Bio-mitigation of organic pollutants using horseradish peroxidase as a promising biocatalytic platform for environmental sustainability.

A wide array of environmental pollutants is often generated and released into the ecosystem from industrial and human activities. Antibiotics, phenolic compounds, hydroquinone, industrial dyes, and Endocrine-Disrupting Chemicals (EDCs) are prevalent pollutants in water matrices. To promote environmental sustainability and minimize the impact of these pollutants, it is essential to eliminate such contaminants. Although there are multiple methods for pollutants removal, many of them are inefficient and environmentally unfriendly. Horseradish peroxidase (HRP) has been widely explored for its ability to oxidize the aforementioned pollutants, both alone and in combination with other peroxidases, and in an immobilized way. Numerous positive attributes make HRP an excellent biocatalyst in the biodegradation of diverse environmentally hazardous pollutants. In the present review, we underlined the major advancements in the HRP for environmental research. Numerous immobilization and combinational studies have been reviewed and summarized to comprehend the degradability, fate, and biotransformation of pollutants. In addition, a possible deployment of emerging computational methodologies for improved catalysis has been highlighted, along with future outlook and concluding remarks.

Biodegradation of environmental pollutants using catalase-based biocatalytic systems.

The synergistic combination of biocatalysts and nanomaterials provides a new interface of a robust biocatalytic system that can effectively remediate environmental pollutants. Enzymes, such as catalase-based constructs, impart the desired candidature for catalytic transformation processes and are potential alternatives to replace conventional remediation strategies that have become laborious and somewhat inefficient. Furthermore, the controlled or uncontrolled discharge of various emerging pollutants (EPs) into water bodies is equally proportional to the fast-growing population and extensive urbanization. EPs affect the entire living being and continuously deteriorate the environmental system, directly or indirectly. The occurrence of EPs (even released after partial treatments, but still in bioactive forms) disturbs ecological integrity. Due to the ineffectiveness of in-practice traditional remediation processes, new and robust treatment measures as effective and sustainable remediation have become a meaningful goal. In this context, special attention has been shifted to engineering an enzyme (catalase)-based biodegradation system with immense prospects in environmental cleanup. The unique synergistic combination of nanomaterials (having multifunctional attributes) with enzymes of interest makes them a state-of-the-art interface that can further ameliorate bio-catalysis and biodegradation performance. This review covers current research and scientific advancement in developing and deploying catalase-based biocatalytic systems to mitigate several EPs from the environment matrices. The biocatalytic features of catalase, along with the mechanistic insight into H2O2 neutralization, several nano-based materials loaded with catalase, including nanoparticles (NPs), carbon nanotubes (CNTs), metal-organic frameworks (MOFs), polymeric-based composites, oxime-functionalized cryo-gel disks, electro-spun nanofibrous membranes, and other hybrid materials have also been discussed with suitable examples.

Bioremediation of various aromatic and emerging pollutants by Bacillus cereus sp. isolated from petroleum sludge.

The accumulation of toxic chemical constituents in sludge and wastewater has fuelled an interest in investigating efficient and eco-friendly wastewater remediation approaches. In this study, a set of bacterial samples were isolated from petroleum sludge and tested for their ability to degrade different aromatic pollutants, including azo dyes and emerging pollutants. Although exhibiting differential specificity, all bacterial isolates were able to degrade different classes of aromatic dyes efficiently. Ribosomal 16S rRNA sequencing of the 12 bacterial isolates showed that they belonged to two different bacterial genera: Bacillus cereus and Pseudomonas guariconensis. Of these 12 strains, MA1 (B. cereus) was the most promising and was chosen for further optimization and biochemical studies. The optimum culture and remediation conditions for MA1 was found to be at pH 7, with 100 ppm dye concentration, and under aerobic condition. In addition to efficiently degrading various aromatic dyes (e.g. Congo Red, Reactive Black 5, PBS, and Toluidine Blue), MA1 was also found to be capable of degrading various emerging pollutants (e.g. prometryn, fluometuron and sulfamethoxazole). Preliminary transcriptome analysis shows that MA1 grown on media containing a mixture of aromatic dyes appears to differentially express a number of genes. Data shown here strongly suggests that petroleum sludge is a rich reservoir of bacteria with powerful remediation abilities.

