Characterization of the four GH12 Endoxylanases from the plant pathogen Fusarium graminearum.
Four putative GH12 genes were found in the Fusarium graminearum genome. The corresponding proteins were expressed in Escherichia coli, purified, and evaluated. FGSG_05851 and FGSG_11037 displayed high activities towards xyloglucan (V(max) of 4 and 11 micronmol/min, respectively), whereas FGSG_07892 and FGSG_16349 were much less active with this substrate (0.081 and 0.004 micronmol/min, respectively). However, all four of these enzymes had a similar binding affinity for xyloglucan. Xyloglucan was the substrate preferred by FGSG_05851, in contrast to the three other enzymes, which preferred beta-glucan or lichenan. Therefore, FGSG_05851 is a xyloglucan-specific glucanase (E.C. 3.2.1.151) rather than an endoglucanase (E.C. 3.2.1.4) with broad substrate specificity. FGSG_11037 displayed a peculiar behavior in that the xyloglucan binding was highly cooperative, with a Hill coefficient of 2.5. Finally, FGSG_05851 essentially degraded xyloglucan into hepta-, octa-, and nonasaccharides, whereas the three other enzymes yielded hepta- and octa-saccharides as well as larger molecules.
Laboratoire d'Ingénierie des Polymères pour les Hautes Technologies, Université de Strasbourg, ECPM, 25 rue Becquerel, 67087 Strasbourg Cedex 2, France.
Olivier
O
Habrylo
Xinghan
X
Song
Anne
A
Forster
Jean-Marc
JM
Jeltsch
Vincent
V
Phalip
D003001:Cloning, Molecular; D043364:Endo-1,4-beta Xylanases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D005936:Glucans / Q000378:metabolism; D010935:Plant Diseases / Q000382:microbiology; D010944:Plants / Q000382:microbiology; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D014990:Xylans / Q000378:metabolism
D005936:Glucans; D011994:Recombinant Proteins; D014990:Xylans; C029353:xyloglucan; D043364:Endo-1,4-beta Xylanases; C015968:lichenin