Efficient remediation of meropenem using Bacillus tropicus EMB20 ß-lactamase immobilized on magnetic nanoparticles.

Reducing antibiotic pollution in the environment in essential to preserve the effectiveness of the available antibiotics. In the present study, ß-lactamase from Bacillus tropicus EMB20 was immobilized onto magnetic nanoparticles (Fe3O4) through covalent coupling method. The nanoconjugate was structurally characterized using SEM, FTIR, UV-spectrometry, and XRD diffraction analyses. The prepared enzyme nanoconjugate was thereafter used for remediation of meropenem (Mer) and showed complete removal of 10 mgL-1 Mer within 3 h of treatment. Moreover, the immobilized enzyme was successfully recovered and reused for up to 5 cycles with 57% removal efficiency. The immobilized preparation was also observed to be effective in the removal of higher Mer concentrations of 25 and 50 mgL-1 with 79% and 75% removal efficiency, respectively. The major hydrolyzed product of Mer was found to be opened-lactam ring structure with m/z 402.16. The hydrolyzed product(s) were observed to be non-toxic as revealed through microbial MTT, confocal microscopy, and growth studies. Under the mixed conditions of 50 mgL-1 ampicillin (Amp), 10 mgL-1 amoxicillin (Amox) and, Mer, the nanoconjugate showed simultaneous complete removal of Amp and Mer, while 49% Amox removal was detected after 3 h of treatment. Moreover, the nanoconjugates also showed concomitant complete removal of antibiotic mixture with in 2 h from aquaculture wastewater. Overall, the study comes out with an efficient approach for remediation of ß-lactam antibiotics from contaminated systems.

Overexpression and repression of key rate-limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa.

Implementing two-way strategies to enhance the lipid production in Rhodotorula mucilaginosa with the help of metabolic engineering was focused on the overexpression of acetyl coenzyme A carboxylase (ACC1 carboxylase) gene and repression of 3-hydroxy 3-methylglutaryl reductase (HMG-CoA reductase). Using an inducer (sodium citrate) and inhibitor (rosuvastatin), the amounts of biomass, lipid, and carotenoid were estimated. In the presence of inhibitor (200 mM), 62% higher lipid concentration was observed, while 44% enhancement was recorded when inducer (3 mM) was used. A combination of both inhibitor and inducer resulted in a 57% increase in lipid concentration by the oleaginous yeast. These results were again confirmed by real-time polymerase chain reaction by targeting the expression of the genes coding for ACC1 carboxylase and 13-fold increase was recorded in the presence of inducer as compared with control. This combined strategy (inducer and inhibitor use) has been reported for the first time as far as the best of our knowledge. The metabolic engineering strategies reported here will be a powerful approach for the enhanced commercial production of lipids.

Alleviation of hexavalent chromium by using microorganisms: insight into the strategies and complications.

Excessive industrialization and anthropogenic activities have resulted in widespread prevalence of heavy metals including hexavalent chromium in the environment. In addition to toxic properties, Cr(VI) possesses high stability and mobility, which in total makes it included in the list of priority heavy metals; thus it needs to be managed urgently. Among different methods available for remediation of Cr(VI), bioremediation is considered as one of the sustainable methods which could effectively be utilized for controlling Cr(VI) pollution. In this aspect, the treatment of Cr(VI)-containing wastewater originating from industries is noteworthy. The present review thus is an attempt to present a systematic overview dealing with studies on remediation of hexavalent chromium by using microorganisms and their application in treatment of Cr(VI)-containing industrial wastewaters. Various factors affecting the Cr(VI) removal and methods to enhance the bio-treatment are highlighted, which might act as a basis for researchers developing Cr(VI) bioremediation techniques.

Efficacy of ureolytic Enterobacter cloacae EMB19 mediated calcite precipitation in remediation of Zn (II).

