Muscle fiber-type specific expression of UCP3-protein is reported here for the firts time, using immunofluorescence microscopy
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In this study we tested 39 Lactococcus lactis strains isolated from diverse habitats for their robustness under heat and oxidative stress, demonstrating high diversity in survival (up to 4 log units). Strains with an L. lactis subsp. lactis phenotype generally displayed more-robust phenotypes than strains with an L. lactis subsp. cremoris phenotype, whereas the habitat from which the strains had been isolated did not appear to influence stress survival. Comparison of the stress survival phenotypes with already available comparative genomic data sets revealed that the absence or presence of specific genes, including genes encoding a GntR family transcriptional regulator, a manganese ABC transporter permease, a cellobiose phosphotransferase system (PTS) component, the FtsY protein, and hypothetical proteins, was associated with heat or oxidative stress survival. Finally, 14 selected strains also displayed diversity in survival after spray drying, ranging from 20% survival for the most robust strains, which appears acceptable for industrial application, to 0.1% survival for the least-tolerant strains. The high and low levels of survival upon spray drying correlated clearly with the combined robustness under heat and oxidative stress. These results demonstrate the relevance of screening culture collections for robustness under heat and oxidative stress on top of the typical screening for acidifying and flavor-forming properties. © 2014, American Society for Microbiology.
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The synthesis of total cellular proteins in Escherichia coli K12 was studied in batch culture following exposure of cells to low concentrations of monochlorophenol, pentachlorophenol and cadmium chloride. Changes in protein patterns were identified after pulse-chase labelling of proteins with [35S]methionine and subsequent two-dimensional gel electrophoresis (2D-PAGE). We demonstrated that besides the induction of some stress proteins, also a transient decrease in the rate of synthesis of other proteins occurred. Two of these proteins were identified as OmpF and aspartate transcarbamoylase (ATCase). Their transient repression appeared to be a general response to stress elicited by different pollutants and may therefore be used as a general and sensitive early warning system for pollutant stress.
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Cell-based production processes in bioreactors and fermenters need to be carefully monitored due to the complexity of the biological systems and the growth processes of the cells. Critical parameters are identified and monitored over time to guarantee product quality and consistency and to minimize over-processing and batch rejections. Sensors are already available for monitoring parameters such as temperature, glucose, pH, and CO2, but not yet for low-concentration substances like proteins and nucleic acids (DNA). An interesting critical parameter to monitor is host cell DNA (HCD), as it is considered an impurity in the final product (downstream process) and its concentration indicates the cell status (upstream process). The Molecular Biosensing group at the Eindhoven University of Technology and Helia Biomonitoring are developing a sensor for continuous biomarker monitoring, based on Biosensing by Particle Motion. With this consortium, we want to explore whether the sensor is suitable for the continuous measurement of HCD. Therefore, we need to set-up a joint laboratory infrastructure to develop HCD assays. Knowledge of how cells respond to environmental changes and how this is reflected in the DNA concentration profile in the cell medium needs to be explored. This KIEM study will enable us to set the first steps towards continuous HCD sensing from cell culture conditions controlling cell production processes. It eventually generates input for machine learning to be able to automate processes in bioreactors and fermenters e.g. for the production of biopharmaceuticals. The project entails collaboration with new partners and will set a strong basis for subsequent research projects leading to scientific and economic growth, and will also contribute to the human capital agenda.