Characterization of pumilacidin, a lipopeptide biosurfactant produced from Bacillus pumilus NITDID1 and its prospect in bioremediation of hazardous pollutants.

Highly hydrophobic compounds like petroleum and their byproducts, once released into the environment, can persist indefinitely by virtue of their ability to resist microbial degradation, ultimately paving the path to severe environmental pollution. Likewise, the accumulation of toxic heavy metals like lead, cadmium, chromium, etc., in the surroundings poses an alarming threat to various living organisms. To remediate the matter in question, the applicability of a biosurfactant produced from the mangrove bacterium Bacillus pumilus NITDID1 (Accession No. KY678446.1) is reported here. The structural characterization of the produced biosurfactant revealed it to be a lipopeptide and has been identified as pumilacidin through FTIR, NMR, and MALDI-TOF MS. The critical micelle concentration of pumilacidin was 120 mg/L, and it showed a wide range of stability in surface tension reduction experiments under various environmental conditions and exhibited a high emulsification index of as much as 90%. In a simulated setup of engine oil-contaminated sand, considerable oil recovery (39.78%) by this biosurfactant was observed, and upon being added to a microbial consortium, there was an appreciable enhancement in the degradation of the used engine oil. As far as the heavy metal removal potential of biosurfactant is concerned, as much as 100% and 82% removal was observed for lead and cadmium, respectively. Thus, in a nutshell, the pumilacidin produced from Bacillus pumilus NITDID1 holds promise for multifaceted applications in the field of environmental remediation.

Removal of lindane in liquid culture using soil bacteria and toxicity assessment in human skin fibroblast and HCT116 cell lines.

The development of effective measures for the remediation of lindane contaminated sites is the need of the hour. In this study, a potent lindane degrading bacteria, identified as Rhodococcus rhodochrous NITDBS9 was isolated from an agricultural field of Odisha that could utilize up to 87% of 100 mg L-1 lindane when grown under liquid culture conditions in mineral salt media in 10 days. The bacteria could produce biofilm in lindane-containing media. Rhodococcus rhodochrous NITDBS9 was further characterized for its plant growth-promoting properties and it was found that the bacteria showed abilities for phytohormone, ammonia and biosurfactant production, etc. This could be beneficial for the bioremediation and improvement of crop production in contaminated sites. Ecotoxicity studies carried out for lindane, and its degradation products in mung bean and mustard seeds showed a reduction in toxicity of lindane after treatment with NITDBS9. NITDBS9 was used with a previously isolated potent lindane degrading strain Paracoccus sp. NITDBR1 in a dual mixed culture for the enhanced removal of lindane in the liquid system i.e. up to 93% in 10 days. Cytotoxicity studies were conducted with lindane before and after treatment with the single and dual mixed cultures on human skin fibroblast and HCT116 cell lines. They revealed a significant reduction in toxicity of lindane after it was bioremediated with the single and dual mixed cultures. Therefore, our proposed strategy could be efficiently used for the detoxification of the lindane-contaminated system, and further work should be done to study the use of these cultures in the contaminated soil system.

Lindane removal in contaminated soil by defined microbial consortia and evaluation of its effectiveness by bioassays and cytotoxicity studies / Eliminación de lindano en suelos contaminados por consorcios microbianos definidos y evaluación de su eficacia mediante bioensayos y estudios de citotoxicidad

