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INTRODUCTION: Withania somnifera (W. somnifera) is a plant with remarkable pharmacological properties. The plant has an impressive profile of medicinal uses in the folk medicine system of several civilizations. AIM: This comprehensive study is aimed to characterize phytochemicals in fruit of W. somnifera and tested for in vitro anticancer potential to find out active candidate in disease prevention and treatment. METHODS: The bioactive components from W. somn-ifera fruit were extracted with polar and non-polar solvents. Anticancer potential of the isolated bioactive was assessed against different cancer cell lines through MTT assay and Incucytes imaging analysis. The extracts were characterized for secondary metabolites using GC-MS (gas chromatography-mass spectrometer), LCMS (liquid chromatography-mass spectrometry)-ESI (electrospray Ionization) and 1H-NMR (electrospray Ionization) techniques. RESULTS: Both freeze-dried and rotary evaporator con-densed extracts exhibited anticancer potential against MDA-MB-231, MCF7- SKOV3 and SKBR3 cell lines. The tested extracts have cell growth inhibition potential ag-ainst mammalian cancer cell line. Hexacosanedioic acid purified from n-hexane extract through HPLC was inves-tigated for its cytotoxicity against breast cancer cell line SKBR3 by using Incucytes imaging analysis. Conclusion: We found that a variety of bioactive compounds existed in this plant. One identified compound that was not investigated for cytotoxicity in previous studies was purified and its application showed cytotoxicity on breast cancer cell lines. A number of bioactive identified from W. somnifera fruit may have an effective potential for development into chemotherapy drugs.

Lipases belong to α/β hydrolases that cause hydrolytic catalysis of triacylglycerols to release monoacylglycerols, diacylglycerols, and glycerol with free fatty acids. Lipases have a common active site that contains three amino acid residues in a conserved Gly-X-Ser-X-Gly motif: a nucleophilic serine residue, an acidic aspartic or glutamic acid residue, and a basic histidine residue. Lipase plays a significant role in numerous industrial and biotechnological processes, including paper, food, oleochemical and pharmaceutical applications. However, its instability and aqueous solubility make application expensive and relatively challenging. Immobilization has been considered as a promising approach to improve enzyme stability, reusability, and survival under extreme temperature and pH environments. Innumerable supporting material in the form of natural polymers and nanostructured materials is a crucial aspect in the procedure of lipase immobilization used to afford biocompatibility, stability in physio-chemical belongings, and profuse binding positions for enzymes. This review outlines the unique structural and functional properties of a large number of polymers and nanomaterials as robust support matrices for lipase immobilization. Given these supporting materials, the applications of immobilized lipases in different industries, such as biodiesel production, polymer synthesis, additives, detergent, textile, and food industry are also discussed.

This study appraises functional and pharmacological attributes and phenolic bioactive compounds profile of various solvent (distilled H2O, 50% C2H5OH, 100% n-hexane, 100% ethyl acetate, 100% CH3OH, 100% chloroform) extracts from fruits and leaves of Natal plum (Carissa grandiflora). The antioxidant activity of the extracts was analyzed by the determinations of total flavonoid contents, total phenolic contents, DPPH radical scavenging capacity and ferric reducing antioxidant power assay. Additionally, anti-inflammatory, anti-cancer, anti-bacterial and individual phenolics composition of the tested extracts were also investigated. The experimental results revealed that the highest percent bioactive compounds extract yield (35.01%) was obtained from fruits with 50% distilled water while lowest (7.2%) from leaves with chloroform. The tested fruit extracts exhibited better antioxidant activity as compared to leaves extracts. Total phenolic and total flavonoid contents in various solvents extracts of fruit and leaves varied from 8.7–143.08, 3.1–96.02 GAE (mg/g) and 6.52–62, 3.6–28.84 CE DW (mg/g) while the levels of total individual phenolic compounds, identified by High Performance Liquid Chromatography, were 400–3500 and 600–4500 mg/g, respectively. Pure (100%) methanolic extract from leaves showed highest activity (10.11 mm inhibition zone diameter) against B. subtilis whereas 50% ethanolic extract from fruits was most potent (8.56 mm) against E. coli. In case of anti-inflammatory activity, 100% CH3OH extract from leaves showed better activity followed by 100% ethyl acetate extract. The high anticancer activity was displayed by 100% chloroform extract from fruits (percent inhibition 89.16%; IC50 2.4 ± 0.09) against MCF-7 cell lines in MTT colorimetric assay. The solvents polarity-based differences were observed in terms of quantifiable yield and activities of the extracted bioactive compounds. In the light of present results, methanol and chloroform extracts from fruits and leaves of natal plum can be explored as potential sources of potent bioactive compounds for nutra-pharmaceutical developments.

