Factors affecting hexose phosphorylation in Acetobacter xylinum.
Fructose was oxidized and converted to cellulose by cells of Acetobacter xylinum grown on fructose or succinate, but not by cells grown on glucose. In resting fructose-grown cells, glucose strongly suppressed fructose utilization. Extracts obtained from fructose- or succinate-grown cells catalyzed the adenosine triphosphate (ATP)-dependent formation of the 6-phosphate esters of glucose and fructose, whereas glucose-grown cell extracts phosphorylated glucose but not fructose. Fructokinase and glucokinase activities were separated and partially purified from cells grown on glucose, fructose, or succinate. Whereas fructokinase phosphorylated fructose only, glucokinase was active towards glucose and less active towards mannose and glucosamine. The optimal pH for the fructokinase was 7.4 and for the glucokinase was 8.5. The K(m) values for the fructokinase were: fructose, 6.2 mm; and ATP, 0.83 mm. The K(m) values for the glucokinase were: glucose, 0.22 mm; and ATP, 4.2 mm. Fructokinase was inhibited by glucose, glucosamine, mannose, and deoxyglucose in a manner competitive with respect to fructose, with K(i) values of 0.1, 0.14, 0.5, and 7.5 mm, respectively. Adenosine diphosphate (ADP) and adenosine monophosphate (AMP) inhibited both kinases noncompetitively with respect to ATP. The K(i) values were: 1.8 mm (ADP) and 2.1 mm (AMP) for fructokinase, and 2.2 mm (ADP) and 9.6 mm (AMP) for glucokinase. Fructose metabolism in A. xylinum appears to be regulated by the synthesis and activity of fructokinase.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000645:Ammonium Sulfate; D002241:Carbohydrates / Q000494:pharmacology; D002245:Carbon Dioxide / Q000096:biosynthesis; D002247:Carbon Isotopes; D002474:Cell-Free System; D002482:Cellulose / Q000096:biosynthesis; D011232:Chemical Precipitation; D003470:Culture Media; D005632:Fructose / Q000378:metabolism; D005941:Glucokinase / Q000378:metabolism; D041822:Gluconacetobacter xylinus / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006600:Hexosephosphates / Q000096:biosynthesis; D006601:Hexoses / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D008365:Manometry; D010085:Oxidative Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D013053:Spectrophotometry; D013386:Succinates / Q000378:metabolism
D002241:Carbohydrates; D002247:Carbon Isotopes; D003470:Culture Media; D006600:Hexosephosphates; D006601:Hexoses; D013386:Succinates; D002245:Carbon Dioxide; D005632:Fructose; D000255:Adenosine Triphosphate; D002482:Cellulose; D010770:Phosphotransferases; D005941:Glucokinase; D008274:Magnesium; D005947:Glucose; D000645:Ammonium Sulfate