Lixi Niu, Shishi Ma
Apr 8, 2018
Citations
0
Influential Citations
0
Citations
Journal
Se pu = Chinese journal of chromatography
Abstract
A high performance liquid chromatography (HPLC) method was established for the detection of 2, 3-pyridinedicarboximide (PDI) and its enzyme reaction products, 3-carbamoyl-α-picolinic acid (α-3CP), using an engineering strain containing the D-hydantoinase gene expression box.The strain pET3a-hyd/BL21(DE3) was collected after induction and added to a PDI saturated aqueous solution.After reacting at 37℃ for 30 min with constant stirring, the supernatant was separated by centrifugation at 13000 r/min and detected by HPLC.The chromatographic conditions were as follows:HypersilTM GOLD C18 column (250 mm×4.6 mm, 5 μm), H2O-acetonitrile (90:10, v/v) containing 0.1%(v/v) trifluoroacetic acid as the mobile phase with a flow rate of 1 mL/min and a detection wavelength of 254 nm.The specific activity of pET3a-hyd/BL21(DE3) was found to be 0.61 U/(mL·10OD600 nm).This study provides a theoretical basis for the preparation of complicated half-amides using biological methods.