In this study, urease mediated calcite precipitation technique was used for remediation of Zn (II). A urease positive Enterobacter cloacae EMB19 was used to produce calcite impregnated with Zn ions. In co-presence of Ca (II), Zn (II) concentrations of 10 and 20 mg L-1 were completely remediated by the bacterium from the media at 72 and 96 h of incubation, respectively. Co-precipitation of Ca (II) and Zn (II) to form calcite-Zn precipitate is one of the major mechanisms of Zn remediation in the present study. Role of urease in calcite-Zn precipitation was substantiated by using urease/carbonate and ammonium enriched cell free culture supernatant (CFS) obtained after sufficient microbial growth. Using CFS, 68% removal of initial 50 mg L-1 Zn (II) was detected. Energy dispersive X-ray spectroscopy (EDX) and X-ray diffraction pattern (XRD) of the precipitate supports calcite mediated remediation of Zn. Remediation of multiple metals viz. Cd, Zn, Cu was also analyzed using CFS laden with urease. The preparation showed 40% Cd, 23% Zn, and 8% Cu reduction from the solution containing initial 25 mg L-1 of each metal. Overall, it can be concluded that, the E. cloacae mediated calcite precipitation technique could effectively be used for alleviation of Zn (II) and other heavy metals from the contaminated environment.

Harnessing the bio-mineralization ability of urease producing Serratia marcescens and Enterobacter cloacae EMB19 for remediation of heavy metal cadmium (II).

In the present study, urease positive Serratia marcescens (NCIM2919) and Enterobacter cloacae EMB19 (MTCC10649) were individually evaluated for remediation of cadmium (II) using ureolysis-induced calcium carbonate precipitation. Both the cultures were observed to efficiently remove cadmium from the media through co-precipitation of Cd (II) and Ca (II). S. marcescens and E. cloacae EMB19, respectively showed 96 and 98% removal of initial 5.0 mg L-1 soluble Cd (II) from the urea and CaCl2 laden media at 96 h of incubation period. At higher Cd (II) concentrations of 10 and 15 mg L-1, cadmium removal efficiency was much higher in case of E. cloacae EMB19 compared to S. marcescens. In-vitro cadmium (II) remediation study using urease containing cell-free culture supernatant of S. marcescens and E. cloacae EMB19 showed respective 98 and 53% removal of initial 50 mg L-1 Cd (II) from the reaction mixtures in co-presence of Ca (II). While in sole presence of Cd (II), only 16 and 8% removal of Cd (II) were detected for S. marcescens and E. cloacae EMB19, respectively. The elemental analysis of the co-precipitated mineral products using Energy Dispersive X-ray spectroscopy (EDX) clearly showed the prevalence of Ca and Cd ions. The morphology Cd-Ca composites formed with respect to both the cultures were observed to be of different shape and size as revealed through Scanning Electron Microscopy (SEM). Entire study hence comes out with a sustainable bioremediation option which could be effectively used to tackle Cd (II) or other heavy metal pollution.

Degradation of azo dye methyl red by alkaliphilic, halotolerant Nesterenkonia lacusekhoensis EMLA3: application in alkaline and salt-rich dyeing effluent treatment.

Effluents from textile industries are highly colored due to vast use of various azo dyes and color is the first visual indicator of pollution. Biological treatment of textile effluent is often hampered due to the alkaline pH and high salinity; a common characteristic of many textile industrial wastewaters. Considering this, the present study explores the potential of a newly isolated halotolerant and alkaliphilic bacterium Nesterenkonia lacusekhoensis EMLA3 for degradation of methyl red (MR) dye under alkaline condition. Strain EMLA3 showed 97% degradation of 50 mg L-1 MR after 16 h at initial pH of 11.5 in nutrient medium. Dye degradation by the isolate is supported by the formation of low-molecular weight metabolites as divulge through GC-MS & FTIR studies Optimum dye degradation was observed in the pH range of 8.0-11.5 and temperature range of 30-35 °C. Significant MR degrading activity of the strain could be achieved in the presence of very high salt level (100-120 g L-1 NaCl) and in co-presence of different heavy metals. Application of strain to alkaline pH, salt, and heavy metals laden-textile effluent resulted in overall 83% dye removal from the effluent after 120 h of treatment under static condition. Furthermore, the property of microbe to drop-down the pH of wastewater from 11.5 to 8.60 after treatment also lowers the need of additional neutralization treatment. The entire study thus comes out with novel application of N. lacusekhoensis-a less explored extremophilic bacterium-for treatment of alkaline and salt-rich azo dye-containing wastewaters.

