Pectobacteria are devastating plant pathogens that infect a large variety of crops, including members of the family Brassicaceae. To infect cabbage crops, these plant pathogens need to overcome the plant's antibacterial defense mechanisms, where isothiocyanates are liberated by hydrolysis of glucosinolates. Here, we found that a Pectobacterium isolate from the gut of cabbage root fly larvae was particularly resistant to isothiocyanate and even seemed to benefit from the abundant Brassica root metabolite 2-phenylethyl isothiocyanate as a nitrogen source in an ecosystem where nitrogen is scarce. The Pectobacterium isolate harbored a naturally occurring mobile plasmid that contained a sax operon. We hypothesized that SaxA was the enzyme responsible for the breakdown of 2-phenylethyl isothiocyanate. Subsequently, we heterologously produced and purified the SaxA protein and characterized the recombinant enzyme. It hydrolyzed 2-phenylethyl isothiocyanate to yield the products carbonyl sulfide and phenylethylamine. It was also active toward another aromatic isothiocyanate but hardly toward aliphatic isothiocyanates. It belongs to the class B metal-dependent beta-lactamase fold protein family but was not, however, able to hydrolyze beta-lactam antibiotics. We discovered that several copies of the saxA gene are widespread in full and draft Pectobacterium genomes and therefore hypothesize that SaxA might be a new pathogenicity factor of the genus Pectobacterium, possibly compromising food preservation strategies using isothiocyanates. P ectobacteria are phytopathogens that cause tuber soft rot and blackleg (stem rot) in many horticulturally and economically important crops during cultivation, transport, or storage. Most Pectobacterium spp., e.g., Pectobacterium carotovorum or P. wasabiae, can be detrimental to many different plants, such as potato, sugar beet, cabbage, wasabi, chicory, and Ornithogalum plants, whereas other Pectobacterium spp., e.g., P. atrosepticum and P. betavasulorum, have a more narrow host range of potato and sugar beet plants, respectively (1).Several pathogenicity factors of soft-rot-causing Pectobacterium spp. have been found during the last decades (2-4). Of these, plant cell wall-degrading enzymes produced by Pectobacterium spp. have a large impact on the progress of the disease, as their production leads to the degradation of invaded plant tissue (2, 3). The production of these enzymes is dependent on cell density and regulated by quorum sensing through N-acetylhomoserine lactone (5) and intracellular regulators (6, 7). Motility and nutrient uptake are also factors influencing phytopathogenicity (4,(8)(9)(10). Besides these general pathogenicity factors, phytopathogens have to overcome the toxicity of plant allelochemicals. Important allelochemicals of members of the family Brassicaceae (cabbages and mustards) are isothiocyanates that are liberated by the glucosinolate-myrosinase defense system (11-13). The antimicrobial effect of isothiocyanates has been mainly attributed to their reactivity w...