Microbial oil-degradation under mild hydrostatic pressure (10 MPa): which pathways are impacted in piezosensitive hydrocarbonoclastic bacteria?

Oil spills represent an overwhelming carbon input to the marine environment that immediately impacts the sea surface ecosystem. Microbial communities degrading the oil fraction that eventually sinks to the seafloor must also deal with hydrostatic pressure, which linearly increases with depth. Piezosensitive hydrocarbonoclastic bacteria are ideal candidates to elucidate impaired pathways following oil spills at low depth. In the present paper, we tested two strains of the ubiquitous Alcanivorax genus, namely A. jadensis KS_339 and A. dieselolei KS_293, which is known to rapidly grow after oil spills. Strains were subjected to atmospheric and mild pressure (0.1, 5 and 10?MPa, corresponding to a depth of 0, 500 and 1000?m, respectively) providing n-dodecane as sole carbon source. Pressures equal to 5 and 10?MPa significantly lowered growth yields of both strains. However, in strain KS_293 grown at 10?MPa CO2 production per cell was not affected, cell integrity was preserved and PO43- uptake increased. Analysis of its transcriptome revealed that 95% of its genes were downregulated. Increased transcription involved protein synthesis, energy generation and respiration pathways. Interplay between these factors may play a key role in shaping the structure of microbial communities developed after oil spills at low depth and limit their bioremediation potential

The De-Ubiquitinylating Enzyme, USP2, Is Associated with the Circadian Clockwork and Regulates Its Sensitivity to Light

We have identified a novel component of the circadian clock that regulates its sensitivity to light at the evening light to dark transition. USP2 (Ubiquitin Specific Protease 2), which de-ubiquitinylates and stabilizes target proteins, is rhythmically expressed in multiple tissues including the SCN. We have developed a knockout model of USP2 and found that exposure to low irradiance light at ZT12 increases phase delays of USP2−/− mice compared to wildtype. We additionally show that USP2b is in a complex with several clock components and regulates the stability and turnover of BMAL1, which in turn alters the expression of several CLOCK/BMAL1 controlled genes. Rhythmic expression of USP2 in the SCN and other tissues offers a new level of control of the clock machinery through de-ubiqutinylation and suggests a role for USP2 during circadian adaptation to environmental day length changes

Reduced TCA cycle rates at high hydrostatic pressure hinder hydrocarbon degradation and obligate oil degraders in natural, deep-sea microbial communities

Petroleum hydrocarbons reach the deep-sea following natural and anthropogenic factors. The process by which they enter deep-sea microbial food webs and impact the biogeochemical cycling of carbon and other elements is unclear. Hydrostatic pressure (HP) is a distinctive parameter of the deep sea, although rarely investigated. Whether HP alone affects the assembly and activity of oil-degrading communities remains to be resolved. Here we have demonstrated that hydrocarbon degradation in deep-sea microbial communities is lower at native HP (10 MPa, about 1000 m below sea surface level) than at ambient pressure. In long-term enrichments, increased HP selectively inhibited obligate hydrocarbon-degraders and downregulated the expression of beta-oxidation-related proteins (i.e., the main hydrocarbon-degradation pathway) resulting in low cell growth and CO2 production. Short-term experiments with HP-adapted synthetic communities confirmed this data, revealing a HP-dependent accumulation of citrate and dihydroxyacetone. Citrate accumulation suggests rates of aerobic oxidation of fatty acids in the TCA cycle were reduced. Dihydroxyacetone is connected to citrate through glycerol metabolism and glycolysis, both upregulated with increased HP. High degradation rates by obligate hydrocarbon-degraders may thus be unfavourable at increased HP, explaining their selective suppression. Through lab-scale cultivation, the present study is the first to highlight a link between impaired cell metabolism and microbial community assembly in hydrocarbon degradation at high HP. Overall, this data indicate that hydrocarbons fate differs substantially in surface waters as compared to deep-sea environments, with in situ low temperature and limited nutrients availability expected to further prolong hydrocarbons persistence at deep sea

Biotechnologies for Marine Oil Spill Cleanup : Indissoluble Ties with Microorganisms

