Mechanisms of Metal Resistance and Homeostasis in Haloarchaea

Haloarchaea are the predominant microflora of hypersaline econiches such as solar salterns, soda lakes, and estuaries where the salinity ranges from 35 to 400 ppt. Econiches like estuaries and solar crystallizer ponds may contain high concentrations of metals since they serve as ecological sinks for metal pollution and also as effective traps for river borne metals. The availability of metals in these econiches is determined by the type of metal complexes formed and the solubility of the metal species at such high salinity. Haloarchaea have developed specialized mechanisms for the uptake of metals required for various key physiological processes and are not readily available at high salinity, beside evolving resistance mechanisms for metals with high solubility. The present paper seeks to give an overview of the main molecular mechanisms involved in metal tolerance in haloarchaea and focuses on factors such as salinity and metal speciation that affect the bioavailability of metals to haloarchaea. Global transcriptomic analysis during metal stress in these organisms will help in determining the various factors differentially regulated and essential for metal physiology

Biostimulation of jarosite and iron oxide-bearing mine waste enhances subsequent metal recovery

Novel resource recovery technologies are required for metals-bearing hazardous wastes in order to achieve circular economy outcomes and industrial symbiosis. Iron oxide and co-occurring hydroxysulphate-bearing wastes are globally abundant and often contain other elements of value. This work addresses the biostimulation of indigenous microbial communities within an iron oxide/ hydroxysulphate-bearing waste and its effect on the subsequent recoverability of metals by hydrochloric, sulphuric, citric acids, and EDTA. Laboratory-scale flow-through column reactors were used to examine the effect of using glycerol (10% w/w) to stimulate the in situ microbial community in an iron oxide/ hydroxysulphate-bearing mine waste. The effects on the evolution of leachate chemistry, changes in microbiological community, and subsequent hydrometallurgical extractability of metals were studied. Results demonstrated increased leachability and selectivity of Pb, Cu, and Zn relative to iron after biostimulation with a total of 0.027kg of glycerol per kg of waste. Biostimulation, which can be readily applied in situ, potentially opens new routes to metal recovery from globally abundant waste streams that contain jarosite and iron oxides

Towards passive bioremediation of dye-bearing effluents using hydrous ferric oxide wastes: Mechanisms, products and microbiology

A novel, circular economy-inspired approach for the “passive” (non-powered and reagent-free) treatment of dye-bearing effluent is presented. The treatment utilises the biogeochemical interaction of dye-bearing wastewater with hydrous ferric oxide (HFO) bearing sludges. The work presented demonstrates for the first time the reuse of HFO-rich waste sludges from potable water and mine water treatment. The waste was used directly without modification or reagent addition, as media/substrate in simple flow-through reactors for the decolourisation and biodegradation of methyl orange (MO) and mixed dyes textile effluent. Three phases of exploratory proof of concept work were undertaken. Columns containing HFO sludges were challenged with solution of MO, and MO amended with glycerol (Phase I), MO in a synthetic textile effluent recipe (Phase II), and real mixed textile effluent containing a mixture of dyes (Phase III). After an initial lag period extensive decolourisation of dye was observed in all cases at rates comparable with pure strains and engineered bioreactor processes, with evidence of biodegradation beyond simple cleavage of the mono azo chromophore and mineralisation. The microbiology of the initial sludge samples in both cases exhibited a diverse range of iron oxidising and reducing bacteria. However, post experiment the microbiology of sludge evolved from being dominated by Proteobacteria to being dominated by Firmicutes. Distinct changes in the microbial community structure were observed in post-treatment MWTS and WTWS where genera capable of iron and sulphate reduction and/or aromatic amine degradation were identified. Average nitrogen removal rates for the columns ranged from 27.8 to 194 g/m3/day which is higher than engineered sequential anaerobic-aerobic bioreactor. Postulated mechanisms for the fast anaerobic decolourisation, biodegradation, and mineralisation of the dyes (as well nitrogen transformations) include various direct and indirect enzymatic and metabolic reactions, as well as reductive attack by continuously regenerated reductants such as Fe(II), HFO bound Fe(II), FeS, and HS−. The ability of iron reducers to degrade aromatic rings is also considered important in the further biodegradation and complete mineralisation of organic carbon. The study reveals that abundant and ubiquitous HFO-rich waste sludges, can be used without amendment, as a substrate in simple flow-through bioremediation system for the decolourisation and partial biodegradation of dyes in textile effluent

