Novel chitosan-based smart bio-nanocomposite films incorporating TiO2 nanoparticles for white bread preservation.

Chitosan (CS)-based bio-nanocomposite food packaging films were prepared via solvent-casting method by incorporating a unique combination of additives and fillers, including polyvinyl alcohol (PVA), glycerol, Tween 80, castor oil (CO), and nano titanium dioxide (TiO2) in various proportions to enhance film properties. For a comprehensive analysis of the synthesized films, Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA), tensile testing, field emission scanning electron microscopy (FESEM), energy dispersive X-ray spectroscopy (EDS), and UV-vis spectrophotometry were employed. Furthermore, the antimicrobial efficacy of the films against S. aureus, E. coli, and A. niger was examined to assess their potential to preserve food from foodborne pathogens. The results claimed that the inclusion of castor oil and TiO2 nanoparticles considerably improved antimicrobial properties, UV-vis light barrier properties, thermal stability, optical transparency, and mechanical strength of the films, while reducing their water solubility, moisture content, water vapor and oxygen permeability. Based on the overall analysis, CS/PVA/CO/TiO2-0.3 film can be selected as the optimal one for practical applications. Furthermore, the practical application of the optimum film was evaluated using white bread as a model food product. The modified film successfully extended the shelf life of bread to 10 days, surpassing the performance of commercial LDPE packaging (6 days), and showed promising attributes for applications in the food packaging sector. These films exhibit superior antimicrobial properties, improved mechanical strength, and extended shelf life for food products, marking a sustainable and efficient alternative to conventional plastic packaging in both scientific research and industrial applications.

Recent Advancements in Sensing of Silver ions by Different Host Molecules: An Overview (2018-2023).

The chemosensors act as powerful tool in the detection of metal ions due to their simplicity, high sensitivity, low cost, low detection limit, rapid photophysical response, and application to the environmental and medical fields. This review article presents an overview for the chemosensing of Ag+ ions based on Calix, MOF, Nanoparticle, COF, Calix, Electrochemical chemosensor published from 2018 to 2023. Here, we have reviewed the sensing of Ag+ ions and summarised the binding response, mechanism, LOD, colorimetric response, adsorption capacity, technique used. The purpose of this review article to provide a detailed summary of the performance of different host chemosensors that are helpful for providing future direction to researchers on Ag+ ion detection and provides path to design effective chemsosensor (simple to synthesize, cost effective, high sensitivity, with more practical application). While studying the related article literature, we came across some challenges and that has been discussed lastly and provided solutions for them.

Rapid arsenite oxidation by Paenarthrobacter nicotinovorans strain SSBW5: unravelling the role of GlpF, aioAB and aioE genes.

A novel arsenite resistant bacterial strain SSBW5 was isolated from the battery waste site of Corlim, Goa, India. This strain interestingly exhibited rapid arsenite oxidation with an accumulation of 5 mM arsenate within 24 h and a minimum inhibitory concentration (MIC) of 18 mM. The strain SSBW5 was identified as Paenarthrobacter nicotinovorans using 16S rDNA sequence analysis. Fourier-transformed infrared (FTIR) spectroscopy of arsenite-exposed cells revealed the interaction of arsenite with several important functional groups present on the cell surface, possibly involved in the resistance mechanism. Interestingly, the whole genome sequence analysis also clearly elucidated the presence of genes, such as GlpF, aioAB and aioE encoding transporter, arsenite oxidase and oxidoreductase enzyme, respectively, conferring their role in arsenite resistance. Furthermore, this strain also revealed the presence of several other genes conferring resistance to various metals, drugs, antibiotics and disinfectants. Further suggesting the probable direct or indirect involvement of these genes in the detoxification of arsenite thereby increasing its tolerance limit. In addition, clumping of bacterial cells was observed through microscopic analysis which could also be a strategy to reduce arsenite toxicity thus indicating the existence of multiple resistance mechanisms in strain SSBW5. In the present communication, we are reporting for the first time the potential of P. nicotinovorans strain SSBW5 to be used in the bioremediation of arsenite via arsenite oxidation along with other toxic metals and metalloids.

