Rational design of a new variant of Reteplase with optimized physicochemical profile and large-scale production in Escherichia coli was written by Seyedhosseini Ghaheh, Hooria;Sajjadi, Shabnam;Shafiee, Fatemeh;Barzegari, Ebrahim;Moazen, Fatemeh;Mir Mohammad Sadeghi, Hamid. And the article was included in World Journal of Microbiology & Biotechnology in 2022.Category: alcohols-buliding-blocks The following contents are mentioned in the article:
Structural engineering of the recombinant thrombolytic drug, Reteplase, and its cost-effective production are important goals in the pharmaceutical industry. In this study, a single-point mutant of the protein was rationally designed and evaluated in terms of physicochem. characteristics, enzymic activity, as well as large-scale production settings. An accurate homol. model of Reteplase was used as the input to appropriate tools to identify the aggregation-prone sites, while considering the structural stability. Selected variants underwent extensive mol. dynamic simulations (total 540 ns) to assess their solvation profile and their thermal stability. The Reteplase-fibrin interaction was investigated by docking. The best variant was expressed in E. coli, and Box-Behnken design was used through response surface methodol. to optimize its expression conditions. M72R mutant demonstrated appropriate stability, enhanced enzymic activity (p < 0.05), and strengthened binding to fibrin, compared to the wild type. The optimal conditions for the variant′s production in a bioreactor was shown to be 37 °C, induction with 0.5 mM IPTG, for 2 h of incubation. Under these conditions, the final amount of the produced enzyme was increased by about 23 mg/L compared to the wild type, with an increase in the enzymic activity by about 2 IU/mL. This study thus offered a new Reteplase variant with nearly all favorable properties, except solubility The impact of temperature and incubation time on its large-scale production were underlined as well. This study involved multiple reactions and reactants, such as (2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1Category: alcohols-buliding-blocks).
(2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1) belongs to alcohols. A strong base can deprotonate an alcohol to yield an alkoxide ion (R―O−). For example, sodamide (NaNH2), a very strong base, abstracts the hydrogen atom of an alcohol. Converting an alcohol to an alkene requires removal of the hydroxyl group and a hydrogen atom on the neighbouring carbon atom. Dehydrations are most commonly carried out by warming the alcohol in the presence of a strong dehydrating acid, such as concentrated sulfuric acid.Category: alcohols-buliding-blocks
Referemce:
Alcohol – Wikipedia,
Alcohols – Chemistry LibreTexts