The present study explores pharmacological potential and phytochemicals profiling of Picrorhiza kurroa extracts against mammalian cancer cell lines and pathogenic microbes. Bioactive extracts from roots of Picrorhiza kurroa were recovered in the methanol, 50% aqueous dichloromethane (50 : 50 v/v) and n-hexane. Antimicrobial activity of the bioactive extracts was assessed against selected strains of bacteria and pathogenic fungi. Aqueous dichloromethane extract showed highest zone of growth inhibition (39.06 ± 1.0 mm) towards Staphylococcus aureus bacteria while methanolic extract showed the lowest inhibition (6.3 ± 4.1 mm) to Escherichia coli bacteria. The tested extracts such as methanol and aqueous dichloromethane exhibited higher inhibition antifungal activity against Aspergillus flavus compared to Fusarium oxysporum. As far as cytotoxicity (MTT assay) of the tested extracts is concerned, n-hexane and aqueous dichloromethane extracts were found to be very active against all cancer cell lines (breast cancer MCF7, MDA-MB-231, SKBR3 and ovarian cancer SKOV3). A preliminary phytochemicals profiling was performed in extracts using GC-MS. Several fractions of active extract were separated with HPLC and analyzed using High Resolution Atmospheric Pressure Chemical Ionization Mass Spectrometry (HR-APCI-MS). Two purified compounds (Dihydromikanolide and 1,3-Dicyclohexyl-4-(cyclohexylimino)-2-(cyclohexylethylamino)-3,4-dihydro-1,3-diazetium) were further evaluated for their anticancer activity against ovarian cancer cell line. Our findings depict that all the tested extracts showed considerable anticancer potential through cell viability assays. The purified compound 1 - Dihydromikanolide from methanolic extract was found to be active against ovarian cancer cells and can be explored as a promising nutra-pharmaceutical candidate against ovarian cancer. However, further studies exploring the molecular pathways and in vivo testing are required.

ancer is the leading cause of death, accounting for approximately one out of six people dying with this disease worldwide. Among all, the breast and ovarian cancers are top-ranked causes of women mortalities compared to other disorders. Although, there is advancement in technologies, but still, there are unresolved concerns to overcome the global disease burden. Currently, plants are being explored as a natural remedy to cure disorders. This research was planned to explore phytochemicals in methanolic extracts of Zizyphus mauritiana and Triticum aestivum, and their pharmacological activities were studied through Agrobacterium tumefaciens bacteria, in vitro breast cancer cell line and ovarian cancer cell line to find out novel candidates in disease control and prevention. Eleven different types of bioactive compounds were analysed in the tested extracts. The highest crude extracts percentage (75±0.02) was observed with Z. mauritiana. The extracts showed promising cell growth inhibition and tumor initiation inhibition in potato disc assay. MTT assay and Incucytes imaging analysis revealed that Z. mauritiana extract had a higher anticancer potential with 40 ± 0.92 cell viability against breast cancer cells (SKBR3) and 45 ±0.29 against ovarian cancer cells (SKOV3). In conclusion, these extracts could be used as chemotherapeutics owing to their cheapness, and easy availability. While detailed study is required for further purification and characterization of bioactives/target compounds and in-vivo activity confirmations.