Biostimulation of jarosite and iron oxide-bearing mine waste enhances subsequent metal recovery

Novel resource recovery technologies are required for metals-bearing hazardous wastes in order to achieve circular economy outcomes and industrial symbiosis. Iron oxide and co-occurring hydroxysulphate-bearing wastes are globally abundant and often contain other elements of value. This work addresses the biostimulation of indigenous microbial communities within an iron oxide/ hydroxysulphate-bearing waste and its effect on the subsequent recoverability of metals by hydrochloric, sulphuric, citric acids and EDTA. Laboratory-scale flow-through column reactors were used to examine the effect of using glycerol (10% w/w) to stimulate the in situ microbial community in an iron oxide/ hydroxysulphate-bearing mine waste. The effects on the evolution of leachate chemistry, changes in microbiological community and subsequent hydrometallurgical extractability of metals were studied. Results demonstrated increased leachability and selectivity of Pb, Cu, and Zn relative to iron after biostimulation with a total of 0.027 kg of glycerol per kg of waste. Biostimulation, which can be readily applied in situ , potentially opens new routes to metal recovery from globally abundant waste streams that contain jarosite and iron oxides

Anti-neoplastic selenium nanoparticles from Idiomarina sp. PR58-8

Selenium nanoparticles (SeNPs) with novel biological activities, cancer cell selectivity, and low toxicity towards normal cells have gained attention for chemo-therapeutic and chemo-preventive applications. These nanoparticles may be synthesized using micro-organisms, which is the green alternative of nanofabrication. Here we report the intracellular synthesis of SeNPs by the moderate halophilic bacterium, Idiomarina sp. PR58-8 using sodium selenite as the precursor. Characterization of SeNPs by XRD exhibited the characteristic Bragg’s peak of hexagonal selenium with a crystallite domain size of 34 nm. Morphological characterization by TEM exhibited spherical nanoparticles with a size distribution of 150–350 nm. The non-protein thiols were found to be involved in resistance/reduction of sodium selenite. The SeNPs exhibited selectivity in exerting cytotoxicity towards human cervical cancer cell line, HeLa, while being non-toxic towards model normal cell line, HaCaT. The SeNPs induced a caspase-dependent apoptosis in HeLa cell lines as exhibited by the ROS assay, apoptotic index assay, and western blot analysis. These results suggest the application of SeNPs synthesized by Idiomarina sp. PR58-8 as potential anti-neoplastic agents

Biosynthesis of nanoparticles from halophiles

Nanobiotechnology is a multidisciplinary branch of nanotechnology which includes fabrication of nanomaterials using biological approaches. Many bacteria, yeast, fungi, algae and viruses have been used for synthesis of various metallic, metal sulfide, metal oxide and alloy nanoparticles, since the first report on biosynthesis of cadmium sulfide quantum dots by Candida glabrata and Schizosaccharomyces pombe in 1989. These nanofactories offer a better size control through compartmentalization in the periplasmic space and vesicles, and are usually capped by stabilizing cellular metabolites. Halophiles depending on their salt requirements may be classified as slight, moderate and extreme halophiles. They are found in marine and/or hypersaline environments. These organisms are known to encounter metals in their environment as the econiches they inhabit serve as ecological sinks for metals. Metal based nanoparticle synthesis by halophilic organisms is in its infancy and has only been reported in few organisms. This chapter aims to shed light on the various halophilic organisms and their by-products that have been exploited for nanomaterial synthesis, the mechanisms that may be involved in the nanomaterial fabrication and the possible applications of the fabricated nanoparticles. A special section would be dedicated for the bioavailability of metals to halophiles under varying salinity conditions

