Characterization of a Salmonella sugar kinase essential for the utilization of fructose-asparagine.
Salmonella can utilize fructose-asparagine (F-Asn), a naturally occurring Amadori product, as its sole carbon and nitrogen source. Conversion of F-Asn to the common intermediates glucose-6-phosphate, aspartate, and ammonia was predicted to involve the sequential action of an asparaginase, a kinase, and a deglycase. Mutants lacking the deglycase are highly attenuated in mouse models of intestinal inflammation owing to the toxic build-up of the deglycase substrate. The limited distribution of this metabolic pathway in the animal gut microbiome raises the prospects for antibacterial discovery. We report the biochemical characterization of the kinase that was expected to transform fructose-aspartate to 6-phosphofructose-aspartate during F-Asn utilization. In addition to confirming its anticipated function, we determined through studies of fructose-aspartate analogues that this kinase exhibits a substrate-specificity with greater tolerance to changes to the amino acid (including the d-isomer of aspartate) than to the sugar.
Department of Chemistry & Biochemistry, The Ohio State University, Columbus, OH 43210, USA.|Department of Chemistry & Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
Pradip K
PK
Biswas
Department of Chemistry & Biochemistry, The Ohio State University, Columbus, OH 43210, USA.|Department of Chemistry & Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
Edward J
EJ
Behrman
Department of Chemistry & Biochemistry, The Ohio State University, Columbus, OH 43210, USA.|Department of Chemistry & Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
Venkat
V
Gopalan
D001216:Asparagine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D001224:Aspartic Acid / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005632:Fructose / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009876:Operon; D010770:Phosphotransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D019779:Salmonella enterica / Q000201:enzymology* / Q000235:genetics; D013237:Stereoisomerism; D013379:Substrate Specificity; D013696:Temperature
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins; D005632:Fructose; D001224:Aspartic Acid; D001216:Asparagine; D010770:Phosphotransferases
6-phosphofructose-aspartate; Salmonella; fructose-asparagine; fructose-aspartate; kinase