Biological effects of culture medium on Tetraselmis chuii and Dunaliella tertiolecta: Implications for emerging pollutants degradation.

In this study, laboratory-scale cultivation of T. chuii and D. tertiolecta was conducted using Conway, F/2, and TMRL media to evaluate their biochemical composition and economic costs. The highest cell density (30.36 × 106 cells/mL) and dry weight (0.65 g/L) for T. chuii were achieved with Conway medium. This medium also produced biomass with maximum lipid content (25.65%), proteins (27.84%), and total carbohydrates (8.45%) compared with F/2 and TMRL media. D. tertiolecta reached a maximum cell density of 17.50 × 106 cells/mL in F/2 medium, which was notably lower than that of T. chuii. Furthermore, the media cost varied from US$0.23 to US$0.74 for each 1 L of media, primarily due to the addition of Na3PO4, KNO3, and cyanocobalamin. Thus, biomass production rates varied between US$38.81 and US$128.80 per kg on a dry weight basis. These findings comprehensively compare laboratory conditions and the costs associated with biomass production in different media. Additionally, this study explored the potential of T. chuii and D. tertiolecta strains, as well as their consortia with bacteria, for the degradation of various emerging pollutants (EPs), including caffeine, salicylic acid, DEET, imidacloprid, MBT, cimetidine, venlafaxine, methylparaben, thiabendazole, and paracetamol. Both microalgal strains demonstrated effective degradation of EPs, with enhanced degradation observed in microalgae-bacterial consortia. These results suggest that the symbiotic relationship between microalgae and bacteria can be harnessed for the bioremediation of EPs, thereby offering valuable insights into the environmental applications of microalgal cultivation.

Biocatalytic degradation of reactive blue 221 and direct blue 297 dyes by horseradish peroxidase immobilized on iron oxide nanoparticles with improved kinetic and thermodynamic characteristics.

In present study, we describe the biodegradation of direct blue (DB) 297 and reactive blue (RB) 221 by immobilizing horseradish peroxidase (HRP) isolated from fresh leaves of Moringa Oliefera on iron oxide nanoparticles. Iron oxide nanoparticles were synthesized by co-precipitation method and showed a maximum immobilization efficiency of 87%. The surface topography of iron oxide nanoparticles was envisaged by scanning electron microscopy (SEM), results showed that magnetic nanoparticles (MNPs) were in the form of aggregates having size of 1 µm. Furthermore, immobilization was confirmed via functional group identification performed by Fourier transformed infrared spectroscopy (FTIR). Immobilization phenomena displaced the optimum temperature from 35 °C to 50 °C moreover, pH optima were altered from 5.0 to 7.0. Vmax and Km for free and immobilized HRP, were 303 U/mg and 1.66 mM and 312 U/mg and 1.94 mM, respectively. Enzymatic thermodynamic measurements (ΔH*, ΔS*, Ea, ΔG*) were also evaluated for immobilized HRP and its free counterpart. Optimum degradation of reactive blue (RB) and direct blue (DB) 297 with free and immobilized HRP was observed at pH 5 and at temperature 40 °C respectively. The removal efficiency of DB 297 and RB 221 with free HRP was 75% and 86% while with immobilized HRP was 81% and 92% respectively. Furthermore, biodegradation of reactive blue (RB) 221 and direct blue (DB) 297 with immobilized and free biocatalyst was also investigated by Fourier transform infrared spectroscopy (FTIR) by identification of groups involved in dye degradation. FTIR results confirmed the 100% degradation of dyes. Immobilized HRP retained significant catalytic activity after five consecutive cycles of dye degradation. In conclusion, Fe3O4 nanoparticles are promising and environmentally friendly media for enzyme immobilization. Moreover, immobilized HRP showed more thermal stability, pH stability and higher dye degradation efficiency as compared to free HRP. Furthermore, the immobilized HRP, economically more convenient and easily removable from reaction media. Owing to its thermal stability, ease of separation from reaction media and reusability, the magnetically separatable immobilized HRP can be exploited successfully for treatment of dye contaminated textile effluents.

