<?xml version="1.0" encoding="UTF-8"?><xml><records><record><source-app name="Biblio" version="7.x">Drupal-Biblio</source-app><ref-type>17</ref-type><contributors><authors><author><style face="normal" font="default" size="100%">E. Kourtoglou</style></author><author><style face="normal" font="default" size="100%">G.E. Anasontzis</style></author><author><style face="normal" font="default" size="100%">D. Mamma</style></author><author><style face="normal" font="default" size="100%">E. Topakas</style></author><author><style face="normal" font="default" size="100%">D.G. Hatzinikolaou</style></author><author><style face="normal" font="default" size="100%">P. Christakopoulos</style></author></authors></contributors><titles><title><style face="normal" font="default" size="100%">Constitutive expression, purification and characterization of a phosphoglucomutase from &lt;em&gt;Fusarium oxysporum&lt;/em&gt;</style></title><secondary-title><style face="normal" font="default" size="100%">Enzyme and Microbial Technology</style></secondary-title></titles><dates><year><style  face="normal" font="default" size="100%">2011</style></year></dates><urls><web-urls><url><style face="normal" font="default" size="100%">http://www.sciencedirect.com/science/article/pii/S0141022910002280</style></url></web-urls></urls><volume><style face="normal" font="default" size="100%">48</style></volume><pages><style face="normal" font="default" size="100%">217-224</style></pages><language><style face="normal" font="default" size="100%">eng</style></language><abstract><style face="normal" font="default" size="100%">&lt;span&gt;The phosphoglucomutase gene from a wild type&amp;nbsp;&lt;/span&gt;&lt;em&gt;Fusarium oxysporum &amp;nbsp;&lt;/em&gt;&lt;span&gt;&amp;nbsp;strain (F3), was homologously expressed, under the control of the constitutive promoter of&amp;nbsp;&lt;/span&gt;&lt;em&gt;gpdA &lt;/em&gt;&lt;span&gt;of&amp;nbsp;&lt;/span&gt;&lt;em&gt;Aspergillus nidulans&lt;/em&gt;&lt;span&gt;. The transformant produced elevated levels of phosphoglucomutase activity compared to the wild type, a fact that facilitated the subsequent purification procedure. The enzyme (&lt;/span&gt;&lt;em&gt;Fo&lt;/em&gt;&lt;span&gt;PGM) 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&amp;nbsp;kDa, while the isoelectric point was 3.5.&amp;nbsp;&lt;/span&gt;&lt;em&gt;Fo&lt;/em&gt;&lt;span&gt;PGM 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&amp;nbsp;°C but the enzyme became unstable at temperatures above 40&amp;nbsp;°C.