This study was undertaken to evaluate the behaviour of vegetative cells and spores of four potent native toxigenic food isolates of Bacillus cereus as affected by selected time-temperature combinations used in processing of Indian traditional foods. The vegetative cells of B. cereus when subjected to sublethal heat treatments, individually, in different heating menstra showed a sigmoidal inactivation pattern, with D-values in the range of 3.45 min at 60°C to 10.6 min at 56°C in saline. Accordingly, the zvalues recorded across the heating menstra ranged from 9.3°C in culture broth to 24°C in whole milk. Similarly, the inactivation pattern for spores for the same isolates was curvilinear with D-values ranging from 4.4 min at 95°C in whole milk to 19.45 min at 85°C in saline. The z-values for spores ranged from 16.6°C in saline to 38.4°C in whole milk. The thermal inactivation pattern observed for vegetative cells and spores indicate that the death rate was not constant during the process of heat treatment.

Zinc Oxide Nanoparticles (ZnONPs) are one of the most widely used metal oxide nanoparticles in biological applications because of their outstanding biocompatibility, affordability, and low toxicity. In biomedicine, ZnONPs have shown promise, particularly in the disciplines of anticancer and antibacterial fields. In comparison to other standard synthesis methods, the environmentally-friendly synthesis of metallic nanoparticles utilizing various plant extracts is a good option. The current research focuses on the synthesis of zinc oxide nanoparticles (ZnONPs) from R. sativus leaf extract under various physical conditions (Precipitation method). Analytical methods were used to confirm and characterize the produced ZnONPs. The spherical nature of the produced nanoparticles was established by SEM analysis. The generation of very pure ZnONPs was confirmed by EDS data. The crystalline nature of the produced nanoparticles, with a particle size of 66.47 nm, was confirmed by XRD. The XRD graphs’ presence of the (100), (002), and (101) planes strongly suggest the production of wurtzite ZnO. The visual and infrared area exhibits transmissions of 84 percent in the pH 10 nanoparticles. The band gap of the nanoparticles increases from 3.34 to 3.38 eV when the pH increases. These nanoparticles were effective against both Gram-positive and Gram-negative bacteria. The effect of several process parameters such as pH and temperature were investigated, and the best conditions were discovered to be pH 12 and 80 °C, respectively. The effect of ZnONPs was tested with human breast cancer cells (MCF-7), and they showed significant cytotoxic results. Collectively, our data suggest that ZnONPs of R. sativus leaf extract inhibit breast cancer cell lines. The ZnONPs are, therefore, a prospective source of chemopreventive drugs that merit additional exploration in order to uncover lead compounds with cancer chemotherapeutic potential.

Pectinolytic enzymes are related enzymes that hydrolyze pectic substances. Pectinolytic enzymes are of great interest in industrial applications for softening fruits, extracting and clarifying juices, extracting olive oil, retting textile fibers, preparing gel, and isolating protoplasts. The current work presents acidic extracellular pectinase production using low-cost agro-industrial waste with the indigenously isolated novel strain Aspergillus cervinus. Two fungal isolates, ARS2 and ARS8, with maximum pectinase activity, 41.88 ± 1.57 IU/mL and 39.27 ± 1.14 IU/mL, respectively, were screened out of 27 isolates from decayed fruit peels (orange, banana, and lemon) and soil containing decomposed vegetables. The isolate ARS2, identified as Aspergillus cervinus by molecular characterization, showed the highest pectinase activity of 43.05 ± 1.38IU/mL during screening and was further used for media component screening and optimization studies. To understand their effect on pectinase activity, one-factor-at-a-time (OFAT) studies were conducted on carbon sources, nitrogen sources, and mineral salts. The OFAT results showed the highest pectinase activity for orange peel (carbon source) at 44.51 ± 1.33 IU/mL, peptone (nitrogen source) at 45.05 ± 1.04 IU/mL, and NaH2PO4 (mineral salts) at 43.21 ± 1.12 IU/mL. The most significant media components screened by the Plackett–Burman (PB) design based on the p-value, Pareto chart, and main effect plot, were orange peel (p < 0.001), peptone (p < 0.001), NaH2PO4 (p < 0.001), and KH2PO4 (p < 0.001), which were further optimized using Response Surface Methodology (RSM) and Central Composite Design (CCD). The optimization results for the media components showed a maximum pectinase activity of 105.65 ± 0.31 IU/mL for 10.63 g orange peel, 3.96 g/L peptone, 2.07 g/L KH2PO4, and 2.10 g/L NaH2PO4. Thus, it was discovered that the indigenously isolated novel strain Aspergillus cervinus ARS2 was able to successfully produce a significant amount of pectinase using agro-industrial waste. Therefore, it can be considered for the large-scale optimized production of pectinase to meet industrial demands.

