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.

Non-magnetic and magnetically responsive support materials immobilized peroxidases for biocatalytic degradation of emerging dye pollutants-A review.

In recent years, the removal of hazardous pollutants from many industries has become a significant challenge for mankind as a growing number of contaminants, including a wide range of organic pollutants, synthetic dyes, and polycyclic aromatic hydrocarbons (PAHs), have inevitably led to an increased anthropogenic impact on the biosphere. Due to the complex aromatic structure, most synthetic dyes show resistance to degrade by the classical approaches, such as coagulation, flotation, adsorption, membrane process, and reverse osmosis. Enzyme-assisted biodegradation of pollutants offers an eco-friendlier and cost-effective alternative to remediate dyes, dyes-based effluents, other toxins, etc. Various plant and microbial oxidoreductase (Horseradish and manganese peroxidase) have recently received more attention for degrading and detoxifying a wide range of dyes either by opening the aromatic ring structure or by precipitation due to their high activity under milder conditions, high substrate specificity, and biodegradable nature. To enhance the efficiency, stability and recyclability, enzymes were immobilized on various support media such as sodium alginate, agarose, chitin/chitosan, polyvinyl alcohol, polyacrylamide, macroporous exchange resins, hydrophobic sol-gels, and nanoporous silica gel, including magnetically separatable media. Among various types of magnetic nanoparticles (MNPs), iron oxide magnetic nanoparticles, such as hematite, magnetite, and maghemite, have gained great attention due to their properties like small size, superparamagnetism, high surface area to volume ratio, and ease of separation for repeated cycles of uses. These carriers can be separated easily and rapidly from the reaction medium by an external magnetic field without being subjected to mechanical stress than centrifugation or filtration. Various methods have been employed for immobilizing oxidoreductase on different media, such as adsorption, covalent binding, entrapment, and encapsulation using different cross-linking agents. Compared to the free enzyme, insolubilized enzymes reduce production costs by enzyme reusability, tolerance to unfavorable environmental conditions, and high catalytic stability. Here, we review various immobilization methods and biocatalytic degradation of emerging dye pollutants, focusing on various non-magnetically and magnetically responsive supports to immobilize peroxidases. Conclusively, magnetically separatable peroxidases show more stability towards extreme temperature and pH conditions and can be used for repeated cycles than free and non-magnetically separatable peroxidase.

Enzyme-assisted bioremediation approach for synthetic dyes and polycyclic aromatic hydrocarbons degradation.

Environmental protection from emerging pollutants has become a significant challenge for mankind as an increasing number of contaminants, including synthetic dyes and polycyclic aromatic hydrocarbons (PAHs), represent a serious risk to ecological and environmental balance. Most synthetic dyes have complex aromatic structures and are resistant to degrade by classical approaches, such as physical and chemical processes, including adsorption, chemical coagulation, flocculation, ion exchange, membrane separation, froth flotation, and reverse osmosis. Enzymes-assisted catalytic transformation of pollutants has become a potential alternative to classical methods because of their ability to react with complex compounds, a quick degradation rate, and producing less harmful by-products. Plant peroxidases, and microbial laccase and lignin-degrading peroxidases (manganese and lignin peroxidase) have gained significant attention for treating aromatic waste due to their capability of oxidizing and detoxifying a wide range of recalcitrant xenobiotics, including PAHs and synthetic dyes. Peroxidases being efficient biocatalysts detoxify an array of toxic compounds by simple free-radical mechanism resulting in the formation of oxidized and depolymerized products of significantly reduced toxicity. Moreover, it is an ecofriendly and economically favorable approach towards the biodegradation of recalcitrant and toxic industrial waste. Among microbial and plant peroxidases, bacterial enzymes have broad substrate specificity and can transform a wide range of recalcitrant substrates. Ligninolytic enzymes oxidize the aromatic ring into quinones and acids by producing free hydroxyl radicals instead of dihydrodiols and mineralize aromatic hydrocarbon in combination with cytochrome P450, monooxygenases, and epoxide hydrolases. In the review, an attempt has been made to provide detailed knowledge about the availability of inexpensive peroxidases sources, their mechanism of action, and degradation potential. The present review summarizes the exploitation of peroxidases from plants, bacteria, and fungus (manganese peroxidase, lignin peroxidase, and laccases) for detoxification and degradation of textile dyes as well as PAHs. Conclusively, peroxidases have great potential to react with almost all classes of synthetic dyes and most PAHs due to broad substrate specificity and transformed them into less harmful metabolites.

Emergence of Haitian variant genotype and altered drug susceptibility in Vibrio cholerae O1 El Tor-associated cholera outbreaks in Solapur, India.

It is evident from previous cholera epidemics/outbreaks in India, Africa and America that isolates of the seventh pandemic Vibrio cholerae El Tor (7PET) with Haitian cholera toxin (HCT) genotype were associated with increased mortality. The present study highlights the emergence of 7PET-HCT isolates causing two cholera outbreaks in Walsang and Wagdari (Solapur, India) in 2016. Molecular analyses revealed that 7PET strains from earlier outbreaks (2010 and 2012) were progenitors of the current 7PET-HCT isolates. Isolates from the 2016 outbreaks carried qnrVC and floR genes and showed reduced susceptibility to tetracycline, ciprofloxacin and azithromycin, drugs recommended by the World Health Organization (WHO) for the treatment of cholera. Remarkably, protein profiling and mass spectrometry revealed disappearance of the outer membrane protein U (OmpU) porin in 7PET-HCT isolates from the second outbreak in 2016. Downregulation of ompU gene expression was also confirmed at the transcriptional level. Strains with downregulated OmpU showed reduced minimum inhibitory concentrations (MICs) for polymyxin B, which is a pore-forming antimicrobial agent. A multipronged approach is of utmost importance to prevent further spread of circulating 7PET-HCT strains. There is a pressing need for the formulation and implementation of international policies to closely monitor the effective use of antibiotics in order to prevent the further rise and spread of antimicrobial resistance.