Magnetic metal-organic frameworks immobilized enzyme-based nano-biocatalytic systems for sustainable biotechnology.

Nanobiocatalysts, in which enzyme molecules are integrated into/onto multifunctional materials, such as metal-organic frameworks (MOFs), have been fascinating and appeared as a new interface of nanobiocatalysis with multi-oriented applications. Among various nano-support matrices, functionalized MOFs with magnetic attributes have gained supreme interest as versatile nano-biocatalytic systems for organic bio-transformations. From the design (fabrication) to deployment (application), magnetic MOFs have manifested notable efficacy in manipulating the enzyme microenvironment for robust biocatalysis and thus assure requisite applications in several areas of enzyme engineering at large and nano-biocatalytic transformations, in particular. Magnetic MOFs-linked enzyme-based nano-biocatalytic systems offer chemo-regio- and stereo-selectivities, specificities, and resistivities under fine-tuned enzyme microenvironments. Considering the current sustainable bioprocesses demands and green chemistry needs, we reviewed synthesis chemistry and application prospects of magnetic MOFs-immobilized enzyme-based nano-biocatalytic systems for exploitability in different industrial and biotechnological sectors. More specifically, following a thorough introductory background, the first half of the review discusses various approaches to effectively developed magnetic MOFs. The second half mainly focuses on MOFs-assisted biocatalytic transformation applications, including biodegradation of phenolic compounds, removal of endocrine disrupting compounds, dye decolorization, green biosynthesis of sweeteners, biodiesel production, detection of herbicides and screening of ligands and inhibitors.

Enzyme-immobilized hierarchically porous covalent organic framework biocomposite for catalytic degradation of broad-range emerging pollutants in water.

Efficient enzyme immobilization is crucial for the successful commercialization of large-scale enzymatic water treatment. However, issues such as lack of high enzyme loading coupled with enzyme leaching present challenges for the widespread adoption of immobilized enzyme systems. The present study describes the development and bioremediation application of an enzyme biocomposite employing a cationic macrocycle-based covalent organic framework (COF) with hierarchical porosity for the immobilization of horseradish peroxidase (HRP). The intrinsic hierarchical porous features of the azacalix[4]arene-based COF (ACA-COF) allowed for a maximum HRP loading capacity of 0.76 mg/mg COF with low enzyme leaching (<5.0 %). The biocomposite, HRP@ACA-COF, exhibited exceptional thermal stability (∼200 % higher relative activity than the free enzyme), and maintained ∼60 % enzyme activity after five cycles. LCMSMS analyses confirmed that the HRP@ACA-COF system was able to achieve > 99 % degradation of seven diverse types of emerging pollutants (2-mercaptobenzothiazole, paracetamol, caffeic acid, methylparaben, furosemide, sulfamethoxazole, and salicylic acid)in under an hour. The described enzyme-COF system offers promise for efficient wastewater bioremediation applications.

Efficient degradation of various emerging pollutants by wild type and evolved fungal DyP4 peroxidases.

The accumulation of emerging pollutants in the environment remains a major concern as evidenced by the increasing number of reports citing their potential risk on environment and health. Hence, removal strategies of such pollutants remain an active area of investigation. One way through which emerging pollutants can be eliminated from the environment is by enzyme-mediated bioremediation. Enzyme-based degradation can be further enhanced via advanced protein engineering approaches. In the present study a sensitive and robust bioanalytical liquid chromatography-tandem mass spectrometry (LCMSMS)-based approach was used to investigate the ability of a fungal dye decolorizing peroxidase 4 (DyP4) and two of its evolved variants-that were previously shown to be H2O2 tolerant-to degrade a panel of 15 different emerging pollutants. Additionally, the role of a redox mediator was examined in these enzymatic degradation reactions. Our results show that three emerging pollutants (2-mercaptobenzothiazole (MBT), paracetamol, and furosemide) were efficiently degraded by DyP4. Addition of the redox mediator had a synergistic effect as it enabled complete degradation of three more emerging pollutants (methyl paraben, sulfamethoxazole and salicylic acid) and dramatically reduced the time needed for the complete degradation of MBT, paracetamol, and furosemide. Further investigation was carried out using pure MBT to study its degradation by DyP4. Five potential transformation products were generated during the enzymatic degradation of MBT, which were previously reported to be produced during different bioremediation approaches. The current study provides the first instance of the application of fungal DyP4 peroxidases in bioremediation of emerging pollutants.

