Electron beam-based fabrication of crosslinked hydrophilic carbon electrodes and their application for capacitive deionization.

In this research, we demonstrated that a crosslinked hydrophilic carbon electrode with better electrochemical performance than hydrophobic counterparts can easily be produced using room-temperature, quick electron-beam irradiation with a hydrophilic methacryloyl-substituted polyvinyl alcohol (SPVA) binder. The SPVA binder was effectively synthesized by trans-esterification of PVA with glycidyl methacrylate. The hydrophilic carbon electrode cast on a graphite sheet from a slurry of activated carbon (AC) and SPVA was irradiated with an electron beam to form a crosslinked structure. The analytical results in terms of the morphology, solvent resistance, chemical composition, and contact angle revealed that the carbon electrode was completely crosslinked by electron-beam irradiation even at the dose of 100 kGy (irradiation time = 180 s). The new electrode exhibited superior water-wettability due to the hydrophilic functionality of SPVA. Furthermore, the hydrophilic carbon electrode with an AC : SPVA composition of 90 : 10 and an absorbed dose of 200 kGy, exhibited a specific capacitance of 127 F g-1 (67% higher than the hydrophobic poly(vinylidene fluoride) (PVDF)-based counterpart with the same composition). The specific capacitance was further improved to 160 F g-1 with an increase in the AC content. The hydrophilic carbon electrode exhibited noticeably better desalination efficiency than the hydrophobic PVDF-based counterpart.

Identification of sigmaB-dependent promoters using consensus-directed search of Streptomyces coelicolor genome.

SigmaB plays an important role in both osmoprotection and proper differentiation in Streptomyces coelicolor A3(2). We searched for candidate members of the sigmaB regulon from the genome database, using the consensus promoter sequence (GNNTN14-16GGGTAC/T). The list consists of 115 genes, and includes all the known sigmaB target genes and many other genes whose functions are related to stress protection and differentiation.

Post-translational regulation of a developmental catalase, CatB, involves a metalloprotease, SmpA and contributes to proper differentiation and osmoprotection of Streptomyces coelicolor.

Streptomyces coelicolor produces at least three different catalases (catalases A, B, and C) under different physiological conditions. Catalase B (CatB) is a developmentally regulated catalase required for proper differentiation and osmoprotection of S. coelicolor. We previously observed that the N-terminal 75(2) amino acids (aa) of CatB are cleaved off, with the remaining 75-kDa processed CatB detectable in the extracellular fraction during sporulation. We here report that either the deletion of the N-terminal 75 aa or the arginine-to-alanine substitution (R75A) at the cleavage site, but not the histidine-to-alanine substitution (H131A) at the catalytic site, impaired both the secretion of CatB proteins and the proper differentiation of S. coelicolor cells. The proteolytic activity responsible for the cleavage of CatB was purified and then identified as a metalloprotease, which was named as SmpA (Streptomyces metalloprotease A). The SmpA protein was newly detected after sporulation, coincident with the intracellular appearance of 75-kDa CatB, which was not detected in the smpA null mutant, confirming that SmpA indeed processes CatB in vivo. The smpA mutant was osmosensitive as catB mutant, but it displayed delayed sporulation, with the 75-kDa CatB still detectable in the extracellular milieu. Based on these results, we propose that the post-translational regulation of CatB, which cleaves the N-terminal 75 aa residues through SmpA is crucial for proper differentiation and osmoprotection of S. coelicolor. In the absence of SmpA, an alternative route for CatB processing may function to allow delayed sporulation.

A master regulator sigmaB governs osmotic and oxidative response as well as differentiation via a network of sigma factors in Streptomyces coelicolor.

The differentiating bacterium Streptomyces coelicolor harbours some 66 sigma factors, which support its complex life cycle. sigma(B), a functional homologue of sigma(S) from Escherichia coli, controls both osmoprotection and differentiation in S. coelicolor A3(2). Microarray analysis revealed sigma(B)-dependent induction of more than 280 genes by 0.2 M KCl. These genes encode several sigma factors, oxidative defence proteins, chaperones, systems to provide osmolytes, cysteine, mycothiol, and gas vesicle. sigma(B) controlled induction of itself and its two paralogues (sigma(L) and sigma(M)) in a hierarchical order of sigma(B)-->sigma(L)-->sigma(M), as revealed by S1 mapping and Western blot analyses. The phenotype of each sigma mutant suggested a sequential action in morphological differentiation; sigma(B) in forming aerial mycelium, sigma(L) in forming spores and sigma(M) for efficient sporulation. sigma(B) was also responsible for the increase in cysteine and mycothiol, the major thiol buffer in actinomycetes, upon osmotic shock, revealing an overlap between protections against osmotic and oxidative stresses. Proteins in sigB mutant were more oxidized (carbonylated) than the wild type. These results support a hypothesis that sigma(B) serves as a master regulator that triggers other related sigma factors in a cascade, and thus regulates differentiation and osmotic and oxidative response in S. coelicolor.

Regulation of sigmaB by an anti- and an anti-anti-sigma factor in Streptomyces coelicolor in response to osmotic stress.

sigmaB, a homolog of stress-responsive sigmaB of Bacillus subtilis, controls both osmoprotection and differentiation in Streptomyces coelicolor A3 (2). Its gene is preceded by rsbA and rsbB genes encoding homologs of an anti-sigma factor, RsbW, and its antagonist, RsbV, of B. subtilis, respectively. Purified RsbA bound to sigmaB and prevented sigmaB-directed transcription from the sigBp1 promoter in vitro. An rsbA-null mutant exhibited contrasting behavior to the sigB mutant, with elevated sigBp1 transcription, no actinorhodin production, and precocious aerial mycelial formation, reflecting enhanced activity of sigmaB in vivo. Despite sequence similarity to RsbV, RsbB lacks the conserved phosphorylatable serine residue and its gene disruption produced no distinct phenotype. RsbV (SCO7325) from a putative six-gene operon (rsbV-rsbR-rsbS-rsbT-rsbU1-rsbU) was strongly induced by osmotic stress in a sigmaB-dependent manner. It antagonized the inhibitory action of RsbA on sigmaB-directed transcription and was phosphorylated by RsbA in vitro. These results support the hypothesis that the rapid induction of sigmaB target genes by osmotic stress results from modulation of sigmaB activity by the kinase-anti-sigma factor RsbA and its phosphorylatable antagonist RsbV, which function by a partner-switching mechanism. Amplified induction could result from a rapid increase in the synthesis of both sigmaB and its inhibitor antagonist.