The role of various parameters like temperature, pH, blood bag concentration, agitation and incubation that influence the production of prodigiosin by Achromobacter denitrificans SP1 was determined. The Plackett-Burman and Box-Behnken experimental designs were employed to statistically optimize and find out the best combinational effect of parameters for the better yield of prodigiosin using blood bag as sole carbon and energy source for the growth of A. denitrificans SP1. The maximum (1.314 mg/ml) prodigiosin production was attained at a temperature of 24 degrees C, pH (8.8), and blood bag (1 g) as optimum; while the predicted value was 1.319 mg/ml with a correlation coefficient of 0.987; which signifies the fitness of the model. Antimicrobial activity of the prodigiosin was also evaluated and found to be an effective agent against bacterial pathogens including Staphylococcus aureus and Proteus mirabilis. Utilization of the plasticizer di (2-ethylhexyl)phthalate (DEHP) in blood bag and the production of antibacterial prodigiosin makes A. denitrificans SP1, an effective competitor toward the pathogenic bacterial disinfection and wastewater treatment processes.

Quorum sensing is a density-dependent chemical process between bacteria, which may be intergenus or intragenus. N-acyl homoserine lactones (HSLs) are a type of small signaling molecules associated with Gram-negative bacteria for monitoring their own population density. The present study unveils the mechanism of HSLs in Achromobacter denitrificans SP1 while transforming di(2-ethylhexyl) phthalate (DEHP) into prodigiosin in a simple basal salt medium. The primary detection of HSLs was done by the colorimetric method. Fourier-transform infrared spectroscopy and liquid chromatography-mass spectrometry-quadrupole time-of-flight confirmed and identified the HSLs. The maximum production of HSLs was observed between 24 and 72 h of incubation, which is noted to be a peak time of DEHP degradation. A total of 57.2% of DEHP was degraded within 30 h and complete degradation was observed within 72 h of incubation. Regulation in the synthesis of various acyl-HSL molecules, viz. 3OC6-HSL in the initial stage of DEHP stress, 3OC8-HSL, and C10-HSL during the time of degradation and 3OC12-HSL on completion of degradation was noticed. The role of HSLs on the production of prodigiosin was confirmed using vanillin as an HSL inhibitor. Through the selective activation of HSL molecules, A. denitrificans SP1 sustain the changing stressful conditions. Supplementation of acyl-HSL signal molecules may boost up the efficacy of A. denitrificans SP1 in both DEHP degradation and prodigiosin production which offers great potential towards the management of DEHP containing plastic wastes.

A novel, ex-situ remediation process for perchlorate contaminated soil is reported in this study. This approach comprises washing the contaminated soil with water, followed by treatment of the wash water in a bioreactor. The treated water reused for the next batch of soil, and the cycle continued. The pilot-scale treatment unit comprising of a soil washing unit (0.75 m(3)) and a fixed-film bioreactor (140 L), both connected in series for continuous operation for a period of three months. The bioreactor was inoculated with a novel perchlorate reducing microbial consortium comprising Serratia marcescens (Gen bank no. HM751096), Bacillus pumilus (Gen bank no. JQ820452) and Micrococcus sp. (Gen bank no. KJ410671). The microbial activity was supported by glucose (glucose/perchlorate ratio = 5), and trace mineral solution. In a typical washing cycle, 2.5 g perchlorate (KCI04) spiked in 670 kg soil was completely removed in three washing cycles, that completed in 6.3 h consuming-360 L water. The pooled wash water containing perchlorate at 8.5 mg/L was treated completely in the bioreactor operated at 4.5 h HRT and -200 mV ORP. Compared with both in-situ and ex-situ remediation methods reported, the present approach has many advantages for treating perchlorate contaminated soil. (C) 2020 Elsevier Ltd. All rights reserved.