Biodegradation of 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) by using Serratia marcescens NCIM 2919.

A solvent tolerant bacterium Serratia marcescens NCIM 2919 has been evaluated for degradation of DDT (1,1,1-trichloro-2,2-bis (4-chlorophenyl) ethane). The bacterium was able to degrade up to 42% of initial 50 mg L-1 of DDT within 10 days of incubation. The highlight of the work was the elucidation of DDT degradation pathway in S. marcescens. A total of four intermediates metabolites viz. 2,2-bis (chlorophenyl)-1,1-dichloroethane (DDD), 2,2-bis (chlorophenyl)-1,1-dichloroethylene (DDE), 2,2-bis (chlorophenyl)-1-chloroethylene (DDMU), and 4-chlorobenzoic acid (4-CBA) were identified by GC-Mass and FTIR. 4-CBA was found to be the stable product of DDT degradation. Metabolites preceding 4-CBA were not toxic to strain as reveled through luxuriant growth in presence of varying concentrations of exogenous DDD and DDE. However, 4-CBA was observed to inhibit the growth of bacterium. The DDT degrading efficiency of S. marcescens NCIM 2919 hence could be used in combination with 4-CBA utilizing strains either as binary culture or consortia for mineralization of DDT. Application of S. marcescens NCIM 2919 to DDT contaminated soil, showed 74.7% reduction of initial 12.0 mg kg-1 of DDT after 18-days of treatment.

Efficacy of Acinetobacter sp. B9 for simultaneous removal of phenol and hexavalent chromium from co-contaminated system.

The present study shows the feasibility of a newly isolated strain Acinetobacter sp. B9 for concurrent removal of phenol and Cr (VI) from wastewater. The experiments were conducted in a batch reactor under aerobic conditions. Initially, when mineral salt solution was used as the culture medium, the strain was found to utilize phenol as sole carbon and energy source while no Cr (VI) removal was observed. However, the addition of glucose as co-carbon source resulted in the removal of both toxicants. This co-removal efficiency of the strain was further improved with nutrient-rich media (NB). Optimum co-removal was determined at 188 mg L(-1) of phenol and 3.5 mg L(-1) of Cr (VI) concentrations at pH 7.0. Strain B9 followed the orthometabolic pathway for phenol degradation. Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR) studies showed sorption of chromium as one of the major mechanisms for Cr (VI) removal by B9 cells. Acinetobacter sp. B9 was later on checked for bioremediation of real tannery wastewater. After 96 h of batch treatment of tannery effluent containing an initial 47 mg L(-1) phenol and 16 mg L(-1) Cr (VI), complete removal of phenol and 87 % reduction of Cr (VI) were attained, showing high efficiency of the bacterial strain for potential application in industrial pollution control.

Deciphering the seasonal dynamics of microplastic morphotypes and associated co-contaminants along the northwest coast of India.

