Nostosins, Trypsin Inhibitors Isolated from the Terrestrial Cyanobacterium Nostoc sp. Strain FSN.

Two new trypsin inhibitors, nostosin A (1) and B (2), were isolated from a hydrophilic extract of Nostoc sp. strain FSN, which was collected from a paddy field in the Golestan Province of Iran. Nostosins A (1) and B (2) are composed of three subunits, 2-hydroxy-4-(4-hydroxyphenyl)butanoic acid (Hhpba), L-Ile, and L-argininal (1) or argininol (2). Nostosins A (1) and B (2) exhibited IC50 values of 0.35 and 55 µM against porcine trypsin, respectively, suggesting that the argininal aldehyde group plays a crucial role in the efficient inhibition of trypsin. Molecular docking of nostosin A (1) (449 Da), leupeptin (426 Da, IC50 0.5 µM), and spumigin E (610 Da, IC50 < 0.1 µM) with trypsin suggested prominent binding similarity between nostosin A (1) and leupeptin but only partial binding similarity with spumigin E. The number of hydrogen bonds between ligands and trypsin increased according to the length and size of the ligand molecule, and the docking affinity values followed the measured IC50 values. Nostosin A (1) is the first highly potent three-subunit trypsin inhibitor with potency comparable to the known commercial trypsin inhibitor leupeptin. These findings expand the known diversity of short-chain linear peptide protease inhibitors produced by cyanobacteria.

Structural and developmental expression of Ss-riok-2, an RIO protein kinase encoding gene of Strongyloides stercoralis.

RIO kinases are essential atypical protein kinases in diverse prokaryotic and eukaryotic organisms, playing significant roles in yeast and humans. However, little is known about their functions in parasitic nematodes. In the present study, we have isolated and characterized the full-length cDNA, gDNA and a putative promoter of a RIOK-2 protein kinase (Ss-RIOK-2) encoding gene (Ss-riok-2) from Strongyloides stercoralis, a medically important parasitic nematode (Order Rhabditida). A three-dimensional structure (3D) model of Ss-RIOK-2 was generated using the Chaetomium thermophilum RIOK-2 protein kinase (Ct-RIOK-2) crystal structure 4GYG as a template. A docking study revealed some critical sites for ATP binding and metal binding. The putative promoter of Ss-riok-2 contains a number of conserved elements. RNAseq analysis revealed the highest levels of the Ss-riok-2 transcript in free-living females and parasitic females. To identify anatomical patterns of Ss-riok-2 expression in S. stercoralis, we observed expression patterns of a transgene construct encoding green fluorescent protein under the Ss-riok-2 promoter in post free-living S. stercoralis. Expression driven by this promoter predominated in intestinal cells. This study demonstrates significant advancement in molecular and cellular biological study of S. stercoralis and of parasitic nematodes generally, and provides a foundation for further functional genomic studies.

Degradation of aniline in Weihe riverbed sediments under denitrification conditions.

Three groups of microcosm tests were conducted to study the possibility of aniline degradation and the effects of organic matter and hydrous metal oxides on the degradation in Weihe riverbed sediments under denitrification conditions. After the riverbed sediments (20 g) and groundwater (800 ml) were put into bottles, aniline, nitrate and other reagents were added, and then the bottles were flushed with N2 for 30 minutes to create microcosms. Samples from the microcosms were employed for the analysis of aniline, nitrate, and chemical oxygen demand (COD). In the first test group, the concentration of aniline remained unchanged when NaN3 (500 mg/L) was added. When there was no nitrate or NaN3, the concentration of aniline also remained unchanged, although COD declined. However, the concentration decreased when nitrate (50 mg/L) was added. Therefore, aniline can be biodegraded under denitrification conditions. In the second test group, when the concentration of nitrate reached 50 mg/L, 300 mg/L or 400 mg/L, either the external or internal organic matter or both of them in Weihe raw sediments inhibited aniline degradation. In the sediments where organic matter alone or organic matter plus hydrous metal oxides were removed, the organic matter still inhibited the degradation when the concentration of nitrate reached 300 mg/L or 400 mg/L, but the external organic matter could accelerate the degradation when the concentration of nitrate was 50 mg/L. The result of the third test group showed that hydrous metal oxides can accelerate degradation. By analyzing the mechanism of the aniline degradation, we conclude that aniline is degradable by microbes in their growth metabolism, in which deamination is involved.