Chemistry is the science of change. But why do chemical reactions take place? Why do chemicals react with each other? The answer is in thermodynamics and kinetics, HPLC of Formula: C10H26N4, 71-44-3, Name is Spermine, SMILES is NCCCNCCCCNCCCN, belongs to amides-buliding-blocks compound. In a document, author is Yuan, Bingbing, introduce the new discover.
High-level expression of nitrile hydratase from Pantoea sp. At-9b in Escherichia coli
Nitrile hydratase (NHase, EC 4.2.1.84) catalyzes the hydration of nitrites to corresponding amides, which was widely used in pharmaceuticals and agrochemicals. In recent years, hetemlogous expression of NHase in Escherichia coli cells has attracted considerable attentions in industrial applications. However, it suffers from drawbacks including inclusion bodies and unbalanced expression of subunits. In this work, a novel NHase (NHasePs) was exploited from Pantoea sp. AT-9b for efficient hetemlogous expression in E. coli cells. Through simulation and optimization of secondary structure of the translation initiation region, the expression level of beta-subunit was significantly improved. The highest overexpression was obtained by NHasePsM2-2 with 2.6-fold higher than that of NHasePs, and the enzyme activity was increased by 8.8 folds. Subsequently, the inclusion body of alpha-subunit was eliminated by introduction of fusion tags. The expression of alpha-subunit of NHasePsM (PGB1 alpha) was 3.2-fold higher than that of NHasePsM2-2. The enzyme activity was improved by 2.1 folds, reaching 558 U/mg DCW. Using a cell loading of 3.0 g/L, accumulation of acrylamide reached 202 g/L. This study is of great significance for promoting the hetemlogous expression of NHase in E. coli system and its practical industrial application.
A reaction mechanism is the microscopic path by which reactants are transformed into products. Each step is an elementary reaction. In my other articles, you can also check out more blogs about 71-44-3. HPLC of Formula: C10H26N4.