Purification and Properties of a Plasmin-like Marine Protease from Clamworm (Perinereis aibuhitensis).

Marine organisms are a rich source of enzymes that exhibit excellent biological activity and a wide range of applications. However, there has been limited research on the proteases found in marine mudflat organisms. Based on this background, the marine fibrinolytic enzyme FELP, which was isolated and purified from clamworm (Perinereis aibuhitensis), has exhibited excellent fibrinolytic activity. We demonstrated the FELP with a purification of 10.61-fold by precipitation with ammonium sulfate, ion-exchange chromatography, and gel-filtration chromatography. SDS-PAGE, fibrin plate method, and LC-MS/MS indicated that the molecular weight of FELP is 28.9 kDa and identified FELP as a fibrinolytic enzyme-like protease. FELP displayed the maximum fibrinolytic activity at pH 9 (407 ± 16 mm2) and 50 °C (724 ± 27 mm2) and had excellent stability at pH 7-11 (50%) or 30-60 °C (60%), respectively. The three-dimensional structure of some amino acid residues of FELP was predicted with the SWISS-MODEL. The fibrinolytic and fibrinogenolytic assays showed that the enzyme possessed direct fibrinolytic activity and indirect fibrinolysis via the activation of plasminogen; it could preferentially degrade Aα-chains of fibrinogen, followed by Bß- and γ-chains. Overall, the fibrinolytic enzyme was successfully purified from Perinereis aibuhitensis, a marine Annelida (phylum), with favorable stability that has strong fibrinolysis activity in vitro. Therefore, FELP appears to be a potent fibrinolytic enzyme with an application that deserves further investigation.

Time-resolved Raman spectroscopy for monitoring the structural evolution of materials during rapid compression.

Rapid compression experiments performed using a dynamic diamond anvil cell (dDAC) offer the opportunity to study compression rate-dependent phenomena, which provide critical knowledge of the phase transition kinetics of materials. However, direct probing of the structure evolution of materials is scarce and so far limited to the synchrotron based x-ray diffraction technique. Here, we present a time-resolved Raman spectroscopy technique to monitor the structural evolutions in a subsecond time resolution. Instead of applying a shutter-based synchronization scheme in previous work, we directly coupled and synchronized the spectrometers with the dDAC, providing sequential Raman data over a broad pressure range. The capability and versatility of this technique are verified by in situ observation of the phase transition processes of three rapid compressed samples. Not only the phase transition pressures but also the transition pathways are reproduced with good accuracy. This approach has the potential to serve as an important complement to x-ray diffraction applied to study the kinetics of phase transitions occurring on time scales of seconds and above.

Identification and Expression Analysis of bZIP Members under Abiotic Stress in Mung Bean (Vigna radiata).

The main aim of this study was to identify the bZIP family members in mung bean and explore their expression patterns under several abiotic stresses, with the overarching goal of elucidating their biological functions. Results identified 75 bZIP members in mung bean, which were unevenly distributed in the chromosomes (1-11), and all had a highly conserved bZIP domain. Phylogenetic analysis divided the members into 10 subgroups, with members in the same subgroup having similar structure and motif. The cis-acting elements in the promoter region revealed that most of the bZIP members might have the connection with abscisic acid, ethylene, and stress responsive elements. The transcriptome data demonstrated that bZIP members could respond to salt stress at different degrees in leaves, but the expression patterns could vary at different time points under stress. Differentially expressed genes (DEGs), such as VrbZIP12, VrbZIP37, and VrZIP45, were annotated into the plant hormone signal transduction pathway, which might be regulated the expression of abiotic stress-related gene (ABF). Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to determine the expression of bZIP members in roots and leaves under drought, alkali, and low-temperature stress. Results showed that bZIP members respond differently to diverse stresses, and their expression was tissue-specific, which suggests that they may have different regulatory mechanism in different tissues. Overall, this study will provide a reference for further research on the functions of bZIP members in mung bean.

Crack propagation in graphene monolayer under tear loading.

Crack propagation in graphene monolayer under tear loading is investigated via an energy-based analytical model and molecular dynamics (MD) simulations. The classical mechanics-based model describes steady-state crack propagation velocity as a function of applied stress, lateral dimension and loading geometry, as well as the critical stress and critical size for initiating steady crack propagation. MD simulations reveal that cracks propagate along the zigzag direction but yield different "fracture surface" roughnesses for different loading geometries. MD simulations and the predictions of the analytical model are in excellent agreement. Our findings lead to an improved fundamental understanding of the mode-III crack of monolayer graphene necessary for the design and fabrication of graphene-based devices.