Molecular cloning and characterization of a novel bifunctional cellobiohydrolase/ß-xylosidase from a metagenomic library of mangrove soil.

A metagenomic library of mangrove soil samples consisting of approximately 11,000 clones was constructed, and a rare bifunctional cellobiohydrolase/ß-xylosidase Cbh2124 was identified by functional screening. Cbh2124 displayed the highest homology (56.43%) with a protein of the glycoside hydrolase 10 (GH10) family from Proteobacteria. Phylogenetic analysis confirmed that Cbh2124 belongs to the GH10 family. The recombinant enzyme showed a strong cellobiohydrolase activity and a relatively high ß-xylosidase activity, and its catalytic efficiency to the cellobiose substrate was as high as 1.27 × 105 s-1·mM-1, the highest efficiency among reported cellobiohydrolases. Of particular interest, some enzymatic properties of the ß-xylosidase activity of Cbh2124 were significantly different from those of the cellobiohydrolase activity. The optimal pH and temperature of the cellobiohydrolase activity of Cbh2124 was 6.4 and 36 °C, and the activity was essentially lost after treatment at 45 °C for 1 h. The optimal pH and temperature of the ß-xylosidase activity of Cbh2124 was 8.0 and 60 °C, and the residual activity was still over 90% after treatment at 80 °C for 6 h. The molecular docking results of the ß-xylosidase activity of Cbh2124 revealed the additional presence of catalytic amino acids Ser175 and Lys420, thus increasing the number of hydrogen bonds involved in the catalytic process, which possibly let to the improved thermostability compared with that of the cellobiohydrolase activity.

Boundary conditions for exhaled airflow from a cough with a surgical or N95 mask.

Wearing surgical or N95 masks is effective in reducing the infection risks of airborne infectious diseases. However, in the literature there are no detailed boundary conditions for airflow from a cough when a surgical or N95 mask is worn. These boundary conditions are essential for accurate prediction of exhaled particle dispersion by computational fluid dynamics (CFD). This study first constructed a coughing manikin with an exhalation system to simulate a cough from a person. The smoke visualization method was used to measure the airflow profile from a cough. To validate the setup of the coughing manikin, the results were compared with measured data from subject tests reported in the literature. The validated coughing manikin was then used to measure the airflow boundary conditions for a cough when a surgical mask was worn and when an N95 mask was worn, respectively. Finally, this study applied the developed airflow boundary conditions to calculate person-to-person particle transport from a cough when masks are worn. The calculated exhaled particle patterns agreed well with the smoke pattern in the visualization experiments. Furthermore, the calculated results indicated that, when the index person wore a surgical and a N95 mask, the total exposure of the receptor was reduced by 93.0% and 98.8%, respectively.

Temporal and spatial far-ultraviolet disinfection of exhaled bioaerosols in a mechanically ventilated space.

Far-UVC with a peak wavelength of 222 nm can potentially be used to inactivate exhaled bioaerosols in an efficient and safe manner. Therefore, this study aimed to experimentally explore the effectiveness of a 222 nm far-UVC light for inactivating bioaerosols, represented by E. coli, exhaled from a manikin in a chamber with mechanical ventilation. The spatial irradiance distribution from the far-UVC light was measured. The susceptibility constant (z-value) for E. coli under the far-UVC light was experimentally obtained. The temporal and spatial concentrations of the bioaerosols exhaled from the manikin were measured under three typical ventilation rates (4, 10, and 36 ACH). According to the results, when the far-UVC light was turned on, the bioaerosol concentrations were lower than those without the far-UVC light under all three ventilation rates, suggesting that far-UVC light can effectively disinfect E. coli under mechanical ventilation. However, the disinfection efficiency of the far-UVC light decreased as the ventilation rate increased, which indicated that the far-UVC light played a more important role in bioaerosol removal under a lower ventilation rate. In general, the results supported the feasibility of using 222 nm far-UVC light for disinfection of exhaled bioaerosols in mechanically ventilated spaces to reduce infection risks.

Characterization of a novel thermostable alkaline lipase derived from a compost metagenomic library and its potential application in the detergent industry.

