Journal of Molecular Catalysis B: Enzymatic p. S60 - S67 (2016)
Update date:2022-08-29
Topics:
Tao, Yanyang
Han, Chao
Wang, Min
Yao, Peiyuan
Yuan, Jing
Feng, Jinhui
Wu, Qiaqing
Zhu, Dunming
An efficient biosynthesis of β-alanine from 3-aminopropionitrile at high concentration has been developed using a one-pot bienzymatic cascade of a nitrilase and an amidase. The nitrilase BjNIT3397 from Bradyrhizobium japonicum strain USDA110 catalyzes the hydrolysis of 3-aminopropionitrile to β-alanine at the concentration up to 3.0 mol/L with the formation 23% of 3-aminopropanamide. In order to eliminate the by-product 3-aminopropanamide, we cloned and characterized a new amidase from Pseudomonas nitroreducens through gene mining. Under the optimal conditions (50 mmol/L Na2HPO4-NaH2PO4 buffer, pH 6.0, 40 °C), 2.0 mol/L (176 g/L) of 3-aminopropanamide was completely hydrolyzed within 12 h. A tandem reaction system was then established to eliminate the by-product 3-aminopropanamide and increase the production of β-alanine to 90% isolated yield with 15.02 g/(L.h) space-time-yield. These results demonstrated that the tandem reaction strategy was an effective method of eliminating the amide by-products in the nitrilase-catalyzed hydrolysis at high substrate concentration.
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