In the present study, the northwest coast of India, bordering the Arabian Sea, was selected to evaluate the microplastic (MP) abundance. This is the first study to emphasize the effects of different seasons on MP distribution. The collected MPs were dried, segregated, and evaluated based on their morphotype, size, color, and polymer type. A total of 1756.6, 7326.6, and 202 particles/kg of sand were estimated in the pre-monsoon, monsoon and post-monsoon seasons, respectively, with a dominance of polypropylene (PP) type of plastic in the pre-monsoon and high-density polyethylene (HDPE) in monsoon and post-monsoon seasons. HDPE and PP collected MPs during the monsoon season were further characterized for associated contaminants. Metal absorbance was detected using SEM-EDX mapping and ICP-MS. The presence of organic compounds (OCs) was analyzed using GC-MS. MPs exhibit distinct associations with metals, among which the HDPE pellet morphotype exhibits a higher range of metal adsorption. Total 61 different OCs were associated with MPs. The HDPE pellets contained the highest amounts of hydrophobic organic compounds. PP pellets were found to contain triglycerides, fatty aldehydes, and alkaloids, along with HOCs. Among morphotypes, pellet forms of MPs were found to adsorb more contaminants. These co-contaminants infiltrate the study area through sewage runoff and shoreline debris deposition, subsequently interacting with MPs. Furthermore, the MP diversity was studied by employing the MP diversity integrated index, which suggests that most of the MP diversity was observed in the pre-monsoon period. The pollution load index employed an MP risk assessment, which presented a low degree of MP contamination. In contrast, the polymer hazard index was calculated as 21650.3 in post-monsoon, placing the area under the extreme danger category. It is evident from the data that the types of MP is more important than their number. Thus, MP morphotypes have importance in the adsorption of co-contaminants.

Halo-alkaliphilic microbes as an effective tool for heavy metal pollution abatement and resource recovery: challenges and future prospects.

The current study presents an overview of heavy metals bioremediation from halo-alkaline conditions by using extremophilic microorganisms. Heavy metal remediation from the extreme environment with high pH and elevated salt concentration is a challenge as mesophilic microorganisms are unable to thrive under these polyextremophilic conditions. Thus, for effective bioremediation of extreme systems, specialized microbes (extremophiles) are projected as potential bioremediating agents, that not only thrive under such extreme conditions but are also capable of remediating heavy metals from these environments. The physiological versatility of extremophiles especially halophiles and alkaliphiles and their enzymes (extremozymes) could conveniently be harnessed to remediate and detoxify heavy metals from the high alkaline saline environment. Bibliometric analysis has shown that research in this direction has found pace in recent years and thus this review is a timely attempt to highlight the importance of halo-alkaliphiles for effective contaminant removal in extreme conditions. Also, this review systematically presents insights on adaptive measures utilized by extremophiles to cope with harsh environments and outlines the role of extremophilic microbes in industrial wastewater treatment and recovery of metals from waste with relevant examples. Further, the major challenges and way forward for the effective applicability of halo-alkaliphilic microbes in heavy metals bioremediation from extremophilic conditions are also highlighted.

Biodegradation of terephthalic acid using Rhodococcus erythropolis MTCC 3951: Insights into the degradation process, applications in wastewater treatment and polyhydroxyalkanoate production.

Terephthalic acid (TPA) is an endocrine disruptor widely used as a plasticizer and as a monomer in the manufacturing of PET bottles. However, because of various harmful effects on humans and the environment, it is now recognized as a priority pollutant whose environmental level needs to be controlled. In the present work, the TPA biodegradation efficacy of the bacterium Rhodococcus erythropolis (MTCC 3951) was studied in mineral salt media with TPA as the sole carbon and energy source. R. erythropolis was observed to degrade 5 mM and 120 mM TPA within 10 h and 84 h of incubation, respectively. The degradation efficiency was further optimized by varying the culture conditions, and the following optimum conditions were obtained: inoculum size- 5% (v/v), temperature- 30 °C, agitation speed- 200 rpm, and pH- 8.0. The bacterium was found to use an ortho-cleavage pathway for TPA degradation determined based on enzymatic and GC-MS studies. Moreover, during the degradation of TPA, the bacterium was observed to produce polyhydroxyalkanoate (PHA)-a biopolymer. Biodegradation of 120 mM TPA resulted in an accumulation of PHA. The PHA granules were visualized using fluorescence and transmission electron microscopy and were later characterized using FTIR spectroscopy. Furthermore, the robustness of the bacterium was demonstrated by its ability to degrade TPA in real industrial wastewater. Overall, R. erythropolis (MTCC 3951) hold the potential for controlling TPA pollution in the environment and vis-à-vis the production of PHA biopolymer.

