Preparation and performance evaluation of novel magnetic porous carriers in fluidized bed bioreactor for wastewater treatment.

Biofilm process is a promising wastewater treatment technology and biofilm carrier (biocarrier) is regarded as the core of this process. However, the traditional commercial biocarriers have their inherent drawbacks, therefore, the development of new-type biocarrier to enhance wastewater treatment efficiency is significantly important to biofilm-based reactors. In this study, based on radical suspension polymerization, a novel kind of magnetic porous carriers (PMCs) was prepared by modifying the porous polymer carriers (PPCs) with inorganic particles, and then applied in a fluidized bed bioreactor (FBBR) with a low packing ratio of 10 % (v/v) to synthetic wastewater treatment. The results showed that this novel biocarrier possesses paramagnetism with saturation magnetization of 1.01emu/g, low density (1.26 g/cm3), excellent hydrophilicity (surface water contact angle approaching zero) and rough surface. Besides, compared with the PPCs, the developed PMCs have larger pores (up to 50 µm or more), in which the larger-sized microbes are able to colonize. Moreover, as compared to the PPCs-based FBBR, the PMCs-based reactor achieved shorter time (7 days) for biofilm formaiton and significantly enhanced NH3-N removal efficiency ( nearly 20 % increase at the level of influent NH3-N concentration about 100 mg/L). High-throughput sequencing (HTS) results indicated that this new biocarrier could promote biodiversity and improve the abundance of Nitrosomonadales (the functional bacteria for ammonia removal in the bio-system), thus enhancing the ammonification process. Therefore, the developed PMCs could be preferable biocarriers for biofilm formation and provide an alternative to the traditional suspended biocarrier, demonstrating a promising potential, even at a lower filling ratio, to enhance the pollutants removal performance.

Anaerobic semi-fixed bed biofilm reactor (An-SFB-BR) for treatment of high concentration p-nitrophenol wastewater under shock loading conditions.

P-nitrophenol (PNP or 4-NP) has been widely used as a biorefractory raw material in chemical industry, whereas been highly concerned for its characteristics of mutagenic/carcinogenic activity and food chain bioaccumulation. In this study, an anaerobic semi-fixed bed biofilm reactor (An-SFB-BR) was constructed and used to treat PNP wastewater which discharged from chemical industries. Experimental results revealed that the An-SFB-BR was successfully cultivated with the gradually increasing of influent PNP from 0 to 540 mg/L (gradually increased 10 mg/L every time in stage II and 30-50 mg/L for stage III), with the observation of an average removal efficiency of 98% for PNP and 80% for chemical oxygen demand (COD), also a biogas production and biogas production rate of 2.1 L/(L·d) and 0.57 m3/kg-COD, respectively. Finally, the conversion rate of P-aminophenol (PAP), the primary intermediate of PNP reached 80% after An-SFB-BR biodegradation. A relatively stable pH was maintained throughout the entire process, and insignificant VFA accumulation. The reactor exhibited a strong toxic shock resistance, and 16S rRNA sequencing results demonstrated that the dominant microbial community changed slightly with the gradually increasing of PNP concentration, which guaranteed the PNP removal efficiency.

Complementary peptide sequence coverage using alternative enzymes for on-line digestion with a triaxial electrospray probe.

Using alternative enzymes for on-line digestion with a triaxial electrospray probe extends sequence coverage. This is the first report of utilization of our triaxial probe for on-line analysis with enzymes other than pepsin, suggesting potential for broader application. The probe allows access to processes occurring on a timescale and/or involving substrate conformations complementary to those for conventional (off-line) digestion. Some of the features observed in application to Abeta fibrils are suggestive of unique reactive intermediates during dissolution. Data obtained with enzyme mixtures suggest synergistic effects.

Oxidation artifacts in the electrospray mass spectrometry of Abeta Peptide.

Gradual corrosion of stainless steel electrospray emitters under conditions of normal use generates surface irregularities that can promote electrical discharge. The increased emission current affects the electrochemical reactions associated with the spray process. When sampling the peptide Abeta(1-40), this is manifest by oxidation of methionine at position 35 to methionine sulfoxide. The resultant mass shift and reduced sensitivity can adversely affect H/D exchange experiments. These effects can be avoided by adding a redox buffer or (preferably) by repolishing the emitter, especially to a rounded geometry.

A triaxial probe for on-line proteolysis coupled with hydrogen/deuterium exchange-electrospray mass spectrometry.

An on-line proteolysis system utilizing a triaxial electrospray probe was developed to aid localization of the hydrogen-bonding interaction sites in hydrogen/deuterium exchange-mass spectrometry (HDX-MS) studies of Abeta (1-40) fibrils. The probe allows delayed introduction of the organic solvent component needed for stable electrospray, thus enhancing hydrolysis performance relative to that of a coaxial probe. Effective on-line digestion was accomplished in approximately 12 s. The probe should be of general utility for HDX-MS studies of amyloid fibrils and other protein aggregates.

