Assessment of alkaline cholesterol oxidase purified from Rhodococcus sp. PKPD-CL for its halo tolerance, detergent and organic solvent stability.

The novel bacterium, Rhodococcus sp. PKPD-CL was isolated and identified from the 'Chilika Lake' located at Odisha state of India, which is a largest brackish water habitat in Asia. Rhodococcus sp. PKPD-CL produces extracellular halo tolerant, detergent and organic solvent stable alkaline cholesterol oxidase. It has apparent molecular weight of 60 kDa and was purified 59 fold by using 60% saturated ammonium sulfate fractionation, anion exchange followed by size exclusion chromatographic techniques with 37% recovery. It showed substrate specificity for 3ß-hydroxysteroids with Km of 1.1 × 10(-4)M for cholesterol. The pH, 8.0 and the temperature, 37 °C were required for its optimum activity. Enzyme is considerably stable at pH 6.0-8.5 and temperature up to 50 °C. At 4 and 30 °C it maintained its 100% activity up to 60 days. The isoelectric point of the enzyme was 9.5. It showed 80% residual activity with 20% NaCl (3.42 M) and 83% relative activity with 12% NaCl (2.05 M) concentration. The metal ions like Zn(2+), Cu(2+), Ag+, Fe(3+), Ba(2+) inhibited the enzyme activity >60% while Hg(2+) served a potent inhibitor whereas Mg(2+) found to be a good enhancer for it. The enzyme was stable in presence of chemical reagents (NaN3, EDTA), detergents (Tween-80, Tween-20, Triton X-100, sodium cholate) and various organic solvents (isopropanol, ethanol, benzene, chloroform, methanol, toluene, ethyl acetate, butanol and dimethylsulfoxide). Such a multi stress tolerant and versatile enzyme produced by Rhodococcus sp. PKPD-CL may serve as a good choice for industrial applications.

Insights into the antioxidant, anti-inflammatory and anti-microbial potential of Nigella sativa essential oil against oral pathogens.

Oral disorders can exert systemic ramifications beyond their localized effects on dental tissues, implicating a wide array of physiological conditions. The utilization of essential oils (EOs) for protection of oral health represents a longstanding practice. Consequently, in this investigation, essential oil derived from Nigella sativa seeds (NSEO) underwent isolation via the hydro-distillation process, followed by a comprehensive evaluation of its antioxidant, anti-inflammatory, anti-fungal, antibacterial activities, and cytocompatibility. The isolated NSEO manifested as a pale-yellow substance and was found to harbor a diverse spectrum of bioactive constituents, including steroids, triterpenoids, flavonoids, phenols, proteins, alkaloids, tannin, sesquiterpenoid hydrocarbons, monoterpenoid alcohol, and monoterpenoid ketone (thymoquinone). Notably, the total phenolic content (TPC) and total flavonoid content (TFC) of NSEO were quantified at 641.23 µg GAE/gm and 442.25 µg QE/g, respectively. Furthermore, NSEO exhibited concentration-dependent inhibition of protein denaturation, HRBC membrane stabilization, and hemolysis inhibition. Comparative analysis revealed that NSEO and chlorhexidine (CHX) 0.2% displayed substantial inhibition of hemolysis compared to aspirin. While NSEO and CHX 0.2% demonstrated analogous antibacterial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, NSEO showcased heightened efficacy against Lactobacillus acidophilus and Candida albicans. Additionally, NSEO exhibited pronounced effects against periodontal pathogens such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Prevotella intermedia. Importantly, no cytotoxicity was observed on human gingival fibroblast cell lines. These findings underscore the potential of NSEO as a potent antibacterial and antifungal agent in the management of oral microbial pathogens, thereby offering avenues for the development of innovative therapies targeting diverse oral inflammatory conditions. Nevertheless, further investigations are imperative to unlock its full therapeutic repertoire.

CogniFiber: Harnessing Biocompatible and Biodegradable 1D Collagen Nanofibers for Sustainable Nonvolatile Memory and Synaptic Learning Applications.