Background: Naringin, pharmaceutically active flavonoid, rapidly metabolizes in liver into naringenin. Both naringin and naringenin have significant biological activity and less toxicity. Objective: In the present study, in silico molecular interactions of naringin and its metabolite naringenin have been evaluated against different human liver fibrosis proteins. Materials and Methods: The major human therapeutic protein targets such as epidermal growth factor receptor (EGFR), vascular endothelial growth factor receptor-2 (1/EGFR-2), fibroblast growth factor receptor-1 (FGFR1), Kelch-like ECH-associated protein-1 (Kaep1), transforming growth factor beta receptor I (TGFBR-1), angiotensin II receptor type-1 (Angio-II-Type-1), Janus kinase-2 (JAK-2), Zeta-chain-associated protein kinase-70 (ZAP-70) have been selected for the docking studies This computational study was performed using Schrodinger Suite Maestro 10.3 Glide software 2015. Results: The studies demonstrated comparable binding affinities of naringin and naringenin with human therapeutic protein targets such as JAK-2, ZAP-70 Kinase, Angio-II-Type 1, TGFBR1, Kaep1, EGFR, VEGFR-2, and FGFR1 when compared to their respective standard drugs such as gefitinib, regorafenib, dovitinib, bardoxolone methyl, SB-431542, olmesartan, and ruxolitinib. Naringin showed better glide score ranging from -8.5 to -13.3 kcal/mol whereas its metabolite Naringenin also showed comparable glide score ranging from -5.4 to -9.3 kcal/mol. The binding of target proteins with respective standard drugs showed -2.2 to -10.12 kcal/mol. Conclusion: The observed in silico human protein interactions of naringin and its metabolite naringenin could be exploited for the anti-liver fibrosis therapy. The results derived from this pioneering virtual study may advance further mechanistic in vitro and preclinical in vivo studies.

The spatio-temporal distribution of perchlorate in water sources around an ammonium perchlorate production unit and its toxicity response on a dominant aquatic plant, Hydrilla verticillata are reported in this study. Samples (n=453) from ground water (open well) and surface water sources within 5km from the production unit over a period of 12 months (2014, June − 2015, May) were screened for ClO4−. During the period, ClO4−concentration in ground water samples close to the production unit increased to >40,000μg/L, and ClO4− was detected at 1740μg/L in well water 1.6km away from the production unit. A community pond in the area also showed an increase in ClO4− level up to 29,000μg/L. In all water sources, ClO4− level was maximum during the rainy season (July, monsoon). A natural degradation of ClO4− was not observed in the area as evident from its persistent level and spreading to more areas. H. verticillata, the dominant vegetation in the pond exhibited severe toxic response like massive decay and loss of photosynthetic pigments such as chlorophyll and carotene due to ClO4− exposure. The plant accumulated ClO4− up to 60±0.8mg/kg wet weight with a Bio-Concentration Factor 2.06±0.005. This is the first report on spatio-temporal distribution of ClO4− at higher levels in a natural environment and its toxicity response to plants under natural condition.

Phthalates are known to cause endocrine disruption in humans and animals. Being lipophilic xenobiotic chemicals, phthalates from the surrounding environments can easily be absorbed into the biological system, thereby causing various health dysfunctions. This molecular docking study evaluates a variety of molecular interactions of 12 commonly used diphthalates and respective monophthalates onto the ligand binding domain (LBD) of the human pregnane X receptor (hPXR), a xenosensor, which would be beneficial for further in vitro and in vivo studies on hazardous phthalates. Out of 12 diphthalates and their monophthalates tested, diisodecyl phthalate (-9.16kcalmol(-1)) showed more affinity toward hPXR whereas diisononyl phthalate (-8.77) and di(2-ethyhexyl)phthalate (-8.56), the predominant plasticizers found in a variety of plastics and allied products, showed comparable binding scores with that of the control ligands such as hyperforine (-9.99) and dexamethasone (-7.36). In addition to the above diphthalates, some of their monophthalates (monoisodecyl phthalate, mono-2-etheylhexyl phthalate, etc.) also established similar interactions with certain crucial amino acids in the LBD, which led to higher G scores. In fact, bisphenol A, a well-studied and proven endocrine disruptor, showed lesser G scores (-6.69) than certain phthalates. Copyright (c) 2016 John Wiley & Sons, Ltd.This molecular docking study using Glide evaluates a variety of molecular interactions of 12 commonly used diphthalates (generally reported as hazardous) and respective monophthalates with the ligand binding domain of the human pregnane X receptor (hPXR). Diisodecyl phthalate showed more affinity towards hPXR; whereas diisononyl phthalate and di(2-ethyhexyl)phthalate, the predominant plasticizers found in a variety of plastics and allied products, showed moderate binding. A higher affinity of certain phthalates to hPXR than its natural ligands and BPA (a proven endocrine disruptor) is alarming.

