Phosphinothricin inhibits plant glutamine synthetase and is used as a herbicide. Streptomyces hygroscopicus and Streptomyces viridochromogenes, which produce phosphinothricin naturally, encode acetyltransferases that confer phosphinothricin resistance. In the Pseudomonas genome database, a number of proteins have been annotated as phosphinothricin acetyltransferases and putative phosphinothricin acetyltransferases. One such protein is PSPTO_3321 from P. syringae, a strain that causes tomato speck. Here, we reveal that PSPTO_3321 from P. syringae, termed syr_pat, is a phosphinothricin acetyltransferase, and also retains a lower level of activity against the structurally similar substrate methionine sulfoximine. We solved a 1.6 Å resolution crystal structure of syr_pat alone and a 2.5 Å resolution structure for a complex with