The current demands of the world’s biotechnological industries are enhancement in enzyme productivity and development of novel techniques for increasing their shelf life. These requirements are inevitable to facilitate large-scale and economic formulation. Enzyme immobilization provides an excellent base for increasing availability of enzyme to the substrate with greater turnover over a considerable period of time. Several natural and synthetic supports have been assessed for their efficiency for enzyme immobilization. Nowadays, immobilized enzymes are preferred over their free counterpart due to their prolonged availability that curtails redundant downstream and purification processes. Future investigations should endeavor at adopting logistic and sensible entrapment techniques along with innovatively modified supports to improve the state of enzyme immobilization and provide new perspectives to the industrial sector.

Casein capped copper nanoparticles at sub inhibitory concentrations function as an efflux pump inhibitor and restores susceptibility to antibiotics in drug resistant bacteria.

In the present study, the syntheses of crystalline silver and gold nanoparticles (NPs) has been demonstrated in a green and environmentally friendly approach using citrus fruit extracts (lemon, Citrus limon (Cl-1) and sweet orange, Citrus limetta (Cl-2)). In addition, potentially hazardous metal ion sensing properties of these NPs in aqueous solution has been explored. Cl-1 and Cl-2 that predominantly contained citric and ascorbic acid, exhibited different reducing abilities towards silver and gold ions into NPs. Cl-1 reduces silver ions into AgNPs only in the presence of sunlight whereas AuNPs from gold ions were formed without exposure to sunlight. In contrast, Cl-2 converts both silver and gold ions into the corresponding NPs in the absence of sunlight. The colorimetric sensor studies of these green synthesized AgNPs (Cl-1-AgNPs) showed selective sensing of the potentially hazardous Hg(2+) ion in water at micromolar concentrations. More importantly, green synthesized Cl-1-AgNPs sensor systems detected Hg(2+) ions in water in a wide pH range (3.2 to 8.5).

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Scutellaria oblonga Benth., a hitherto phytochemically unexplored Indian medicinal folklore plant was extracted with acetone and subjected to chromatography to yield nine flavonoids, for the first time from this plant. Antimicrobial assays were performed against 11 foodborne pathogens, and three molecules (Techtochrysin, Negletein and Quercitin-3-glucoside) depicted significant activity. These molecules were assessed for their rate of antibacterial action using time-kill curves which depicted complete inhibition of most of the bacteria within 12-16 h. The significant biofilm-reducing capability exhibited by these three molecules formed a significant finding of the current study. In most of the experiments, a 90-95% reduction in biofilms was observed. Thus, flavonoids as natural molecules from S. oblonga could be further researched to be used as potent antimicrobial and antibiofilm agents.

Multi drug resistant (MDR) pathogens pose a serious threat to public health since they can easily render most potent drugs ineffective. Efflux pump inhibitors (EPI) can be used to counter the MDR phenotypes arising due to increased efflux. In the present study, a series of dithiazole thione derivatives were synthesized and checked for its antibacterial and efflux pump inhibitory (EPI) activity. Among 10 dithiazole thione derivatives, real-time efflux studies revealed that seven compounds were potent EPIs relative to CCCP. Zebrafish toxicity studies identified four non-toxic putative EPIs. Both DTT3 and DTT9 perturbed membrane potential and DTT6 was haemolytic. Among DTT6 and DTT10, the latter was less toxic as evidenced by histopathology studies. Since DTT10 was non-haemolytic, did not affect the membrane potential, and was least toxic, it was chosen further for in vivo study, wherein DTT10 potentiated effect of ciprofloxacin against clinical strain of MRSA and reduced bacterial burden in muscle and skin tissue of infected zebrafish by ~ 1.7 and 2.5 log fold respectively. Gene expression profiling of major efflux transport proteins by qPCR revealed that clinical isolate of MRSA, in the absence of antibiotic, upregulated NorA, NorB and MepA pump, whereas it downregulates NorC and MgrA relative to wild-type strain of Staphylococcus aureus. In vitro studies with NorA mutant strains and substrate profiling revealed that at higher concentrations DTT10 is likely to function as a competitive inhibitor of NorA efflux protein in S. aureus, whereas at lower concentrations it might inhibit ciprofloxacin efflux through NorB and MepA as implied by docking studies. A novel non-toxic, non-haemolytic dithiazole thione derivative (DTT10) was identified as a potent competitive inhibitor of NorA efflux pump in S. aureus using in silico, in vitro and in vivo studies. This study also underscores the importance of using zebrafish infection model to screen and evaluate putative EPI for mitigating MDR strains of S. aureus.

Nutraceutical pinostrobin displayed synergy with ciprofloxacin against diverse bacteria causing drastic 128 fold reduction in ciprofloxacin MIC in MRSA and significant antibiofilm effect against E. coli and P. aeruginosa at subinhibitory concentrations. Studies with NorA mutants of S.aureus revealed efflux inhibition of pinostrobin is mediated by efflux pumps other than NorA.

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