The present study describes the culture characteristics, isolation and partial purification of a yellow pigment produced by a novel bacterium, Micrococcus luteus strain BAA2. The growth conditions of M. luteus strain BAA2 were optimized. It showed the optimum growth (OD: 0.55 at λ660) at 28oC (pH 7, 24 h), while the maximum pigment (OD: 0.50 at λ466) production was at 36 h (pH 7). The yellow pigment secreted in the medium was extracted in methanol, and purified using silica column chromatography, and the pigment was characterized using thin layer chromatography (2 spots: Rf 0.38 and 0.43), UV-visible and IR spectroscopic techniques; and both spectroscopic profiles showed the characteristic peaks of carotenoid pigment.

Phthalic acid esters or phthalates are ubiquitous environmental pollutants known for their adverse health effects in test animals and, of late, in humans. Thus, in this molecular docking study - using Glide (Schrodinger) - the molecular interactions of 31 ligands, including 12 diphthalates, their monophthalates and phthalic acid with selected human ketosteroid receptors, i.e., androgen (hAR), progesterone (hPR) and glucocorticoid (hGR) receptors were explored and their binding affinities were compared with that of corresponding natural steroids and a known endocrine disrupting xenobiotic, bisphenol A (BPA). Mostly, diphthalates and monophthalates showed the potential for antisteroidal activity by interacting with hAR, hPR and hGR. Of them, diphenyl phthalate showed the highest G score (-7.70 kcalmol(-1)) with hAR, and the crucial amino acid (aa) residues in the ligand binding domain (LBD) of this receptor involved in the molecular interactions were Phe 764, Leu 704, Asn 705 and Thr 877. The mono-iso-decyl phthalate showed the highest G score (-8.36) with the hPR, and the crucial aa residues in the LBD interactions were Arg 766 Gln 725 and Phe 778. The mono-iso-decyl phthalate also showed more affinity (-8.44) towards hGR than the natural ligand, and the aa residues in the LBD interactions were Gln 570 and Met 604. In addition to these, some other phthalates established comparable interactions with certain aa residues located in the LBD of these receptors, which resulted in higher G scores. Contrastingly, BPA and some natural ligands tested in this study showed lower G scores with these receptors than certain phthalates reported herein, i.e., certain phthalates are more toxic than the proven toxic BPA. Copyright (c) 2015 John Wiley & Sons, Ltd.Phthalates are ubiquitous environmental pollutants, known for their adverse health effects in test animals, and of late in humans. Molecular interactions of diphthalates, respective monophthalates, phthalic acid and the known endocrine disruptor, bisphenol A with human ketosteroid receptors were explored using Glide (Schrodinger); and their binding efficiencies were compared with that of the natural steroids. From the in silico evidences, most of these diphthalates and their monophthalates showed potentials for anti-steroidal activity by interacting with human ketosteroid receptors.

Phthalates or phthalic acid esters are a group of xenobiotic and hazardous compounds blended in plastics to enhance their plasticity and versatility. Enormous quantities of phthalates are produced globally for the production of plastic goods, whose disposal and leaching out into the surroundings cause serious concerns to the environment, biota and human health. Though in silico computational, in vitro mechanistic, pre-clinical animal and clinical human studies showed endocrine disruption, hepatotoxic, teratogenic and carcinogenic properties, usage of phthalates continues due to their cuteness, attractive chemical properties, low production cost and lack of suitable alternatives. Studies revealed that microbes isolated from phthalate-contaminated environmental niches efficiently bioremediate various phthalates. Based upon this background, this review addresses the enumeration of major phthalates used in industry, routes of environmental contamination, evidences for health hazards, routes for in situ and ex situ microbial degradation, bacterial pathways involved in the degradation, major enzymes involved in the degradation process, half-lives of phthalates in environments, etc. Briefly, this handy module would enable the readers, environmentalists and policy makers to understand the impact of phthalates on the environment and the biota, coupled with the concerted microbial efforts to alleviate the burden of ever increasing load posed by phthalates.

