Identification of the covalently bound flavins of D-gluconate dehydrogenases from Pseudomonas aeruginosa and Pseudomonas fluorescens and of 2-keto-D-gluconate dehydrogenase from Gluconobacter melanogenus.
An improved method is presented for the purification of 8 alpha-(N1-histidyl)riboflavin, 8 alpha-(N3-histidyl)riboflavin and their 2',5'-anhydro forms, which permits the isolation of sizeable quantities of each of these compounds from a synthetic mixture in pure form. Flavin peptides were isolated from the D-gluconate dehydrogenases of Pseudomonas aeruginosa and Pseudomonas fluorescens and from the 2-keto-D-gluconate dehydrogenase of Gluconobacter melanogenus. After conversion into the aminoacyl-riboflavin, the flavin in all three enzymes was identified as 8 alpha-(N3-histidyl)riboflavin. By sequential treatment with nucleotide pyrophosphatase and alkaline phosphatase, the flavin in each enzyme was shown to be in the dinucleotide form.
D002237:Carbohydrate Dehydrogenases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005415:Flavins / Q000302:isolation & purification / Q000378:metabolism*; D006088:Gram-Negative Aerobic Bacteria / Q000201:enzymology*; D007525:Isoelectric Focusing; D011549:Pseudomonas / Q000201:enzymology; D012256:Riboflavin / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D005415:Flavins; C016491:N(3)-histidylriboflavin; D002237:Carbohydrate Dehydrogenases; C020577:gluconate 2-dehydrogenase; C047452:dehydrogluconate dehydrogenase; D012256:Riboflavin