Three-Step Purification and Characterization of Organic Solvent-Tolerant and Alkali-Thermo-Tolerant Xylanase from <i>Bacillus paramycoides</i> T4 [MN370035]

In the present study, an extracellular alkali-thermo-tolerant xylanase from Bacillus paramycoides was produced in the presence of an organic solvent. The enzyme was purified by ammonium sulphate precipitation, gel filtration, and ion exchange chromatography, with an overall recovery of 25.9%. The purified enzyme hada 70 kDa molecular weight (MW) confirmed by SDS-PAGE gel analysis. The maximum enzyme activity was reported at 55 °C and pH 7.0. Xylanase activity and stability were improved in the presence of 30% (v/v) n-dodecane, iso-octane, n-decane, and cyclohexane (7 days). The enzyme activity was improved by Co2+, EDTA, and Triton-X-100 while vigorously repressed by Hg2+ and Cu2+. The purified enzyme showed 1.473 mg/mL Km and 654.017 µg/mL/min Vmax values. The distinctive assets of the isolate verified the potential application in the field of biomass conversion into fuel and other industrial processes. Organic solvent-tolerant xylanases can be used for concurrent saccharification and bioethanol production, the amplification of intoxicating beverages, and the fermenting industry

Click Biotinylation of PLGA Template for Biotin Receptor Oriented Delivery of Doxorubicin Hydrochloride in 4T1 Cell-Induced Breast Cancer

PLGA was functionalized with PEG and biotin using click chemistry to generate a biotin receptor targeted copolymer (biotinylated–PEG-PLGA) which in turn was used to fabricate ultrafine nanoparticles (BPNP) of doxorubicin hydrochloride (DOX) for effective delivery in 4T1 cell induced breast cancer. However, adequate entrapment of a hydrophilic bioactive like DOX in a hydrophobic polymer system made of PLGA is not usually possible. We therefore modified a conventional W/O/W emulsion method by utilizing NH₄Cl in the external phase to constrain DOX in dissolved polymer phase by suppressing DOX’s inherent aqueous solubility as per common ion effect. This resulted in over 8-fold enhancement in entrapment efficiency of DOX inside BPNP, which otherwise is highly susceptible to leakage due to its relatively high aqueous solubility. TEM and DLS established BPNP to be sized below 100 nm, storage stability studies showed that BPNP were stable for one month at 4 °C, and <i>in vitro</i> release suggested significant control in drug release. Extensive <i>in vitro</i> and <i>in vivo</i> studies were conducted to propound anticancer and antiproliferative activity of BPNP. Plasma and tissue distribution study supplemented by pertinent <i>in vivo</i> fluorescence imaging mapped the exact fate of DOX contained inside BPNP once it was administered intravenously. A comparative safety profile via acute toxicity studies in mice was also generated to out rightly establish usefulness of BPNP. Results suggest that BPNP substantially enhance anticancer activity of DOX while simultaneously mitigating its toxic potential due to altered spatial and temporal presentation of drug and consequently deserve further allometric iteration