Constructing arabinofuranosidases for dual arabinoxylan debranching activity

Weijun Wang, Nikola Andric, Cody Sarch, Bruno Teixeira Silva, Tiina Maija Tenkanen, Emma R. Master

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Enzymatic conversion of arabinoxylan requires -L-arabinofuranosidases able to remove -L-arabinofuranosyl residues (-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 -L-arabinofuranosidase with AXH-m activity) and BadAbf43A (a family GH43 -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 -(13)-L-Araf and -(12)-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 -L-Araf from WAX after a 20hr treatment. H-1 NMR analyses further revealed differences in SthAbf62A_BadAbf43 rate of removal of specific -L-Araf substituents from WAX, where 9.4 times higher activity was observed toward d--(13)-L-Araf compared to m--(13)-L-Araf positions.
Original languageEnglish
JournalBiotechnology and Bioengineering (Print)
Volume115
Issue number1
Pages (from-to)41-49
Number of pages9
ISSN0006-3592
DOIs
Publication statusPublished - Jan 2018
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 1183 Plant biology, microbiology, virology
  • 220 Industrial biotechnology
  • α-L-arabinofuranohydrolase
  • activity synergy
  • arabinoxylan fusion enzyme
  • dual α-L-Araf debranching activity
  • ALPHA-L-ARABINOFURANOSIDASE
  • WHEAT ARABINOXYLAN
  • BY-PRODUCT
  • XYLAN
  • BIOMASS
  • ARABINOFURANOHYDROLASE
  • HEMICELLULOSES
  • BIOCHEMISTRY
  • RESIDUES
  • ENZYMES
  • alpha-L-arabinofuranohydrolase
  • activity synergy
  • arabinoxylan fusion enzyme
  • dual-L-Araf debranching activity
  • ALPHA-L-ARABINOFURANOSIDASE
  • WHEAT ARABINOXYLAN
  • BY-PRODUCT
  • XYLAN
  • BIOMASS
  • ARABINOFURANOHYDROLASE
  • HEMICELLULOSES
  • BIOCHEMISTRY
  • RESIDUES
  • ENZYMES

Cite this

Wang, Weijun ; Andric, Nikola ; Sarch, Cody ; Teixeira Silva, Bruno ; Tenkanen, Tiina Maija ; Master, Emma R. / Constructing arabinofuranosidases for dual arabinoxylan debranching activity. In: Biotechnology and Bioengineering (Print). 2018 ; Vol. 115, No. 1. pp. 41-49.
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title = "Constructing arabinofuranosidases for dual arabinoxylan debranching activity",
abstract = "Enzymatic conversion of arabinoxylan requires -L-arabinofuranosidases able to remove -L-arabinofuranosyl residues (-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 -L-arabinofuranosidase with AXH-m activity) and BadAbf43A (a family GH43 -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 -(13)-L-Araf and -(12)-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 -L-Araf from WAX after a 20hr treatment. H-1 NMR analyses further revealed differences in SthAbf62A_BadAbf43 rate of removal of specific -L-Araf substituents from WAX, where 9.4 times higher activity was observed toward d--(13)-L-Araf compared to m--(13)-L-Araf positions.",
keywords = "1183 Plant biology, microbiology, virology, 220 Industrial biotechnology, α-L-arabinofuranohydrolase, activity synergy, arabinoxylan fusion enzyme, dual α-L-Araf debranching activity, ALPHA-L-ARABINOFURANOSIDASE, WHEAT ARABINOXYLAN, BY-PRODUCT, XYLAN, BIOMASS, ARABINOFURANOHYDROLASE, HEMICELLULOSES, BIOCHEMISTRY, RESIDUES, ENZYMES, alpha-L-arabinofuranohydrolase, activity synergy, arabinoxylan fusion enzyme, dual-L-Araf debranching activity, ALPHA-L-ARABINOFURANOSIDASE, WHEAT ARABINOXYLAN, BY-PRODUCT, XYLAN, BIOMASS, ARABINOFURANOHYDROLASE, HEMICELLULOSES, BIOCHEMISTRY, RESIDUES, ENZYMES",
author = "Weijun Wang and Nikola Andric and Cody Sarch and {Teixeira Silva}, Bruno and Tenkanen, {Tiina Maija} and Master, {Emma R.}",
year = "2018",
month = "1",
doi = "10.1002/bit.26445",
language = "English",
volume = "115",
pages = "41--49",
journal = "Biotechnology and Bioengineering (Print)",
issn = "0006-3592",
publisher = "John Wiley & Sons Ltd.",
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Constructing arabinofuranosidases for dual arabinoxylan debranching activity. / Wang, Weijun; Andric, Nikola; Sarch, Cody; Teixeira Silva, Bruno; Tenkanen, Tiina Maija; Master, Emma R.

