viral Aerosols

Woo

Grippin

Anwar

et al. 2012

Appl Environ Microbiol

View full text Add to dashboard Cite

Although respirators and filters are designed to prevent the spread of pathogenic aerosols, a stockpile shortage is anticipated during the next flu pandemic. Contact transfer and reaerosolization of collected microbes from used respirators are also a concern. An option to address these potential problems is UV irradiation, which inactivates microbes by dimerizing thymine/uracil in nucleic acids. The objective of this study was to determine the effects of transmission mode and environmental conditions on decontamination efficiency by UV. In this study, filters were contaminated by different transmission pathways (droplet and aerosol) using three spraying media (deionized water . UV irradiation at constant intensity was applied for two time intervals at each relative humidity condition. The highest inactivation efficiency (IE), around 5.8 logs, was seen for DI aerosols containing MS2 on filters at LRH after applying a UV intensity of 1.0 mW/cm 2 for 30 min. The IE of droplets containing MS2 was lower than that of aerosols containing MS2. Absorption of UV by high water content and shielding of viruses near the center of the aggregate are considered responsible for this trend. Across the different media, IEs in AS and in BE were much lower than in DI for both aerosol and droplet transmission, indicating that solids present in AS and BE exhibited a protective effect. For particles sprayed in a protective medium, RH is not a significant parameter.

show abstract

“…BE (0.3 vol %), which was used to provide nutrients, can contribute encasement. BE is a mixture of peptides, amino acids, nucleotide fractions, organic acids, minerals and some vitamins, derived from infusion of beef (Cote 1999). AS (0.6 vol %), 0.3% mucin from porcine stomach (Sigma-Aldrich, M1778) plus salts in DI water as a mucus simulant, was used to mimic human respiratory fluid.…”

Section: Effects Of Relative Humidity and Spraying Medium On Uv Deconmentioning

confidence: 99%

Evaluation of the Performance of Iodine-Treated Biocidal Filters Under the Influence of Environmental Parameters

Woo¹,

Li²,

Wu³

2013

View full text Add to dashboard Cite

Media Composition, Microbial, Laboratory Scale

Cited by 8 publications

References 55 publications

Use of a stress-minimisation paradigm in high cell density fed-batch Escherichia coli fermentations to optimise recombinant protein production

Use of a stress-minimisation paradigm in high cell density fed-batch Escherichia coli fermentations to optimise recombinant protein production

Effects of Relative Humidity and Spraying Medium on UV Decontamination of Filters Loaded with Viral Aerosols

Evaluation of the Performance of Iodine-Treated Biocidal Filters Under the Influence of Environmental Parameters

Contact Info

Product

Resources

About