@article {2248, title = {Constitutive expression, purification and characterization of a phosphoglucomutase from Fusarium oxysporum}, journal = {Enzyme and Microbial Technology}, volume = {48}, year = {2011}, pages = {217-224}, abstract = {The phosphoglucomutase gene from a wild type\ Fusarium oxysporum \ \ strain (F3), was homologously expressed, under the control of the constitutive promoter of\ gpdA of\ Aspergillus nidulans. The transformant produced elevated levels of phosphoglucomutase activity compared to the wild type, a fact that facilitated the subsequent purification procedure. The enzyme (FoPGM) was purified to homogeneity applying three anion exchange and one gel filtration chromatography steps. The native enzyme revealed a monomeric structure with a molecular mass of 60\ kDa, while the isoelectric point was 3.5.\ FoPGM was active in pH ranged from 6.0 to 8.0, with an optimum using 3-(N-morpholino)propanesulfonic acid buffer at 7.0, while loss of activity was observed when phosphate buffer was used in the above mentioned pH range. The optimal temperature for activity was 45\ {\textdegree}C but the enzyme became unstable at temperatures above 40\ {\textdegree}C.\ FoPGM requires the presence of a divalent cation for its function with maximum activity being obtained with Co2+. The apparent\ Km\ for Co2+\ was found to be 10\ μM. The enzyme was also active with other divalent metal ions such as Mn2+, Mg2+, Ni2+\ and Ca2+\ but to a lesser extent. The following kinetic constants were determined:\ vmax, 0.74\ μmol\ mgprotein-1\ min-1;kcat, 44.2\ min-1;\ Km(G1P), 0.10\ mM;Km(G1,6diP), 1.03\ μM;\ kcat/Km(G1P), 443\ mM-1\ min-1\ and\ kcat/Km(G1,6diP), 42,860\ mM-1\ min-1. The enzyme was considered to follow a Ping Pong substituted enzyme or enzyme isomerization mechanism.}, url = {http://www.sciencedirect.com/science/article/pii/S0141022910002280}, author = {E. Kourtoglou and G.E. Anasontzis and D. Mamma and E. Topakas and D.G. Hatzinikolaou and P. Christakopoulos} }