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- Title
Simultaneous Determination of Bulky Imides and Their Hydrolyzed Products by D-Hydantoinase with Cyclic-imide-hydrolyzing Activity using HPLC.
- Authors
Niu, L.-X.; Xie, L.-L.
- Abstract
A novel HPLC method for simultaneous detecting bulky imides and their hydrolyzed products by D-hydantoinase was established. Phthalimide (PI) and 2,3-pyridinedicarboximide (PDI) can be regiospecifically hydrolyzed by engineered strain pET3a-hyd/Escherichia coli BL21(DE3) expressed D-hydantoinase into phthalamic acid (PA) and 3-carbamoyl-α-picolinic acid (α-3CP). The processes were analyzed by HPLC with H2O-acetonitrile as mobile phase containing 0.1% (v/v) trifluoroacetic acid at 254 nm (PDI, α‑3CP and β-2CP) and 220 nm (PI and PA) detector wavelength, respectively. The specific activities of the engineered strain account for 0.15 U/(mL × 10 OD600) to PI and 0.61 U/(mL × 10 OD600) to PDI.
- Subjects
HIGH performance liquid chromatography; IMIDES; TRIFLUOROACETIC acid
- Publication
Applied Biochemistry & Microbiology, 2022, Vol 58, Issue 1, p57
- ISSN
0003-6838
- Publication type
Article
- DOI
10.1134/S0003683822010070