Production and characterization of Komagataeibacter xylinus SGP8 nanocellulose and its calcite based composite for removal of Cd ions.

In the present study, fermentative production of bacterial nanocellulose (BNC) by using Komagataeibacter xylinus strain SGP8 and characterization of nanocellulose is presented. The bacterium was able to produce 1.82 g L-1 of cellulose in the form of pellicle in standard Hestrin-Schramn (HS) medium. The morpho-structural characterization of the BNC using scanning electron microscopy (SEM) and X-ray diffraction (XRD) studies, respectively revealed nanofibrillar structure and high crystallinity index (~86%). The thermogravimetric analysis (TGA) showed the stability of BNC up to 280 °C, further rise in temperature to 350 °C results in depolymerization of the sample. In order to show the applicability of produced BNC, it was modified first using calcite (CaCO3) and thereafter characterized using SEM, XRD, FTIR, and TGA studies. The BNC-CaCO3 composites as a sorbent resulted in >99% removal of initial 10 mg L-1 of Cd (II) at pH 5, 7 and 9 after 12 h of treatment. Moreover, the composite was also found to be competent in removing high concentrations of Cd (25 and 50 mg L-1) from the solution (69-70%). Overall, the above results suggest that cellulose produced by K. xylinus strain SGP8 showed excellent material properties, and modified BNC (BNC-CaCO3 composite) could effectively be used for remediation of toxic levels of Cd from the contaminated system.

Ecological and toxicological manifestations of microplastics: current scenario, research gaps, and possible alleviation measures.

Microplastics (MPs) and associated contaminants have become a major environmental concern. From available literature, their ubiquitous presence is now well established. However, the kind and level of toxicological impacts these MPs accomplish on various life forms are not well understood. Nevertheless, the environmental toxicity of MP is now being revealed gradually with supporting studies involving groups of lower organisms. Additionally, the presence of microplastics also disturbs the functions of ecosystem through affecting the vulnerable life forms, thus ecological manifestations of MPs also need to be analyzed. The present review encompasses an overview of toxicological effects mediated by various types of MPs present in the environment; it covers the types of toxicity they may cause and other effects on humans and other species. In this review, aquatic systems are used as primary models to describe various eco-toxicological effects of MPs. Various research gaps as well as methods to alleviate the level of MPs, and future strategies are also comprehensively highlighted in the review.

Enzymatic Remediation of Polyethylene Terephthalate (PET)-Based Polymers for Effective Management of Plastic Wastes: An Overview.

Globally, plastic-based pollution is now recognized as one of the serious threats to the environment. Among different plastics, polyethylene terephthalate (PET) occupies a pivotal place, its excess presence as a waste is a major environmental concern. Mechanical, thermal, and chemical-based treatments are generally used to manage PET pollution. However, these methods are usually expensive or generate secondary pollutants. Hence, there is a need for a cost-effective and environment-friendly method for efficient management of PET-based plastic wastes. Considering this, enzymatic treatment or recycling is one of the important methods to curb PET pollution. In this regard, PET hydrolases have been explored for the treatment of PET wastes. These enzymes act on PET and end its breakdown into monomeric units and subsequently results in loss of weight. However, various factors, specifically PET crystallinity, temperature, and pH, are known to affect this enzymatic process. For effective hydrolysis of PET, high temperature is required, which facilitates easy accessibility of substrate (PET) to enzymes. However, to function at this high temperature, there is a requirement of thermostable enzymes. The thermostability could be enhanced using glycosylation, immobilization, and enzyme engineering. Furthermore, the use of surfactants, additives such as Ca2+, Mg2+, and hydrophobins (cysteine-rich proteins), has also been reported to enhance the enzymatic PET hydrolysis through facilitating easy accessibility of PET polymers. The present review encompasses a brief overview of the use of enzymes toward the management of PET wastes. Various methods affecting the treatment process and different constraints arising thereof are also systematically highlighted in the review.

