title: Substrate preference of 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase in Burkholderia thailandensis.
abstract: 5'-Methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTAN) plays crucial roles in the production of autoinducers and methionine metabolism. Putative genes encoding MTAN and AdoHcyase from Burkholderia thailandensis were cloned and characterized. The K(m)  values of MTAN for 5'-methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH) were 19 and 58 μM, respectively. The catalytic efficiency of MTAN for SAH was only 0.004% of the value for MTA, indicating an almost complete substrate preference of MTAN for MTA. The results of autoinducer-2 assay of B. thailandensis and recombinants indicated that LuxS enzyme activity was lacking in Burkholderia species. Instead, AdoHcyase hydrolysed SAH directly to homocysteine and adenosine in the activated methyl cycle. Meanwhile, the K(m)  value of AdoHcyase for SAH was determined to be 40 μM. Sequence analysis revealed that MTAN had much higher diversity than AdoHcyase, which likely contributes to its substrate preference for MTA. Furthermore, the phylogenetic tree of MTAN sequences revealed that LuxS(+)  bacteria could be discriminated from LuxS(-)  bacteria. These results suggested that the substrate preference of MTAN for MTA and SAH degradation pathway evolved with the bacterial-activated methyl cycle.
authors.affiliation: Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Gao
authors.forename: Dasheng
authors.initials: D
authors.lastname: Zheng
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Yuan
mesh_terms: D019117:Burkholderia / Q000201:enzymology*; D003001:Cloning, Molecular; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D003839:Deoxyadenosines / Q000378:metabolism*; D014644:Genetic Variation; D009699:N-Glycosyl Hydrolases / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D012435:S-Adenosylhomocysteine / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013872:Thionucleosides / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D003839:Deoxyadenosines; D013872:Thionucleosides; C008500:5'-methylthioadenosine; D012435:S-Adenosylhomocysteine; D009699:N-Glycosyl Hydrolases; C019606:adenosylhomocysteine nucleosidase