title: YtjE from Lactococcus lactis IL1403 Is a C-S lyase with alpha, gamma-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 alpha,gamma-elimination activity toward L-methionine. In addition, gas chromatographic-mass spectrometry analysis showed that YtjE activity resulted in the formation of H2S 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.
authors.affiliation: Department of Dairy Science and Technology, Instituto del Frío, C/José Antonio Novais 10, Ciudad Universitaria, Madrid 28040, Spain. ifrcm94@if.csic.es
authors.forename: M Carmen
authors.initials: MC
authors.lastname: Martínez-Cuesta
authors.forename: Carmen
authors.initials: C
authors.lastname: Peláez
authors.forename: John
authors.initials: J
authors.lastname: Eagles
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Gasson
authors.forename: Teresa
authors.initials: T
authors.lastname: Requena
authors.forename: Sean B
authors.initials: SB
authors.lastname: Hanniffy
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013437:Carbon-Sulfur Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002611:Cheese / Q000382:microbiology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D013294:Lactococcus lactis / Q000201:enzymology* / Q000235:genetics; D008715:Methionine / Q000378:metabolism*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013457:Sulfur Compounds / Q000378:metabolism; D014835:Volatilization
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D013457:Sulfur Compounds; D008715:Methionine; D013437:Carbon-Sulfur Lyases