Understanding the Interactions Between FXR1 and PLC��1

The aim of this project is to understand the interactions between the proteins FXR1 and PLCβ1. It is known that the protein PLCβ1 will bind to FXR1, as FXR1 is a stress granule protein, but not much more of that relationship is known. Using neuronal PC12 cells, different techniques of gene overexpression and fluorescent imaging were used to observe both protein activity and localization. It was hypothesized that PLCβ1 may have specific interactions with FXR1 when subjected to a stressful environment. It was found through these experiments that FXR1 forms stress granules in response to specific stresses. Furthermore, PLCβ1 and FXR1 may not interact with one another directly, but possibly through other protein or RNA interactions

Biogas from source separated organic waste within a circular and life cycle perspective. A case study in Ontario, Canada

Abstract The appropriate transformation and valorisation of biogas offers environmental and economic opportunities in a future with restrictions upon fossil-based fuels and materials. The LCA method was used to quantify and compare the potential environmental impacts of an AD plant incorporating biogas co-generation and upgrading options, namely AD-CHP and AD-RNG. Using an average Anaerobic Digestion facility in Ontario, Canada, modelled after real facilities, as a case study, electricity and steel were identified as potential hotspot input materials carrying a disproportionate environmental burden for biogas production. With a system expansion approach, the biogas was subsequently utilized to produce (1) both heat and electricity using a Combined Heat and Power (CHP) system, or (2) upgraded to renewable natural gas (also called biomethane) through chemical amine scrubbing, respectively. In comparing the biogas co-generation and upgrading options, the AD-CHP alternative resulted in a lesser environmental load, two times lower when compared to the AD-RNG biomethane recovery option. Furthermore, the avoided burden of producing fossil-based electricity, natural gas, and chemical fertilizer was analyzed and compared against their renewable counterparts. Significant reductions in emissions and in the depletion of fossil fuels were achieved, thus confirming the positive efforts of diverting organic waste from landfills to reduce organic waste disposal impacts and improve the management of organic waste. The analysis has provided useful insights to bioenergy project developers, policy makers and the scientific community regarding the processing of source separated organic waste, biogas production, and its upgrading alternatives in a circular economy perspective

Enzymatic Antioxidant Signatures in Hyperthermophilic Archaea

To fight reactive oxygen species (ROS) produced by both the metabolism and strongly oxidative habitats, hyperthermophilic archaea are equipped with an array of antioxidant enzymes whose role is to protect the biological macromolecules from oxidative damage. The most common ROS, such as superoxide radical (O2-.) and hydrogen peroxide (H2O2), are scavenged by superoxide dismutase, peroxiredoxins, and catalase. These enzymes, together with thioredoxin, protein disulfide oxidoreductase, and thioredoxin reductase, which are involved in redox homeostasis, represent the core of the antioxidant system. In this review, we offer a panorama of progression of knowledge on the antioxidative system in aerobic or microaerobic (hyper)thermophilic archaea and possible industrial applications of these enzymes

A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from Thermus thermophilus

Arsenic (As) pollution is a widespread problem worldwide. In recent years, biosensors based on enzymatic inhibition have been developed for arsenic detection, making the study of the effect of inhibitors on the selected enzymatic activity crucial for their setup. The arsenate reductase of Thermus thermophilus HB27, TtArsC, reduces As(V) into As(III), but is also endowed with phosphatase activity. This work investigates the inhibitory effects of As(V) and As(III) on phosphatase activity by taking advantage of a simple colorimetric assay; the results show that both of them are noncompetitive inhibitors affecting the Vmax but not the KM of the reaction. However, their Ki values are different from each other (15.2 ± 1.6 µm for As(V) and 394.4 ± 40.3 µm with As(III)), indicating a higher inhibitory effect by As(V). Moreover, the inhibition-based biosystem results to be selective for As(V) since several other metal ions and salts do not affect TtArsC phosphatase activity; it exhibits a sensitivity of 0.53 ± 0.03 mU/mg/µm and a limit of detection (LOD) of 0.28 ± 0.02 µm. The good sensitivity and specificity for As(V) point to consider inhibition of TtArsC phosphatase activity for the setup of a novel biosensor for the detection of As(V)

Developing a procedure for the integration of Life Cycle Assessment and Emergy Accounting approaches. The Amalfi paper case study

