Insights into the discovery and intervention of metalloproteinase in marine hazardous jellyfish.

The proliferation of toxic organisms caused by changes in the marine environment, coupled with the rising human activities along the coastal lines, has resulted in an increasing number of stinging incidents, posing a serious threat to public health. Here, we evaluated the systemic toxicity of the venom in jellyfish Chrysaora quinquecirrha at both cellular and animal levels, and found that jellyfish tentacle extract (TE) has strong lethality accompanied by abnormal elevation of blood biochemical indicators and pathological changes. Joint analysis of transcriptome and proteome indicated that metalloproteinases are the predominant toxins in jellyfish. Specially, two key metalloproteinases DN6695_c0_g3 and DN8184_c0_g7 were identified by mass spectrometry of the red blood cell membrane and tetracycline hydrochloride (Tch) inhibition models. Structurally, molecular docking and kinetic analysis are employed and observed that Tch could inhibit the enzyme activity by binding to the hydrophobic pocket of the catalytic center. In this study, we demonstrated that Tch impedes the metalloproteinase activity thereby reducing the lethal effect of jellyfish, which suggests a potential strategy for combating the health threat of marine toxic jellyfish.

Comparison of the structure-function of five newly members of the calcin family.

Calcins are a group of scorpion toxin peptides specifically binding to ryanodine receptors (RyRs) with high affinity, and have the ability to activate and stabilize RyR in a long-lasting subconductance state. Five newly calcins synthesized compounds exhibit typical structural characteristics of a specific family through chemical synthesis and virtual analysis. As the calcins from the same species, Petersiicalcin1 and Petersiicalcin2, Jendekicalcin2 and Jendekicalcin3, have only one residue difference. Both Petersiicalcin1 and Petersiicalcin2 exhibited different affinities in stimulating [3H]ryanodine binding, but the residue mutation resulted in a 2.7 folds difference. Other calcins also exhibited a stimulatory effect on [3H]ryanodine binding to RyR1, however, their affinities were significantly lower than that of Petersiiicalcin1 and Petersiiicalcin2. The channel domain of RyR1 was found to be capable of binding with the basic residues of these calcins, which also exhibited interactions with the S6 helices on RyR1. Dynamic simulations were conducted for Petersiicalcin1 and Petersiicalcin2, which demonstrated their ability to form a highly stable conformation and resulting in an asymmetric tetramer structure of RyR1. The discovery of five newly calcins further enriches the diversity of the natural calcin family, which provides more native peptides for the structure-function analysis between calcin and RyRs.

The Effect of Acidic Residues on the Binding between Opicalcin1 and Ryanodine Receptor from the Structure-Functional Analysis.

Calcin is a group ligand with high affinity and specificity for the ryanodine receptors (RyRs). Little is known about the effect of its acidic residues on the spacial structure as well as the interaction with RyRs. We screened the opicalcin1 acidic mutants and investigated the effect of mutation on activity. The results indicated that all acidic mutants maintained the structural features, but their surface charge distribution underwent significant changes. Molecular docking and dynamics simulations were used to analyze the interaction between opicalcin1 mutants and RyRs, which demonstrated that all opicalcin1 mutants effectively bound to the channel domain of RyR1. This stable binding induced a pronounced asymmetry in the structure of the RyR tetramer, exhibiting a high degree of structural dissimilarity. [3H]Ryanodine binding to RyR1 was enhanced in D2A and D15A, which was similar to opicalcin1, but that effect was suppressed in E12A and E29A and reversed for the DE-4A, thereby inhibiting ryanodine binding. Opicalcin1 and DE-4A also exhibited the ability to form stable docking structures with RyR2. Acidic residues play a crucial role in the structure of calcin and its functional interaction with RyRs that is beneficial for the calcin optimization to develop more active peptide lead compounds for RyR-related diseases.

Bioaugmentation of intertidal sludge enhancing the development of salt-tolerant aerobic granular sludge.