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The survival and growth of vegetative cells and spores of toxigenic Bacillus cereus CFR 1534 was investigated in a dehydrated (dry mix) blend of flour (roasted finger millet and Bengal gram) and chocolate milk. In the case of dry mix, there was a marginal initial increase of 2 logs in 2 days of storage at 20 and 30C, respectively. In chocolate milk, there was an increase of 1 log with certain inoculum levels introduced and storage temperatures. The survival pattern of vegetative cells/spores was primarily influenced by the initial numbers and, to a slight extent, by the water activity of food matrices. Although stored samples were negative for toxigenic trait of phosphatidylinositol phospholipase C in polymerase chain reaction, associated lecithinase activity was detected in plate assay. Response surface plots for vegetative cells and spores of B. cereus CFR 1534 did indicate the potential risk associated with post‐processing contamination of B. cereus. Practical Applications The ability of vegetative cells and spores of a toxigenic culture of Bacillus cereus to survive and grow in two food matrices was studied. Often, the bacterial species tend to remain in the product and increase in their numbers in a slow paced manner. The product with no visual changes, but harbors viable population of toxigenic Bacillus cereus can be of serious public health concern. The understanding of product profile would be of help in assessing the probable risk factors that could be associated with opportunistic foodborne pathogen like B. cereus in the food chain establishment.

Xylanases are enzymes that convert xylan into xylose, xylobiose, and xylotriose. The present study deals with the production and optimization of xylanase through Solid-State Fermentation (SSF) using different agricultural wastes by   Aspergillus spp. The Plackett Burman (PB) design was used to screen significant media components affecting the xylanase production. The carbon sources screened were wheat bran, rice bran, sugarcane bagasse, corn cob, and orange peel. The nitrogen sources screened were yeast extract, peptone, (NH4)2SO4, Na2NO3, and urea. Also, nine different salts such as KCl, MgSO4, Na2HPO4, CaCl2, FeSO4, ZnSO4, Na2CO3, KH2PO4, and NaH2PO4 which act as trace elements were screened. The results showed that wheat bran, yeast extract, Na2NO3 and KCl are the significant factors that affect xylanase production. A 33 Full Factorial Design (FFD) was performed to optimize the significant media components (wheat bran, KCl, yeast extract) obtained from PB design using Response Surface Methodology (RSM). Statistical analysis of results showed that wheat bran, KCl, yeast extract, and interaction between wheat bran and yeast extract were found to be significant. The optimum concentration of wheat bran, KCl, yeast extract was 8 g/L, 0.1 g/L and 3 g/L. The Partial purification of xylanase was carried out using ammonium salt precipitation and dialysis. Gel filtration chromatography was performed to optimize the elution time, which was found to be 6 minutes. Application of xylanase in orange juice clarification was studied at 40 °C, 50 °C, and 60 °C. The optimum temperature obtained was 60 ºC.

Microbial lipases are the biocatalyst of choice for the present and future because of their characteristics, including their ability to remain active as an enzyme throughout a broad pH, temperature, and substrate range. The goal of the current investigation was to find novel sources of substrates and isolates from soil contaminated by oil for the synthesis of lipase. On tributyrin media, 10 lipolytic bacterial strains that were isolated from oil-contaminated soil were grown. Using the zone of clearance, it was possible to identify the isolates with the highest activity. Following phylogenetic tree analysis, molecular characterization of the 16S rRNA sequence of the bacterial isolates revealed that it was Bacillus halotolerans (VSH 09). The enzyme was purified to near homogeneity. The enzyme activity was found to be optimum at a pH of 7.0 and a temperature of 35 °C. While Ni2+ and Cu2+ had no effect, the presence of Mg2+ and Ca2+ exhibited the highest levels of enzyme activity. At 1%, tributyrin as a substrate exhibited its highest level of activity. The molecular weight, as determined by SDS-PAGE, was found to be 38 kDa. The kinetics of the enzyme were found to be 41.66 and 9.37 mg/mL for Vmax and Km, respectively. The high yield of lipase produced by this method suggests that it holds potential for production on a large scale and could be used for various biotechnological applications.

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