Biology subjects

Teuta Pilizota

Publications and source records attributed to Teuta Pilizota.

3 recordsLinked to original sources

bioRxiv2016

Characterization of the effects of n-butanol on the cell envelope of E. coli

Biofuel alcohols have severe consequences on the microbial hosts used in their biosynthesis, which limits the productivity of the bioconversion. The cell envelope is one of the most strongly affected structures, in particular, as the external concentration of biofuels rises during biosynthesis. Damage to the cell envelope can have severe consequences, such as impairment of transport into and out of the cell; however the nature of butanol-induced envelope damage has not been well characterized. In the present study, the effects of n-butanol on the cell envelope of Escherichia coli were investigated. Using enzyme and fluorescence-based assays, we observed that 1% v/v n-butanol resulted in release of lipopolysaccharides from the outer membrane of E. coli and caused leakiness in both outer and inner membranes. Higher concentrations of n-butanol, within the range of 2% - 10% (v/v), resulted in inner membrane protrusion through the peptidoglycan observed by characteristic blebs. The findings suggest that strategies for rational engineering of butanol-tolerant bacterial strains should take into account all components of the cell envelope.

Microbiology↗

bioRxiv2016

General calibration of microbial growth in microplate readers

Optical density (OD) measurements of microbial growth are one of the most common techniques used in microbiology, with applications ranging from antibiotic efficacy studies, studies of growth under different nutritional or stress environments, to studies of different mutant strains, including those harbouring synthetic circuits. OD measurements are performed under the assumption that the OD value obtained is proportional to the cell number, i.e. the concentration of the sample. However, the assumption holds true in a limited range of conditions and calibration techniques that determine that range are currently missing. Here we present a set of calibration procedures and considerations that are necessary to successfully estimate the cell concentration from OD measurements.

Microbiology↗

bioRxiv2016

Inferring time-derivatives, including cell growth rates, using Gaussian processes

Often the time-derivative of a measured variable is of as much interest as the variable itself. For a growing population of biological cells, for example, the population's growth rate is typically more important than its size. Here we introduce a non-parametric method to infer first and second time-derivatives as a function of time from time-series data. Our approach is based on established properties of Gaussian processes and therefore applies to a wide range of data. In tests, the method is at least as accurate as others, but has several advantages: it estimates errors both in the inference and in any summary statistics, such as lag times, allows interpolation with the corresponding error estimation, and can be applied to any number of experimental replicates. As illustrations, we infer growth rate from measurements of the optical density of populations of microbial cells and estimate the rate of in vitro assembly of an amyloid fibril and both the speed and acceleration of two separating spindle pole bodies in a single yeast cell. Being accessible through both a GUI and from scripts, our algorithm should have broad application across the sciences.

Microbiology↗