Selenite bioreduction with concomitant green synthesis of selenium nanoparticles by a selenite resistant EPS and siderophore producing terrestrial bacterium.

Environmental bacterial isolates play a very important role in bioremediation of metals and toxic metalloids. A bacterial strain with high selenite (SeO32-) tolerance and reducing capability was isolated from electronic waste dump site in Banaras Hindu University, Varanasi, India. Based on 16 S rRNA sequencing and BLAST search, this bacterial isolate was identified as Bacillus paramycoides and designated as strain MF-14. It tolerated Sodium selenite up to 110 mM when grown aerobically in LB broth and reduced selenite into elemental selenium (Se0) significantly within 24 h with concomitant biosynthesis of selenium nanoparticles as clearly revealed by brick red precipitate and specific surface plasmon resonance peak at 210 nm using UV-Visible spectrophotometer. Scanning electron microscopy (SEM) analysis of this bacterial strain exposed to 1mM and 5 mM selenite also demonstrated morphological alterations as cell enlargement due to accumulation and bioprecipitation of elemental selenium (Se0). The FTIR analysis clearly demonstrated that functional groups present on the surface of biogenic selenium nanoparticles (SeNPs) play a significant role in the stabilization and capping of SeNPs. Furthermore, these SeNPs were characterized using spectroscopic analysis involving Dynamic light scattering, zeta potential, XPS, FTIR, XRD and Raman spectroscopy which clearly revealed particle size 10-700 nm, amorphous nature, stability as well as it's oxidation state. The biochemical studies have demonstrated that membrane bound reductase enzyme may be responsible for significant reduction of selenite into elemental selenium. Therefore, we may employ Bacillus paramycoides strain MF-14 successfully for bioremediation of selenite contaminated environmental sites with concomitant green synthesis of SeNPs.

Cyanobacteria: miniature factories for green synthesis of metallic nanomaterials: a review.

Nanotechnology is one of the most promising and advanced disciplines of science that deals with synthesis, characterization and applications of different types of Nanomaterials (NMs) viz. nanospheres, nanoparticles, nanotubes, nanorods, nanowires, nanocomposites, nanoalloys, carbon dots and quantum dots. These nanosized materials exhibit different physicochemical characteristics and act as a whole unit during its transport. The unique characteristics and vast applications of NMs in diverse fields viz. electronics, agriculture, biology and medicine have created huge demand of different type of NMs. Conventionally physical and chemical methods were adopted to manufacture NMs which are expensive and end up with hazardous by-products. Therefore, green synthesis exploiting biological resources viz. algae, bacteria, fungi and plants emerged as a better and promising alternative due to its cost effective and ecofriendly approach and referred as nanobiotechnology. Among various living organisms, cyanobacteria have proved one of the most favourable bioresources for NMs biosynthesis due to their survival in diverse econiches including metal and metalloid contaminated sites and capability to withstand high levels of metals. Biosynthesis of metallic NMs is accomplished through bioreduction of respective metal salts by various capping agents viz. alkaloids, pigments, polysaccharides, steroids, enzymes and peptides present in the biological systems. Advancement in the field of Nanobiotechnology has produced large number of diverse NMs from cyanobacteria which have been used as antimicrobial agents against Gram positive and negative human pathogens, anticancer agents, luminescent nanoprobes for imaging of cells, antifungal agents against plant pathogens, nanocatalyst and semiconductor quantum dots in industries and in bioremediation in toxic pollutant dyes. In the present communication, we have reviewed cyanobacteria mediated biosynthesis of NMs and their applications in various fields.

Biogenesis of selenium nanospheres using Halomonas venusta strain GUSDM4 exhibiting potent environmental applications.

