Extremophilic microorganisms often produce novel bioactive compounds to survive under harsh environmental conditions. Exopolysaccharides (EPSs), a constitutive part of bacterial biofilm, are functional biopolymers that act as a protecting sheath to the extremophilic bacteria and are of high industrial value. In this study, we elucidate a new EPS produced by thermophilic Bacillus haynesii CamB6 from a slightly acidic (pH 5.82) Campanario hot spring (56.4 °C) located in the Central Andean Mountains of Chile. Physicochemical properties of the EPS were characterized by different techniques: Scanning electron microscopy- energy dispersive X-ray spectroscopy (SEM-EDS), Atomic Force Microscopy (AFM), High-Performance Liquid Chromatography (HPLC), Gel permeation chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), 1D and 2D Nuclear Magnetic Resonance (NMR), and Thermogravimetric analysis (TGA). The EPS demonstrated amorphous surface roughness composed of evenly distributed macromolecular lumps. GPC and HPLC analysis showed that the EPS is a low molecular weight heteropolymer composed of mannose (66%), glucose (20%), and galactose (14%). FTIR analysis demonstrated the polysaccharide nature (–OH groups, Acetyl groups, and pyranosic ring structure) and the presence of different glycosidic linkages among sugar residues, which was further confirmed by NMR spectroscopic analyses. Moreover, D-mannose α-(1→2) and α-(1→4) linkages prevail in the CamB6 EPS structure. TGA revealed the high thermal stability (240 °C) of the polysaccharide. The functional properties of the EPS were evaluated for food industry applications, specifically as an antioxidant and for its emulsification, water-holding (WHC), oil-holding (OHC), and flocculation capacities. The results suggest that the study EPS can be a useful additive for the food-processing industry.

In the present study, Chryseobacterium geocarposphaerae DD3 isolated from textile industry dye effluent in West Bengal, India, displayed significant tolerance to sulfonated di-azo dye Congo red (CR), up to 500 ppm. The optimum decolourisation revealed that C. geocarposphaerae DD3 was capable of 96.52% decolourisation of 0.2 g L⁻¹ CR within 12 h of treatment in the presence of 5 g L⁻¹ glucose as supplementary carbon source. Biodegradation analysis of decolourised CR containing water was investigated by FTIR, MS and ¹H NMR, which confirmed the absence of azo bond as well as the toxic aromatic amines. Further, phytotoxicity analysis was performed to assess the toxicity of CR before and after bacterial treatment. Growth indexes of Vigna radiata L. seed confirmed that the biodegraded water was non-phytotoxic in comparison to the control CR solution. Multivariate analyses confirmed the same, showing significant differences between measured plant health indicators for CR solutions, whereas no significant differences were found between distilled and treated water. This study is novel as it is the first report of dye degradation by C. geocarposphaerae and may lead to a sustainable way of treating dye-contaminated water in the near future.

Glycosyltransferase (GTs) is a wide class of enzymes that transfer sugar moiety, playing a key role in the synthesis of bacterial exopolysaccharide (EPS) biopolymer. In recent years, increased demand for bacterial EPSs has been observed in pharmaceutical, food, and other industries. The application of the EPSs largely depends upon their thermal stability, as any industrial application is mainly reliant on slow thermal degradation. Keeping this in context, EPS producing GT enzymes from three different bacterial sources based on growth temperature (mesophile, thermophile, and hyperthermophile) are considered for in silico analysis of the structural–functional relationship. From the present study, it was observed that the structural integrity of GT increases significantly from mesophile to thermophile to hyperthermophile. In contrast, the structural plasticity runs in an opposite direction towards mesophile. This interesting temperature-dependent structural property has directed the GT–UDP-glucose interactions in a way that thermophile has finally demonstrated better binding affinity (−5.57 to −10.70) with an increased number of hydrogen bonds (355) and stabilizing amino acids (Phe, Ala, Glu, Tyr, and Ser). The results from this study may direct utilization of thermophile-origin GT as best for industrial-level bacterial polysaccharide production.

Xylose Isomerase (XI) is an intramolecular oxidoreductase enzyme and catalyzes the reversible conversion of ketoses and aldoses in addition to the bioconversion of ethanol from xylose in the production of bioethanol from hemicellulose. It has a broad range of industrial applications in the food and pharmaceutical sectors, particularly in the production of the sweetener high fructose corn syrup (HFCS). It is one of the most widely used industrial enzymes after protease. Taking this into consideration, four bacterial XI sources were selected based on growth temperature, i.e., psychrophile, mesophile, thermophile, and hyperthermophile, for analyzing Xylose Isomerase’s structure-function characteristics. It was found that thermophilic XI was structurally less stable than mesophilic and hyperthermophilic XI, whereas structural plasticity ran opposite towards mesophiles. The interaction of xylose isomerase (XI) with two ligands, namely Amino-2-Hydroxymethyl-Propane-1,3-Diol and (4R)-2-Methylpentane-2,4- Diol, was also studied. Mesophilic XI demonstrated better binding affinity with structurally stabilizing amino acids (Ala, Asp, Gly, Leu, and Arg). In comparison, Thermophilic XI showed nearly similar binding affinity with both Amino-2-Hydroxymethyl-Propane-1,3-Diol and (4R)-2-Methylpentane-2,4-Diol. The results of this investigation suggest that thermophilic XI, followed by mesophilic XI, would be the most appropriate for establishing process stability and sustainability in the food industry.

The keratinase serves as a catalyst with potential application in various industries, including textiles, leather, pharmaceuticals, cosmetics, and biotechnological applications. Previous studies have predominantly focused on fungal keratinase; however, our present work focuses on bacterial keratinase only. The novelty of using bacterial keratinase over fungal keratinase is because of its faster enzymatic activity, broader substrate specificity, increased stability and cost-effective production, making it efficient and adaptable for industrial applications. However, our present research investigates bacterial keratinase through in silico structural (physicochemical properties, primary, secondary, and tertiary structures) as well as functional analyses (protein–protein interaction networks, subcellular localization, and domain/motif presence), coupled with molecular docking against standard anti-acne drugs in use (adapalene, erythromycin, tretinoin, isotretinoin, and benzoyl peroxide). In this study, two bacterial keratinases, one from mesophilic Pseudomonas aeruginosa (6FZX) and another from thermophilic Meiothermus taiwanensis WR-220 (5WSL) have been studied to demonstrate comparative structural stability and functional activity. These enzymes are intracellular, facilitating proteolysis with metalloendopeptidase activity. Notably, keratinase exhibited a strong binding affinity for adapalene (binding energies of − 8.5 kcal/mol and − 9.6 kcal/mol for 5WSL and 6FZX, respectively) and lowest affinity to erythromycin (binding energy − 6.4 kcal/mol and − 5.9 kcal/mol for 5WSL and 6FZX respectively) among other studied anti-acne drugs like, tretinoin, isotretinoin, and benzoyl peroxide. In the case of de novo designing of anti-acne compounds, enhanced binding affinities are the key factor for selection of potential components. Our in silico analyses indicate that thermophilic keratinase displays better structural stability and functionality, particularly under high pH and high-temperature conditions, making it a promising candidate for cosmeceutical applications. Thermophilic keratinase may effectively operate in demanding skin environments and may offer advantages over mesophilic keratinase. These insights from comparative computational analysis will pave the way for further wet lab validation and industrial experiments to guide the selection of optimal candidates in antiacne formulations.