In: Biotechnology and Bioengineering (Print), Vol. 115, No. 1, 01.2018, p. 41-49.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Constructing arabinofuranosidases for dual arabinoxylan debranching activity

AU - Wang, Weijun

AU - Andric, Nikola

AU - Sarch, Cody

AU - Teixeira Silva, Bruno

AU - Tenkanen, Tiina Maija

AU - Master, Emma R.

PY - 2018/1

Y1 - 2018/1

N2 - Enzymatic conversion of arabinoxylan requires -L-arabinofuranosidases able to remove -L-arabinofuranosyl residues (-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 -L-arabinofuranosidase with AXH-m activity) and BadAbf43A (a family GH43 -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 -(13)-L-Araf and -(12)-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 -L-Araf from WAX after a 20hr treatment. H-1 NMR analyses further revealed differences in SthAbf62A_BadAbf43 rate of removal of specific -L-Araf substituents from WAX, where 9.4 times higher activity was observed toward d--(13)-L-Araf compared to m--(13)-L-Araf positions.

AB - Enzymatic conversion of arabinoxylan requires -L-arabinofuranosidases able to remove -L-arabinofuranosyl residues (-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 -L-arabinofuranosidase with AXH-m activity) and BadAbf43A (a family GH43 -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 -(13)-L-Araf and -(12)-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 -L-Araf from WAX after a 20hr treatment. H-1 NMR analyses further revealed differences in SthAbf62A_BadAbf43 rate of removal of specific -L-Araf substituents from WAX, where 9.4 times higher activity was observed toward d--(13)-L-Araf compared to m--(13)-L-Araf positions.

KW - 1183 Plant biology, microbiology, virology

KW - 220 Industrial biotechnology

KW - α-L-arabinofuranohydrolase

KW - activity synergy

KW - arabinoxylan fusion enzyme

KW - dual α-L-Araf debranching activity

KW - ALPHA-L-ARABINOFURANOSIDASE

KW - WHEAT ARABINOXYLAN

KW - BY-PRODUCT

KW - XYLAN

KW - BIOMASS

KW - ARABINOFURANOHYDROLASE

KW - HEMICELLULOSES

KW - BIOCHEMISTRY

KW - RESIDUES

KW - ENZYMES

KW - alpha-L-arabinofuranohydrolase

KW - activity synergy

KW - arabinoxylan fusion enzyme

KW - dual-L-Araf debranching activity

KW - ALPHA-L-ARABINOFURANOSIDASE

KW - WHEAT ARABINOXYLAN

KW - BY-PRODUCT

KW - XYLAN

KW - BIOMASS

KW - ARABINOFURANOHYDROLASE

KW - HEMICELLULOSES

KW - BIOCHEMISTRY

KW - RESIDUES

KW - ENZYMES

U2 - 10.1002/bit.26445

DO - 10.1002/bit.26445

M3 - Article

VL - 115

SP - 41

EP - 49

JO - Biotechnology and Bioengineering (Print)

JF - Biotechnology and Bioengineering (Print)

SN - 0006-3592

IS - 1

ER -