Selective Bacterial Growth Inactivation by pH-Sensitive Sulfanilamide Functionalized Carbon Dots.

Carbon dots (CDs) were synthesized hydrothermally by mixing citric acid (CA) and an antifolic agent, sulfanilamide (SNM), employed for pH sensing and bacterial growth inactivation. Sulfanilamide is a prodrug; aromatic hetero cyclization of the amine moiety along with other chemical modifications produces an active pharmacological compound (chloromycetin and miconazole), mostly administered for the treatment of various microbial infections. On the other hand, the efficacy of the sulfanilamide molecule as a drug for antimicrobial activity was very low. We anticipated that the binding of the sulfanilamide molecule on the carbon dot (CD) surface may form antibacterial CDs. Citric acid was hybridized with sulfanilamide during the hydrothermal preparation of the CDs. The molecular fragments of bioactivated sulfanilamide molecule play a crucial role in bacterial growth inactivation for Gram-positive and Gram-negative bacteria. The functional groups of citric acid and sulfanilamide were conserved during the CD formation, facilitating the zwitterionic behavior of CDs associated with its photophysical activity. At low concentrations of CDs, the antibacterial activity was apparent for Gram-positive bacteria only. This Gram-positive bacteria selectivity was also rationalized by zeta potential measurement.

Thermostable Bacterial Laccase: Catalytic Properties and Its Application in Biotransformation of Emerging Pollutants.

Laccases have been predominantly reported in fungi, and primarily, fungal laccases are currently exploited in industrial applications. However, extremophilic bacterial laccases possess immense potential, as they can withstand extreme temperatures, pH, and salt concentrations. In addition, unlike fungal laccases, the production of bacterial laccases is cost-effective. Therefore, bacterial laccases are gaining significant attention for their large-scale applications. Previously, we reported a novel thermostable laccase (LacT) from Brevibacillus agri. Herein, we have confirmed that LacT shares a high sequence similarity with CotA laccase from Bacillus amyloliquefaciens. Peptide mass fingerprinting of LacT was conducted via matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF/MS-MS). Inductively coupled plasma-optical emission spectroscopic (ICP-OES) analysis revealed the presence of ∼3.95 copper ions per protein molecule. Moreover, the secondary and tertiary structure of LacT was studied using circular dichroism (CD) and fluorescence spectroscopy. The absence of notable shifts in CD and fluorescence spectra with an increase in temperature established that LacT remains intact even at elevated temperatures. Analysis of the thermal denaturation profile of LacT by thermogravimetric analysis (TGA) also confirmed its temperature stability. Thereafter, we exploited LacT in its application for the bioremediation of phenolic endocrine disruptors, namely, triclosan, 4,4'-dihydroxybiphenyl, and dienestrol. LacT oxidizes 4,4'-dihydroxybiphenyl and triclosan but no LacT activity was detected with dienestrol. The rate of biotransformation of 4,4'-dihydroxybiphenyl and triclosan increased in the presence of CuSO4 and a redox mediator, ABTS. Transformation of dienestrol was observed only with LacT in the presence of ABTS. This study establishes the application of LacT for the bioremediation of phenolic compounds.

Recent Advances in the Development of Oxidoreductase-Based Biosensors for Detection of Phenolic Antioxidants in Food and Beverages.

Antioxidants are known to exhibit a protective effect against reactive oxygen species (ROS)-related oxidative damage. As a result, inclusion of exogenous antioxidants in the diet has greatly increased. In this sense, detection and quantification of such antioxidants in various food and beverage items are of eminent importance. Monophenols and polyphenols are among the most prominent natural antioxidants. In this regard, biosensors have emerged as a simple, fast, and economical method for determination of such antioxidants. Owing to the fact that majority of the phenolic antioxidants are electroactive, oxidoreductase enzymes are the most extensively availed bioreceptors for their detection. Herein, the different types of oxidoreductases that have been utilized in biosensors for the biorecognition and quantification of natural phenolic compounds commonly present in foods and beverages are discussed. Apart from the most accustomed electrochemical biosensors, this review sheds light on the alternative transduction systems for the detection of phenolic antioxidants. Recent advances in the strategies involved in enzyme immobilization and surface modification of the biosensing platform are analyzed. This review aims to provide a brief overview of the latest developments in biosensor technology for phenolic antioxidant analysis in foodstuffs and future directions in this field.