Production of an esterase by Achromobacter denitrifi cans strain SP1 a di(2-ethylhexyl)phthalate (DEHP) degrading novel bacterium in a modified basal salt medium supplemented with DEHP as an inducercum-additional carbon source was studied. The Plackett Burman and Box Behnken designs were applied to statistically optimize the production parameters, which resulted in an increase of esterase production by 24%. For the production of the maximum (30.5 U) intracellular esterase, 10 mM DEHP and 72 h incubation at pH 8.0 were found as optimum conditions; while the predicted value was 28.8 U with a correlation coefficient of 0.932; which signifies the fitness of the model. The optimum activity of the 2.5 folds purified esterase was 89.5 U with 20 mM para-nitrophenyl acetate as substrate (50 degrees C, pH 8.0 and for 30 min), and various metal ions were found to retarde the esterase activity. The approximate MW of partially purified esterase was 53 kDa, and the activity of esterase was also confirmed by native-PAGE. The Km and Vmax values of esterase were 1.308 mM and 62.52 mu mol min(-1) mg(-1), respectively. Briefly, this was the first report on an enzyme from the DEHP degrading A. denitrificans SP1, which in comparison with esterase from other phthalate degrading bacteria and fungi showed better Km and Vmax. (C) 2015 Elsevier Ltd. All rights reserved.

This study describes how Achromobacter denitrificans strain SP1, a novel isolate from heavily plastics-contaminated sewage sludge efficiently consumed the hazardous plasticizer, di(2-ethylhexyl)phthalate (DEHP) as carbon source supplemented in a simple basal salt medium (BSM). Response surface methodology was employed for the statistical optimization of the process parameters such as temperature (32 °C), agitation (200 rpm), DEHP concentration (10 mM), time (72 h) and pH (8.0). At these optimized conditions, experimentally observed DEHP degradation was 63%, while the predicted value was 59.2%; and the correlation coefficient between them was 0.998, i.e., highly significant and fit to the predicted model. Employing GC–MS analysis, the degradation pathway was partially deduced with intermediates such as mono(2-ethylhexyl)phthalate and 2-ethyl hexanol. Briefly, this first report describes A. denitrificans strain SP1 as a highly efficient bacterium for completely remediating the hazardous DEHP (10 mM) in 96 h in BSM (50% consumed in 60 h), which offers great potentials for efficiently cleaning the DEHP-contaminated environments such as soil, sediments and water upon its deployment.

This first report describes the purification and identification of an orange-red pigment produced by Achromobacter denitrificans strain SP1 (isolated from sewage sludge heavily contaminated with plastics) during its growth in a simple basal salt medium supplemented with the hazardous di(2-ethylhexyl)phthalate (DEHP) blended in PVC blood bag (in situ) or free DEHP (ex situ) as carbon source. The cell-bound pigment was elucidated, characterized at molecular level, and described as an unusual 25C prodigiosin analog for the first time. At laboratory conditions (in flasks), the dry cell mass was 75.2mg/g blood bag, which upon extraction yielded 7.1mg prodigiosin; at this stage the pH of the medium was dropped from 7.2 to 3.5. Considering its pharmaceutical importance, taking 10 known prodigiosins as controls, this 25C prodigiosin was subjected to molecular docking studies, showed comparable and promising binding efficiencies with the crucial molecular human targets like cycloxygenase-2, ZAP-70 kinase and Jak-3 kinase.