Using composted soil samples, a metagenomic library consisting of 36,000 clones was constructed. Then, a novel lipase, Lip54q, which belongs to the VIII family of lipolytic enzymes, was identified from the metagenomic library by functional screening. To explore the enzymatic properties of Lip54q, lip54q was heterologous expressed in Escherichia coli with a high expression level of recombinant protein up to 720 mg/L. The recombinant enzyme showed the highest activity (28,160 U/mg) against a C10 substrate at pH 9.0 and 47°C, and was stable at temperatures ≤50°C and pH 8.0-11.0. Of particular interest, the surfactants, Tween-20, Tween-80 and Tritonx-100, exhibited strong promoting effects on Lip54q activities regardless of whether low concentrations (0.1%) or high concentrations (10%) were used. Application studies of Lip54q using six commercial detergents indicated that the enzyme had strong tolerance and immersion resistance to all six detergents. The results of oil-stain removal experiments suggested that addition of the enzyme to various commercial detergents could significantly improve the abilities of these detergents to remove oil-stains. Furthermore, the results of a molecular docking analysis of Lip54q showed that both the C10 substrate and linoleic acid molecules could form hydrogen bond interactions with the catalytic amino acids, Ser-268, Glu-168, and Asp-192, in the catalytic center of the enzyme, and the hydrogen bond distances were shorter. The electrostatic attraction between the enzyme and the substrate formed by the hydrogen bond with a shorter distance is stronger, which is conducive to the formation of a more stable complex between the enzyme and the substrate, thus increasing the activity of the enzyme to such substrate. These results 1ay a good foundation for application of this enzyme in the detergent industry in the future.

Similar seed dispersal systems by local frugivorous birds in native and alien plant species in a coastal seawall forest.

Frugivorous birds play an important role in seed dispersal. Alien plant species' seeds are dispersed by local birds in order to establish populations in new habitats. Alien plant species that produce fruits similar to that of native species have the potential to attract local birds, creating new mutualistic systems that are similar to the local ones. In autumn 2018 and 2019, we studied the seed dispersal systems of an alien plant species, Phytolacca americana, and a native species, Cayratia japonica, in a coastal seawall forest. Both plant species' fruit, frugivorous bird foraging behaviors, seed germination rates, and seedling microhabitats were examined to determine whether the alien species had a similar seed dispersal system to that of the native species. Our results showed that P. americana and C. japonica had similar fruit type, color, and ripening period. There was a positive correlation between the percentage rate of fruit ripening and the percentage rate of fruit missing for both plant species, indicating that local frugivorous birds have the potential to sufficiently disperse the alien seeds to enable its spread in the coastal seawall forest (simple linear regression, P. americana: ß = 0.863 ± 0.017, R2 adj = 0.978, P < 0.01; C. japonica: ß = 0.787 ± 0.034, R2 adj = 0.898, P < 0.01). Eleven bird species consumed the fruits of the alien species or native species during the study period. Similar results were shown across alien and native species in bird foraging behavior (feeding frequency, feeding duration and first stop distance) indicating that a similar seed dispersal relationship had been established between local frugivorous and both plant species. The alien plant had a higher number of fruits carried by birds, suggesting that P. americana had a slightly higher fruit consumption than that of C. japonica (t-test, P < 0.01). Alien plant seedlings grow more abundant in forest gap microhabitat (t-test, P < 0.01). Our results confirmed that bird digestion promotes seed germination success in both plant species. Our study suggests that in a narrow coastal seawall forest, alien plant species can successfully establish their populations by relying on similar seed dispersal systems as the local species.

Modeling transient particle transport in transient indoor airflow by fast fluid dynamics with the Markov chain method.

It is crucial to accurately and efficiently predict transient particle transport in indoor environments to improve air distribution design and reduce health risks. For steady-state indoor airflow, fast fluid dynamics (FFD) + Markov chain model increased the calculation speed by around seven times compared to computational fluid dynamics (CFD) + Eulerian model and CFD + Lagrangian model, while achieving the same level of accuracy. However, the indoor airflow could be transient, if there were human behaviors involved like coughing or sneezing and air was supplied periodically. Therefore, this study developed an FFD + Markov chain model solver for predicting transient particle transport in transient indoor airflow. This investigation used two cases, transient particle transport in a ventilated two-zone chamber and a chamber with periodic air supplies, for validation. Case 1 had experimental data for validation and the results showed that the predicted particle concentration by FFD + Markov chain model matched well with the experimental data. Besides, it had similar accuracy as the CFD + Eulerian model. In the second case, the prediction by large eddy simulation (LES) was used for validating the FFD. The simulated particle concentrations by the Markov chain model and the Eulerian model were similar. The simulated particle concentrations by the Markov chain model and the Eulerian model were similar. The computational time of the FFD + Markov chain model was 7.8 times less than that of the CFD + Eulerian model.