Defining functional diversity for lignocellulose degradation in a microbial community using multi-omics studies

Abstract\ud \ud Background\ud Lignocellulose is one of the most abundant forms of fixed carbon in the biosphere. Current industrial approaches to the degradation of lignocellulose employ enzyme mixtures, usually from a single fungal species, which are only effective in hydrolyzing polysaccharides following biomass pre-treatments. While the enzymatic mechanisms of lignocellulose degradation have been characterized in detail in individual microbial species, the microbial communities that efficiently breakdown plant materials in nature are species rich and secrete a myriad of enzymes to perform “community-level” metabolism of lignocellulose. Single-species approaches are, therefore, likely to miss important aspects of lignocellulose degradation that will be central to optimizing commercial processes.\ud \ud \ud Results\ud Here, we investigated the microbial degradation of wheat straw in liquid cultures that had been inoculated with wheat straw compost. Samples taken at selected time points were subjected to multi-omics analysis with the aim of identifying new microbial mechanisms for lignocellulose degradation that could be applied in industrial pre-treatment of feedstocks. Phylogenetic composition of the community, based on sequenced bacterial and eukaryotic ribosomal genes, showed a gradual decrease in complexity and diversity over time due to microbial enrichment. Taxonomic affiliation of bacterial species showed dominance of Bacteroidetes and Proteobacteria and high relative abundance of genera Asticcacaulis, Leadbetterella and Truepera. The eukaryotic members of the community were enriched in peritrich ciliates from genus Telotrochidium that thrived in the liquid cultures compared to fungal species that were present in low abundance. A targeted metasecretome approach combined with metatranscriptomics analysis, identified 1127 proteins and showed the presence of numerous carbohydrate-active enzymes extracted from the biomass-bound fractions and from the culture supernatant. This revealed a wide array of hydrolytic cellulases, hemicellulases and carbohydrate-binding modules involved in lignocellulose degradation. The expression of these activities correlated to the changes in the biomass composition observed by FTIR and ssNMR measurements.\ud \ud \ud Conclusions\ud A combination of mass spectrometry-based proteomics coupled with metatranscriptomics has enabled the identification of a large number of lignocellulose degrading enzymes that can now be further explored for the development of improved enzyme cocktails for the treatment of plant-based feedstocks. In addition to the expected carbohydrate-active enzymes, our studies reveal a large number of unknown proteins, some of which may play a crucial role in community-based lignocellulose degradation.This work was funded by Biotechnology and Biological Sciences Research\ud Council (BBSRC) Grants BB/1018492/1, BB/K020358/1 and BB/P027717/1, the\ud BBSRC Network in Biotechnology and Bioenergy BIOCATNET and São Paulo\ud Research Foundation (FAPESP) Grant 10/52362-5. ERdA thanks EMBRAPA\ud Instrumentation São Carlos and Dr. Luiz Alberto Colnago for providing the\ud NMR facility and CNPq Grant 312852/2014-2. The authors would like to thank\ud Deborah Rathbone and Susan Heywood from the Biorenewables Develop‑\ud ment Centre for technical assistance in rRNA amplicon sequencing

Reversible doping of graphene field effect transistors by molecular hydrogen: The role of the metal/graphene interface

In this work, we present an investigation regarding how and why molecular hydrogen (H2) changes the electronic properties of graphene field effect transistors (GFETs). We demonstrate that interaction with H2 leads to local doping of graphene near of the graphene-contact heterojunction. We also show that such interaction is strongly dependent on the characteristics of the metal-graphene interface. By changing the type of metal in the contact, we observe that Ohmic contacts can be strongly or weakly electrostatically coupled with graphene. For strongly coupled contacts, the signature of the charge transfer effect promoted by the contacts results on asymmetric ambipolar conduction, and such asymmetry can be tunable under interaction with H2. On the other hand, for contacts weakly coupled with graphene, the hydrogen interaction has a more profound effect. In such a situation, the devices show a second charge neutrality point (CNP) in graphene transistor transfer curves (a double-peak response) upon H2 exposure. We propose that this double-peak phenomenon arises from the decoupling of the work function of graphene and that of the metallic electrodes induced by the H2 molecules. We also show that the gas-induced modifications at the metal-graphene interface can be exploited to create a controlled graphene p-n junction, with considerable electron transfer to graphene layer and significant variation in the graphene resistance. These effects can pave the way for a suitable metallic contact engineering providing great potential for the application of such devices as gas sensors

Enhancing the response of NH<inf>3</inf> graphene-sensors by using devices with different graphene-substrate distances

Graphene (G) is a two-dimensional material with exceptional sensing properties. In general, graphene gas sensors are produced in field effect transistor configuration on several substrates. The role of the substrates on the sensor characteristics has not yet been entirely established. To provide further insight on the interaction between ammonia molecules (NH3) and graphene devices, we report experimental and theoretical studies of NH3 graphene sensors with graphene supported on three substrates: SiO2, talc and hexagonal boron nitride (hBN). Our results indicate that the charge transfer from NH3 to graphene depends not only on extrinsic parameters like temperature and gas concentration, but also on the average distance between the graphene sheet and the substrate. We find that the average distance between graphene and hBN crystals is the smallest among the three substrates, and that graphene-ammonia gas sensors based on a G/hBN heterostructure exhibit the fastest recovery times for NH3 exposure and are slightly affected by wet or dry air environment. Moreover, the dependence of graphene-ammonia sensors on different substrates indicates that graphene sensors exhibit two different adsorption processes for NH3 molecules: one at the top of the graphene surface and another at its bottom side close to the substrate. Therefore, our findings show that substrate engineering is crucial to the development of graphene-based gas sensors and indicate additional routes for faster sensors

Probing the Electronic Properties of Monolayer MoS<inf>2</inf> via Interaction with Molecular Hydrogen

This work presents a detailed experimental investigation of the interaction between molecular hydrogen (H2) and monolayer MoS2 field effect transistors (MoS2 FET), aiming for sensing application. The MoS2 FET exhibits a response to H2 that covers a broad range of concentration (0.1–90%) at a relatively low operating temperature range (300–473 K). Most important, H2 sensors based on MoS2 FETs show desirable properties such as full reversibility and absence of catalytic metal dopants (Pt or Pd). The experimental results indicate that the conductivity of MoS2 monotonically increases as a function of the H2 concentration due to a reversible charge transferring process. It is proposed that such process involves dissociative H2 adsorption driven by interaction with sulfur vacancies in the MoS2 surface (VS). This description is in agreement with related density functional theory studies about H2 adsorption on MoS2. Finally, measurements on partially defect-passivated MoS2 FETs using atomic layer deposited aluminum oxide consist of an experimental indication that the VS plays an important role in the H2 interaction with the MoS2. These findings provide insights for future applications in catalytic process between monolayer MoS2 and H2 and also introduce MoS2 FETs as promising H2 sensors