Selenite reducing bacterial strain (GUSDM4) isolated from Mandovi estuary of Goa, India was identified as Halomonas venusta based on 16S rRNA gene sequence analysis. Its maximum tolerance level for sodium selenite (Na2SeO3) was 100 mM. The 2, 3-diaminonaphthalene-based spectroscopic analysis demonstrated 96 and 93% reduction of 2 and 4 mM Na2SeO3 respectively to elemental selenium (Se0) during the late stationary growth phase. Biosynthesis of Se nanoparticles (SeNPs) commenced within 4 h during the log phase, which was evident from the brick red color in the growth medium and a characteristic peak at 265 nm revealed by UV-Vis spectrophotometry. The intracellular periplasmic synthesis of SeNPs in GUSDM4 was confirmed by transmission electron microscopy (TEM). Characterization of SeNPs by X-ray crystallography, TEM and energy-dispersive X-ray analysis (EDAX) clearly demonstrated spherical SeNPs of 20-80 nm diameter with hexagonal crystal lattice. SeNPs (0.8 and 1 mg/L) primed seeds under arsenate [As(V)] stress showed increase in shoot length, root length and biomass by 1.4-, 1.5- and 1.1-fold respectively, as compared to As(V) primed seeds alone. The proline and phenolic content in seeds primed with SeNPs under arsenate stress showed alleviated levels proving its ameliorative potential. SeNPs also demonstrated anti-biofilm activity at 20 µg/mL against human pathogens which was evident by scanning electron microscopic (SEM) analysis. SeNPs interestingly revealed mosquito larvicidal activity also. Therefore, these studies have clearly demonstrated amazing potential of the marine bacterium, Halomonas venusta in biosynthesis of SeNPs and their applications as ameliorative, anti-biofilm and mosquito larvicidal agents which is the first report of its kind.

Biogenic synthesis and characterization of selenium nanoparticles and their applications with special reference to antibacterial, antioxidant, anticancer and photocatalytic activity.

Oxyanions of selenium, selenite (SeO3)2- and selenate (SeO4)2- are toxic to terrestrial and aquatic biota but few microorganisms including cyanobacteria are resistant to high levels of selenite. Cyanobacteria evade selenite toxicity through bioreduction and synthesis of selenium nanoparticles (SeNPs). In this study, extracellular biosynthesis of SeNPs (Se0) using cyanobacterium, Anabaena sp. PCC 7120 on exposure to sodium selenite and characterization was done by using UV-visible spectroscopy, SEM-EDX, TEM and FTIR analyses which confirmed spherical shape with size range of 5-50 nm diameter. These biogenic SeNPs demonstrated significant antibacterial and anti-biofilm activity against bacterial pathogens. Furthermore, these SeNPs showed high antioxidant activity at minimum concentration of 50 µg/mL and significant anti-proliferative activity against HeLa cell line with IC50 value of 5.5 µg/mL. The SeNPs also induced accumulation of cancer cells in the sub-G1 phase which was clearly observed in cellular and nuclear morphology. These biofabricated SeNPs also reduced and decolorized toxic methylene blue dye significantly through photocatalytic degradation. Therefore Anabaena sp. PCC 7120 may be employed as a green bioresource to synthesize SeNPs with potential applications in medicine and environmental bioremediation.

Biological characterization of Bacillus flexus strain SSAI1 transforming highly toxic arsenite to less toxic arsenate mediated by periplasmic arsenite oxidase enzyme encoded by aioAB genes.

Bacillus flexus strain SSAI1 isolated from agro-industry waste, Tuem, Goa, India displayed high arsenite resistance as minimal inhibitory concentration was 25 mM in mineral salts medium. This bacterial strain exposed to 10 mM arsenite demonstrated rapid arsenite oxidation and internalization of 7 mM arsenate within 24 h. The Fourier transformed infrared (FTIR) spectroscopy of cells exposed to arsenite revealed important functional groups on the cell surface interacting with arsenite. Furthermore, scanning electron microscopy combined with electron dispersive X-ray spectroscopy (SEM-EDAX) of cells exposed to arsenite revealed clumping of cells with no surface adsorption of arsenite. Transmission electron microscopy coupled with electron dispersive X-ray spectroscopic (TEM-EDAX) analysis of arsenite exposed cells clearly demonstrated ultra-structural changes and intracellular accumulation of arsenic. Whole-genome sequence analysis of this bacterial strain interestingly revealed the presence of large number of metal(loid) resistance genes, including aioAB genes encoding arsenite oxidase responsible for the oxidation of highly toxic arsenite to less toxic arsenate. Enzyme assay further confirmed that arsenite oxidase is a periplasmic enzyme. The genome of strain SSAI1 also carried glpF, aioS and aioE genes conferring resistance to arsenite. Therefore, multi-metal(loid) resistant arsenite oxidizing Bacillus flexus strain SSAI1 has potential to bioremediate arsenite contaminated environmental sites and is the first report of its kind.