Lindane contamination in different environmental matrices has been a global concern for long. Bacterial consortia consisting of Paracoccus sp. NITDBR1, Rhodococcus rhodochrous NITDBS9, Ochrobactrum sp. NITDBR3, NITDBR4 and NITDBR5 were used for the bioremediation of soil artificially contaminated with lindane. The bacteria, Paracoccus sp. NITDBR1 and Rhodococcus rhodochrous NITDBS9, have been selected based on their lindane degrading capacity in liquid culture conditions (~80-90 %). The remaining three bacteria were chosen for their auxiliary properties for plant growth promotion, such as nitrogen fixation, phosphate solubilization, indole-3-acetic acid production and ammonia production under in vitro conditions. In this study, market wastes, mainly vegetable wastes, were added to the soil as a biostimulant to form a biomixture for assisting the degradation of lindane by bioaugmentation. Residual lindane was measured at regular intervals of 7 days to monitor the biodegradation process. It was observed that the consortium could degrade ~80% of 50 mg kg-1 lindane in soil which was further increased in the biomixture after six weeks of incubation. Bioassays performed on plant seeds and cytotoxicity studies performed on human skin fibroblast and HCT116 cell lines revealed that the groups contaminated with lindane and treated with the bacterial consortium showed lower toxicity than their respective controls without any bacteria. Hence, the use of both pesticide degrading and plant growth-promoting bacteria in a consortium can be a promising strategy for improved bioremediation against chemical pesticides, particularly in soil and agricultural fields, simultaneously enhancing crop productivity in those contaminated soil(AU)

Lindane removal in contaminated soil by defined microbial consortia and evaluation of its effectiveness by bioassays and cytotoxicity studies.

Lindane contamination in different environmental matrices has been a global concern for long. Bacterial consortia consisting of Paracoccus sp. NITDBR1, Rhodococcus rhodochrous NITDBS9, Ochrobactrum sp. NITDBR3, NITDBR4 and NITDBR5 were used for the bioremediation of soil artificially contaminated with lindane. The bacteria, Paracoccus sp. NITDBR1 and Rhodococcus rhodochrous NITDBS9, have been selected based on their lindane degrading capacity in liquid culture conditions (~80-90 %). The remaining three bacteria were chosen for their auxiliary properties for plant growth promotion, such as nitrogen fixation, phosphate solubilization, indole-3-acetic acid production and ammonia production under in vitro conditions. In this study, market wastes, mainly vegetable wastes, were added to the soil as a biostimulant to form a biomixture for assisting the degradation of lindane by bioaugmentation. Residual lindane was measured at regular intervals of 7 days to monitor the biodegradation process. It was observed that the consortium could degrade ~80% of 50 mg kg-1 lindane in soil which was further increased in the biomixture after six weeks of incubation. Bioassays performed on plant seeds and cytotoxicity studies performed on human skin fibroblast and HCT116 cell lines revealed that the groups contaminated with lindane and treated with the bacterial consortium showed lower toxicity than their respective controls without any bacteria. Hence, the use of both pesticide degrading and plant growth-promoting bacteria in a consortium can be a promising strategy for improved bioremediation against chemical pesticides, particularly in soil and agricultural fields, simultaneously enhancing crop productivity in those contaminated soil.

Fate of GdF3 nanoparticles-loaded PEGylated carbon capsules inside mice model: a step toward clinical application.

The successful translation of nanostructure-based bioimaging and/or drug delivery system needs extensive in vitro and in vivo studies on biocompatibility, biodistribution, clearance, and toxicity for its diagnostic applications. Herein, we have investigated the in vitro cyto-hemocompatibility, in vivo biodistribution, clearance, and toxicity in mice after systemic administration of GdF3 nanoparticles loaded PEGylated mesoporous carbon capsule (GdF3-PMCC)-based theranostic system. In vitro cyto-hemocompatibility study showed a very good biocompatibility up to concentration of 500 µg/ml. Biodistribution studies carried out from 1 h to 8 days showed that GdF3-PMCC was found in major organs, such as liver, kidney, spleen, and muscle till 4th day and it was negligible in any tissue after 8th day. The clearance study was carried out for a period of 8 days and it was observed that the urinary system is the main route of excretion of GdF3-PMCC. The tissue toxicity study was done for 15 days and histopathological analysis indicated that the GdF3-PMCC based theranostic system does not have any adverse effect in tissues. Thus, PMCCs are nontoxic and can be applied as theranostic agents in contrast to the other carbon-based systems (PEGylated carbon nanotubes and PEGylated graphene oxide) which showed significant toxicity.