Evaluation of Acinetobacter sp. B9 for Cr (VI) resistance and detoxification with potential application in bioremediation of heavy-metals-rich industrial wastewater.

Present work demonstrates Cr (VI) detoxification and resistance mechanism of a newly isolated strain (B9) of Acinetobacter sp. Bioremediation potential of the strain B9 is shown by simultaneous removal of major heavy metals including chromium from heavy-metals-rich metal finishing industrial wastewater. Strain B9 tolerate up to 350 mg L(-1) of Cr (VI) and also shows level of tolerance to Ni (II), Zn (II), Pb (II), and Cd (II). The strain was capable of reducing 67 % of initial 7.0 mg L(-1) of Cr (VI) within 24 h of incubation, while in presence of Cu ions 100 % removal of initial 7.0 and 10 mg L(-1) of Cr (VI) was observed with in 24 h. pH in the range of 6.0-8.0 and inoculum size of 2 % (v/v) were determined to be optimum for dichromate reduction. Fourier transform infrared spectroscopy and transmission electron microscopy studies suggested absorption or intracellular accumulation and that might be one of the major mechanisms behind the chromium resistance by strain B9. Scanning electron microscopy showed morphological changes in the strain due to chromium stress. Relevance of the strain for treatment of heavy-metals-rich industrial wastewater resulted in 93.7, 55.4, and 68.94 % removal of initial 30 mg L(-1) Cr (VI), 246 mg L(-1) total Cr, and 51 mg L(-1) Ni, respectively, after 144 h of treatment in a batch mode.

Effectiveness of sal deoiled seed cake as an inducer for protease production from Aeromonas sp. S1 for its application in kitchen wastewater treatment.

The present study is an attempt to demonstrate the feasibility of sal (Shorea robusta) deoiled cake--a forest-based industrial by-product--as a cheaper media supplement for augmented protease production from Aeromonas sp. S1 and application of protease in the treatment of kitchen wastewater. Under optimized conditions, protease production could successfully be enhanced to 5.13-fold (527.5 U mL(-1)) on using sal deoiled seed cake extract (SDOCE), as medium additive, compared to an initial production of 102.7 U mL(-1) in its absence. The culture parameters for optimum production of protease were determined to be incubation time (48 h), pH (7.0), SDOCE concentration (3 % (v/v)), inoculum size (0.3-0.6 % (v/v)), and agitation rate (100 rpm). The enzyme was found to have an optimum pH and temperature of 8.0 and 60 °C, respectively. The protease preparation was tested for treatment of organic-laden kitchen wastewater. After 96 h of wastewater treatment under static condition, enzyme preparation was able to reduce 74 % biological oxygen demand, 37 % total suspended solids, and 41 % oil and grease. The higher and improved level of protease obtained using sal deoiled seed cake-based media hence offers a new approach for value addition to this underutilized biomass through industrial enzyme production. The protease produced using this biomass could also be used as pretreatment tool for remediation of organic-rich food wastewater.

Novel application of Mahua (Madhuca sp.) flowers for augmented protease production from Aeromonas sp. S1.

The present study explored the utilization of Mahua (Madhuca sp.) flowers, a major non-timber forest product (NTFP) of India, as a low-cost, natural substrate for protease production under submerged fermentation. Bacterial strain Aeromonas sp. Si1, previously reported by us, was used as the protease producer. Using Mahua flower extract (MFE) as the medium additive, the protease production could successfully be enhanced by 5.6-fold (564.5 UmL-1) after 24 h of fermentation under optimized conditions compared with initial production of 99.9 UmL' in the absence of MFE. The cultural parameters for optimum production of protease were determined to be: incubation time-24 h; pH-7.0; MFE concentration-5% (v/v); inoculum size-0.3% (v/v) and agitation rate-200 rpm. The results obtained demonstrate the potential of cheaper and abundantly available Mahua flowers for induction of proteases, and thus offer a new approach for value addition to this biomass through industrial enzyme production.