Computational Study on Temperature Driven Structure–Function Relationship of Polysaccharide Producing Bacterial Glycosyl Transferase Enzyme

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.

EPS364, a Novel Deep-Sea Bacterial Exopolysaccharide, Inhibits Liver Cancer Cell Growth and Adhesion

The prognosis of liver cancer was inferior among tumors. New medicine treatments are urgently needed. In this study, a novel exopolysaccharide EPS364 was purified from Vibrio alginolyticus 364, which was isolated from a deep-sea cold seep of the South China Sea. Further research showed that EPS364 consisted of mannose, glucosamine, gluconic acid, galactosamine and arabinose with a molar ratio of 5:9:3.4:0.5:0.8. The relative molecular weight of EPS364 was 14.8 kDa. Our results further revealed that EPS364 was a β-linked and phosphorylated polysaccharide. Notably, EPS364 exhibited a significant antitumor activity, with inducing apoptosis, dissipation of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS) in Huh7.5 liver cancer cells. Proteomic and quantitative real-time PCR analyses indicated that EPS364 inhibited cancer cell growth and adhesion via targeting the FGF19-FGFR4 signaling pathway. These findings suggest that EPS364 is a promising antitumor agent for pharmacotherapy.

Biosorption of Heavy Metals by the Bacterial Exopolysaccharide FucoPol

Despite the efforts for minimizing the usage of heavy metals, anthropogenic activities still generate high amounts of wastewater containing these contaminants that cause significant health and environmental problems. Given the drawbacks of the conventional physical and chemical methods currently used, natural biosorbents (microbial cells or their products) arise as promising environmentally friendly alternatives. In this study, the binding efficiency of the polysaccharide secreted by Enterobacter A47, FucoPol, towards lead (Pb2+), cobalt (Co2+), copper (Cu2+) and zinc (Zn2+) cations was demonstrated. FucoPol revealed a higher performance for the biosorption of Pb2+, with a maximum overall metal removal of 93.9 ± 5.3% and a specific metal uptake of 41.1 ± 2.3 mg/gEPS, from a Pb2+ solution with an initial concentration of 10 mg/L, by a 5 g/L FucoPol solution. The overall metal removal decreased considerably (≤31.3 ± 1.6%) for higher Pb2+ concentrations (48 and 100 mg/L) probably due to the saturation of FucoPol’s binding sites. Pb2+ removal was also less efficient (66.0 ± 8.2%) when a higher FucoPol concentration (10 g/L) was tested. Pb2+ removal efficiency of FucoPol was maximized at pH 4.3, however, it was affected by lower pH values (2.5–3.3). Moreover, the FucoPol’s sorption performance was unaffected (overall metal removal: 91.6–93.9%) in the temperature range of 5–40 °C. These findings demonstrate FucoPol’s great potential for utilization as a biodegradable and safe biosorbent for treating waters and wastewaters contaminated with Pb2+.

Kingella kingaeSurface Polysaccharides Promote Resistance to Neutrophil Phagocytosis and Killing

ABSTRACTBacterial pathogens have evolved strategies that enable them to evade neutrophil-mediated killing. The Gram-negative coccobacillusKingella kingaeis an emerging pediatric pathogen and is increasingly recognized as a common etiological agent of osteoarticular infections and bacteremia in young children.K. kingaeproduces a polysaccharide capsule and an exopolysaccharide, both of which are important for protection against complement-mediated lysis and are required for full virulence in an infant rat model of infection. In this study, we examined the role of theK. kingaepolysaccharide capsule and exopolysaccharide in protection against neutrophil killing. In experiments with primary human neutrophils, we found that the capsule interfered with the neutrophil oxidative burst response and prevented neutrophil binding ofK. kingaebut had no effect on neutrophil internalization ofK. kingae. In contrast, the exopolysaccharide resisted the bactericidal effects of antimicrobial peptides and efficiently blocked neutrophil phagocytosis ofK. kingae. This work demonstrates that theK. kingaepolysaccharide capsule and exopolysaccharide promote evasion of neutrophil-mediated killing through distinct yet complementary mechanisms, providing additional support for theK. kingaesurface polysaccharides as potential vaccine antigens. In addition, these studies highlight a novel interplay between a bacterial capsule and a bacterial exopolysaccharide and reveal new properties for a bacterial exopolysaccharide, with potential applicability to other bacterial pathogens.IMPORTANCEKingella kingaeis a Gram-negative commensal in the oropharynx and represents a leading cause of joint and bone infections in young children. The mechanisms by whichK. kingaeevades host innate immunity during pathogenesis of disease remain poorly understood. In this study, we established that theK. kingaepolysaccharide capsule and exopolysaccharide function independently to protectK. kingaeagainst reactive oxygen species (ROS) production, neutrophil phagocytosis, and antimicrobial peptides. These results demonstrate the intricacies ofK. kingaeinnate immune evasion and provide valuable information that may facilitate development of a polysaccharide-based vaccine againstK. kingae.