Immunomodulating Hydrogels as Stealth Platform for Drug Delivery Applications.

Non-targeted persistent immune activation or suppression by different drug delivery platforms can cause adverse and chronic physiological effects including cancer and arthritis. Therefore, non-toxic materials that do not trigger an immunogenic response during delivery are crucial for safe and effective in vivo treatment. Hydrogels are excellent candidates that can be engineered to control immune responses by modulating biomolecule release/adsorption, improving regeneration of lymphoid tissues, and enhancing function during antigen presentation. This review discusses the aspects of hydrogel-based systems used as drug delivery platforms for various diseases. A detailed investigation on different immunomodulation strategies for various delivery options and deliberate upon the outlook of such drug delivery platforms are conducted.

Expanding the Biocatalytic Scope of Enzyme-Loaded Polymeric Hydrogels.

In recent years, polymeric hydrogels have appeared promising matrices for enzyme immobilization to design, signify and expand bio-catalysis engineering. Therefore, the development and deployment of polymeric supports in the form of hydrogels and other robust geometries are continuously growing to green the twenty-first-century bio-catalysis. Furthermore, adequately fabricated polymeric hydrogel materials offer numerous advantages that shield pristine enzymes from denaturation under harsh reaction environments. For instance, cross-linking modulation of hydrogels, distinct rheological behavior, tunable surface entities along with elasticity and mesh size, larger surface-volume area, and hydrogels' mechanical cushioning attributes are of supreme interest makes them the ideal candidate for enzyme immobilization. Furthermore, suitable coordination of polymeric hydrogels with requisite enzyme fraction enables pronounced loading, elevated biocatalytic activity, and exceptional stability. Additionally, the unique catalytic harmony of enzyme-loaded polymeric hydrogels offers numerous applications, such as hydrogels as immobilization matrix, bio-catalysis, sensing, detection and monitoring, tissue engineering, wound healing, and drug delivery applications. In this review, we spotlight the applied perspective of enzyme-loaded polymeric hydrogels with recent and relevant examples. The work also signifies the combined use of multienzyme systems and the future directions that should be attempted in this field.

Efficient Degradation of 2-Mercaptobenzothiazole and Other Emerging Pollutants by Recombinant Bacterial Dye-Decolorizing Peroxidases.

In recent years, concerns are being raised about the potential harmful effects of emerging pollutants (EPs) on human and aquatic lives. Extensive research is being conducted on developing efficient remediation strategies to target this new class of toxic pollutants. Studies focused on biological (enzyme-based) methods have shown potential as greener and possibly more economical alternatives to other treatment approaches, such as chemical methods. The current study focused on the use of recombinantly produced novel bacterial peroxidases, namely dye-decolorizing peroxidases (DyPs), to study their effectiveness in degrading a number of diverse EPs. In this context, a sensitive bioanalytical Liquid chromatography-tandem mass spectrometry (LCMSMS)-based method was developed to simultaneously detect a mixture of 31 EPs and to examine their degradability by a panel of seven different recombinant bacterial DyPs (rDyPs). We show that up to 9 of the 31 tested EPs could be degraded by at least one of the DyPs tested. The results also indicated that not all rDyPs behaved similarly in their abilities to degrade EPs, as some rDyPs (such as SviDyP and CboDyP) showed a promising potential to degrade EPs while others (such as ScDyP) were almost ineffective. Additionally, the role of redox mediators for effective emerging pollutant degradation by rDyPs was also examined, which showed dramatic improvement in the DyP-mediated degradation of five different EPs. Detailed analysis of 2-mercaptobenzothiazole degradation by SviDyP showed that six distinct breakdown products were generated. The present study showed for the first time that recombinant bacterial DyPs can be used for wastewater remediation by degrading a range of different EPs.

Robust nanocarriers to engineer nanobiocatalysts for bioprocessing applications.

