Identification and characterization of an amidase from Leclercia adecarboxylata for efficient biosynthesis of L-phosphinothricin

Abstract

L-phosphinothricin (L-PPT) is an important broad-spectrum herbicide with expanding utilization because it is environmentally benign. A strain Leclercia adecarboxylata ZJB-17008 with capability of catalyzing rac-4-(hydroxy(methyl)phosphoryl)-2-(2-phenylacetamido) butanoic acid (rac-S) to L-PPT was screened and identified, from which an amidase (La-Ami) was cloned and secretory expressed in Bacillus subtilis WB 800 for the bioproduction of L-PPT. The recombinant La-Ami exhibited an excellent enantioselectivity (99.9% ee) and remarkable thermostability with a half-life of 19.8 h at 50 °C. Furthermore, La-Ami displaying a high space-time yield of 787.2 g L−1 d−1 at 50 °C and pH 8.5 under the rac-S concentration of 500 mM (150 g L−1). The finally refined L-PPT was obtained with a purity of 99% and a total yield reached 90%. These results implying that this secretory expressed amidase La-Ami is possible to be applied in the large-scale bioproduction of L-PPT.