RT Journal Article SR Electronic(1) A1 Alonso, Jorge A1 Garc�a, Jos� L.YR 1996 T1 Proline iminopeptidase gene from Xanthomonas campestris pv. citri JF Microbiology, VO 142 IS 10 SP 2951 OP 2957 DO https://doi.org/10.1099/13500872-142-10-2951 PB Microbiology Society, SN 1465-2080, AB The pip gene coding for the proline iminopeptidase (Pip) of Xanthomonas campestris pv. citri was cloned in an Escherichia coli leuB strain using a selective medium containing the dipeptide d-Ala-l-Leu as the sole source of l-leucine. Nucleotide sequencing of this gene revealed a 939 bp open reading frame encoding a 312 amino acid protein (35 126 Da). The deduced amino acid sequence showed 47% identity with the Pip from Neisseria gonorrhoeae. A lacZ-pip fusion gene was overexpressed in E. coli under the control of the Plac promoter. The Pip of X. campestris hydrolysed l-prolyl-p-nitroanilide with the highest efficiency, but was also able to hydrolyse l-alanyl-p-nitroanilide and d-alanyl-p-nitroanilide. The molecular mass of Pip was found to be 35 kDa by SDS-PAGE and 120 kDa by gel filtration, suggesting that the active enzyme is a multimer., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/13500872-142-10-2951