Purpose: Recent advances in nanotechnology have given rise to the potential utilization of nanoparticles as food, nano-medicine/biomedicines. Patient: The study aimed to investigate the effects of nano-zinc oxide (nano-zinc) on the bio-assimilation of mineral (Zn) in mice, aged 3– 6 weeks. Methods: ZnO nanoparticles were added to the basal diet as a supplement at amounts of 0.07, 0.14 and 0.21 mg/kg. The synthesized material was characterized by Fourier transform infrared spectrophotometer, particle size, scanning electron microscope, Thermogravimetric Analysis Thermal, X-ray diffraction spectrophotometer and Zeta potential. Results: In-vitro bioavailability of synthesized group ZnO (120 nm) was 43%, whereas for standard group ZnO (50 nm) was reported as 55%. In-vivo bioavailability of zinc oxide illustrated the maximum absorption level compared with the control. In-vivo toxicity was characterized as damage done to the liver and spleen tissues with a high dose of 0.21 mg/kg, while smaller doses indicated no toxic effects. Conclusion: The study provided important insights on the toxicological effects of ZnO nanoparticles, depending on dose rate and bio-assimilation, as well as particles, under various conditions (in-vitro and in-vivo). These findings will motivate further detailed research on nano-based medicine for alleviating malnutrition conditions.

Elaeagnus umbellata, a member of the Elaeagnaceae family, is native to Pakistan, China, India, Korea, and Japan. It is found commonly at altitudes ranging from 1200 to 2100 m and thrives on eroded and degraded land due to its ability to fix nitrogen. The plant also grows under variable pH (4–8) and drought, and is used locally as fuel wood, fencing, fodder, basket making, and shelterbelts. The fruit of the plant is well known for its essential nutrients and medicinal compounds such as vitamins, minerals, essential fatty acids, carotenoids (lycopene), soluble solids, and sugars. Medicinally, it is widely believed to protect against myocardial infections, pulmonary infections, and various forms of cancers. Ten ecotypes from variable microclimatic conditions were investigated for their morphological, molecular and biochemical diversity improvement and commercialization purposes. Comparisons and disabilities indicated significant variability in terms of morphological (plant height, number of branches, thorn size and number, leaf area, fruit size, 100 fruit weight, and yield), molecular (SDS-PAGE), and micro- and macronutrient (vitamin C, Fe, mg, P, Na, K, essential oils, and sugar) bases among the ecotypes. This variability will be helpful in developing commercial varieties of the plant utilizing the conventional techniques of selection and hybridization for economic activities. The plant has ample quantities of multiple micronutrients, thus indicating their expression through a powerful promoter at one place (fruit mesocarp). Efforts to identify and isolate the micronutrient genes (vitamin A, C, E, and Fe), the deficiency of which causes malnutrition and disabilities within the population of developing countries. Micronutrient genes have also been initiated for their characterization and future transformation into staple food crops for stable bio-fortification.