&amp;nbsp;&lt;/span&gt;&lt;em&gt;FoP&lt;/em&gt;&lt;span&gt;GM requires the presence of a divalent cation for its function with maximum activity being obtained with Co&lt;/span&gt;&lt;sup&gt;2+&lt;/sup&gt;&lt;span&gt;. The apparent&amp;nbsp;&lt;/span&gt;&lt;em&gt;K&lt;/em&gt;&lt;sub&gt;&lt;em&gt;m&lt;/em&gt;&lt;/sub&gt;&lt;span&gt;&amp;nbsp;for Co&lt;/span&gt;&lt;sup&gt;2+&lt;/sup&gt;&lt;span&gt;&amp;nbsp;was found to be 10&amp;nbsp;μM. The enzyme was also active with other divalent metal ions such as Mn&lt;/span&gt;&lt;sup&gt;2+&lt;/sup&gt;&lt;span&gt;, Mg&lt;/span&gt;&lt;sup&gt;2+&lt;/sup&gt;&lt;span&gt;, Ni&lt;/span&gt;&lt;sup&gt;2+&lt;/sup&gt;&lt;span&gt;&amp;nbsp;and Ca&lt;/span&gt;&lt;sup&gt;2+&lt;/sup&gt;&lt;span&gt;&amp;nbsp;but to a lesser extent. The following kinetic constants were determined:&amp;nbsp;&lt;/span&gt;&lt;span id=&quot;mmlsi5&quot; class=&quot;mathmlsrc&quot;&gt;&lt;span class=&quot;formulatext stixSupport mathImg&quot; data-mathurl=&quot;/science?_ob=MathURL&amp;amp;_method=retrieve&amp;amp;_eid=1-s2.0-S0141022910002280&amp;amp;_mathId=si5.gif&amp;amp;_user=111111111&amp;amp;_pii=S0141022910002280&amp;amp;_rdoc=1&amp;amp;_issn=01410229&amp;amp;md5=5e74e5d58a775a70e27bf86d5435cbbc&quot;&gt;v&lt;sub&gt;max&lt;/sub&gt;&lt;/span&gt;&lt;span class=&quot;mathContainer hidden&quot;&gt;&lt;span class=&quot;mathCode&quot;&gt;&lt;/span&gt;&lt;/span&gt;&lt;/span&gt;&lt;span&gt;, 0.74&amp;nbsp;μmol&amp;nbsp;mg&lt;/span&gt;&lt;sub&gt;protein&lt;/sub&gt;&lt;sup&gt;−1&lt;/sup&gt;&lt;span&gt;&amp;nbsp;min&lt;/span&gt;&lt;sup&gt;−1&lt;/sup&gt;;&lt;span&gt;&amp;nbsp;&lt;/span&gt;&lt;em&gt;k&lt;/em&gt;&lt;sub&gt;cat&lt;/sub&gt;&lt;span&gt;, 44.2&amp;nbsp;min&lt;/span&gt;&lt;sup&gt;−1&lt;/sup&gt;&lt;span&gt;;&amp;nbsp;&lt;/span&gt;&lt;em&gt;K&lt;/em&gt;&lt;sub&gt;&lt;em&gt;m&lt;/em&gt;&lt;/sub&gt;&lt;span&gt;(G1P), 0.10&amp;nbsp;mM;&lt;/span&gt;&lt;em&gt;K&lt;/em&gt;&lt;sub&gt;&lt;em&gt;m&lt;/em&gt;&lt;/sub&gt;&lt;span&gt;(G1,6diP), 1.03&amp;nbsp;μM;&amp;nbsp;&lt;/span&gt;&lt;em&gt;k&lt;/em&gt;&lt;sub&gt;cat&lt;/sub&gt;&lt;span&gt;/&lt;/span&gt;&lt;em&gt;K&lt;/em&gt;&lt;sub&gt;&lt;em&gt;m&lt;/em&gt;&lt;/sub&gt;&lt;span&gt;(G1P), 443&amp;nbsp;mM&lt;/span&gt;&lt;sup&gt;−1&lt;/sup&gt;&lt;span&gt;&amp;nbsp;min&lt;/span&gt;&lt;sup&gt;−1&lt;/sup&gt;&lt;span&gt;&amp;nbsp;and&amp;nbsp;&lt;/span&gt;&lt;em&gt;k&lt;/em&gt;&lt;sub&gt;cat&lt;/sub&gt;&lt;span&gt;/&lt;/span&gt;&lt;em&gt;K&lt;/em&gt;&lt;sub&gt;&lt;em&gt;m&lt;/em&gt;&lt;/sub&gt;&lt;span&gt;(G1,6diP), 42,860&amp;nbsp;mM&lt;/span&gt;&lt;sup&gt;−1&lt;/sup&gt;&lt;span&gt;&amp;nbsp;min&lt;/span&gt;&lt;sup&gt;−1&lt;/sup&gt;&lt;span&gt;. The enzyme was considered to follow a Ping Pong substituted enzyme or enzyme isomerization mechanism.&lt;/span&gt;</style></abstract></record></records></xml>