Homologue expression of a β-xylosidase from native Aspergillus niger.
Xylan constitutes the second most abundant source of renewable organic carbon on earth and is located in the cell walls of hardwood and softwood plants in the form of hemicellulose. Based on its availability, there is a growing interest in production of xylanolytic enzymes for industrial applications. β-1,4-xylan xylosidase (EC 3.2.1.37) hydrolyses from the nonreducing end of xylooligosaccharides arising from endo-1,4-β-xylanase activity. This work reports the partial characterization of a purified β-xylosidase from the native strain Aspergillus niger GS1 expressed by means of a fungal system. A gene encoding β-xylosidase, xlnD, was successfully cloned from a native A. niger GS1 strain. The recombinant enzyme was expressed in A. niger AB4.1 under control of A. nidulans gpdA promoter and trpC terminator. β-xylosidase was purified by affinity chromatography, with an apparent molecular weight of 90 kDa, and showed a maximum activity of 4,280 U mg protein(-1) at 70°C, pH 3.6. Half-life was 74 min at 70°C, activation energy was 58.9 kJ mol(-1), and at 50°C optimum stability was shown at pH 4.0-5.0. β-xylosidase kept residual activity >83% in the presence of dithiothreitol (DTT), β-mercaptoethanol, sodium dodecyl sulfate (SDS), ethylenediaminetetraacetate (EDTA), and Zn(2+). Production of a hemicellulolytic free xylosidase showed some advantages in applications, such as animal feed, enzymatic synthesis, and the fruit-juice industry where the presence of certain compounds, high temperatures, and acid media is unavoidable in the juice-making process.
DIPA, PROPAC. Facultad de Química, Universidad Autónoma de Querétaro, C.U. Cerro de las Campanas s/n. Col. Las Campanas, 76010 Qro., Querétaro, Mexico.
A
A
Amaro-Reyes
B E
BE
García-Almendárez
D G
DG
Vázquez-Mandujano
S
S
Amaya-Llano
E
E
Castaño-Tostado
R G
RG
Guevara-González
O
O
Loera
C
C
Regalado
D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D043364:Endo-1,4-beta Xylanases / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D014990:Xylans / Q000378:metabolism
D014990:Xylans; D043364:Endo-1,4-beta Xylanases