In this study, we report the biochemical characterization of a novel serine protease from seeds of Cucumis maderaspatensis, aimed with assessing the anticoagulant and antiplatelet activities. The purified serine protease was obtained by subjecting the seed extract to ammonium sulphate precipitation followed by anion exchange and gel filtration chromatography. Twenty seven-fold purification with the specific activity of 884.2 U/mg of protease activity was obtained. The characterization of the novel protease enzyme activity for optimum temperature, pH and effect of different protease inhibitors and metal ions were measured using caseinolytic assay and casein zymogram. The relative molecular mass of the novel neutral serine protease (CmSP) is ~ 32 kDa. Its anticoagulant was determined by assessing the delay in plasma re-calcification time in both platelet-rich and platelet-poor plasma. The antiplatelet activity of serine protease was demonstrated by inhibition of agonists induced platelet aggregation; it was in the order of Epinephrine > Adenosine tri phosphate. Further studies would decipher the mechanism of action to understand its therapeutic potential as an antiplatelet and anticoagulant molecule.

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Cassia siamea is a nonedible legume belonging to Fabaceae. The seed of C. siamea contains ~16% of protein. The study reports the biochemical characterization of purified novel serine protease inhibitor from seeds of C. siamea, aimed with assessing the anti-inflammatory activity. The seed extract was subjected to ammonium sulfate precipitation followed by fast protein liquid chromatography (FPLC)-anion exchange chromatography and affinity-chromatography to obtain a relative pure protease inhibitor. Thirty-fivefold purification with the specific activity of 250 U/mg of trypsin inhibitory unit was obtained. The characterization of protease inhibitor for optimum temperature, pH, and metal ions were measured using N-α-benzoyl-DL-arginine-pnitroanilide (BAPNA) assay and casein zymogram. The C. siamea trypsin inhibitor (CsTI) has a relative molecular mass of 25.540 kDa. Purified CsTI and Dolichos biflorus were tested for anti-inflammatory efficacy against A549 and RAW264.7 cell lines.The inhibitory activity of both purified inhibitors are comparable and are potent toward anti-inflammatory activity. The purified inhibitor shows to be a promising candidate as anti-inflammatory agent by targeting the serine proteases.

IntroductionHoney bee gut microbiota have an important role in host health, nutrition, host-symbiont interaction, and interaction behavior with the surrounding environment. Recent discoveries of strain-level variation, characteristics of protective and nutritional capabilities, and reports of eco-physiological significance to the microbial community have emphasized the importance of honey bee gut microbiota. Many regions of Asia and Africa are inhabited by the dwarf honey bee, Apis florea. Studying its microflora and potential for pollination is therefore of foremost importance.MethodsIn the present investigation, we aimed to explore the gut bacteriobiome composition of two distinct honey bee species, Apis florea and Apis cerana indica using high throughput sequencing. Functional predictions of bee gut bacterial communities using PICRUSt2 was carried out.Results and discussionThe phylum Proteobacteria dominated the bacterial community in both A. cerana indica (50.1%) and A. florea (86.7%), followed by Firmicutes (26.29 and 12.81%), Bacteroidetes (23.19 and 0.04%) and Actinobacteria (0.4 and 0.02%) respectively. The gut bacteria of A. cerana indica was more diverse than that of A. florea. The observed variations in bacterial genomic diversity among these critical pollinator species may have been influenced by the apiary management techniques, ecological adaptation factors or habitat size. These variations can have a significant effect in understanding host-symbiont interactions and functioning of gut microbiota highlighting the importance of metagenomic survey in understanding microbial community ecology and evolution. This is the first comparative study on variation in bacterial diversity between two Asian honey bees.

Introduction and Aim:Immobilization of enzyme on silver nanoparticles (AgNPs) is one way to improve their stability and activity and can be reused for large-scaleapplications. The present study was aimed to evaluateand characterizethetrypsin enzyme immobilized on biogenicallysynthesized silver nanoparticles.Materials and Methods: Immobilization of trypsin enzyme was optimized with time and varying concentration of silver nanoparticles, which were greensynthesized using avocado seedextracts. The residual activity of trypsin enzyme after immobilization was estimated. The reusabilityand temperature stability of the immobilized enzyme were studied. Results:The immobilized enzyme showed maximum activity of 1006.0 U during the time duration of 18 hr of incubation with AgNPs. Reusability of immobilized enzyme in avocado AgNPs was assessed under optimal conditions, the activity of immobilized enzyme was loss after 5 repeated cycles. The enzyme captured on AgNPswas released in the presence of anionic detergent. Temperaturestability was assessed for immobilized enzyme on AgNPs and the immobilized enzyme was stable up to55ºC and later it loss their activity. Conclusion: The study reports a feasible method of enzyme immobilization on biogenically synthesized silver nanoparticles. The enzyme immobilized on silver nanoparticles is mainly through non-covalent forces. Further studies are required to improve the reusability of enzyme. Keywords: Silver nanoparticles;enzyme immobilization;trypsin;temperature stability;reusability;detergent.