The synergistic integration of bio-catalysis engineering with nanostructured materials, as unique multifunctional carrier matrices, has emerged as a new interface of nanobiocatalysis (NBC). NBC is an emerging innovation that offers significant considerations to expand the designing and fabrication of robust catalysts at the nanoscale with improved catalytic characteristics for multipurpose bioprocessing applications. In addition, nanostructured materials with unique structural, physical, chemical, and functional entities have manifested significant contributions in mimicking the enzyme microenvironment. A fine-tuned enzyme microenvironment with an added-value of NBC offers chemo- regio- and stereo- selectivities and specificities. Furthermore, NBC is growing rapidly and will become a powerful norm in bio-catalysis with much controlled features, such as selectivity, specificity, stability, resistivity, induce activity, reaction efficacy, multi-usability, improved mass transfer efficiency, high catalytic turnover, optimal yield, ease in recovery, and cost-effectiveness. Considering the above critics and unique structural, physicochemical, and functional attributes, herein, we present and discuss advances in NBC and its bioprocessing applications in different fields. Briefly, this review is focused on four parts, i.e., (1) NBC as a drive towards applied nanobiocatalysts (as an introduction with opportunities), (2) promising nanocarriers to develop nanobiocatalysts, (3) applications in the fields of biotransformation, biofuel production, carbohydrate hydrolysis, bio-/nanosensing, detergent formulations, and extraction and purification of value-added compounds, and (4) current challenges, concluding remarks, and future trends.

Challenges and Recent Advances in Enzyme-Mediated Wastewater Remediation-A Review.

Different classes of artificial pollutants, collectively called emerging pollutants, are detected in various water bodies, including lakes, rivers, and seas. Multiple studies have shown the devastating effects these emerging pollutants can have on human and aquatic life. The main reason for these emerging pollutants in the aquatic environment is their incomplete removal in the existing wastewater treatment plants (WWTPs). Several additional treatments that could potentially supplement existing WWTPs to eliminate these pollutants include a range of physicochemical and biological methods. The use of enzymes, specifically, oxidoreductases, are increasingly being studied for their ability to degrade different classes of organic compounds. These enzymes have been immobilized on different supports to promote their adoption as a cost-effective and recyclable remediation approach. Unfortunately, some of these techniques have shown a negative effect on the enzyme, including denaturation and loss of catalytic activity. This review focuses on the major challenges facing researchers working on the immobilization of peroxidases and the recent progress that has been made in this area. It focuses on four major areas: (1) stability of enzymes upon immobilization, enzyme engineering, and evolution; (2) recyclability and reusability, including immobilization on membranes and solid supports; (3) cost associated with enzyme-based remediation; and (4) scaling-up and bioreactors.

Immobilized Soybean Peroxidase Hybrid Biocatalysts for Efficient Degradation of Various Emerging Pollutants.

In the present study, soybean peroxidase (SBP) was covalently immobilized onto two functionalized photocatalytic supports (TiO2 and ZnO) to create novel hybrid biocatalysts (TiO2-SBP and ZnO-SBP). Immobilization caused a slight shift in the pH optima of SBP activity (pH 5.0 to 4.0), whereas the free and TiO2-immobilized SBP showed similar thermal stability profiles. The newly developed hybrid biocatalysts were used for the degradation of 21 emerging pollutants in the presence and absence of 1-hydroxy benzotriazole (HOBT) as a redox mediator. Notably, all the tested pollutants were not equally degraded by the SBP treatment and some of the tested pollutants were either partially degraded or appeared to be recalcitrant to enzymatic degradation. The presence of HOBT enhanced the degradation of the pollutants, while it also inhibited the degradation of some contaminants. Interestingly, TiO2 and ZnO-immobilized SBP displayed better degradation efficiency of a few emerging pollutants than the free enzyme. Furthermore, a combined enzyme-chemical oxidation remediation strategy was employed to degrade two recalcitrant pollutants, which suggest a novel application of these novel hybrid peroxidase-photocatalysts. Lastly, the reusability profile indicated that the TiO2-SBP hybrid biocatalyst retained up to 95% degradation efficiency of a model pollutant (2-mercaptobenzothiazole) after four consecutive degradation cycles.

