Flavin adenine dinucleotide-dependent 4-phospho-D-erythronate dehydrogenase is responsible for the 4-phosphohydroxy-L-threonine pathway in vitamin B6 biosynthesis in Sinorhizobium meliloti.
The vitamin B6 biosynthetic pathway in Sinorhizobium meliloti is similar to that in Escherichia coli K-12; in both organisms this pathway includes condensation of two intermediates, 1-deoxy-D-xylulose 5-phosphate and 4-phosphohydroxy-L-threonine (4PHT). Here, we report cloning of a gene designated pdxR that functionally corresponds to the pdxB gene of E. coli and encodes a dye-linked flavin adenine dinucleotide-dependent 4-phospho-D-erythronate (4PE) dehydrogenase. This enzyme catalyzes the oxidation of 4PE to 3-hydroxy-4-phosphohydroxy-alpha-ketobutyrate and is clearly different in terms of cofactor requirements from the pdxB gene product of E. coli, which is known to be an NAD-dependent enzyme. Previously, we revealed that in S. meliloti IFO 14782, 4PHT is synthesized from 4-hydroxy-l-threonine and that this synthesis starts with glycolaldehyde and glycine. However, in this study, we identified a second 4PHT pathway in S. meliloti that originates exclusively from glycolaldehyde (the major pathway). Based on the involvement of 4PE in the 4PHT pathway, the incorporation of different samples of 13C-labeled glycolaldehyde into pyridoxine molecules was examined using 13C nuclear magnetic resonance spectroscopy. On the basis of the spectral analyses, the synthesis of 4PHT from glycolaldehyde was hypothesized to involve the following steps: glycolaldehyde is sequentially metabolized to D-erythrulose, D-erythrulose 4-phosphate, and D-erythrose 4-phosphate by transketolase, kinase, and isomerase, respectively; and D-erythrose 4-phosphate is then converted to 4PHT by the conventional three-step pathway elucidated in E. coli, although the mechanism of action of the enzymes catalyzing the first two steps is different.
Tazoe
Masaaki
M
Department of applied Microbiology, Nippon Roche Research Center, Kamakura, Kanagawa, Japan. m.tazoe@lab.tamagawa.ac.jp
Ichikawa
Keiko
K
Hoshino
Tatsuo
T
D000079:Acetaldehyde / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism*; D005798:Genes, Bacterial; D007535:Isomerases / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010755:Organophosphates / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D010770:Phosphotransferases / Q000378:metabolism; D016415:Sequence Alignment; D016962:Sinorhizobium meliloti / Q000378:metabolism*; D013912:Threonine / Q000031:analogs & derivatives / Q000378:metabolism; D014174:Transketolase / Q000378:metabolism; D025101:Vitamin B 6 / Q000096:biosynthesis*
C098563:4-phosphothreonine; D001426:Bacterial Proteins; D010755:Organophosphates; D005182:Flavin-Adenine Dinucleotide; D013912:Threonine; D025101:Vitamin B 6; D010088:Oxidoreductases; D014174:Transketolase; D010770:Phosphotransferases; D007535:Isomerases; D000079:Acetaldehyde; C010972:glycolaldehyde