Proteases or peptidases constitute the largest group of enzymes in bio-industry with a long array of uses.They play an invincible role in industrial biotechnology, especially in detergent, food and pharmaceutical arena.This focused review encompasses an overview on alkaline proteases, mainly of microbial sources in a handy module.Following an introduction and general classification with evolutionary insight, major sources of proteases (animal, plant and microbial including fungal, bacterial), their general properties with mechanism of action and molecular masses are discussed.Proteases from Bacillus spp.have been given special attention.In addition to this, an overview on the applications of proteases in detergent, tannery, food, metal recovery and waste treatment industries is also addressed briefly.

This unique study describes how Aspergillus japonicus, Penicillium brocae and Purpureocillium lilacinum, three novel isolates of our laboratory from heavily plastics-contaminated soil completely utilized the plasticizer di(2-ethylhexyl)phthalate (DEHP) bound to PVC blood storage bags (BB) in simple basal salt medium (BSM) by static submerged growth (28 °C). Initial quantification as well as percentage utilization of DEHP blended to BB were estimated periodically by extracting it into n-hexane. A two-stage cultivation strategy was employed for the complete mycoremediation of DEHP from BB in situ. During the first growth stage, about two-third parts of total (33.5 % w/w) DEHP bound to BB were utilized in two weeks, accompanied by increased fungal biomass (~0.15–0.32 g per g BB) and sharp declining (to ~3) of initial pH (7.2). At this stagnant growth state (low pH), spent medium was replaced by fresh BSM (pH, 7.2), and thus in the second stage the remaining DEHP (one-third) in BB was utilized completely. The ditches and furrows seen from the topology of the BB as seen by the 3D AFM image further confirmed the bioremediation of DEHP physically bound to BB in situ. Of the three mycelial fungi employed, P. lilacinum independently showed highest efficiency for the complete utilization of DEHP bound to BB, whose activity was comparable to that of the consortium comprising all the three fungi described herein. To sum up, the two-stage cultivation strategy demonstrated in this study shows that a batch process would efficiently remediate the phthalic acid esters blended in plastics on a large scale, and thus it offers potentials for the management of plastics wastes.

ABSTRACTThis exhaustive in silico study looks into the molecular interactions of phthalates and their metabolites with human peroxisome proliferator‐activated receptor (hPPAR) and retinoid X receptor (hRXR) α, β and γ subtypes – the nuclear receptor proteins function as transcription factors by regulating the expression of downstream genes. Apart from the much discussed plasticizer bisphenol A, we examined the binding affinities of 15 common diphthalates and their monophthalates, natural (linoleic acid, conjugated linoleic acid) and synthetic (bezafibrate, pioglitazone, GW 50156) ligands with hPPARs. In addition to these phthalates, specific natural (retinoic and phytanic acids) and synthetic (bexarotene, rosiglitazone) ligands were examined with hRXRs. The Maestro, Schrödinger Suite 2012 was used for the molecular docking study. In general, natural ligands of hPPAR showed less binding efficiencies than phthalic acid esters and drugs. The diphthalate di‐iso‐decyl phthalate showed the highest G score (–9.99) with hPPAR (γ), while its monophthalate (mono‐iso‐decyl phthalate) showed a comparatively less G score (–9.56). Though the PPAR modulator GW 50156 showed strong affinity with all hPPAR subtypes, its highest G score (–12.43) was with hPPARβ. Hazardous di(2‐ethylhexyl)phthalate generally showed a greater preference to hRXRs than hPPARs, but its highest G score (–10.87) was with hRXRα; while its monophthalate (Mono(2‐ethylhexyl)phthalate) showed a lesser G score (–8.59). The drug bexarotene showed the highest G score (–13.32) with hRXRβ. Moreover, bisphenol A showed more affinity towards hRXR. Briefly, this study gives an overview on the preference of phthalic acid esters, natural and synthetic ligands on to hPPAR and hRXR subtypes, which would lead to further in vitro mechanistic as well as in vivo preclinical and clinical studies. Copyright © 2013 John Wiley & Sons, Ltd.

