YtjE from Lactococcus lactis IL1403 Is a C-S Lyase with α,γ-Elimination Activity toward Methionine

Abstract

Cheese microbiota and the enzymatic conversion of methionine to volatile sulfur compounds (VSCs) are important factors in flavor formation during cheese ripening and the foci in biotechnological approaches to flavor improvement. The product of ytjE of Lactococcus lactis IL1403, suggested to be a methionine-specific aminotransferase based on genome sequence analysis, was therefore investigated for its role in methionine catabolism. The ytjE gene from Lactococcus lactis IL1403 was cloned in Escherichia coli and overexpressed and purified as a recombinant protein. When tested, the YtjE protein did not exhibit a specific methionine aminotransferase activity. Instead, YtjE exhibited C-S lyase activity and shared homology with the MalY/PatC family of enzymes involved in the degradation of l -cysteine, l -cystine, and l -cystathionine. YtjE was also shown to exhibit α,γ-elimination activity toward l -methionine. In addition, gas chromatographic-mass spectrometry analysis showed that YtjE activity resulted in the formation of H ₂ S from l -cysteine and methanethiol (and its oxidized derivatives dimethyl disulfide and dimethyl trisulfide) from l -methionine. Given their significance in cheese flavor development, VSC production by YtjE could offer an additional approach for the development of cultures with optimized aromatic properties.