Export-deficient monoubiquitinated PEX5 triggers peroxisome removal in SV40 large T antigen-transformed mouse embryonic fibroblasts

Peroxisomes are ubiquitous cell organelles essential for human health. To maintain a healthy cellular environment, dysfunctional and superfluous peroxisomes need to be selectively removed. Although emerging evidence suggests that peroxisomes are mainly degraded by pexophagy, little is known about the triggers and molecular mechanisms underlying this process in mammalian cells. In this study, we show that PEX5 proteins fused to a bulky C-terminal tag trigger peroxisome degradation in SV40 large T antigen-transformed mouse embryonic fibroblasts. In addition, we provide evidence that this process is autophagy-dependent and requires monoubiquitination of the N-terminal cysteine residue that marks PEX5 for recycling. As our findings also demonstrate that the addition of a bulky tag to the C terminus of PEX5 does not interfere with PEX5 monoubiquitination but strongly inhibits its export from the peroxisomal membrane, we hypothesize that such a tag mimics a cargo protein that cannot be released from PEX5, thus keeping monoubiquitinated PEX5 at the membrane for a sufficiently long time to be recognized by the autophagic machinery. This in turn suggests that monoubiquitination of the N-terminal cysteine of peroxisomeassociated PEX5 not only functions to recycle the peroxin back to the cytosol, but also serves as a quality control mechanism to eliminate peroxisomes with a defective protein import machinery.This work was supported by grants from the ’Fonds voor Wetenschappelijk Onderzoek-Vlaanderen (Onderzoeksprojecten G.0754.09 and G095315N)’ (to MF and PVV), the KU Leuven (OT/09/045, OT/14/100, and DBOF/10/059) (to MF and PVV), and by FEDER funds through the Operational Competi-tiveness Program, COMPETE, and by national funds through FCT, Fundação para a Ciência e a Tecnologia, under the projects FCOMP-01–0124-FEDER-019731 (PTDC/BIA-BCM/118577/2010) and FCOMP-01–0124-FEDER-022718 (PEst-C/SAU/LA0002/2011) (to JEA). MN was supported by a FLOF fellow-ship from the Department of Cellular and Molecular Medicine (KU Leuven). TF was supported by Fundação para a Ciência e aTecnologia, Programa Operacional Potencial Humano do QREN, and Fundo Social Europeu

Biotechnologies for critical raw material recovery from primary and secondary sources: R&D priorities and future perspectives

Europe is confronted with an increasing supply risk of critical raw materials. These can be defined as materials of which the risks of supply shortage and their impacts on the economy are higher compared to most of other raw materials. Within the framework of the EU Innovation Partnership on raw materials Initiative, a list of 14 critical materials was defined, including some bulk metals, industrial minerals, the platinum group metals and rare earth elements. To tackle the supply risk challenge, innovation is required with respect to sustainable primary mining, substitution of critical metals, and urban mining. In these three categories, biometallurgy can play a crucial role. Indeed, microbe–metal interactions have been successfully applied on full scale to win materials from primary sources, but are not sufficiently explored for metal recovery or recycling. On the one hand, this article gives an overview of the microbial strategies that are currently applied on full scale for biomining; on the other hand it identifies technologies, currently developed in the laboratory, which have a perspective for large scale metal recovery and the needs and challenges on which bio-metallurgical research should focus to achieve this ambitious goal

Modular advanced oxidation process enabled by cathodic hydrogen peroxide production.

Hydrogen peroxide (H2O2) is frequently used in combination with ultraviolet (UV) light to treat trace organic contaminants in advanced oxidation processes (AOPs). In small-scale applications, such as wellhead and point-of-entry water treatment systems, the need to maintain a stock solution of concentrated H2O2 increases the operational cost and complicates the operation of AOPs. To avoid the need for replenishing a stock solution of H2O2, a gas diffusion electrode was used to generate low concentrations of H2O2 directly in the water prior to its exposure to UV light. Following the AOP, the solution was passed through an anodic chamber to lower the solution pH and remove the residual H2O2. The effectiveness of the technology was evaluated using a suite of trace contaminants that spanned a range of reactivity with UV light and hydroxyl radical (HO(•)) in three different types of source waters (i.e., simulated groundwater, simulated surface water, and municipal wastewater effluent) as well as a sodium chloride solution. Irrespective of the source water, the system produced enough H2O2 to treat up to 120 L water d(-1). The extent of transformation of trace organic contaminants was affected by the current density and the concentrations of HO(•) scavengers in the source water. The electrical energy per order (EEO) ranged from 1 to 3 kWh m(-3), with the UV lamp accounting for most of the energy consumption. The gas diffusion electrode exhibited high efficiency for H2O2 production over extended periods and did not show a diminution in performance in any of the matrices

