A Novel Sulfatase for Acesulfame Degradation in Wastewater Treatment Plants as Evidenced from Strains.

The artificial sweetener acesulfame is a persistent pollutant in wastewater worldwide. So far, only a few bacterial isolates were recently found to degrade acesulfame efficiently. In and strains, a Mn-dependent metallo-β-lactamase-type sulfatase and an amidase signature family enzyme catalyze acesulfame hydrolysis via acetoacetamide--sulfonate to acetoacetate. Here, we describe a new acesulfame sulfatase in strains isolated from wastewater treatment plants in Germany. Their genomes do not encode the Mn-dependent sulfatase. Instead, a formylglycine-dependent sulfatase gene was found, together with the acetoacetamide--sulfonate amidase gene on a plasmid shared by all known acesulfame-degrading strains. Heterologous expression, proteomics, and size exclusion chromatography corroborated the physiological function of the sulfatase in acesulfame hydrolysis. Since both acesulfame sulfatase types are absent in other bacterial genomes or metagenome-assembled genomes, we surveyed 73 tera base pairs of wastewater-associated metagenome raw data sets. / sulfatase gene signatures were regularly found from 2013, particularly in North America, Europe, and East Asia, whereas sulfatase gene signatures were first detected in 2020. Moreover, signatures for the sulfatase and amidase genes co-occur only in six data sets from China, Finland, and Mexico, suggesting that the genes were enriched or introduced quite recently in wastewater treatment facilities.