Qudsia Tabassam,1 Tahir Mehmood,1,2 Abdul Rauf Raza,1 Azmat Ullah,3 Farhan Saeed,4 Faqir Muhammad Anjum5 1Department of Chemistry, University of Sargodha, Sargodha, Pakistan; 2Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences-UVAS, Lahore, Pakistan; 3Department of Food Science and Human Nutrition, University of Veterinary and Animal Sciences-UVAS, Lahore, Pakistan; 4Institute of Home & Food Sciences, Government College University Faisalabad, Faisalabad, Pakistan; 5Vice Chancellor Secretariat, University of the Gambia, Banjul, The GambiaCorrespondence: Tahir Mehmood; Faqir Muhammad Anjum Email tahir.mehmoodbiochem@uvas.edu.pk; dranjum@utg.edu.gmBackground: Nanotechnology is gaining emerging interest in advanced drug discovery therapeutics due to their tremendous properties including enhanced delivery of therapeutic payload, extensive surface to volume ratio, high permeability, retention behaviors, etc. The gold nanoparticles (AuNPs) are favored due to their advanced features, such as biogenic, tunable physiochemical response, ease in synthesis, and wide range of biomedical applications. The phytochemicals have been focused to design Au nano-carrier-based conjugation for active-targeting drug delivery due to their nano conjugation ability.Aim: The present study describes the facile synthesis of 20nm spherical AuNPs and their conjugation with reported anti-cancer phytocompound Withanolide-A (1).Methods: The 20nm sAuNPs were synthesized chemically and characterized their phytochemical gold nanoconjugates through UV-visible spectroscopy, dynamic light scattering (DLS) and transmission electron microscopy (TEM) imaging techniques. The anti-cancer therapeutic potentials were tested with both nanoconjugates and pure WithanolideA (1) by using SKBR3 breast cancer cells line.Results: The synthesized sAuNPs showed significant conjugation with Withanolide-A and showed stability. Furthermore, these Au nanoconjugates with Withanolide-A (1) significantly induce blockage of SKBR3 cell growth at half maximal active concentration that compared to pure Withanolide-A (1).Conclusion: Our findings provide a foundation to further progress how they can overcome cancer drug resistance by conjugating active drugs in combination with AuNPs through optimizing the effective drug concentration and removing the surface barrier.Keywords: Withania somnifera, Withanolide-A, gold nanoparticle; AuNP, breast cancer, anti-cancer and incucyte

Pectinase is an important group of industrial enzymes. Pectinase manufacturing occupies about 10% of the overall enzyme production world over. The aim of this study is to produce pectin lyase from Schizophyllum commune using the mosambi (sweet lime) fruit peels as substrate in solid state fermentation. The cultural parameters optimized through response surface methodology showed maximum pectin lyase production of 480.45 U/mL at initial medium pH of 6, incubation temperature of 35 °C, time period of 1 day, substrate concentration of 15 g and 3 mL of inoculum size. A purification fold of 3.08 with 355 U/mg specific activity and 4.16% yield was obtained after purification. Enzyme immobilization was done by entrapment with sodium alginate and adsorption with chitosan. Chitosan immobilized enzyme exhibited best thermal stability in the range of 45–55 °C and pH 8.0–9.0. Enzyme activity was stimulated in the presence of Ca2+ and Mg2+ while EDTA inhibited the enzyme activity. Chitosan immobilized pectin lyase was stable up to six cycles of reuse. The pH and thermal stability of S. commune pectin lyase makes it an important enzyme for industrial use. The results showed that pectin lyase produced from S. commune has significant potential for applications in the detergent and fruit juice industry. The enzyme produced from citrus agro waste via the proposed optimized biotechnological process can be explored for multiple industrial applications.

The aim of this study was to use the under-utilized vegetable seed oil and produce high and better yield of biodiesel. The response surface methodology (RSM) was used, based on central composite rotatable design (CCRD), to optimize four trans-esterification reaction variables for getting high yield: catalyst concentration (CC; 0.25–1.0 %), reaction temperature (RT; 45–65 °C), CH3OH-to-oil molar ratio (6:1–12:1) and reaction time (30–90 min). The 24 full factorial CCRD design was applied, using four different parameters at five levels, each lead to 30 experiments to produce Pongamia pinnata oil-methyl esters (POMEs). The molar ratio of CH3OH to oil and RT were the most significant (p