Modular Advanced Oxidation Process Enabled by Cathodic Hydrogen Peroxide Production

Hydrogen peroxide (H₂O₂) is frequently used in combination with ultraviolet (UV) light to treat trace organic contaminants in advanced oxidation processes (AOPs). In small-scale applications, such as wellhead and point-of-entry water treatment systems, the need to maintain a stock solution of concentrated H₂O₂ increases the operational cost and complicates the operation of AOPs. To avoid the need for replenishing a stock solution of H₂O₂, a gas diffusion electrode was used to generate low concentrations of H₂O₂ directly in the water prior to its exposure to UV light. Following the AOP, the solution was passed through an anodic chamber to lower the solution pH and remove the residual H₂O₂. The effectiveness of the technology was evaluated using a suite of trace contaminants that spanned a range of reactivity with UV light and hydroxyl radical (HO^•) in three different types of source waters (i.e., simulated groundwater, simulated surface water, and municipal wastewater effluent) as well as a sodium chloride solution. Irrespective of the source water, the system produced enough H₂O₂ to treat up to 120 L water d^–1. The extent of transformation of trace organic contaminants was affected by the current density and the concentrations of HO^• scavengers in the source water. The electrical energy per order (<i>E</i>_EO) ranged from 1 to 3 kWh m^–3, with the UV lamp accounting for most of the energy consumption. The gas diffusion electrode exhibited high efficiency for H₂O₂ production over extended periods and did not show a diminution in performance in any of the matrices

Biosupported bimetallic Pd-Au nanocatalysts for dechlorination of environmental contaminants

Biologically produced monometallic palladium nanoparticles (bio-Pd) have been shown to catalyze the dehalogenation of environmental contaminants, but fail to efficiently catalyze the degradation of other important recalcitrant halogenated compounds. This study represents the first report of biologically produced bimetallic Pd/Au nanoparticle catalysts. The obtained catalysts were tested for the dechlorination of diclofenac and trichlorethylene. When aqueous bivalent Pd(II) and trivalent Au(III) ions were both added to concentrations of 50 mg L(-1) and reduced simultaneously by Shewanella oneidensis in the presence of H(2), the resulting cell-associated bimetallic nanoparticles (bio-Pd/Au) were able to dehalogenate 78% of the initially added diclofenac after 24 h; in comparison, no dehalogenation was observed using monometallic bio-Pd or bio-Au. Other catalyst-synthesis strategies did not show improved dehalogenation of TCE and diclofenac compared with bio-Pd. Synchrotron-based X-ray diffraction, (scanning) transmission electron microscopy and energy dispersive X-ray spectroscopy indicated that the simultaneous reduction of Pd and Au supported on cells of S. oneidensis resulted in the formation of a unique bimetallic crystalline structure. This study demonstrates that the catalytic activity and functionality of possibly environmentally more benign biosupported Pd-catalysts can be improved by coprecipitation with Au

Concomitant microbial generation of palladium nanoparticles and hydrogen to immobilize chromate.

&lt;p&gt;The catalytic properties of various metal nanoparticles have led to their use in environmental remediation. Our aim is to develop and apply an efficient bioremediation method based on in situ biosynthesis of bio-Pd nanoparticles and hydrogen. C. pasteurianum BC1 was used to reduce Pd(II) ions to form Pd nanoparticles (bio-Pd) that primarily precipitated on the cell wall and in the cytoplasm. C. pasteurianum BC1 cells, loaded with bio-Pd nanoparticle in the presence of glucose, were subsequently used to fermentatively produce hydrogen and to effectively catalyze the removal of soluble Cr(VI) via reductive transformation to insoluble Cr(III) species. Batch and aquifer microcosm experiments using C. pasteurianum BC1 cells loaded with bio-Pd showed efficient reductive Cr(VI) removal, while in control experiments with killed or viable but Pd-free bacterial cultures no reductive Cr(VI) removal was observed. Our results suggest a novel process where the in situ microbial production of hydrogen is directly coupled to the catalytic bio-Pd mediated reduction of chromate. This process offers significant advantages over the current groundwater treatment technologies that rely on introducing preformed catalytic nanoparticles into groundwater treatment zones and the costly addition of molecular hydrogen to above ground pump and treat systems.&lt;/p&gt;</p