Fluorescent Lead(IV) sulfide nanoparticles synthesized by Idiomarina sp. strain PR58-8 for bioimaging applications

The fabrication of nanoparticles by microorganisms presents a “green” method for generating biocompatible nanomaterials. We discovered the intracellular biosynthesis of fluorescent lead(IV) sulfide nanoparticles by the moderate halophile, Idiomarina sp. strain PR58-8. The bacterium tolerated up to 8 mM Pb(NO3)2 during growth. Non-protein thiols dose-dependently increased in response to metal exposure, which suggests they are involved in the growth of PbS2 crystals and lead detoxification. Using X-ray diffraction, transmission electron microscopy (TEM), high-resolution TEM, and energy dispersive analysis of X-rays, the nanoparticles were characterized as spherical β-PbS2 nanoparticles (PbS2NPs) with a tetragonal crystal lattice, a crystallite domain size of 2.38 nm, and an interplanar distance of 0.318 nm. A narrow symmetric emission spectrum with a Gaussian distribution and an emission maximum at 386 nm was obtained when the particles were excited at 570 nm. The PbS2NPs exhibited a large Stokes' shift (8,362 cm−1) and a relatively high quantum yield (67%). These properties, along with fluorescence that was maintained in various microenvironments and their biocompatibility, make these nanoparticles excellent candidates for bioimaging. The particles were internalized by HeLa cells and evenly distributed within the cytoplasm, exhibiting their potential for in situ bioimaging applications. The “as-synthesized” lead(IV) sulfide nanoparticles may provide expanded opportunities for targeted bioimaging via modifying the surface of the particles. IMPORTANCE This article reports the intracellular synthesis of fluorescent lead(IV) sulfide nanoparticles (PbS2NPs) by a microorganism. All previous reports on the microbial synthesis of lead-based nanoparticles are on lead(II) sulfide that exhibits near-infrared fluorescence, requiring expensive instrumentation for bioimaging. Bioimaging using PbS2NPs can be achieved using routine epifluorescence microscopes, as it fluoresces in the visible range. The research on PbS2 nanoparticles to date is on their chemical synthesis employing toxic precursors, extreme pH, pressure, and temperature, resulting in cytotoxic products. In this context, the synthesis of PbS2 nanoparticles by Idiomarina sp. strain PR58-8, described in this work, occurs at ambient temperature and pressure and results in the generation of biocompatible nanoparticles with no hazardous by-products. The excellent fluorescence properties that these particles exhibit, as well as their abilities to easily penetrate the cells and evenly distribute within the cytoplasm, make them exceptional candidates for bioimaging applications. This study demonstrated the synthesis and fluorescence bioimaging application of microbially synthesized PbS2 nanoparticles

In vivo synthesis of selenium nanoparticles by Halococcus salifodinae BK18 and their anti-proliferative properties against HeLa cell line

Nanoparticles synthesis by bacteria and yeasts has been widely reported, however, synthesis using halophilic archaea is still in a nascent stage. This study aimed at the intracellular synthesis of selenium nanoparticles (SeNPs) by the haloarchaeon Halococcus salifodinae BK18 when grown in the presence of sodium selenite. Crystallographic characterization of SeNPs by X-ray diffraction, Selected area electron diffraction, and transmission electron microscopy exhibited rod shaped nanoparticles with hexagonal crystal lattice, a crystallite domain size of 28 nm and an aspect ratio (length:diameter) of 13:1. Energy disruptive analysis of X-ray analysis confirmed the presence of selenium in the nano-preparation. The nitrate reductase enzyme assay and the inhibitor studies indicated the involvement of NADH-dependent nitrate reductase in SeNPs synthesis and metal tolerance. The SeNPs exhibited good anti-proliferative properties against HeLa cell lines while being non-cytotoxic to normal cell line model HaCat, suggesting the use of these SeNPs as cancer chemotherapeutic agent. This is the first study on selenium nanoparticles synthesis by haloarchaea. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1480–1487, 201