The ubiquitous exploitation of petroleum hydrocarbons (HCs) has been accompanied by accidental spills and chronic pollution in marine ecosystems, including the deep ocean. Physicochemical technologies are available for oil spill cleanup, but HCs must ultimately be mineralized by microorganisms. How environmental factors drive the assembly and activity of HC-degrading microbial communities remains unknown, limiting our capacity to integrate microorganism-based cleanup strategies with current physicochemical remediation technologies. In this review, we summarize recent findings about microbial physiology, metabolism and ecology and describe how microbes can be exploited to create improved biotechnological solutions to clean up marine surface and deep waters, sediments and beaches. Cleaning up oil spills in marine environments ultimately relies on microbial metabolism of HC, which complements the current chemicophysical techniques used in emergency response.Consolidated biotechnologies include microbial communities biostimulation, biosurfactant supplementation and bioaugmentation HC-degrading microbial cells.The effectiveness of biotechnologies is limited by our understanding of the microbial ecology of polluted marine systems. We lack knowledge on how environmental factors, such as hydrostatic pressure, temperature and dispersant toxicity, affect microbial successions.The recent availability of meta-omics data and the improved understanding of microbial metabolism are leading to novel biotechnologies for marine oil spill cleanup, such as slow-release particles for efficient biostimulation and bioelectrochemical approaches for sediment cleanup

A multi-step physicochemical-biotechnological approach for the valorization of olive mill wastewaters

Waste valorization processes carried out through integrated multi-step biorefinery approaches can allow a massive exploitation of the waste organic matter. Olive mill wastewaters (OMWs) are agro-industrial wastes of a high environmental concern. A relevant part of their high COD is typically due to polyphenolic compounds, which are known to be toxic if concentrated to such extents. On the other hands, polyphenols are natural antioxidants of special relevance for several industrial sectors. Therefore, their recovery from OMWs provides the double opportunity to obtain high-added value biomolecules and to reduce the phytotoxicity of the effluent. To such an aim, an effective solid phase extraction process was recently developed [1]. The first aim of the present work was to define a protocol for the recovery and reuse of both the adsorbent (Amberlite XAD16 non-polar resin) and extraction solvent (ethanol), in order to verify the feasibility of a possible process scale-up. Very encouraging results were obtained: ethanol was recovered by means of a rotary evaporator, thus obtaining a concentrated phenolic mixture, whose antioxidant properties were demonstrated via ORAC and DPPH assays; furthermore, after its employment, the resin was washed with a sulphuric acid solution and regenerated: no significant losses of the resin adsorption capabilities were observed after 10 operation cycles. The exploitation of the OMW organic matter was further addressed toward the biotechnological production of biobased chemicals, such as H2 and volatile fatty acids (VFAs), which represent a feasible substrate for aerobic bacteria able to produce and store biopolymers such as polyhydroxyalkanoates (PHAs) [3]. A non conventional anaerobic digestion process carried out under acidogenic conditions for the obtainment of VFAs from dephenolized OMWs was recently developed [4]. The second aim of the present study was a further assessment of that process, with the aim of minimizing the process HRT. At a HRT = 5 days, a stable process capable of an effective bioconversion of the OMW organic matter into VFAs was obtained, with a VFA final concentration of about 19.7 gCOD/L, representing about 83% of the overall effluent COD. References [1] Bertin, L., Ferri, F., Scoma, A., Marchetti, L., Fava, F.: Recovery of high added value natural polyphenols from actual olive mill wastewater through solid phase extraction. Chem. Eng. J. 171, 1287-1293 (2011) [2] Beccari, M., Bertin, L., Dionisi, D., Fava, F., Lampis, S., Majone, M., Valentino, F., Vallini, G., Villano, M.,: Exploiting olive oil mill effluents as a renewable resource for production of biodegradable polymers through a combined anaerobiceaerobic process. J. Chem. Technol. Biotechnol. 84, 901-908 (2009) [3] Scoma, A., Bertin, L., Zanaroli, G., Fraraccio, S., Fava, F.: A physicochemical\u2013biotechnological approach for an integrated valorization of olive mill wastewater. Biores. Technol. 102, 10273-10279 (2011