Structural differences in Abeta amyloid protofibrils and fibrils mapped by hydrogen exchange--mass spectrometry with on-line proteolytic fragmentation.

We report here structural differences between Abeta(1-40) protofibrils and mature amyloid fibrils associated with Alzheimer's disease as determined using hydrogen-deuterium exchange-mass spectrometry (HX-MS) coupled with on-line proteolysis. Specifically, we have identified regions of the Abeta(1-40) peptide containing backbone amide hydrogen atoms that are protected from HX or exposed when this peptide is incorporated into protofibrils or amyloid fibrils formed in phosphate-buffered saline without stirring at 37 degrees C. Study of protofibrils was facilitated by use of the protofibril-stabilizing agent calmidazolium chloride. Our data clearly show that both the C-terminal segment 35-40 and the N-terminal segment 1-19 are highly exposed to HX in both fibrils and protofibrils. In contrast, the internal fragment 20-34 is highly protected from exchange in fibrils but much less so in protofibrils. The data suggest that the beta-sheet elements comprising the amyloid fibril are already present in protofibrils, but that they are expanded into some adjacent residues upon the formation of mature amyloid. The N-terminal approximately ten residues appear to be unstructured in both protofibrils and fibrils. The 20-30 segment of Abeta(1-40) is more ordered in fibrils than in protofibrils, suggesting that, if protofibrils are a mechanistic precursor of fibrils, the transition from protofibril to fibril involves substantial ordering of this region of the Abeta peptide.

Structural properties of Abeta protofibrils stabilized by a small molecule.

Metastable oligomeric and protofibrillar forms of amyloidogenic proteins have been implicated as on-pathway assembly intermediates in amyloid formation and as the major toxic species in a number of amyloid diseases including Alzheimer's disease. We describe here a chemical biology approach to structural analysis of Abeta protofibrils. Library screening yielded several molecules that stimulate Abeta aggregation. One of these compounds, calmidazolium chloride (CLC), rapidly and efficiently converts Abeta(1-40) monomers into clusters of protofibrils. As monitored by electron microscopy, these protofibrils persist for days when incubated in PBS at 37 degrees C, with a slow transition to fibrillar structures apparent only after several weeks. Like normal protofibrils, the CLC-Abeta aggregates exhibit a low thioflavin T response. Like Abeta fibrils, the clustered protofibrils bind the anti-amyloid Ab WO1. The CLC-Abeta aggregates exhibit the same protection from hydrogen-deuterium exchange as do protofibrils isolated from a spontaneous Abeta fibril formation reaction: approximately 12 of the 39 Abeta(1-40) backbone amide protons are protected from exchange in the protofibril, compared with approximately twice that number in amyloid fibrils. Scanning proline mutagenesis analysis shows that the Abeta molecule in these protofibrillar assemblies exhibits the same flexible N and C termini as do mature amyloid fibrils. The major difference in Abeta conformation between fibrils and protofibrils is added structural definition in the 22-29 segment in the fibril. Besides aiding structural analysis, compounds capable of facilitating oligomer and protofibril formation might have therapeutic potential, if they act to sequester Abeta in a form and/or location that cannot engage the toxic pathway.

Characterization of the rutin-metal complex by electrospray ionization tandem mass spectrometry.

According to the strong application background of bioflavonoid and metal-flavonoid complexes, novel electrospray ionization tandem mass spectrometry (ESI-MSn) was applied to investigate the structure and fragmentation mechanism of transition metal-rutin complexes. In the full-scan mass spectra, different stoichiometric ratios of rutin-metal complexes were found. In the reaction between rutin and Cu, four kinds of complexes with four different stoichiometric ratios were produced. In the reaction between rutin and Zn, Mn(II), and Fe(II), only two kind of complexes with stoichiometric ratios of 1:1 and 1:2 occured. In further tandem mass spectrometric experiments of different rutin-metal complexes, product fragments came from the neutral loss of the external rhamnose and the internal glucose unit, oligosaccharide chain, aglycone, and small organic molecules. According to the MSn data, we proposed a mechanism for all fragments of the rutin-Cu complex A and the structure of two rutin-Cu complexes, C and D.

Analysis of flavonoid constituents from leaves of Acanthopanax senticosus harms by electrospray tandem mass spectrometry.

Three known flavonoids, quercetin, quercitrin (quercetin-3-0-rhamnoside) and rutin (quercetin-3-0 rutinoside), have been identified for the first time in the leaves of Acanthopanax senticosus Harms by using electrospray tandem mass spectrometry techniques (ESI-MS(n)). The flavonoid hyperin (quercetin-3-0-beta-galactoside), already known to be present, was also investigated. The diagnostic fragment ions of the aglycone quercetin were obtained in the ESI-MS(n) experiments, and a fragmentation mechanism proposed.