Arsenite biotransformation and bioaccumulation by Klebsiella pneumoniae strain SSSW7 possessing arsenite oxidase (aioA) gene.

Arsenite oxidizing Klebsiella pneumoniae strain SSSW7 isolated from shipyard waste Goa, India showed a minimum inhibitory concentration of 21 mM in mineral salts medium. The strain possessed a small supercoiled plasmid and PCR amplification of arsenite oxidase gene (aioA) was observed on plasmid as well as chromosomal DNA. It was confirmed that arsenite oxidase enzyme was a periplasmic protein with a 47% increase in arsenite oxidase activity at 1 mM sodium arsenite. Scanning electron microscopy coupled with electron dispersive X-ray spectroscopic (SEM-EDS) analysis of 15 mM arsenite exposed cells revealed long chains of cells with no surface adsorption of arsenic. Transmission electron microscopy combined with electron dispersive X-ray spectroscopic (TEM-EDS) analysis demonstrated plasma membrane disruption, cytoplasmic condensation and periplasmic accumulation of arsenic. The bacterial strain oxidized 10 mM of highly toxic arsenite to less toxic arsenate after 24 h of incubation. Fourier transformed infrared (FTIR) spectroscopy confirmed the interaction of arsenite with functional groups present on the bacterial cell surface. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of 5 mM arsenite exposed cells demonstrated over-expression of 87 kDa and 14 kDa proteins of two subunits aioA and aioB of heterodimer arsenite oxidase enzyme as compared to control cells. Therefore, this bacterial strain might be employed as a potential candidate for bioremediation of arsenite contaminated environmental sites.

Tellurite biotransformation and detoxification by Shewanella baltica with simultaneous synthesis of tellurium nanorods exhibiting photo-catalytic and anti-biofilm activity.

Tellurite reducing bacterial strain was isolated from Zuari estuary, Goa India which could tolerate 5.5 mM potassium tellurite with a minimum inhibitory concentration of 6 mM. This strain was designated as GUSDZ9 and was identified as Shewanella baltica (accession number: MF350629) based on 16S rRNA gene sequencing and BLAST analysis. The Diethyl-dithiocarbamate based colorimetric analysis clearly demonstrated a complete reduction of 2 mM tellurite to elemental tellurium during the late stationary phase. Te Nanoparticles (TeNPs) biosynthesis which initiated at early log phase (i.e. 4 h) was evidently monitored through colour change and a peak due to surface plasmon resonance at 210 nm using UV-Vis spectroscopic analysis. X-ray crystallographic studies and transmission electron microscopy revealed unique nano-rods with a diameter ranging from 8 to 75 nm. Energy dispersive X-ray analysis further confirmed the presence of pure tellurium. The biogenic TeNPs at 10 and 5 µg/mL evidently demonstrated 90% degradation of methylene blue dye and anti-biofilm activity against potential Gram-positive and Gram-negative human pathogens respectively. The alkaline comet assay revealed time and dose-dependent genotoxicity at concentrations higher than 15 µg/mL of TeNPs. This study clearly demonstrated the potential of Shewanella baltica strain GUSDZ9 in bioremediation of toxic tellurite through bio-reduction into elemental tellurium and involvement of biogenic TeNPs in the photo-catalytic reduction of methylene blue and anti-biofilm activity. This is the first report of its kind on the synthesis of biogenic TeNPs from Shewanella baltica demonstrating photo-catalytic, anti-biofilm activity as well as genotoxicity.

Enhanced exopolysaccharide production and biofilm forming ability in methicillin resistant Staphylococcus sciuri isolated from dairy in response to acyl homoserine lactone (AHL).

