With the decrease of fossil based fuels and the environmental impact of them over the planet, it seems necessary to seek the sustainable sources of clean energy. Biofuels, is becoming a worldwide...

In this study a strategy based on using signal sequence and statistically optimized medium was applied for efficient extracellular production of the recombinant form of the therapeutic enzyme l-asparaginase. l-Asparaginase from E. coli is an important enzyme used in the cancer treatment, which has been produced and studied previously by other researchers. Nevertheless, to date no study has investigated the extracellular production of recombinant l-asparaginase in the culture medium optimized using response surface methodology. For this purpose, in this study at first, a complete gene of l-asparaginase II with its own signal sequence from a locally isolated E. coli was cloned and expressed in E. coli BL21 (DE3). Subsequently, the production of the active form of recombinant l-asparaginase was evaluated by asparaginase activity assay in the culture media 4 h after induction with IPTG in 250 mL shake flasks to evaluate the effect of eleven nutrient factors on the extracellular l-asparaginase production. Statistical design of experiments was employed in the screening stage and the most effective nutrients were selected for further optimization using central composite face design. Analysis of the results revealed a mature protein with correct N-terminal amino acids of l-asparaginase II in the culture medium. The highest enzyme activity of 17,386 U/L was resulted in the optimized medium consisting of 7.75 g/L tryptone, 9 g/L yeast extract, 5.25 g/L peptone and 0.6 g/L calcium chloride at shake flask level.

The pharmaceutical research sector has been facing the challenge of neurotherapeutics development and its inherited high-risk and high-failure-rate nature for decades. This hurdle is partly attributable to the presence of brain barriers, considered both as obstacles and opportunities for the entry of drug substances. The blood–cerebrospinal fluid (CSF) barrier (BCSFB), an under-studied brain barrier site compared to the blood–brain barrier (BBB), can be considered a potential therapeutic target to improve the delivery of CNS therapeutics and provide brain protection measures. Therefore, leveraging robust and authentic in vitro models of the BCSFB can diminish the time and effort spent on unproductive or redundant development activities by a preliminary assessment of the desired physiochemical behavior of an agent toward this barrier. To this end, the current review summarizes the efforts and progresses made to this research area with a notable focus on the attribution of these models and applied techniques to the pharmaceutical sector and the development of neuropharmacological therapeutics and diagnostics. A survey of available in vitro models, with their advantages and limitations and cell lines in hand will be provided, followed by highlighting the potential applications of such models in the (neuro)therapeutics discovery and development pipelines.