Tyrosinase and laccase-producing Bacillus aryabhattai TFG5 and its role in the polymerization of phenols.

BACKGROUND: Tyrosinases and laccases are oxidoreductase enzymes that are used widely in the food, feed, textile, and biofuel industries. The rapidly growing industrial demand for bacterial oxido-reductases has encouraged research on this enzyme worldwide. These enzymes also play a key role in the formation of humic substances (HS) that are involved in controlling the biogeochemical carbon cycle, providing nutrients and bio-stimulants for plant growth, and interacting with inorganic and organic pollutants besides increasing carbon sequestration and mitigating greenhouse gas emission in the environment. The present study aimed to screen and characterize extracellular tyrosinase and laccase-producing soil bacteria that could be utilized in the polymerization of phenols. RESULTS: Twenty isolates from different soil samples collected from forest ecosystems were characterized through ARDRA using restriction digestion with AluI, HpaII, and HaeIII restriction enzymes. The results of Hierarchical Cluster Analysis (HCA) revealed a 60 % similarity coefficient among 13 out of 20 isolates, of which, the isolate TFG5 exhibited only 10 % similarity when compared to all the other isolates. The isolate TFG5 exhibited both tyrosinase (1.34 U.mL- 1) and laccase (2.01 U.mL- 1) activity and was identified as Bacillus aryabhattai. The increased polymerization activity was observed when B. aryabhattai TFG5 was treated with phenols. The monomers such as catechol, p-Hydroxy benzoic acid, ferulic acid, and salicylic acid were polymerized efficiently, as evidenced by their FT-IR spectra depicting increased functional groups compared to the standard mushroom tyrosinase. CONCLUSIONS: The polymerization ability of B. aryabhattai TFG5 could be applied to phenol-rich wastewater treatment for efficient precipitation of phenols. Furthermore, tyrosinases can be used for enhancing the synthesis of HS in soil.

Bacillus aryabhattai TFG5-mediated synthesis of humic substances from coir pith wastes.

BACKGROUND: Humic substances (HS) form the largest proportion among all the constituents of soil organic matter and are a key component of the terrestrial ecosystem. HS plays a multifunctional role in the environment by controlling the biogeochemical carbon cycle, providing nutrients and bio-stimulants for plant growth, and interacting with inorganic and organic pollutants. The rate of formation of HS in soils determines its productivity and carbon sequestration capacity. Enhancement of HS synthesis in the soil through the microbial route not only increases CO2 sequestration but also mitigates the greenhouse gas emissions in the environment. RESULT: In this study, we attempted to understand the mechanism of formation and enhancement of HS from coir pith wastes using the tyrosinase produced by Bacillus aryabhattai TFG5. The bacterium TFG5 isolated from the termite garden produced the tyrosinase (1.34 U mL-1) and laccase (2.1 U mL-1) at 48 h and 60 h of fermentation, respectively. The extracellular tyrosinase from B. aryabhattai TFG5 was designated as TyrB. Homology modeling of TyrB revealed a structure with a predicted molecular mass of 35.23 kDa and two copper ions in the active center with its conserved residues required for the tyrosinase activity. TyrB efficiently transformed and polymerized standard phenols, such as p-cresol, p-hydroxyl benzoic acid, Levo DOPA, and 2,6 DMP, besides transforming free phenols in coir pith wash water (CWW). Additionally, UV-Vis and FT-IR spectra of the degradation products of the coir pith treated with TyrB revealed the formation of HS within 3 days of incubation. Furthermore, the E472/664 ratio of the degradation products revealed a higher degree of condensation of the aromatic carbons and the presence of more aliphatic structures in the HS. CONCLUSION: The results confirmed the influence of TyrB for the effective synthesis of HS from coir pith wastes. The results of the present study also confirm the recently accepted theory of humification proposed by the International Humic Substances Society.

Nano-ellagic acid: inhibitory actions on aldose reductase and α-glucosidase in secondary complications of diabetes, strengthened by in silico docking studies.

