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Biology subjects

Brown, R. C.

Publications and source records attributed to Brown, R. C..

3 recordsLinked to original sources

Adaptive evolution of Candida maltosa improves the bioconversion of depolymerized plastic feedstock by targeting biosurfactant production

Thermal oxo-degradation (TOD) of plastic can transform plastic waste into fermentable feedstocks. Bioconversion of the TOD products could be used as feedstocks in a biorefinery concept and lead to new avenues for plastic waste upcycling. Previous work demonstrated this concept with high density polyethylene (HDPE), the most abundant type of plastic, and identified the nonconventional yeast Candida maltosa as a promising candidate for this application. Here we describe the evolution of an improved strain of C. maltosa and characterize the uptake mechanisms of TOD products from HDPE (TOD_HDPE). Batch cultures in series passaged at the mid-exponential growth phase applied a selective pressure for faster growth and resulted in a >100% increase in specific growth rate when using TOD_HDPE as a carbon source. The evolved strain was compared to the parent strain to identify the cellular and biochemical changes associated with the improved phenotype and the uptake mechanisms involved in the bioconversion of TOD_HDPE. This comparison found that C. maltosa secretes biosurfactants capable of solubilizing hydrocarbons. The adaptive evolution resulted in changes in biosurfactant production that translated to improved emulsification of alkanes and increased solubilization of fatty alcohols and alkanes. In addition to the changes in metabolites, the study identified increases in membrane permeability associated with a reduction in ergosterol that may also play a role in the improved phenotype. These findings support the development of C. maltosa and other potential microbial cell factories for plastic biorefineries and may inform future design strategies.

microbiology↗

An integrated platform to systematically identify causal variants and genes for polygenic human traits.

Genome-wide association studies (GWAS) have identified over 150,000 links between common genetic variants and human traits or complex diseases. Over 80% of these associations map to polymorphisms in non-coding DNA. Therefore, the challenge is to identify disease-causing variants, the genes they affect, and the cells in which these effects occur. We have developed a platform using ATAC-seq, DNaseI footprints, NG Capture-C and machine learning to address this challenge. Applying this approach to red blood cell traits identifies a significant proportion of known causative variants and their effector genes, which we show can be validated by direct in vivo modelling.

genetics↗

Drug reformulation for a neglected disease. The NANOHAT project to develop a safer more effective sleeping sickness drug.

Human African trypanosomiasis (HAT or sleeping sickness) is caused by the parasite Trypanosoma brucei sspp. The disease has two stages, a haemolymphatic stage after the bite of an infected tsetse fly, followed by a central nervous system stage where the parasite penetrates the brain, causing death if untreated. Treatment is stage-specific, due to the blood-brain barrier, with less toxic drugs such as pentamidine used to treat stage 1. The objective of our research programme was to develop an intravenous formulation of pentamidine which increases CNS exposure by some 10-100 fold, leading to efficacy against a model of stage 2 HAT. This target candidate profile is in line with drugs for neglected diseases inititative recommendations. To do this, we evaluated the physicochemical and structural characteristics of formulations of pentamidine with Pluronic micelles (triblock-copolymers of polyethylene-oxide and polypropylene oxide), selected candidates for efficacy and toxicity evaluation in vitro, quantified pentamidine CNS delivery of a sub-set of formulations in vitro and in vivo, and progressed one pentamidine-Pluronic formulation for further evaluation using an in vivo single dose brain penetration study. Screening pentamidine against 40 CNS targets did not reveal any major neurotoxicity concerns, however, pentamidine had a high affinity for the imidazoline2 receptor. The reduction in insulin secretion in MIN6 {beta}-cells by pentamidine maybe secondary to pentamidine-mediated activation of {beta}-cell imidazoline receptors and impairment of cell viability. Pluronic F68 (0.01%w/v)-pentamidine formulation had a similar inhibitory effect on insulin secretion as pentamidine alone and an additive trypanocidal effect in vitro. However, all Pluronics tested (P85, P105 and F68) did not significantly enhance brain exposure of pentamidine. These results are relevant to further developing block-copolymers as nanocarriers, improving BBB drug penetration and understanding the side effects of pentamidine.

pharmacology and toxicology↗