Study of nano-hydroxyapatite tagged alkaline protease isolated from Himalayan sub-alpine Forest soil bacteria and role in recalcitrant feather waste degradation.

Purified calcium serine metalloprotease from Stenotrophomonas maltophilia strain SMPB12 exhibits highest enzyme activity at pH 9 and temperature range between 15 °C-25 °C. Enzyme supplemented with 40 µM Ca-Hap-NP (NP-protease) showed maximum elevated activity of 17.29 µmole/min/ml (1.9-fold of original protease activity). The thermostability of the enzyme was maintained for 1 h at 60 °C over an alkaline pH range 7.5-10, as compared to the NP untreated enzyme whose activity was of 8.97 µmole/min/ml. A significant loss of activity with EDTA (1.05 µmole/min/ml, 11.75 %), PMSF (0.93 µmole/min/ml, 10.46 %) and Hg2+ (3.81 µmole/min/ml, 42.49 %) was also observed. Kinetics study of NP-protease showed maximum decreases in Km (28.11 %) from 0.28 mM (NP untreated enzyme) to 0.22 mM (NP-protease) along with maximum increase in Vmax (42.88 %) from 1.25 µmole/min/ml to 1.79 µmole/min/ml at varying temperatures. The enhanced activity of NP-protease was able to efficiently degrade recalcitrant solid wastes like feather to produce value-added products like amino acids and helps in declogging recalcitrant solid wastes. The nano-enabled protease may be utilized in a smaller amount for degrading in bulk recalcitrant solid proteinaceous waste at 15 °C temperature as declogging agents providing an eco-friendly efficient process.

Quorum quenching activity of pentacyclic triterpenoids leads to inhibition of biofilm formation by Acinetobacter baumannii.

The quorum quenching (QQ) potential of three pentacyclic triterpenoids, glycyrrhetinic acid (GRA), ursolic acid (UA) and betulinic acid (BA), representing distinct groups of compounds, was evaluated. Violacein production by Chromobacterium violaceum and pyocyanin production by Pseudomonas aeruginosa were severely affected by GRA, UA and BA, suggesting a perturbation of N-acyl homoserine lactone (ASL) based signaling. Molecular docking analysis revealed a possible interaction between ASL-synthase and ASL-dependent transcriptional activator homologs from P. aeruginosa and Acinetobacter baumannii with common binding pockets for GRA, UA and BA. The triterpenoids inhibited biofilm formation by A. baumannii and affected the overall structure of biofilms. When administered in combination, two of the three molecules fostered antibiotic action against A. baumannii biofilms, widening the scope for developing novel combinations against the pathogen.

A comprehensive and comparative study on the action of pentacyclic triterpenoids on Vibrio cholerae biofilms.

While serving as environmental reservoir for V. cholerae infection, biofilms are also crucial for intestinal colonization of the pathogen. Triterpenoids, a group of bioactive phytochemicals, have been tested for antibiofilm activity against model biofilm forming bacteria in recent times. In this context, glycyrrhetinic acid (GRA), ursolic acid (UA) and betulinic acid (BA), representing three categorically distinct groups of pentacyclic triterpenoids, are targeted for profiling their impact on Vibrio cholerae C6709 biofilms. The triterpenoids substantially affected biofilm associated attributes like formation, substratum adherence and dispersion from preformed biofilms. Though at variable degree, the compounds decreased cell surface hydrophobicity and composition in terms of macromolecular content. Not only EPS-associated extracellular enzyme activities were estimated to be reduced by triterpenoid exposure, ultra structural analysis also revealed that GRA, UA and BA can affect extracellular polymeric substance (EPS) content. Albeit total extracellular proteolytic activity remained unaffected by the triterpenoids, GRA treatment resulted in considerable reduction of extracellular gelatinase activity. Molecular docking analysis indicated potential interaction with cyclic di-GMP sensor VpsT, autoinducer-2 sensor kinase LuxP-LuxQ and transcriptional activator HapR, components of complex quorum sensing networks modulating biofilm formation. Comprehensive analysis of antibiotic action revealed accentuation of cephalosporin antibiotics with GRA and UA while BA potentiated action of fluoroquinolones against biofilmed bacteria, widening the scope of combinatorial therapeutic strategy.

Inhibition of biofilm formation of Pseudomonas aeruginosa by caffeine: a potential approach for sustainable management of biofilm.

Pseudomonas aeruginosa is a potent biofilm forming organism causing several diseases on host involving biofilm. Several natural and synthetic molecules have been explored towards inhibiting the biofilm formation of Pseudomonas aeruginosa. In the current report, the role of a natural molecule namely caffeine was examined against the biofilm forming ability of P. aeruginosa. We have observed that caffeine shows substantial antimicrobial activity against P. aeruginosa wherein the minimum inhibitory concentration (MIC) of caffeine was found to be 200 µg/mL. The antibiofilm activity of caffeine was determined by performing a series of experiments using its sub-MIC concentrations (40 and 80 µg/mL). The results revealed that caffeine can significantly inhibit the biofilm development of P. aeruginosa. Caffeine has been found to interfere with the quorum sensing of P. aeruginosa by targeting the swarming motility. Molecular docking analysis further indicated that caffeine can interact with the quorum sensing proteins namely LasR and LasI. Thus, the result indicated that caffeine could inhibit the formation of biofilm by interfering with the quorum sensing of the organism. Apart from biofilm inhibition, caffeine has also been found to reduce the secretion of virulence factors from Pseudomonas aeruginosa. Taken together, the results revealed that in addition to biofilm inhibition, caffeine can also decrease the spreading of virulence factors from Pseudomonas aeruginosa.

Capsaicin-induced activation of p53-SMAR1 auto-regulatory loop down-regulates VEGF in non-small cell lung cancer to restrain angiogenesis.

Lung cancer is the leading cause of cancer-related deaths worldwide. Despite decades of research, the treatment options for lung cancer patients remain inadequate, either to offer a cure or even a substantial survival advantage owing to its intrinsic resistance to chemotherapy. Our results propose the effectiveness of capsaicin in down-regulating VEGF expression in non-small cell lung carcinoma (NSCLC) cells in hypoxic environment. Capsaicin-treatment re-activated p53-SMAR1 positive feed-back loop in these cells to persuade p53-mediated HIF-1α degradation and SMAR1-induced repression of Cox-2 expression that restrained HIF-1α nuclear localization. Such signal-modulations consequently down regulated VEGF expression to thwart endothelial cell migration and network formation, pre-requisites of angiogenesis in tumor micro-environment. The above results advocate the candidature of capsaicin in exclusively targeting angiogenesis by down-regulating VEGF in tumor cells to achieve more efficient and cogent therapy of resistant NSCLC.

Restoration of p53/miR-34a regulatory axis decreases survival advantage and ensures Bax-dependent apoptosis of non-small cell lung carcinoma cells.

Tumor-suppressive miR-34a, a direct target of p53, has been shown to target several molecules of cell survival pathways. Here, we show that capsaicin-induced oxidative DNA damage culminates in p53 activation to up-regulate expression of miR-34a in non-small cell lung carcinoma (NSCLC) cells. Functional analyses further indicate that restoration of miR-34a inhibits B cell lymphoma-2 (Bcl-2) protein expression to withdraw the survival advantage of these resistant NSCLC cells. In such a proapoptotic cellular milieu, where drug resistance proteins are also down-regulated, p53-transactivated Bcl-2 associated X protein (Bax) induces apoptosis via the mitochondrial death cascade. Our results suggest that p53/miR-34a regulatory axis might be critical in sensitizing drug-resistant NSCLC cells.