Constructing arabinofuranosidases for dual arabinoxylan debranching activity

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A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
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Date
2018-01
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Language
en
Pages
41-49
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BIOTECHNOLOGY AND BIOENGINEERING, Volume 115, issue 1
Abstract
Enzymatic conversion of arabinoxylan requires α-L-arabinofuranosidases able to remove α-L-arabinofuranosyl residues (α-L-Araf) from both mono- and double-substituted D-xylopyranosyl residues (Xylp) in xylan (i.e., AXH-m and AXH-d activity). Herein, SthAbf62A (a family GH62 α-L-arabinofuranosidase with AXH-m activity) and BadAbf43A (a family GH43 α-L-arabinofuranosidase with AXH-d3 activity), were fused to create SthAbf62A_BadAbf43A and BadAbf43A_SthAbf62A. Both fusion enzymes displayed dual AXH-m,d and synergistic activity toward native, highly branched wheat arabinoxylan (WAX). When using a customized arabinoxylan substrate comprising mainly α-(1 → 3)-L-Araf and α-(1 → 2)-L-Araf substituents attached to disubstituted Xylp (d-2,3-WAX), the specific activity of the fusion enzymes was twice that of enzymes added as separate proteins. Moreover, the SthAbf62A_BadAbf43A fusion removed 83% of all α-L-Araf from WAX after a 20 hr treatment. 1H NMR analyses further revealed differences in SthAbf62A_BadAbf43 rate of removal of specific α-L-Araf substituents from WAX, where 9.4 times higher activity was observed toward d-α-(1 → 3)-L-Araf compared to m-α-(1 → 3)-L-Araf positions.
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| openaire: EC/H2020/648925/EU//BHIVE
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Wang , W , Andric , N , Sarch , C , Silva , B T , Tenkanen , M & Master , E R 2018 , ' Constructing arabinofuranosidases for dual arabinoxylan debranching activity ' , Biotechnology and Bioengineering , vol. 115 , no. 1 , pp. 41-49 . https://doi.org/10.1002/bit.26445