Engineering of a multifunctional hemicellulase.
To engineer a multifunctional xylan-degrading enzyme, a chimera was created by fusing the xylanase domain of the Clostridium thermocellum xylanase (xynZ) and a dual functional arabinofuranosidase/xylosidase (DeAFc; from a compost starter mixture) through a flexible peptide linker. The xylanase domain of xynZ possesses previously unreported endoglucanase activity. The chimera, possessing the activities of xylanase, endoglucanase, arabinofuranosidase and xylosidase, was expressed in E. coli and purified. The chimera closely resembled the parental enzymes in pH, temperature optima and kinetics, and was more active than the parental enzyme mixture in the hydrolysis of natural xylans and corn stover.
Fan
Zhanmin
Z
Department of Plant and Soil Sciences and Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY 40546, USA.
Werkman
Joshua R
JR
Yuan
Ling
L
D048013:Acetivibrio thermocellus / Q000201:enzymology / Q000235:genetics; D043364:Endo-1,4-beta Xylanases / Q000235:genetics / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature; D014990:Xylans / Q000378:metabolism*; D014995:Xylosidases / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D014990:Xylans; D006026:Glycoside Hydrolases; D014995:Xylosidases; C023305:hemicellulase; C019713:alpha-N-arabinofuranosidase; D043364:Endo-1,4-beta Xylanases