Owing to the production of alpha, beta and gamma amylase subtypes; starch degrading microbes, especially bacteria have an invincible role in the food, fermentation, textile and paper industries. Of them, α-amylases from Bacillus spp. have contributed tremendous advancements in bio-industry, especially in starch, detergent and pharmaceutical arena. Though general reviews are seen in literature on amylases, no focused review is available yet solely on α-amylases produced by Bacillus spp. Hence, this focused review on α-amylases from the genus Bacillus is designed in such a way that it should give a vivid picture on most of the aspects on bacillial α-amylases in a handy module with an industrial perspective. With a short introduction on amylases in general, α-amylases from various species of Bacillus reviewed herein encompasses production of α-amylases by submerged and solid-state fermentations; nutrients and other factors required for maximizing production; immobilization strategies for whole cells or purified enzyme; an overview on the molecular weight of the enzyme; followed by distinct sections for purification, characterisation, stability and crystal structure; and concluded with a section on industrial applications of the α-amylases from Bacillus spp.

Being lipophilic xenobiotic chemicals, phthalates from the surrounding environments can easily be absorbed into the biological system, thereby causing various health problems including cancer and endocrine disruption in test animals and also in humans. In the present in silico study employing Glide, Schrodinger Suite 2012, we analysed in detail the binding affinities of 12 commonly used diphthalates and their metabolites (corresponding mono ester and phthalic acid) onto the ligand-binding domain (LBD) of the human estrogen receptor (hER), human estrogen receptor (hER) and human estrogen related receptor (hERR). Natural ligand 17 estradiol (E2), known xenoestrogen bisphenol A, the phytoestrogen genistein, the agonists/antagonists 4-hydroxy tamoxifen and raloxifene were also docked onto these receptors as positive controls for comparing the binding efficiencies with that of phthalates and their metabolites. Results revealed that E2 had less binding affinity to the receptors in comparison to certain phthalates, i.e. maximum binding scores (G score, kcal/mol) were diisononyl phthalate (-9.44) to hER, monophenyl phthalate (-8.66) to hER and di(2-ethylhexyl)phthalate (-9.38) to hERR. The most concerned monophthalates established additional H bonds with certain surrounding crucial amino acid residues in the LBD, and thus showed more affinity to all the receptors than even the natural ligand and other well-characterised xenoestrogens as demonstrated in this study. Briefly, this study gives an insight into the virtual binding behaviours of commonly used phthalates and their metabolites onto hERs and hERR, which would accelerate further in vitro mechanistic, preclinical and clinical studies on real in vitro or in vivo platforms.

In this study, we explored the efficacy of raw potato flour (PF) as supplement to the conventional LB medium (LB control, designated as M1) for enhancing the concomitant production of endospores and delta-endotoxin from Bacillus thuringiensis subsp. kurstaki by solid-state fermentation (SSF). Of different concentrations and combinations of media tested, 10% (wilt) PF supplemented LB medium (M2) was found as the best source for the maximum yield of toxin. After 12 h submerged fermentation (SmF) at 37 degrees C and 125 rpm, M2 was made into a wet-solid matter for SSF by removing the supernatant (1000 xg, 10 min); the resultant pellet subsequently incubated statically (37 C) for the production of B. thuringiensis subsp. lcurstaki toxin (Btk-toxin). In comparison to Ml, yield of 3-endotoxin purified by sucrose density gradient centrifugation method from M2 was about 6-fold higher (53% recovery). This maximum yield from M2 was obtained at 48 h (as against 72 h from M1), thus the gestation period of M2 was reduced by 24 h with higher yield. In addition to the quantitative data, qualitative photomicrographs taken by image analyzer, scanning electron and fluorescent microscopes and digital camera showed physical evidences for the upper hand of SSF over conventional SmF for the enhanced production of Btk-toxin. SOS-PAGE image of the purified d-endotoxin showed three major fractions with apparent MWs 66, 45 and 30 liDa. Briefly, if low-cost agricultural products like PF is used as supplement to LB, by SSF strategy, production of Btk-toxin could be enhanced to 6-fold in short gestation time without losing its entomotoxicity efficiency. (C) 2013, The Society for Biotechnology, japan. All rights reserved.