Isolation and characterization of a lindane degrading bacteria Paracoccus sp. NITDBR1 and evaluation of its plant growth promoting traits

Lindane contamination in different environmental compartments is still posing a serious threat to our environment and effective measures need to be taken for the detoxification of lindane. Soil bacteria isolated from agricultural fields are known to possess certain plant growth promoting traits like the production of phytohormones, production of ammonia, nitrogen fixation and solubilization of phosphorus, etc. In the present study, an indigenous bacterial strain Paracoccus sp. NITDBR1 have been isolated from an agricultural field in Manipur, India which could grow on 100 mg L−1 lindane as the sole source of carbon and could degrade up to 90% of lindane in mineral salt media under liquid culture conditions in 8 days. The strain NITDBR1 was able to form biofilm in lindane media and the addition of substrate like glucose and sucrose enhanced the biofilm formation by 1.3 and 1.17-fold respectively in 3 days. The strain NITDBR1 could produce glycolipid and glycoprotein based biosurfactants. It was also found to possess plant growth promoting traits like nitrogen fixation and indole-3-acetic acid production to assist crop production. The phytotoxicity studies carried out on mustard seeds revealed that the degradation products formed after treatment with NITDBR1 could lower the toxicity of lindane for root elongation by 1.3-fold. Therefore, strain NITDBR1 could be useful for the bioremediation of soil contaminated with lindane with lesser damage to the environment, biofilm forming ability may help the bacteria survive under stressed environmental conditions, and biosurfactant production will help in increasing the bioavailability of contaminants. The plant growth promoting traits can be beneficial for agriculture. With such soil friendly activities coupled with pesticide degradation, this strain can be used for environmental as well as agricultural applications

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Isolation and characterization of a lindane degrading bacteria Paracoccus sp. NITDBR1 and evaluation of its plant growth promoting traits.

Lindane contamination in different environmental compartments is still posing a serious threat to our environment and effective measures need to be taken for the detoxification of lindane. Soil bacteria isolated from agricultural fields are known to possess certain plant growth promoting traits like the production of phytohormones, production of ammonia, nitrogen fixation and solubilization of phosphorus, etc. In the present study, an indigenous bacterial strain Paracoccus sp. NITDBR1 have been isolated from an agricultural field in Manipur, India which could grow on 100 mg L-1 lindane as the sole source of carbon and could degrade up to 90% of lindane in mineral salt media under liquid culture conditions in 8 days. The strain NITDBR1 was able to form biofilm in lindane media and the addition of substrate like glucose and sucrose enhanced the biofilm formation by 1.3 and 1.17-fold respectively in 3 days. The strain NITDBR1 could produce glycolipid and glycoprotein based biosurfactants. It was also found to possess plant growth promoting traits like nitrogen fixation and indole-3-acetic acid production to assist crop production. The phytotoxicity studies carried out on mustard seeds revealed that the degradation products formed after treatment with NITDBR1 could lower the toxicity of lindane for root elongation by 1.3-fold. Therefore, strain NITDBR1 could be useful for the bioremediation of soil contaminated with lindane with lesser damage to the environment, biofilm forming ability may help the bacteria survive under stressed environmental conditions, and biosurfactant production will help in increasing the bioavailability of contaminants. The plant growth promoting traits can be beneficial for agriculture. With such soil friendly activities coupled with pesticide degradation, this strain can be used for environmental as well as agricultural applications.

Modelling the growth kinetics of Kocuria marina DAGII as a function of single and binary substrate during batch production of ß-Cryptoxanthin.