Staphylococcus sciuri is an emerging human pathogen widely found in dairy industries. In this study, we have isolated methicillin resistant Staphylococcus sp. from biofilm formed on utensil used in the dairy society situated at Raia, Goa and was designated as NN14. The isolate NN14 was identified through 16S rRNA sequencing as S. sciuri (GenBank accession number MF621976). This report reveals that the S. sciuri strain NN14 responds positively to the, acyl-homoserine lactone (AHL) having 6-carbon long acyl chain i.e. N-hexanoyl-homoserine lactone molecule (C6-HSL) with gradual rise in their biofilm establishing potential as the concentration of AHL was increased from 250 nM, 500 nM to 1 µM when compared to control (without C6-HSL) by performing crystal violet assay using 48 well microtiter plate. Also, exopolysaccharide (EPS) production was found to increase with gradual increase in C6-HSL concentration from 250 nM, 500 nM to 1 µM proving potential role of EPS in biofilm formation. These results were further proved by scanning electron microscopy where increased in biofilm and EPS production with increase in C6-HSL concentration was observed. The biofilm forming capability of S. sciuri strain NN14 was found to decreased significantly when it was subjected to 10 µg/ml of (R)-2-(2-hydroxynaphthalen-1-yl)-thiazolidine-4-carboxylic acid, however with the addition of 250 and 500 nM, C6-HSL in presence of the antimicrobial compound (R)-2-(2-hydroxynaphthalen-1-yl)-thiazolidine-4-carboxylic acid, the biofilm development in bacterial strain NN14 was increased when compared with control. Our results demonstrated that the C6-HSL molecule neutralize the effect of antibacterial compound and enhances EPS production and biofilm development in S. sciuri.

Phosphatase mediated bioprecipitation of lead as pyromorphite by Achromobacter xylosoxidans.

Achromobacter xylosoxidans strain SJ11, tolerating up to 4.0 mM lead nitrate, in a defined minimal medium was isolated from the waste of a battery manufacturing industry, Goa, India. Interestingly, it formed white precipitate on exposure to lead nitrate which was also evident from scanning electron micrograph (SEM). Energy dispersive X-ray spectroscopic analysis revealed the presence of lead (48.5% by weight) along with phosphorus and chlorine in the precipitate. Transmission electron microscopy (TEM) of bacterial cells clearly refuted the possibility of intracellular lead uptake confirming extracellular precipitation as a predominant mechanism of lead resistance in this bacterium. The extracellular precipitate was further identified as pyromorphite [Pb5(PO4)3Cl] by X-ray diffraction analysis. This was also corroborated by fourier transformed infrared spectroscopy (FTIR) indicating a significant involvement of phosphate groups. Atomic absorption spectroscopic analysis clearly demonstrated that 465.8 mg g-1 lead was precipitated by the bacterial cells. There was remarkable increase of 160% in phosphatase activity suggesting it's important role in lead precipitation. This was further substantiated by significant up-regulation of phosphatase, CheZ using LC-MS/MS. Therefore phosphatase mediated extracellular precipitation of lead as pyromorphite by A. xylosoxidans strain SJ11 clearly demonstrated it's potential in bioremediation of lead contaminated environmental sites.

Rapid and efficient method to extract metagenomic DNA from estuarine sediments.

Metagenomic DNA from sediments of selective estuaries of Goa, India was extracted using a simple, fast, efficient and environment friendly method. The recovery of pure metagenomic DNA from our method was significantly high as compared to other well-known methods since the concentration of recovered metagenomic DNA ranged from 1185.1 to 4579.7 µg/g of sediment. The purity of metagenomic DNA was also considerably high as the ratio of absorbance at 260 and 280 nm ranged from 1.88 to 1.94. Therefore, the recovered metagenomic DNA was directly used to perform various molecular biology experiments viz. restriction digestion, PCR amplification, cloning and metagenomic library construction. This clearly proved that our protocol for metagenomic DNA extraction using silica gel efficiently removed the contaminants and prevented shearing of the metagenomic DNA. Thus, this modified method can be used to recover pure metagenomic DNA from various estuarine sediments in a rapid, efficient and eco-friendly manner.

Metallothionein assisted periplasmic lead sequestration as lead sulfite by Providencia vermicola strain SJ2A.

