Children learn through play, and they learn more than adults might expect. Science experiments are a great way to spark their curiosity, Recommanded Product: 31643-49-931643-49-9, Name is 4-Nitrophthalonitrile, SMILES is C1=C(C(=CC=C1[N+](=O)[O-])C#N)C#N, belongs to nitriles-buliding-blocks compound. In a article, author is Wu, Zheming, introduce new discover of the category.
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.
Note that a catalyst decreases the activation energy for both the forward and the reverse reactions and hence accelerates both the forward and the reverse reactions. you can also check out more blogs about 31643-49-9. Recommanded Product: 31643-49-9.