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Biology subjects

Sandrelli, F.

Publications and source records attributed to Sandrelli, F..

3 recordsLinked to original sources

Drosophila melanogaster as a platform for the functional expression of engineered PET-degrading enzymes

Insects offer promising opportunities for organic waste bioconversion; however, they cannot efficiently degrade synthetic polymers such as polyethylene terephthalate (PET). Here, we generated transgenic Drosophila melanogaster lines to express in vitro-evolved variants of two PET-degrading enzymes with distinct biochemical properties: an engineered Ideonella sakaiensis PETase variant (TS-{Delta}IsPET) and a leaf-branch compost cutinase variant (TA-{Delta}LCC). Both enzymes, fused to a Drosophila gut-derived secretory signal, were produced and secreted by both Drosophila cultured S2R+ cells and transgenic larvae. Both enzymes were glycosylated upon secretion, a post-translational modification that did not abolish their catalytic activity. Notably, TA-{Delta}LCC displayed [~]6-fold higher esterase activity than TS-{Delta}IsPET in larval extracts and TA-{Delta}LCC-containing extracts depolymerised PET nanoparticles in vitro under enzyme-favourable conditions. Transgenic flies showed normal development, fertility and survival. Morphological and biochemical analysis confirmed that TA-{Delta}LCC expression did not alter midgut structure and function. Together, these results establish Drosophila melanogaster as a model for functional expression and comparative evaluation of engineered PET-degrading enzymes and identify TA-{Delta}LCC as a promising candidate for exploitation in insect species relevant to plastic contaminated waste bioconversion. HighlightsO_LITransgenic D. melanogaster enables in vivo study of engineered PET enzymes C_LIO_LIEngineered TS-{Delta}IsPET and TA-{Delta}LCC are functional in larval extracts C_LIO_LITA-{Delta}LCC was selected for PET nanoparticle assays due to higher pNPA activity C_LIO_LID. melanogaster model enables comparative evaluation of PET-degrading enzymes C_LI

biochemistry↗

A functional circadian clock regulates composition and daily bacterial load of the gut microbiome in Drosophila melanogaster

While Drosophila melanogaster serves as a crucial model for investigating both the circadian clock and gut microbiome, our understanding of their relationship in this organism is still limited. Recent analyses suggested that the Drosophila gut microbiome modulates the host circadian tran-scriptome to minimize rapid oscillations in response to changing environments. To delve deeper into the potential relationship between the gut microbiota and circadian clock in Drosophila, we examined the composition and abundance of the gut microbiota in wild-type and arrhythmic per01 flies, under 12 h:12 h light: dark (12:12 LD) and constant darkness (DD) conditions. The gut microbiota of wild-type and per01 flies showed differences in composition, suggesting that the D. melanogaster circadian clock has a role in shaping the gut microbiome. In 12:12 LD and DD conditions, per01 mutants showed significant daily variations in gut bacterial quantity, unlike wild-type flies. This suggests that the circadian clock in D. melanogaster plays a role in maintaining daily stability in gut microbiome load. Finally, some gut bacteria exhibited significant 24 h fluctuations in their relative abundance, which appeared independent from the fly circadian clock, suggesting that certain gut commensal bacteria in Drosophila may possess a host-independent circadian clock.

microbiology↗

Light affects behavioral despair involving the clock gene Period 1

Light at night has strong effects on physiology and behavior of mammals. It affects mood in humans, which is exploited as light therapy, and has been shown to reset the circadian clock in the suprachiasmatic nuclei (SCN). This resetting is paramount to align physiological and biochemical timing to the environmental light-dark cycle. Here we provide evidence that light affects mood-related behaviors also in mice by activating the clock gene Period1 (Per1) in the lateral habenula (LHb), a brain region known to modulate mood-related behaviors. A light pulse given at ZT22 to wild type mice caused profound changes of gene expression in the mesolimbic dopaminergic system including the nucleus accumbens (NAc). Sensory perception of smell and G-protein coupled receptor signaling was affected the most in this brain region. Interestingly, most of these genes were not affected in Per1 knock-out animals, indicating that induction of Per1 by light serves as a filter for light-mediated gene induction in the brain. Taken together we show that light affects mood-related behavior in mice at least in part via induction of Per1 in the LHb with consequences on signaling mechanisms in the mesolimbic dopaminergic system.

genetics↗