Three parallel bioreactors were operated with different inoculation of activated sludge (R1), intertidal sludge (ItS) (R2), and ItS-added AS (R3), respectively, to explore the effects of ItS bioaugmentation on the formation of salt-tolerant aerobic granular sludge (SAGS) and the enhancement of COD removal performance. The results showed that compared to the control (R1-2), R3 promoted a more rapid development of SAGS with a cultivation time of 25 d. Following 110-day cultivation, R3 exhibited a higher granular diameter of 1.3 mm and a higher hydrophobic aromatic protein content than that in control. Compared to the control, the salt-tolerant performance in R3 was also enhanced with the COD removal efficiency of 96.4% due to the higher sludge specific activity of 14.4 g·gVSS-1·d-1 and the salinity inhibition constant of 49.3 gL-1. Read- and genome-resolved metagenomics together indicated that a higher level of tryptophan/tyrosine synthase gene (trpBD, tyrBC) and enrichment of the key gene hosts Rhodobacteraceae, Marinicella in R3, which was about 5.4-fold and 1.4-fold of that in control, could be the driving factors of rapid development of SAGS. Furthermore, the augmented salt-tolerant potential in R3 could result from that R1 was dominated by Rhodospirillaceae, Bacteroidales, which carried more trehalose synthase gene (otsB, treS), while the dominant members Rhodobacteraceae, Marinicella in R3 were main contributors to the glycine betaine synthase gene (ectC, betB, gbsA). This study could provide deeper insights into the rapid development and improved salt-tolerant potential of SAGS via bioaugmentation of intertidal sludge, which could promote the application of hypersaline wastewater treatment.

Performance and granular characteristics of salt-tolerant aerobic granular reactors response to multiple hypersaline wastewater.

Aerobic granular sludge (AGS) technology has been recognized as a promising alternative to alleviate the osmotic stress of hypersaline wastewater. However, the response of AGS process to composite hypersaline wastewater on removal performance and populations was yet to be understood. In this work, two sequenced batch reactors were operated in parallel in absence (R0) and presence (R1) of high concentration sulfate as proxy for single and mixed salts (30 g salt·L-1) respectively. Results demonstrated that the presence of sulfate in hypersaline wastewater enhanced chemical oxygen demand (COD) and total nitrogen (TN) removals of 95.3% and 65.5% respectively with lower accumulations of nitrite. High-throughput 16 S rRNA gene sequencing technique elucidated that Denitromonas (31.6%) and Xanthomarina (17.0%) were the more dominant genera in AGS response to mixed salts with high sulfate and laid the biological basis for strengthening removal performance. The enrichment of halophilic Luteococcus (23.5%) in the AGS surface indicated the potential role of mixed salts in shaping the physical properties and surface population structure of AGS. Our work could facilitate the potential applications of AGS technology for industrial hypersaline wastewater treatment with complicated compositions.

Metagenomics revealed the phase-related characteristics during rapid development of halotolerant aerobic granular sludge.

Efforts to produce aerobic granular sludge (AGS) for high-efficient and stable nutrient removal in high saline wastewaters have gained much attention recently. This study was undertaken to describe the phase-related characteristics of the rapid formation of glucose-fed salt-tolerant AGS (SAGS) generated from common municipal activated sludge using metagenomic approaches. The time needed for SAGS formation is about 11 days in a multi-ion matrix salinity of 3%. There were three distinct developmental phases during sludge maturation which were designated: I) the salinity adaptation phase (days 1-2), II) the particle-size transition phase (days 3-5) and III) the maturation and steady-state phase (days 6-11), respectively. Genome-based analysis revealed that during the phase I, members of the genus Mangrovibacter, which has the potential to secrete extracellular polymeric substances (EPS), dominated during the formation of initial SAGS aggregates. During phase II, fungi of the class Saccharomycetes, in particular the genus Geotrichum, became dominant and provided a matrix for bacterial attachment. This mutualistic interaction supported the rapid development and maintenance of mature SAGS. This work characterizes a robust approach for the rapid development of SAGS for efficient saline sewage treatment and provides unique insight into the granulation mechanism occurring during the development process.