Background The diffusion of antibiotics through the outer membrane is primarily affected by the porin super family, changes contribute to antibiotic resistance. Recently we demonstrated that the CpxAR two-component signaling system alters the expression of an uncharacterized porin OmpC KP , to mediate antimicrobial resistance in K. pneumoniae . Principal Findings In this study, functional characterization of the putative porin OmpC KP (denoted kpnO ) with respect to antimicrobial susceptibility and virulence was evaluated by generating an isogenic mutant, Δ kpnO in a clinical isolate of K. pneumoniae . Estimation of uronic acid content confirmed that Δ kpnO produced ∼2.0 fold lesser capsular polysaccharide than the wild-type. The Δ kpnO displayed higher sensitivity to hyper osmotic and bile conditions. Disruption of kpnO increased the susceptibility of K. pneumoniae to oxidative and nitrostative stress by ∼1.6 fold and >7 fold respectively. The loss of the Klebsiella porin led to an increase in the minimum inhibitory concentration of tetracycline (3-fold), nalidixic acid (4-fold), tobramycin (4-fold), streptomycin (10-fold), and spectinomycin (10-fold), which could be restored following complementation. The single deletion of kpnO reduced the survival of the pathogen by 50% when exposed to disinfectants. In Caenorhabditis elegans model, the kpnO mutant exhibited significantly (P<0.01) lower virulence. To dissect the role of PhoBR signaling system in regulating the expression of the kpnO , a phoB KP isogenic mutant was constructed. The phoB KP mutant exhibited impaired gastrointestinal stress response and decreased antimicrobial susceptibility. The mRNA levels of kpnO were found to be 4-fold less in phoB KP mutant compared to wild type. A regulatory role of PhoB KP for the expression of kpnO was further supported by the specific binding of PhoB KP to the putative promoter of kpnO . Conclusions and Significance Loss of PhoBR regulated porin KpnO resulted in increased antimicrobial resistance, increased susceptibility to gastrointestinal stress, and reduced virulence in K. pneumoniae NTUH-K2044.

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Abstract:This article aims to study the codigestion of food waste (FW) and three different lignocellulosic wastes (LW) (Corn stover (CS), Prairie cordgrass (PCG), and Unbleached paper (UBP)) for thermophilic anaerobic digestion to overcome the limitations of digesting food waste alone (volatile fatty acids accumulation and low C:N ratio). Using an enriched thermophilic methanogenic consortium, all the food and lignocellulosic waste mixtures showed positive synergistic effects of codigestion. After 30 days of incubation at 60 • C (100 rpm), the highest methane yield of 305.45 L·kg −1 volatile solids (VS) was achieved with a combination of FW-PCG-CS followed by 279.31 L·kg −1 VS with a mixture of FW-PCG. The corresponding volatile solids reduction for these two co-digestion mixtures was 68% and 58%, respectively. This study demonstrated a reduced hydraulic retention time for methane production using FW and LW.

Various microorganisms thrive under extreme environments, like hot springs, hydrothermal vents, deep marine ecosystems, hyperacid lakes, acid mine drainage, high UV exposure, and more. To survive against the deleterious effect of these extreme circumstances, they form a network of biofilm where exopolysaccharides (EPSs) comprise a substantial part. The EPSs are often polyanionic due to different functional groups in their structural backbone, including uronic acids, sulfated units, and phosphate groups. Altogether, these chemical groups provide EPSs with a negative charge allowing them to (a) act as ligands toward dissolved cations as well as trace, and toxic metals; (b) be tolerant to the presence of salts, surfactants, and alpha-hydroxyl acids; and (c) interface the solubilization of hydrocarbons. Owing to their unique structural and functional characteristics, EPSs are anticipated to be utilized industrially to remediation of metals, crude oil, and hydrocarbons from contaminated wastewaters, mines, and oil spills. The biotechnological advantages of extremophilic EPSs are more diverse than traditional biopolymers. The present review aims at discussing the mechanisms and strategies for using EPSs from extremophiles in industries and environment bioremediation. Additionally, the potential of EPSs as fascinating biomaterials to mediate biogenic nanoparticles synthesis and treat multicomponent water contaminants is discussed.

A novel, thermostable, alkalophilic β-D-galactosidase (Mbgl) was isolated from a metagenome of geothermal springs in northern Himalayan region of India. Mbgl was 447 amino acids in size and had conserved catalytic residues E170 and E358, indicating that it belonged to family 1 of glycosyl hydrolases showing maximum homology (89 %) with uncharacterized β-galactosidase of Eubacterium, Meiothermus ruber DSM1279. Temperature and pH optima of Mbgl were 65 °C and 8.0 respectively, and it retained 80 % activity even at pH 10.0. Mbgl was active as a homotetramer, recognized β-(1,4)-D-galactoside as the preferred glycosidic bond, and preferentially hydrolyzed pNPgal with K(m) 3.33 mM and k(cat) 2,000 s(-1). It displayed high transglycosylation activity with wide acceptor specificity including hexoses and pentoses leading to the formation of prebiotic galacto-oligosaccharides whereas its lactose hydrolysis potential was low.