In the present investigation, growth kinetics of Kocuria marina DAGII during batch production of ß-Cryptoxanthin (ß-CRX) was studied by considering the effect of glucose and maltose as a single and binary substrate. The importance of mixed substrate over single substrate has been emphasised in the present study. Different mathematical models namely, the Logistic model for cell growth, the Logistic mass balance equation for substrate consumption and the Luedeking-Piret model for ß-CRX production were successfully implemented. Model-based analyses for the single substrate experiments suggested that the concentrations of glucose and maltose higher than 7.5 and 10.0 g/L, respectively, inhibited the growth and ß-CRX production by K. marina DAGII. The Han and Levenspiel model and the Luong product inhibition model accurately described the cell growth in glucose and maltose substrate systems with a R 2 value of 0.9989 and 0.9998, respectively. The effect of glucose and maltose as binary substrate was further investigated. The binary substrate kinetics was well described using the sum-kinetics with interaction parameters model. The results of production kinetics revealed that the presence of binary substrate in the cultivation medium increased the biomass and ß-CRX yield significantly. This study is a first time detailed investigation on kinetic behaviours of K. marina DAGII during ß-CRX production. The parameters obtained in the study might be helpful for developing strategies for commercial production of ß-CRX by K. marina DAGII.

Statistical Optimization of Media Components for Production of Fibrinolytic Alkaline Metalloproteases from Xenorhabdus indica KB-3.

Xenorhabdus indica KB-3, a well-known protease producer, was isolated from its entomopathogenic nematode symbiont Steinernema thermophilum. Since medium constituents are critical to the protease production, the chemical components of the selected medium (soya casein digest broth) were optimized by rotatable central composite design (RCCD) using response surface methodology (RSM). The effects of all five chemical components (considered as independent variables), namely tryptone, soya peptone, dextrose, NaCl, and dipotassium phosphate, on protease production (dependent variable) were studied, and it was found that tryptone and dextrose had maximum influence on protease production. The protease production was increased significantly by 66.31% under optimal medium conditions (tryptone-5.71, soya peptone-4.9, dextrose-1.45, NaCl-6.08, and dipotassium phosphate-0.47 in g/L). To best of knowledge, there are no reports on optimization of medium component for protease production by X. indica KB-3 using RSM and their application in fibrinolysis. This study will be useful for industrial processes for production of protease enzyme from X. indica KB-3 for its application in the field of agriculture and medicine.

Experimental and modeling studies on microwave-assisted extraction of mangiferin from Curcuma amada.

Mangiferin, a bioactive compound having potent nutraceutical, strong antioxidant and pharmacological significance has been extracted using microwave-assisted extraction (MAE) technique from Curcuma amada, commonly known as mango ginger. The extraction solvent ethanol is eco-friendly, nontoxic and reduces the risk of environmental hazards. The influence of several independent variables such as microwave power, ethanol concentration, extraction (irradiation) time and pre-leaching time has been studied under varying conditions using one-factor-at-a-time analysis to obtain an optimal extraction ratio. The maximum mangiferin content of 1.1156 mg/g is obtained at microwave power of 550 W and extraction time of 50 s with 80 % ethanol as a solvent and pre-leaching time of 20 min. The results indicate that microwave power and ethanol concentration have the most significant effect on the yield of mangiferin content. The presence of mangiferin in final Curcuma amada extract is confirmed through high-performance liquid chromatography and the functional groups are identified through Fourier transform infrared spectroscopy analyses using standard mangiferin. The experimental profiles are fitted into a two-parameter modified first-order kinetic model and a three-parameter modified logistic model and checked using the goodness-of-fit criterion. The Curcuma amada retained its antioxidant activity after MAE treatment and the antioxidant activity of mangiferin obtained after extraction using DPPH free radical scavenging assay is studied.

Microwave assisted extraction of mangiferin from Curcuma amada.

Mangiferin present in Curcuma amada was extracted with the help of microwave assisted extraction (MAE). The extraction solvent used was ethanol, which is eco-friendly and reduced the risk of environmental hazard. The mangiferin content was found to increase until 500 W, but decreased as the microwave power was increased further. A similar threshold was also obtained for microwave irradiation time. Following a mathematical analysis, an optimal mangiferin yield of 41 µg/mL was obtained from an extraction time of 15.32 s for a microwave power of 500 W.

Optimization of process parameters by response surface methodology and kinetic modeling for batch production of canthaxanthin by Dietzia maris NIT-D (accession number: HM151403).

