Phosphomannose isomerase of Xanthomonas campestris: a zinc activated enzyme.
Phosphomannose isomerase (pmi, EC 5.3.1.8) was purified to homogeneity from a wild strain of Xanthomonas campestris. The apparent molecular weight as determined by SDS-PAGE and Sephadex G-100 Superfine was found to be 58 kDa. The purified enzyme showed a single band on acrylamide gel electrophocusing with pI = 5.25. The optimum pH was 7.0 and the Km for D-mannose-6-phosphate was 2 mM. Pmi can be activated by bivalent cations with the order of Co2+>Zn2+>Mn2+>Ni2+>Ca2+. Addition of low concentration of ZnCl2 (2 x 10[-7] M) in the growth medium resulted in the enhancement of pmi activity to around 2.5 x fold. The half life of pmi, as it was measured by the addition of chloramphenicol, was 110 min, whereas in the medium supplemented with ZnCl2 was 270 min. Chemical modification experiments implied the existence of one histidyl residue located at or near the active site.
Papoutsopoulou
S V
SV
Department of Chemistry, Aristotle University of Thessaloniki, Greece.
Kyriakidis
D A
DA
D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability / Q000187:drug effects; D006207:Half-Life; D007526:Isoelectric Point; D007700:Kinetics; D008359:Mannose-6-Phosphate Isomerase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D019163:Reducing Agents / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D016959:Xanthomonas campestris / Q000201:enzymology* / Q000235:genetics; D015032:Zinc / Q000494:pharmacology*
D019163:Reducing Agents; D008359:Mannose-6-Phosphate Isomerase; D015032:Zinc; D004047:Diethyl Pyrocarbonate