Here, resistive switching (RS) devices are fabricated using naturally abundant, nontoxic, biocompatible, and biodegradable biomaterials. For this purpose, 1D chitosan nanofibers (NFs), collagen NFs, and chitosan-collagen NFs are synthesized by using an electrospinning technique. Among different NFs, the collagen-NFs-based device shows promising RS characteristics. In particular, the optimized Ag/collagen NFs/fluorine-doped tin oxide RS device shows a voltage-tunable analog memory behavior and good nonvolatile memory properties. Moreover, it can also mimic various biological synaptic learning properties and can be used for pattern classification applications with the help of the spiking neural network. The time series analysis technique is employed to model and predict the switching variations of the RS device. Moreover, the collagen NFs have shown good cytotoxicity and anticancer properties, suggesting excellent biocompatibility as a switching layer. The biocompatibility of collagen NFs is explored with the help of NRK-52E (Normal Rat Kidney cell line) and MCF-7 (Michigan Cancer Foundation-7 cancer cell line). Additionally, the biodegradability of the device is evaluated through a physical transient test. This work provides a vital step toward developing a biocompatible and biodegradable switching material for sustainable nonvolatile memory and neuromorphic computing applications.

Pharmaceutical and nutraceutical potential of natural bioactive pigment: astaxanthin.

Astaxanthin (3,3'-dihydroxy-ß,ß-carotene-4,4'-dione) is an orange-red, lipophilic keto-carotenoid pigment. It is majorly found in marine ecosystems particularly in aquatic animals such as salmon, shrimp, trout, krill, crayfish, and so on. It is also synthesized in microalgae Heamatococcus pluvialis, Chlorococcum, Chlorella zofingiensis, red yeast Phaffia rhodozyma and bacterium Paracoccus carotinifaciens. Some aquatic and terrestrial creatures regarded as a primary and secondary sources of the astaxanthin producing and accumulating it through their metabolic pathways. Astaxanthin is the powerful antioxidant, nutritional supplement as well as promising therapeutic compound, observed to have activities against different ravaging diseases and disorders. Researchers have reported remarkable bioactivities of astaxanthin against major non-communicable chronic diseases such as cardiovascular diseases, cancer, diabetes, neurodegenerative, and immune disorders. The current review discusses some structural aspects of astaxanthin. It further elaborates its multiple potencies such as antioxidant, anti-inflammatory, anti-proliferative, anti-cancer, anti-obese, anti-diabetic, anti-ageing, anti-TB, anti-viral, anti-COVID 19, neuro-protective, nephro-protective, and fertility-enhancing properties. These potencies make it a more precious entity in the preventions as well as treatments of prevalent systematic diseases and/or disorders. Also, the review is acknowledging and documenting its powerful bioactivities in relation with the pharmaceutical as well as nutraceutical applicability.

Statistical media optimization for the production of clinical uricase from Bacillus subtilis strain SP6.

In this study, a potent uricase producing organism was isolated by a thorough screening and identified as Bacillus subtilis strain SP6 by using 16s rDNA sequencing. Response surface methodological optimization was employed for the enhanced production of uricase from newly isolated Bacillus subtilis strain SP6. In media optimization studies, Plackett Burman (PB) design was used for the selection of the critical media components; which were further optimized using central composite design (CCD). Lactose, soya peptone, uric acid and FeSO4.7H2O were found to be the critical factors influencing the enzyme production. Optimum uricase production with these factors was deduced using central composite design. Significant level of the factors were 12.2 g/L of lactose, 12.79 g/L of soya peptone, 2.55 g/L of uric acid and 0.00325 g/L FeSO4.7H2O. Use of statistical optimization upsurges uricase yield from 1.2 U/ml to 15.87 U/ml enhancing the overall production by 13.23 fold; which confirms that the model is effective for process optimization.

Use of statistical experimental methods for optimization of collagenolytic protease production by Bacillus cereus strain SUK grown on fish scales.