Abstract The analysis of complex systems requires an integrated application of different assessment methods also taking into account different scales and points of view to gain a systemic understanding of the investigated case study. Life Cycle Assessment (LCA) and Emergy Accounting (EMA) are both environmental assessment methods, showing many similarities in the way they are performed, especially with respect to the inventory construction and to the interpretation of results. They also show great differences, the main residing in the different perspectives they give. LCA applies a consumer side perspective, and its space and time scales are set at a boundary capable to include all the process phases in terms of location and durability and their direct impacts on the investigated areas. On the other hand, throughout its donor side perspective, EMA expands the boundaries of the system over the entire biosphere space and time scales. Differences and similarities between LCA and EMA may gain added value by their implementation within a procedural framework which exploits the characteristics of the two methods. The present work proposes a methodological procedure based on the sequential and integrated application of LCA and EMA methods, called LEAF (LCA & EMA Applied Framework). The traditional Amalfi paper production is used as a test case study. The procedure stems include: (i) an ex-ante LCA analysis, to identify the hotspots of the investigated case study; (ii) the assessment of the environmental performance of the system through the development of different EMA-based improvement scenarios built around the chosen hotspots; and (iii) an ex-post LCA application built on each scenario results in order to detect the different environmental burdens. The application of LEAF to the traditional Amalfi paper production shows that the use of a more sustainable energy source is an effective solution (among the set of proposed options) to increase the sustainability of the investigated system

Power generation from slaughterhouse waste materials : an emergy accounting assessment

Unidad de excelencia María de Maeztu MdM-2015-0552The linear path "extraction-production-consumption-waste", imposed by humans to natural ecosystems, where all material flows are instead circular, has become unsustainable. Understanding the potential value of some of these "by-products", in order to exploit them effectively in a biorefinery perspective, may help overcoming resource shortages and decrease environmental impacts. This study investigates energy and resource restoration from animal by-products. The slaughterhouse waste undergoes a rendering process to separate residual meal and fat. The latter is combusted in a co-generation plant to produce electricity and heat. The process is carefully assessed using Emergy Accounting approach with the aim of evaluating benefits and environmental load of the process considering the advantages achieved compared with the demand for ecosystem services and natural capital depletion. Moreover, the case aims at exploring three different methodological assumptions referring to the upstream burdens carried by the waste management system, proposing a modified exergy-based allocation rule. The electricity generated shows performances in terms of Unit Emergy Values ranging between 2.7E+05 sej/J, 2.2E+06 sej/J and 3.1E+07 sej/J among the different cases investigated, comparable to power from fossil fuels and renewables sources, and it provides an environmentally sound alternative to conventional waste disposal

Insight into CAZymes of Alicyclobacillus mali FL18: Characterization of a New Multifunctional GH9 Enzyme

In the bio-based era, cellulolytic and hemicellulolytic enzymes are biocatalysts used in many industrial processes, playing a key role in the conversion of recalcitrant lignocellulosic waste biomasses. In this context, many thermophilic microorganisms are considered as convenient sources of carbohydrate-active enzymes (CAZymes). In this work, a functional genomic annotation of Alicyclobacillus mali FL18, a recently discovered thermo-acidophilic microorganism, showed a wide reservoir of putative CAZymes. Among them, a novel enzyme belonging to the family 9 of glycosyl hydrolases (GHs), named AmCel9, was identified; in-depth in silico analyses highlighted that AmCel9 shares general features with other GH9 members. The synthetic gene was expressed in Escherichia coli and the recombinant protein was purified and characterized. The monomeric enzyme has an optimal catalytic activity at pH 6.0 and has comparable activity at temperatures ranging from 40 °C to 70 °C. It also has a broad substrate specificity, a typical behavior of multifunctional cellulases; the best activity is displayed on β-1,4 linked glucans. Very interestingly, AmCel9 also hydrolyses filter paper and microcrystalline cellulose. This work gives new insights into the properties of a new thermophilic multifunctional GH9 enzyme, that looks a promising biocatalyst for the deconstruction of lignocellulose

Bioprospecting of Extremophilic Microorganisms to Address Environmental Pollution

Geothermal springs are rich in various metal ions due to the interaction between rock and water that takes place in the deep aquifer. Moreover, due to seasonality variation in pH and temperature, fluctuation in element composition is periodically observed within these extreme environments, influencing the environmental microbial communities. Extremophilic microorganisms that thrive in volcanic thermal vents have developed resistance mechanisms to handle several metal ions present in the environment, thus taking part to complex metal biogeochemical cycles. Moreover, extremophiles and their products have found an extensive foothold in the market, and this holds true especially for their enzymes. In this context, their characterization is functional to the development of biosystems and bioprocesses for environmental monitoring and bioremediation. To date, the isolation and cultivation under laboratory conditions of extremophilic microorganisms still represent a bottleneck for fully exploiting their biotechnological potential. This work describes a streamlined protocol for the isolation of thermophilic microorganisms from hot springs as well as their genotypical and phenotypical identification through the following steps: (1) Sampling of microorganisms from geothermal sites ("Pisciarelli", a volcanic area of Campi Flegrei in Naples, Italy); (2) Isolation of heavy metal resistant microorganisms; (3) Identification of microbial isolates; (4) Phenotypical characterization of the isolates. The methodologies described in this work might be generally applied also for the isolation of microorganisms from other extreme environments