A newly discovered arabinoxylan-specific arabinofuranohydrolase. Synergistic action with xylanases from different glycosyl hydrolase families.
Arabinofuranosidase Abf43A from Bacillussp. BP-7 i s a newly discovered arabinoxylan arabinofuranohydrolase (AXH). It is a modular enzyme comprised of a GH43 catalytic domain and a carbohydrate-binding module of family CBM6. Recombinant Abf43A showed high activity on arabinoxylans, being rye arabinoxylan the preferred substrate on which the purified enzyme exhibited a Km of 10.6 ± 3.3 mg/ml and a Vmax of 29.2 ± 3.4 U/mg. Thin-layer chromatography analysis of hydrolysis products showed arabinose as the only sugar released by the enzyme from its substrates. The GH43 and CBM6 modules of the enzyme were individually cloned and expressed in Escherichia coli. While the isolated catalytic GH43 module did not show hydrolytic activity, the purified CBM6 bound to soluble arabinoxylan in affinity gel electrophoresis analysis. Evaluation of cooperative activity of arabinofuranosidase Abf43A with xylanases from families GH10, GH11, andGH30, (Xyn10A, Xyn11E, and Xyn30D from Paenibacillus barcinonensis) on arabinoxylan depolymerization revealed that the studied enzyme showed synergism with Xyn11E, a 2.54-fold increase in the amount of sugars released. On the contrary, Abf43A did not show synergism with the xylanases of families GH10 or GH30 evaluated. The enzyme characterized contributes to understanding the role of this class of enzymes in the catalytic depolymerization of arabinoxylans and their potential for the production of valuable xylooligosaccharides from these abundant plant polymers.
Department of Microbiology, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028, Barcelona, Spain.
Adrià
A
Valls
Department of Microbiology, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028, Barcelona, Spain.
Pilar
P
Diaz
Department of Microbiology, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028, Barcelona, Spain.
F I Javier
FIJ
Pastor
Department of Microbiology, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028, Barcelona, Spain. susanavalenzuela@ub.edu.
Susana V
SV
Valenzuela
D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001711:Biotransformation; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D006868:Hydrolysis; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D014990:Xylans / Q000378:metabolism; D014995:Xylosidases / Q000378:metabolism*
D011994:Recombinant Proteins; D014990:Xylans; C085118:arabinoxylan; D006026:Glycoside Hydrolases; D014995:Xylosidases; C019713:alpha-N-arabinofuranosidase