Enzyme-Loaded Flower-Shaped Nanomaterials: A Versatile Platform with Biosensing, Biocatalytic, and Environmental Promise.

As a result of their unique structural and multifunctional characteristics, organic-inorganic hybrid nanoflowers (hNFs), a newly developed class of flower-like, well-structured and well-oriented materials has gained significant attention. The structural attributes along with the surface-engineered functional entities of hNFs, e.g., their size, shape, surface orientation, structural integrity, stability under reactive environments, enzyme stabilizing capability, and organic-inorganic ratio, all significantly contribute to and determine their applications. Although hNFs are still in their infancy and in the early stage of robust development, the recent hike in biotechnology at large and nanotechnology in particular is making hNFs a versatile platform for constructing enzyme-loaded/immobilized structures for different applications. For instance, detection- and sensing-based applications, environmental- and sustainability-based applications, and biocatalytic and biotransformation applications are of supreme interest. Considering the above points, herein we reviewed current advances in multifunctional hNFs, with particular emphasis on (1) critical factors, (2) different metal/non-metal-based synthesizing processes (i.e., (i) copper-based hNFs, (ii) calcium-based hNFs, (iii) manganese-based hNFs, (iv) zinc-based hNFs, (v) cobalt-based hNFs, (vi) iron-based hNFs, (vii) multi-metal-based hNFs, and (viii) non-metal-based hNFs), and (3) their applications. Moreover, the interfacial mechanism involved in hNF development is also discussed considering the following three critical points: (1) the combination of metal ions and organic matter, (2) petal formation, and (3) the generation of hNFs. In summary, the literature given herein could be used to engineer hNFs for multipurpose applications in the biosensing, biocatalysis, and other environmental sectors.

Never let a crisis go to waste: Repurposing independent research projects to enhance students' critical thinking skills.

The recent lockdown of laboratories in the COVID-19 era has negatively impacted many lab-based courses for undergraduate students; however, research project classes have been hardest hit. Unlike the lab-based lecture courses, where many of the lab instructors have resorted to virtual experiments and videos to substitute for actual lab experiments, senior year independent research courses, which are supposed to involve real lab-based research, are struggling to find an appropriate solution. A possible solution to cases where the coronavirus pandemic lockdown has led to no or little student-generated lab data are to shift the focus of these research project courses such that they are taught as mentor-guided critical thinking exercises, using the primary literature. As an example, the lack of complete understanding for the molecular basis of protein thermostability can be used to promote such higher-level thinking skills.

An innovative bioanalytical research project course to train undergraduate students on liquid chromatography-mass spectrometry.

Active-learning strategies such as undergraduate research courses and course-based undergraduate research experiences (CUREs), which engage students in the practical experiments, have been reported as efficient ways for effective teaching and training of graduating students. Recently, many practical examples have been published in various fields to develop critical skills needed by undergraduate students for graduate school or the workforce; however, very few examples have been published for bioanalytical topics, specifically, Liquid Chromatography-Mass Spectrometry (LC-MS-MS). Herein, we describe an innovative undergraduate research course that was used to train well-prepared graduating seniors on LC-MS-MS and in developing a sensitive method for detecting vitamin B9 (folic acid) levels in different milk samples. During this semester-long exercise, the students were exposed to literature review, basic UV-Vis spectroscopy, and HPLC and LC-MS-MS techniques. Additionally, as part of this laboratory-based research course, they researched published approaches on analyzing folic acid in food samples and devised a method to extract and quantify folic acid levels in different brands of milk using LC-MS-MS instruments. Feedback from students and faculty members was very positive as the students felt that this opportunity helped them to become more comfortable using HPLC and LC-MS-MS instruments and felt empowered to answer real-life analytical problems. This laboratory-based undergraduate research course can also be modified and used as an advanced bioanalytical laboratory exercise in biochemistry or analytical chemistry courses. © 2019 International Union of Biochemistry and Molecular Biology, 47(3):228-238, 2019.

Biocatalytic degradation/redefining "removal" fate of pharmaceutically active compounds and antibiotics in the aquatic environment.