Lead resistant Providencia vermicola strain SJ2A was isolated from the waste of a battery manufacturing industry which could tolerate upto 3.0mM lead nitrate in the minimal medium. Interestingly, this isolate showed presence of a plasmid borne metallothionein gene, bmtA that matched significantly (96%) with that of Pseudomonas aeruginosa. Scanning electron micrographs of bacterial cells exposed to lead revealed a unique alteration in the cell morphology from rods to long inter-connected filaments. On the other hand, electron dispersive X-ray spectroscopy (EDX) clearly indicated no significant lead adsorption therefore, we speculated intracellular sequestration in this bacterial strain. Transmission electron micrographs of the bacterial cells exposed to lead evidently demonstrated periplasmic sequestration of lead which was also supported by Fourier transformed infrared spectroscopic (FTIR) analysis. The bacterium internalised 155.12mg Pb2+/g biomass as determined by atomic absorption spectroscopy. Subsequently, the accumulated lead was identified as lead sulfite by X-ray diffraction studies. Therefore P. vermicola strain SJ2A has potential to bioremediate lead contaminated environmental sites.

Biotransformation of Tributyltin chloride by Pseudomonas stutzeri strain DN2

A bacterial isolate capable of utilizing tributyltin chloride (TBTCl) as sole carbon source was isolated from estuarine sediments of west coast of India and identified as Pseudomonas stutzeri based on biochemical tests and Fatty acid methyl ester (FAME) analysis. This isolate was designated as strain DN2. Although this bacterial isolate could resist up to 3 mM TBTCl level, it showed maximum growth at 2 mM TBTCl in mineral salt medium (MSM). Pseudomonas stutzeri DN2 exposed to 2 mM TBTCl revealed significant alteration in cell morphology as elongation and shrinkage in cell size along with roughness of cell surface. FTIR and NMR analysis of TBTCl degradation product extracted using chloroform and purified using column chromatography clearly revealed biotransformation of TBTCl into Dibutyltin dichloride (DBTCl2) through debutylation process. Therefore, Pseudomonas stutzeri strain DN2 may be used as a potential bacterial strain for bioremediation of TBTCl contaminated aquatic environmental sites.

Biotransformation of Tributyltin chloride by Pseudomonas stutzeri strain DN2.

A bacterial isolate capable of utilizing tributyltin chloride (TBTCl) as sole carbon source was isolated from estuarine sediments of west coast of India and identified as Pseudomonas stutzeri based on biochemical tests and Fatty acid methyl ester (FAME) analysis. This isolate was designated as strain DN2. Although this bacterial isolate could resist up to 3 mM TBTCl level, it showed maximum growth at 2 mM TBTCl in mineral salt medium (MSM). Pseudomonas stutzeri DN2 exposed to 2 mM TBTCl revealed significant alteration in cell morphology as elongation and shrinkage in cell size along with roughness of cell surface. FTIR and NMR analysis of TBTCl degradation product extracted using chloroform and purified using column chromatography clearly revealed biotransformation of TBTCl into Dibutyltin dichloride (DBTCl2) through debutylation process. Therefore, Pseudomonas stutzeri strain DN2 may be used as a potential bacterial strain for bioremediation of TBTCl contaminated aquatic environmental sites.

Lead resistant bacteria: lead resistance mechanisms, their applications in lead bioremediation and biomonitoring.

Lead (Pb) is non-bioessential, persistent and hazardous heavy metal pollutant of environmental concern. Bioremediation has become a potential alternative to the existing technologies for the removal and/or recovery of toxic lead from waste waters before releasing it into natural water bodies for environmental safety. To our best knowledge, this is a first review presenting different mechanisms employed by lead resistant bacteria to resist high levels of lead and their applications in cost effective and eco-friendly ways of lead bioremediation and biomonitoring. Various lead resistant mechanisms employed by lead resistant bacteria includes efflux mechanism, extracellular sequestration, biosorption, precipitation, alteration in cell morphology, enhanced siderophore production and intracellular lead bioaccumulation.

Isolation and identification of Aeromonas caviae strain KS-1 as TBTC- and lead-resistant estuarine bacteria.

