Genomics- and Transcriptomics-Guided Discovery of Clavatols from Arctic Fungi Penicillium sp. MYA5.

Clavatols exhibit a wide range of biological activities due to their diverse structures. A genome mining strategy identified an A5cla cluster from Penicillium sp. MYA5, derived from the Arctic plant Dryas octopetala, is responsible for clavatol biosynthesis. Seven clavatols, including one new clavatol derivate named penicophenone F (1) and six known clavatols (2-7), were isolated from Penicillium sp. MYA5 using a transcriptome mining strategy. These structures were elucidated by comprehensive spectroscopic analysis. Antibacterial, aldose reductase inhibition, and siderophore-producing ability assays were conducted on compounds 1-7. Compounds 1 and 2 demonstrated inhibitory effects on the ALR2 enzyme with inhibition rates of 75.3% and 71.6% at a concentration of 10 µM, respectively. Compound 6 exhibited antibacterial activity against Staphylococcus aureus and Escherichia coli with MIC values of 4.0 µg/mL and 4.0 µg/mL, respectively. Additionally, compounds 1, 5, and 6 also showed potential iron-binding ability.

Characterization and high-yield production of non-N-glycosylated recombinant human BCMA-Fc in Pichia pastoris.

B-cell maturation antigen (BCMA) fused at the C-terminus to the Fc portion of human IgG1 (BCMA-Fc) blocks B-cell activating factor (BAFF) and proliferation-inducing ligand (APRIL)-mediated B-cell activation, leading to immune disorders. The fusion protein has been cloned and produced by several engineering cell lines. To reduce cost and enhance production, we attempted to express recombinant human BCMA-Fc (rhBCMA-Fc) in Pichia pastoris under the control of the AOX1 methanol-inducible promoter. To produce the target protein with uniform molecular weight and reduced immunogenicity, we mutated two predicted N-linked glycosylation sites. The secretory yield was improved by codon optimization of the target gene sequence. After fed-batch fermentation under optimized conditions, the highest yield (207 mg/L) of rhBCMA-Fc was obtained with high productivity (3.45 mg/L/h). The purified functional rhBCMA-Fc possessed high-binding affinity to APRIL and dose-dependent inhibition of APRIL-induced proliferative activity in vitro through three-step purification. Thus, this yeast-derived expression method could be a low-cost and effective alternative to the production of rhBCMA-Fc in mammalian cell lines.

An electrochemical immunosensor for ultrasensitive detection of carbohydrate antigen 199 based on Au@Cu(x)OS yolk-shell nanostructures with porous shells as labels.

A novel and sensitive electrochemical immunosensor for ultrasensitive detection of pancreatic cancer biomarker carbohydrate antigen 199 (CA199) was proposed by using Au@Cu(x)OS yolk-shell nanostructures with porous shells as labels for signal amplification. Au@Cu(x)OS yolk-shell nanostructures exhibit high electrocatalytic activity toward the reduction of hydrogen peroxide (H2O2) as analytical signal. Moreover, secondary antibody (Ab2) can adsorb on the surface of Au@Cu(x)OS with porous shells which has large surface area and could greatly increase the probability of Ab2-antigen interactions thereby leading to higher sensitivity. Reduced graphene oxide-tetraethylene pentamine (rGO-TEPA), containing abundant amine groups, was supported Au nanoparticles as a support platform to immobilize the primary antibody (Ab1). The resulting sensing interface of rGO-TEPA/AuNPs could provide a large electroconductive surface area, allowing high loadings of the biological recognition elements as well as the occurrence of electrocatalytic and electron-transfer processes. Under optimal conditions, the immunosensor exhibited a wide linear response to CA199 ranging from 0.001 to 12 U/mL with a low detection limit of 0.0005 U/mL. The designed immunosensor displayed good precision, high sensitivity, acceptable stability and reproducibility, and has been applied to the analysis of serum with satisfactory results. The proposed method provides a new promising platform of clinical immunoassay for other biomolecules.

Effect of contact to the atmosphere and dilution on phosphorus recovery from human urine through struvite formation.

Phosphorus (P) in hydrolysed urine can be recovered through struvite formation. In the present study, batch experiments were conducted to investigate the effects of contact to the atmosphere (i.e. open and closed) and dilution [Vurine/(Vurine + Vwater)] (i.e. 100%, 50% and 25%) on P recovery from fresh urine through struvite formation with the addition of magnesium chloride (molar ratios of Mg/P = 1.3 and 2.0) after 32 d of storage. The P loss mainly occurred during the initial stages of precipitation with calcium and magnesium (5 d). The precipitates formed at the bottom of the jars were identified by X-ray diffraction to be struvite, hydroxyapatite and calcite. The results showed that the P recovery efficiency from urine solutions in open jars was lower than that in closed jars. It caused significant ammonia volatilization in open jars, resulting in higher nitrogen loss, lower pH values and lower supersaturation. The P recovery efficiency decreased with dilution, which is related to lower pH and lower supersaturation resulting from water dilution. An increase in the Mg/P ratio from 1.3 to 2.0 enhanced P recovery to some extent in urine solutions with different dilutions. The largest P recovery efficiency was 93.7% and 97.3% at an Mg/P ratio of 1.3 and 2.0 for the 100% urine solutions in closedjars, respectively. Scanning electron microscopy revealed smaller struvite particle sizes at lower dilutions (100% and 50% urine) compared with higher dilutions (25% urine).

Influence of process parameters on phosphorus recovery by struvite formation from urine.

Batch experiments were conducted to examine the influence of various process parameters on phosphorus (P) recovery by struvite formation from urine. The results showed that the Mg/P molar ratio is one of the most important parameters affecting P recovery. The Mg/P molar ratio of 1.3 was found the most reasonable for struvite formation, and the P removal efficiency reached more than 96.6%. An increase in pH of urine solutions from 8.7 to 10.0 did not significantly affect P removal, but the quality of crystal formed at pH 10.0 was poor based on scanning electron microscopy analysis. A longer mixing time positively affected struvite formation, and compared to without mixing, the P removal efficiency increased from 72.7 to 97.3% after 5 min of mixing. The addition of seed material had no influence on the P removal efficiency, but contributed to the formation of struvite clusters.

The load dependence of the physical properties of a molecular motor.

The physical properties of a molecular motor with load changing in a wide range will be discussed in this study, in particular the mean velocity, output power and energy efficiency. The main difficulty of this study is that both the states of the molecular motor and the energy barriers between them change with the loading force. Moreover, with the change of load, the number of motor states may also change, so different models should be used to calculate the corresponding physical quantities in different ranges of load. The results show that, in contrast to the usual intuition, the mean velocity and output power of the molecular motor do not change continuously with load.