In this study, novel and cheap sources like fish scales and molasses were used for the production of collagenolytic protease. Statistical optimization of different parameters for the production of collagenolytic protease by Bacillus cereus strain SUK has been carried out using response surface methodology (RSM). Three most significant medium components identified by Plackett-Burman (PB) were fish scales, molasses, and incubation time, which were further optimized using central composite design (CCD). The medium having fish scales 9.38 g l-1, molasses 2.42 g l-1, and incubation time of 67.34 h was found to be optimum for maximum collagenolytic protease production. B. cereus strain SUK has shown multiple plant growth-promoting traits, whereas degraded fish scale hydrolysates (FSHs) were having antimicrobial as well as plant growth-promoting abilities. The collagenolytic efficiency of this isolate can be exploited in an eco-friendly process of bioconversion of fish waste, representing an alternative way of waste management that could be used to produce various value-added products, such as collagenolytic protease, microbial biomass, amino acids, protein hydrolysates, and collagen peptides.

Ag Nanoparticles Connected to the Surface of TiO2 Electrostatically for Antibacterial Photoinactivation Studies.

Supported silver nanoparticles (Ag NPs) were prepared by chemical reduction method with a sol-gel method. The structure, morphology, and interconnectivity of Ag/TiO2 nanocomposites (NCs) were analyzed using different instrumental techniques. Transmission electron microscopy reveals the Ag NPs have uniformly distributed and anchored on the surface of TiO2 . The reduction in electron-hole recombination was measured by Photoluminescence measurements lead, to an increased photocatalytic inactivation of bacteria. Increase in the amount of Ag NPs on TiO2 resulted in a slight decrease in optical band gap energy of TiO2 . The effect of Ag NPs content on the photocatalytic properties of TiO2 for inhibition of bacteria in visible light irradiation was studied. Furthermore, the antibacterial activity of Ag/TiO2 NCs in the presence of UVA light was studied against gram-positive Staphylococcus aureus and gram-negative Escherichia coli bacterial strain by plate count method. Lower values of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the catalysts were observed and used to determine the tolerance factor which is shown bactericidal nature of the NCs. Subsequently, time-killing assay of Ag/TiO2 NCs was shown dynamics of antimicrobial activity. These multifold antibacterial studies exhibited potent antibacterial nature of the NCs and employed in the wider range of biomedical fields.

Response surface methodology-based optimization of production media and purification of α-galactosidase in solid-state fermentation by Fusarium moniliforme NCIM 1099.

Response surface methodology was used to enhance the production of α-galactosidase from Fusarium moniliforme NCIM 1099 in solid-state fermentation. Plackett-Burman design was employed for selection of critical media constituents which were optimized by central composite rotatable design. Wheat bran, peptone and FeSO4·7H2O were identified as significant medium components using PB design. Further CCRD optimized medium components as wheat bran; 4.62 µg, peptone; 315.42 µg, FeSO4·7H2O; 9.04 µg. RSM methodological optimization increased the enzyme production from 13.17 to 207.33 U/g showing 15.74-fold enhancement. The α-galactosidase was purified by 70% fractionation followed by DEAE anion exchange column chromatography which yields 23.33% with 28.68-fold purification. The molecular weight of α-galactosidase was 57 kDa which was determined by SDS-PAGE analysis. Purified enzyme has optimum pH of 4.0 and was found to be stable in wide pH range of 3.0-9.0. Its optimum temperature was 50 °C, whereas its activity remains above 50% up to 2 h at 75 °C. Hg2+ was found to be a potent inhibitor and Mg2+ acted as an activator of enzyme. No significant change was observed in enzyme activity for galactose concentration, ranging from 1 to 100 mM. The K m values of enzyme for substrates p-nitrophenyl-α-D-galactopyranoside, melibiose and raffinose were 0.20, 1.36, and 3.66 mM, respectively. Low K m and stability to various physiological conditions of enzyme represents its potential which can be exploited in various industrial applications.