Recently, the increasing concentration and persistent appearance of antibiotics traces in the water streams are considered an issue of high concern. In this context, an array of antibiotics has been categorized as pollutants of emerging concern due to their complex and highly stable bioactivity, indiscriminate usage with ultimate release into water bodies, and notable persistence in environmental matrices. Moreover, antibiotics traces containing household sewage/drain waste and pharmaceutical wastewater effluents contain a range of bioactive/toxic organic compounds, inorganic salts, pharmaceutically-active ingredients, or a mixture of all, which possesses negative influences ranging from ecological pollution to damage biodiversity. Moreover, their uncontrolled and undesirable bioaccumulation also poses a potential threat to target and non-target organisms in the environment. Aiming to tackle this issue effectively, various detection, quantification, degradation, and redefining "removal" processes have been proposed and investigated based on physical, chemical, and biological strategies. Though both useful and side effects of antibiotics on humans and animals are usually investigated thoroughly following safety and toxicity measures, however, their direct or indirect environmental impacts are not well reviewed yet. Owing to the considerable research gap, the environmental perfectives of antibiotics traces and their effects on target and non-target populations have now become the topic of research interest. Based on literature evidence, over the past several years, numerous individual studies have been performed and published covering various aspects of antibiotics. However, a comprehensive compilation on enzyme-based degradation of antibiotics is still lacking and requires careful consideration. Hence, this review summarizes up-to-date literature on enzymes as biocatalytic systems, explicitly, free as well as immobilized forms and their effective exploitation for the degradation of various antibiotics traces and other pharmaceutically-active compounds present in the water bodies. It is further envisioned that the enzyme-based strategies, for antibiotics degradation or removal, discussed herein, will help readers for a better understanding of antibiotics persistence in the environment along with the associated risks and removal measures. In summary, the current research thrust presented in this review will additionally evoke researcher to engineer robust and sustainable processes to effectively remediate antibiotics-contaminated environmental matrices.

Expression, purification, and characterization of biologically active full-length Mason-Pfizer monkey virus (MPMV) Pr78Gag.

MPMV precursor polypeptide Pr78Gag orchestrates assembly and packaging of genomic RNA (gRNA) into virus particles. Therefore, we have expressed recombinant full-length Pr78Gag either with or without His6-tag in bacterial as well as eukaryotic cultures and purified the recombinant protein from soluble fractions of the bacterial cultures. The recombinant Pr78Gag protein has the intrinsic ability to assemble in vitro to form virus like particles (VLPs). Consistent with this observation, the recombinant protein could form VLPs in both prokaryotes and eukaryotes. VLPs formed in eukaryotic cells by recombinant Pr78Gag with or without His6-tag can encapsidate MPMV transfer vector RNA, suggesting that the inclusion of the His6-tag to the full-length Pr78Gag did not interfere with its expression or biological function. This study demonstrates the expression and purification of a biologically active, recombinant Pr78Gag, which should pave the way to study RNA-protein interactions involved in the MPMV gRNA packaging process.

Differential Degradation and Detoxification of an Aromatic Pollutant by Two Different Peroxidases.

Enzymatic degradation of organic pollutants is a new and promising remediation approach. Peroxidases are one of the most commonly used classes of enzymes to degrade organic pollutants. However, it is generally assumed that all peroxidases behave similarly and produce similar degradation products. In this study, we conducted detailed studies of the degradation of a model aromatic pollutant, Sulforhodamine B dye (SRB dye), using two peroxidases-soybean peroxidase (SBP) and chloroperoxidase (CPO). Our results show that these two related enzymes had different optimum conditions (pH, temperature, H2O2 concentration, etc.) for efficiently degrading SRB dye. High-performance liquid chromatography and liquid chromatography -mass spectrometry analyses confirmed that both SBP and CPO transformed the SRB dye into low molecular weight intermediates. While most of the intermediates produced by the two enzymes were the same, the CPO treatment produced at least one different intermediate. Furthermore, toxicological evaluation using lettuce (Lactuca sativa) seeds demonstrated that the SBP-based treatment was able to eliminate the phytotoxicity of SRB dye, but the CPO-based treatment did not. Our results show, for the first time, that while both of these related enzymes can be used to efficiently degrade organic pollutants, they have different optimum reaction conditions and may not be equally efficient in detoxification of organic pollutants.