Pectinolytic enzymes or pectinases are synthesized naturally by numerous microbes and plants. These enzymes degrade various kinds of pectin which exist as the major component of the cell wall in plants. A pectinase gene encoding endo-polygalacturonase (endo-PGase) enzyme was isolated from Pectobacterium carotovorum a plant pathogenic strain of bacteria and successfully cloned into a secretion vector pHT43 having σ^A-dependent promoter for heterologous expression in Bacillus subtilis (WB800N).The desired PCR product was 1209bp which encoded an open reading frame of 402 amino acids. Recombinant proteins showed an estimated molecular weight of 48 kDa confirmed by sodium dodecyl sulphate–polyacrylamide-gel electrophoresis. Transformed B. subtilis competent cells harbouring the engineered pHT43 vector with the foreign endo-PGase gene were cultured in 2X-yeast extract tryptone medium and subsequently screened for enzyme activity at various temperatures and pH ranges. Optimal activity of recombinant endo-PGase was found at 40°C and pH 5.0. To assay the catalytic effect of metal ions, the recombinant enzyme was incubated with 1 mM concentration of various metal ions. Potassium chloride increased the enzyme activity while EDTA, Zn⁺⁺ and Ca⁺⁺, strongly inhibited the activity. The chromatographic analysis of enzymatic hydrolysates of polygalacturonic acid (PGA) and pectin substrates using HPLC and TLC revealed tri and tetra-galacturonates as the end products of recombinant endo-PGase hydrolysis. Conclusively, endo-PGase gene from the plant pathogenic strain was successfully expressed in Bacillus subtilis for the first time using pHT43 expression vector and could be assessed for enzyme production using a very simple medium with IPTG induction. These findings proposed that the Bacillus expression system might be safer to escape endotoxins for commercial enzyme production as compared to yeast and fungi. Additionally, the hydrolysis products generated by the recombinant endo-PGase activity offer their useful applications in food and beverage industry for quality products.

SweetpotatoIpomoea batatasis known as a hexaploid species. Here, we analyzed carotenoids contained in the leaves and tubers of sweetpotato cultivars ‘White Star’ (WS) and W71. These cultivars were found to contain several carotenoids unique to sweetpotato tubers such as β-carotene-5,6,5′,8′-diepoxide and β-carotene-5,8-epoxide. Next, we isolated two kinds of carotene cyclase genes that encode lycopene β- and ε-cyclases from the WS and W71 leaves, by RT-PCR and subsequent RACE. Two and three lycopene β-cyclase gene sequences were, respectively, isolated from WS, namedIbLCYb1,2, and from W71,IbLCYb3,4,5. Meanwhile, only a single lycopene ε-cyclase gene sequence, designatedIbLCYe, was isolated from both WS and W71. These genes were separately introduced into a lycopene-synthesizingEscherichia colitransformed with thePantoea ananatis crtE,crtBandcrtIgenes, followed by HPLC analysis. β-Carotene was detected inE. colicells that carriedIbLCYb1-4, indicating that theIbLCYb1-4genes encode lycopene β-cyclase. Meanwhile, the introduction ofIbLCYeinto the lycopene-synthesizingE. coliled to efficient production of δ-carotene with a monocyclic ε-ring, providing evidence that theIbLCYegene codes for lycopene ε-(mono)cyclase. Expression of the β- and ε-cyclase genes was analyzed as well.

Endopolygalacturonase (EndoPGase), EC: 3.2.1.15. is one of the crucial pectinases belonging to the class of carbohydrase. The catalytic action of EndoPGase captivates the attention of the production of this extremely valuable catalyst in the industrial sector. The main focus was to ascertain a potential bacterial candidate for endoPGase production. The isolated bacterial strain was further identified by 16S rRNA gene sequencing. The parameters for enzyme biosynthesis were optimized by a single and multiple factor approach at a time. The results of our investigation led to the identification of a potent strain of Bacillus subtilis NR2 [strain 168]. The sequence of 16S rRNA amplified from the isolated bacterium has been submitted to GenBank under accession number ON738697. The strain was found active for pectic enzyme activity under shaking- flask fermentation at pH 5.0 and 50 °C temperature of incubation. Among all monomeric and polymeric substrates (inducer-substrates), citrus pectin, followed by potato starch and pectin (Sigma) were considered the best enzyme inducers at 1% concentration. In comparison, an increased wheat bran concentration at 5% was proved to be ideal for EndoPGase biosynthesis Moreover, an increasing trend in enzyme activity was observed with the increasing concentration of inducer. The combined effect of three variables (pH, inducer-substrates, and inducer-substrate concentration) was explored by response surface methodology (RSM) involving a Box–Behnken design (BBD). Based on the results, we concluded that the soil-isolated B. subtilis can be effectively utilized for commercial-scale pectinase enzyme biosynthesis.