Dietzia maris NIT-D, a canthaxanthin producer, was isolated during routine screening of pigment-producing bacteria. Response surface methodology was applied for statistical designing of process parameters for biomass and canthaxanthin production. The effects of four process parameters (considered as independent variables), namely temperature (10-30 °C), pH (4.75-5.75), shaker speed (75-135 rpm) and percentage inoculum (0.5-2.5 %) on the biomass and canthaxanthin yield (considered as dependent variables) were studied. As much as 122 mg L(-1) of canthaxanthin was obtained when Dietzia maris NIT-D was incubated for 120 h at 25 °C and 120 rpm, initial pH and percentage inoculum being 5.5 and 2 % respectively. The pigment yield is the highest reported till date, with Dietzia maris as the test organism. The maximum biomass yield was 7.39 g L(-1) under optimized process parameters. The predicted values were also verified by validation experiments in 5-day fermentation. Different mathematical models were used to describe growth and production, considering the effect of glucose in batch mode. The kinetic constants were calculated by fitting the experimental data to the models. Cell growth was inhibited beyond a glucose concentration of 15 g L(-1). Andrews' model gave the best fit with a R (2) value of 0.9993. During the exponential growth phase, the specific growth rate was found to remain fairly constant with respect to time. There was no inhibitory effect due to intracellular product accumulation for all concentrations of glucose. This observation is the first of its kind, as previous studies have reported that increasing accumulation of intracellular carotenoid exerts greater degree of inhibition on growth.

Hydrogen production by Cyanobacteria.

The limited fossil fuel prompts the prospecting of various unconventional energy sources to take over the traditional fossil fuel energy source. In this respect the use of hydrogen gas is an attractive alternate source. Attributed by its numerous advantages including those of environmentally clean, efficiency and renew ability, hydrogen gas is considered to be one of the most desired alternate. Cyanobacteria are highly promising microorganism for hydrogen production. In comparison to the traditional ways of hydrogen production (chemical, photoelectrical), Cyanobacterial hydrogen production is commercially viable. This review highlights the basic biology of cynobacterial hydrogen production, strains involved, large-scale hydrogen production and its future prospects. While integrating the existing knowledge and technology, much future improvement and progress is to be done before hydrogen is accepted as a commercial primary energy source.

Inactive enzymatic mutant proteins (phosphoglycerate mutase and enolase) as sugar binders for ribulose-1,5-bisphosphate regeneration reactors.

BACKGROUND: Carbon dioxide fixation bioprocess in reactors necessitates recycling of D-ribulose1,5-bisphosphate (RuBP) for continuous operation. A radically new close loop of RuBP regenerating reactor design has been proposed that will harbor enzyme-complexes instead of purified enzymes. These reactors will need binders enabling selective capture and release of sugar and intermediate metabolites enabling specific conversions during regeneration. In the current manuscript we describe properties of proteins that will act as potential binders in RuBP regeneration reactors. RESULTS: We demonstrate specific binding of 3-phosphoglycerate (3PGA) and 3-phosphoglyceraldehyde (3PGAL) from sugar mixtures by inactive mutant of yeast enzymes phosphoglycerate mutase and enolase. The reversibility in binding with respect to pH and EDTA has also been shown. No chemical conversion of incubated sugars or sugar intermediate metabolites were found by the inactive enzymatic proteins. The dissociation constants for sugar metabolites are in the micromolar range, both proteins showed lower dissociation constant (Kd) for 3-phosphoglycerate (655-796 muM) compared to 3-phosphoglyceraldehyde (822-966 muM) indicating higher affinity for 3PGA. The proteins did not show binding to glucose, sucrose or fructose within the sensitivity limits of detection. Phosphoglycerate mutase showed slightly lower stability on repeated use than enolase mutants. CONCLUSIONS: The sugar and their intermediate metabolite binders may have a useful role in RuBP regeneration reactors. The reversibility of binding with respect to changes in physicochemical factors and stability when subjected to repeated changes in these conditions are expected to make the mutant proteins candidates for in-situ removal of sugar intermediate metabolites for forward driving of specific reactions in enzyme-complex reactors.