No abstract

Three coronaviruses (CoVs) have threatened the world population by causing outbreaks in the last two decades. In late 2019, the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) emerged and caused the coronaviruses to disease 2019 (COVID-19), leading to the ongoing global outbreak. The other pandemic coronaviruses, SARS-CoV and Middle East respiratory syndrome CoV (MERS-CoV), share a considerable level of similarities at genomic and protein levels. However, the differences between them lead to distinct behaviors. These differences result from the accumulation of mutations in the sequence and structure of spike (S) glycoprotein, which plays an essential role in coronavirus infection, pathogenicity, transmission, and evolution. In this review, we brought together many studies narrating a sequence of events and highlighting the differences among S proteins from SARS-CoV, MERS-CoV, and SARS-CoV-2. It was performed here, analysis of S protein sequences and structures from the three pandemic coronaviruses pointing out the mutations among them and what they come through. Additionally, we investigated the receptor-binding domain (RBD) from all S proteins explaining the mutation and biological importance of all of them. Finally, we discuss the mutation in the S protein from several new isolates of SARS-CoV-2, reporting their difference and importance. This review brings into detail how the variations in S protein that make SARS-CoV-2 more aggressive than its relatives coronaviruses and other differences between coronaviruses.

A cowpea class I chitinase (VuChiI) was expressed in the methylotrophic yeast P. pastoris. The recombinant protein was secreted into the culture medium and purified by affinity chromatography on a chitin matrix. The purified chitinase migrated on SDS-polyacrylamide gel electrophoresis as two closely-related bands with apparent molecular masses of 34 and 37 kDa. The identity of these bands as VuChiI was demonstrated by mass spectrometry analysis of tryptic peptides and N-terminal amino acid sequencing. The recombinant chitinase was able to hydrolyze colloidal chitin but did not exhibit enzymatic activity toward synthetic substrates. The highest hydrolytic activity of the cowpea chitinase toward colloidal chitin was observed at pH 5.0. Furthermore, most VuChiI activity (approximately 92%) was retained after heating to 50 °C for 30 min, whereas treatment with 5 mM Cu caused a reduction of 67% in the enzyme's chitinolytic activity. The recombinant protein had antifungal activity as revealed by its ability to inhibit the spore germination and mycelial growth of Penicillium herquei. The three-dimensional structure of VuChiI was resolved at a resolution of 1.55 Å by molecular replacement. The refined model had 245 amino acid residues and 381 water molecules, and the final R-factor and R values were 14.78 and 17.22%, respectively. The catalytic domain of VuChiI adopts an α-helix-rich fold, stabilized by 3 disulfide bridges and possessing a wide catalytic cleft. Analysis of the crystallographic model and molecular docking calculations using chito-oligosaccharides provided evidences about the VuChiI residues involved in sugar binding and catalysis, and a possible mechanism of antifungal action is suggested.

As the addition of low concentrations of oxygen can favor the initial degradation of benzene, toluene, ethylbenzene, and xylenes (BTEX) compounds, this work verified the applicability of the microaerobic technology to enhance BTEX removal in an anaerobic bioreactor supplemented with high and low co-substrate (ethanol) concentrations. Additionally, structural alterations on the bioreactor microbiota were assessed throughout the experiment. The bioreactor was fed with a synthetic BTEX-contaminated water (~ 3 mg L of each compound) and operated at a hydraulic retention time of 48 h. The addition of low concentrations of oxygen (1.0 mL min of atmospheric air at 27 °C and 1 atm) assured high removal efficiencies (> 80%) for all compounds under microaerobic conditions. In fact, the applicability of this technology showed to be viable to enhance BTEX removal from contaminated waters, especially concerning benzene (with a 30% removal increase), which is a very recalcitrant compound under anaerobic conditions. However, high concentrations of ethanol adversely affected BTEX removal, especially benzene, under anaerobic and microaerobic conditions. Finally, although bacterial community richness decreased at low concentrations of ethanol, in general, the bioreactor microbiota could deal with the different operational conditions and preserved its functionality during the whole experiment.

We studied the feasibility of the microaerobic process, in comparison with the traditional chemical absorption process (NaOH), on HS removal in order to improve the biogas quality. The experiment consisted of two systems: R1, biogas from an anaerobic reactor was washed in a NaOH solution, and R2, headspace microaeration with atmospheric air in a former anaerobic reactor. The microaeration used for low sulfate concentration wastewater did not affect the anaerobic digestion, but even increased system stability. Methane production in the R2 was 14 % lower compared to R1, due to biogas dilution by the atmospheric air used. The presence of oxygen in the biogas reveals that not all the oxygen was consumed for sulfide oxidation in the liquid phase indicating mass transfer limitations. The reactor was able to rapidly recover its capacity on HS removal after an operational failure. Bacterial and archaeal richness shifted due to changes in operational parameters, which match with the system functioning. Finally, the microaerobic system seems to be more advantageous for both technical and economical reasons, in which the payback of microaerobic process for HS removal was 4.7 months.

This study aimed to evaluate different air dosing strategies such as microaeration flow rates and air dosing points to enhance HS removal in microaerobic systems treating low-strength wastewaters. Efficiency and stability of the reactors, as well as biogas quality, were assessed, and microbial community changes were evaluated using the PCR-DGGE technique. The results showed that the air dosing point affected the HS concentration and that air dosing at the headspace promoted the highest HS removal efficiency. The airflow rate also affected the process, since HS concentration in the biogas was higher at 0.1 mL air.min than at 0.3 mL air.min. The methane concentration in the biogas was also affected by both air dosing point and flow rate, since the lowest value was observed at the highest airflow rate of the headspace dosing point, due to dilution by the N influx applied to the system. The highest productivity and operational efficiency were observed at this air dosing point, with this airflow (HD), which corroborates with the operational results and the ecological parameters, since the microaeration at this stage promoted high bacterial and archaeal species richness and diversity, optimum functional organization, high COD and HS removal efficiencies.