Increased blood sugar levels in prolonged diabetes lead to secondary complications such as retinopathy, neuropathy, and nephropathy, which gradually end in death. Synthesis of nano-phytomedicines from active phytoconstituents for novel emerging applications in the field of pharmaceuticals is of huge interest among researchers. In the present investigation, encapsulated ellagic acid (NEA) was synthesized at four different concentrations (0.2%, 0.3%, 0.4%, 0.5%) using ZnO nanoparticles as encapsulating agent. The surface morphology (fiber-like structures) of the nanoparticles were determined by scanning electron microscopy (SEM) and particle size (161-297 nm) and zeta potential (- 54.9-38.4 mV) were determined by dynamic light scattering technique. Further, the α-glucosidase and aldose reductase enzymes were significantly inhibited by the 0.4% of NEA compared to the other concentrations which strengthened our studies in overcoming the secondary complications of diabetes. The interaction analysis between ellagic acid and insulin receptor found Hit 1 among 10 executed ∆G score and energy of - 5.76, - 4.63 kcal/mol and formed polar bond with Arg 113 with - 1.175 Å distance. The residues Arg115, Lys116, Phe118, Ile115, Arg1131, Arg1155, Ile1157, Lys1165 and Phe1186 were found in ligand-protein interactions. ADME/T analysis of hit 1 was within the acceptable range without any toxic functional groups, providing a framework for developing novel therapeutics.

The use of slow releasing nanoparticle encapsulated Azadirachtin formulations for the management of Caryedon serratus O. (groundnut bruchid).

Nanobiotechnology is one of the emerging fields and its interventions in agriculture is been attracting the scientific community. Herein, the authors first to report on control of groundnut bruchid (Caryedon serratus O.) using nanoscale zinc oxide (ZnONPs) particles and nanoscale chitosan (CNPs) particles-based Azadirachtin formulations. ZnONPs and CNPs were prepared using sol-gel and ion tropic gelation techniques, respectively. Neem seed kernel extract (NSKE) 5% and Neem oil (3000 and 1000 ppm) were encapsulated using the prepared nanoscale materials and characterised using the techniques such as dynamic light scattering, high-resolution transmission electron microscopy. Spherical-shaped nanoparticles were formed after encapsulation with the required bio-materials (ZnONPs 33.1 nm; CNPs 78.8 nm; neem oil encapsulated (3000 ppm) ZnONPs 182.9 nm; NSKE encapsulated ZnONPs 84.9 nm) and observed that the particles are stable (52.3 mV for ZnONPs, -36.2 mV for CNPs, -43.0 mV for neem oil encapsulated (3000 ppm) ZnONPs and -39.4 mV for NSKE encapsulated ZnONPs). NSKE encapsulated CNPs were able to contain groundnut bruchid up to 180 days with 54.61% weight loss compared to other formulations tested. Thus biomaterial encapsulated nanoscale material formulations are proved to be effective in controlling stored grain pests to reduce huge economic losses.

Biosorption of lead, copper and cadmium using the extracellular polysaccharides (EPS) of Bacillus sp., from solar salterns.

Extracellular Polysaccharides (EPS) from both prokaryotes and eukaryotes have a great deal of research interest as they protect the producer from different stresses including antibiotics, ionic stress, desiccation and assist in bio-film formation, pathogenesis, adhesion, etc. In this study haloalkaliphilic Bacillus sp., known to cope with osmophilic stress, was selected and screened for EPS production. The EPS were isolated, partially purified and chemical characteristics were documented using liquid FT-IR followed by assessment of heavy metal biosorption (lead, copper and cadmium) using Atomic Absorption Spectroscopy (AAS). The EPS extracted from three isolates B. licheniformis NSPA5, B. cereus NSPA8 and B. subtilis NSPA13 showed maximum biosorption of Lead followed by Copper and Cadmium. Of the tested isolates, the EPS from isolate B. cereus NSPA8 showed maximum (90 %) biosorption of the lead.

Lantana camara leaf extract mediated silver nanoparticles: Antibacterial, green catalyst.

Silver nanoparticles (AgNPs) have been synthesized by Lantana camara leaf extract through simple green route and evaluated their antibacterial and catalytic activities. The leaf extract (LE) itself acts as both reducing and stabilizing agent at once for desired nanoparticle synthesis. The colorless reaction mixture turns to yellowish brown attesting the AgNPs formation and displayed UV-Vis absorption spectra. Structural analysis confirms the crystalline nature and formation of fcc structured metallic silver with majority (111) facets. Morphological studies elicit the formation of almost spherical shaped nanoparticles and as AgNO3 concentration is increased, there is an increment in the particle size. The FTIR analysis evidences the presence of various functional groups of biomolecules of LE is responsible for stabilization of AgNPs. Zeta potential measurement attests the higher stability of synthesized AgNPs. The synthesized AgNPs exhibited good antibacterial activity when tested against Escherichia coli, Pseudomonas spp., Bacillus spp. and Staphylococcus spp. using standard Kirby-Bauer disc diffusion assay. Furthermore, they showed good catalytic activity on the reduction of methylene blue by L. camara extract which is monitored and confirmed by the UV-Vis spectrophotometer.