Tributyltin chloride (TBTC)- and lead-resistant estuarine bacterium from Mandovi estuary, Goa, India was isolated and identified as Aeromonas caviae strain KS-1 based on biochemical characteristics and FAME analysis. It tolerates TBTC and lead up to 1.0 and 1.4 mM, respectively, in the minimal salt medium (MSM) supplemented with 0.4 % glucose. Scanning electron microscopy clearly revealed a unique morphological pattern in the form of long inter-connected chains of bacterial cells on exposure to 1 mM TBTC, whereas cells remained unaltered in presence of 1.4 mM Pb(NO3)2 but significant biosorption of lead (8 %) on the cell surface of this isolate was clearly revealed by scanning electron microscopy coupled with energy dispersive X-ray spectroscopy. SDS-PAGE analysis of whole-cell proteins of this lead-resistant isolate interestingly demonstrated three lead-induced proteins with molecular mass of 15.7, 16.9 and 32.4 kDa, respectively, when bacterial cells were grown under the stress of 1.4 mM Pb (NO3)2. This clearly demonstrated their possible involvement exclusively in lead resistance. A. caviae strain KS-1 also showed tolerance to several other heavy metals, viz. zinc, cadmium, copper and mercury. Therefore, we can employ this TBTC and lead-resistant bacterial isolate for lead bioremediation and also for biomonitoring TBTC from lead and TBTC contaminated environment.

Biological characterization of lead-enhanced exopolysaccharide produced by a lead resistant Enterobacter cloacae strain P2B.

A lead resistant bacterial strain isolated from effluent of lead battery manufacturing company of Goa, India has been identified as Enterobacter cloacae strain P2B based on morphological, biochemical characters, FAME profile and 16S rDNA sequence data. This bacterial strain could resist lead nitrate up to 1.6 mM. Significant increase in exopolysaccharide (EPS) production was observed as the production increased from 28 to 108 mg/L dry weight when exposed to 1.6 mM lead nitrate in Tris buffered minimal medium. Fourier-transformed infrared spectroscopy of this EPS revealed presence of several functional groups involved in metal binding viz. carboxyl, hydroxyl and amide groups along with glucuronic acid. Gas chromatography coupled with mass spectrometry analysis of alditol-acetate derivatives of acid hydrolysed EPS produced in presence of 1.6 mM lead nitrate demonstrated presence of several neutral sugars such as rhamnose, arabinose, xylose, mannose, galactose and glucose, which contribute to lead binding hydroxyl groups. Scanning electron microscope coupled with energy dispersive X-ray spectrometric analysis of this lead resistant strain exposed to 1.6 mM lead nitrate interestingly revealed mucous EPS surrounding bacterial cells which sequestered 17 % lead (as weight %) extracellularly and protected the bacterial cells from toxic effects of lead. This lead resistant strain also showed multidrug resistance. Thus these results significantly contribute to better understanding of structure, function and environmental application of lead-enhanced EPSs produced by bacteria. This lead-enhanced biopolymer can play a very important role in bioremediation of several heavy metals including lead.

Pseudomonas aeruginosa strain WI-1 from Mandovi estuary possesses metallothionein to alleviate lead toxicity and promotes plant growth.

A bacterial isolate from Mandovi estuary Goa, India, which can resist 0.6mM lead nitrate in Tris-buffered minimal medium was identified as Pseudomonas aeruginosa and designated as strain WI-1. PCR amplification clearly revealed presence of bmtA gene encoding bacterial metallothionein responsible for metal sequestration and AAS analysis proved intracellular bioaccumulation of 26.5mg lead/gram dry weight of cells. SDS-PAGE analysis confirmed lead induced bacterial metallothionein with molecular weight 11 kDa, which corresponds to the predicted bmtA gene. Significant growth inhibition of phytopathogenic fungi Fusarium oxysporum NCIM 1008 by siderophore-rich culture supernatant was also observed. Pot experiment with Pisum sativum L inoculated with this strain revealed higher seed germination percentage and significant growth promotion than uninoculated seeds in a soil amended with 7.704 g/kg lead, which indicates amelioration of lead toxicity. This lead resistant strain showed cross tolerance to cadmium, mercury and Tributyltin chloride (TBTC) along with resistance to multiple antibiotics.