TiO2 with Tandem Fractionation (TAFT): An Approach for Rapid, Deep, Reproducible, and High-Throughput Phosphoproteome Analysis.

Mass-spectrometry-based phosphoproteomic workflows traditionally require efficient prefractionation and enrichment of phosphopeptides to gain an in-depth, global, and unbiased systematic investigation of phosphoproteome. Here we present TiO2 with tandem fractionation (TAFT) approach, which combines titanium dioxide (TiO2) enrichment and tandem high-pH reverse-phase (HpRP) for phosphoproteome analysis in a high-throughput manner; the entire workflow takes only 3 h to complete without laborious phosphopeptide preparation. We applied this approach to HeLa and HepG2.2.15 cells to characterize the capability of TAFT approach, which enables deep identification and quantification of more than 14 000 unique phosphopeptides in a single sample from 1 mg of protein as starting materials in <4 h of MS measurement. In total, we identified and quantified 21 281 phosphosites in two cell lines with >91% selectivity and high quantitative reproducibility (average Pearson correlation is 0.90 between biological replicates). More generally, the presented approach enables rapid, deep, and reproducible phosphoproteome analysis in a high-throughput manner with low cost, which should facilitate our understanding of signaling networks in a wide range of biological systems or the process of clinical applications.

Fabrication of high performance TFN membrane incorporated with graphene oxide via support-free interfacial polymerization.

A high-performance thin film nanocomposite (TFN) membrane containing graphene oxide (GO) nanosheets was constructed using a support-free interfacial polymerization (SFIP) technique. In this study, an ultrathin composited polyamide (PA) nanofilm was synthesized at the free piperazine (PIP)-GO suspension/trimesoyl chloride (TMC) interface, followed by transfer onto a polysulfone (PSf) UF substrate. The impact of GO loading (0, 0.1, 0.5, or 1 mg/mL) on the physiochemical properties, surface morphology, and hydrophilicity of the composited PA layer and membrane separation performance was investigated. It was found that the GO-modified TFN membranes showed ultra-high hydrophilicity due to the increase in the number of carboxyl and hydroxyl groups in the PA layer. We propose that GO nanosheets play a key role in improving membrane permeability because a strong hydration layer is formed between the water molecules and GO (embedded in the PA layer), acting as a protective film and minimizing the chance of foulants contacting the membrane surface. Compared with TFC, TFN-GO-0.5 simultaneously exhibited a higher water permeability of up to 12.8 L·m-2·h-1·bar-1 (58.1% higher than the TFC membrane) and a higher Na2SO4 rejection of approximately 98.4%. Moreover, the introduction of GO nanosheets into TFN membrane resulted in an improved antifouling performance. This facile SFIP method reveals the potential of GO nanosheets for the development of high performance TFN membranes.

Preparation of low carbon olefins on a core-shell K-Fe5C2@ZSM-5 catalyst by Fischer-Tropsch synthesis.

In this study, a core-shell catalyst based on Fe5C2@ZSM-5 (ZSM-5 capped Fe5C2 as active phase) is prepared by the coating-carbonization method for Fischer-Tropsch synthesis (FTS). Further, the designed ZSM-5 zeolites are utilized to screen the low carbon hydrocarbons from the products generated on the iron carbide active centre, and for catalytic disassembly of the long-chain hydrocarbons into low carbon olefins. Prior to utilization, the physical-chemical properties of the prepared catalysts are systematically characterized by various techniques of X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET), Fourier transform infrared (FT-IR), and scanning electron microscopy (SEM) as well as transmission electron microscopy (TEM) observations, in addition to the effects of coating-carbonization, molecular sieve coating amount, and K-doping on core-shell iron-based catalysts. Next, the performance of Fischer-Tropsch synthesis is investigated in a micro-fixed bed reactor. The results manifest that, comparing with Fe5C2 and a supported Fe/ZSM-5 catalyst prepared by the traditional impregnation method, the core-shell Fe5C2@ZSM-5 catalysts show higher CO conversion rate, reaction activity and selectivity to low-carbon olefins. Comparatively, the Fe5C2@ZSM-5C catalyst prepared by carbonization after the coating method exhibited more surface area, smaller average pore size, and more reactive active sites, resulting in the improvement of screening of high carbon hydrocarbons and the enhancement of selectivity to low carbon olefins, in comparison to those prepared by the carbonization-coating method. In conclusion, the K-doping catalyst had significantly improved the reactive activity of the core-shell Fe5C2@ZSM-5 catalyst and the selectivity to low carbon olefins, while the CO conversion on K-Fe5C2@ZSM-20C still remained good.

Genome-based analysis to understanding rapid resuscitation of cryopreserved anammox consortia via sequential supernatant addition.

Simple cryopreservation of anaerobic ammonium-oxidation (anammox) consortia has become a promising preservation technology for the fast start-up of the anammox process. Here, we use genome-resolved metagenomics and metatranscriptomics to understand of the microbial interaction in a simple and effective resuscitation process for long-term cryopreserved anammox consortia by sequential addition of anammox SBR supernatant. Performance results showed that sequential addition of anammox supernatant significantly reduced the resuscitation time of the granule-based anammox process from 40 to 20 days. Genome-centric metagenomics were used to recover 19 high-quality draft genomes of anammox and heterotrophic bacteria. Comparative metatranscriptomic analysis was conducted to examine the gene expression of Candidatus Kuenenia stuttgartiensis, the dominant anammox bacterium, and heterotrophic bacteria to better understand their potential interactions. Proteobacteria-affiliated bacteria found in the supernatant were highly active in producing the secondary metabolites molybdopterin cofactor and folate which are needed for growth of the auxotrophic anammox bacteria. In addition, the significantly higher expression levels of hzsA and CO2-fixtion genes in the Candidatus Kuenenia genome indicated the anammox bacteria were likely more active and growing faster after sequential anammox supernatant addition during the resuscitation process. The resuscitation treatment pulse assays confirmed that sequential addition of supernatant was an effective way for the rapid resuscitation of anammox consortia. Our findings offer the first evidence of cross-feeding during the rapid resuscitation of cryopreserved anammox consortia.

Photophysical properties and singlet oxygen generation of three sets of halogenated corroles.

The luminescence, excited-state absorption, and singlet oxygen generation measurements were performed on three kinds of halogenated corroles: monohydroxyl halogenated corroles (Corrole-F, Corrole-Cl, Corrole-I), peripherally fluorine-substituted corroles (F0, F5, F10, F15), and gallium complexes (F10-Ga, F15-Ga). The fluorescence intensities progressively decrease whereas the triplet quantum yields, oxygen quenching rates, and singlet oxygen quantum yields increase with the increasing of the monohydroxyl halogen atomic weight. Replacing hydrogen atoms of meso-phenyl groups with fluorine atoms induces the blue-shifts of the emission spectra, higher triplet quantum yield, and smaller oxygen quenching rates. Of all peripherally fluorine-substituted corroles, F10 exhibited the highest singlet oxygen quantum yield. In comparison with the free base corroles, both gallium corrole complexes display much stronger fluorescence with the large blue-shifts of emission peaks and slightly higher triplet quantum yields but smaller oxygen quenching rates and singlet oxygen quantum yields. The reasons for the different photophysical behaviors of these corroles are discussed.