Structure of the AliC GH13 α−amylase from Alicyclobacillus sp, reveals accommodation of starch branching points in the α−amylase family

Jon Agirre, Olga V. Moroz, Sebastian Meier, Jesper Braak, Astrid Munch, Tine Hoff, Carsten Anderson, Keith S. Wilson*, Gideon J. Davies

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

251 Downloads (Pure)

Abstract

α−amylases are glycoside hydrolases that break the α−1,4 bonds in starch and related glycans. The degradation of starch is rendered difficult by varying degrees of α−1,6 branch points and their possible accommodation within the active centre of α−amylase enzymes. Given the myriad industrial uses for starch and thus also for α−amylase-catalysed starch degradation and modification, there is considerable interest in how different α−amylases might accommodate these branches thus impacting on the potential limit dextrins and societal applications. Here, we sought to probe the branch-point accommodation of the Alicyclobacillus sp. CAZy family GH13 α−amylase, prompted by our observation of a molecule of glucose in the position that may represent a branch point in an acarbose complex solved at 2.1 Å resolution. Limit digest analysis, by 2D NMR, using both pullulan (a regular linear polysaccharide of α−1,4, α−1,4, α−1,6 repeating trisaccharides) and amylopectin starch showed how the Alicyclobacillus sp enzyme could accept α−1,6 branches in, at least, -2, +1 and +2 subsites consistent with 3-D structures with glucosyl moieties in +1 and +2 subsites and the solvent exposure of the -2 6-hydroxyl group. Together the work provides a rare insight into branch point acceptance in these industrial catalysts.
Original languageEnglish
JournalActa crystallographica Section D: Structural biology
Volume75
Issue number1
Pages (from-to)1-7
Number of pages7
ISSN2059-7983
DOIs
Publication statusPublished - 2019

Keywords

  • AliC GH13 α-analyse
  • Starch brancing points
  • Glycoside hydrolases
  • Pullulan
  • Carbohydrate-active enzymes
  • Alicyclobacillus

Cite this

Agirre, Jon ; Moroz, Olga V. ; Meier, Sebastian ; Braak, Jesper ; Munch, Astrid ; Hoff, Tine ; Anderson, Carsten ; Wilson, Keith S. ; Davies, Gideon J. / Structure of the AliC GH13 α−amylase from Alicyclobacillus sp, reveals accommodation of starch branching points in the α−amylase family. In: Acta crystallographica Section D: Structural biology . 2019 ; Vol. 75, No. 1. pp. 1-7.
@article{dfbb3925f0474216a31f0173d82d8f8a,
title = "Structure of the AliC GH13 α−amylase from Alicyclobacillus sp, reveals accommodation of starch branching points in the α−amylase family",
abstract = "α−amylases are glycoside hydrolases that break the α−1,4 bonds in starch and related glycans. The degradation of starch is rendered difficult by varying degrees of α−1,6 branch points and their possible accommodation within the active centre of α−amylase enzymes. Given the myriad industrial uses for starch and thus also for α−amylase-catalysed starch degradation and modification, there is considerable interest in how different α−amylases might accommodate these branches thus impacting on the potential limit dextrins and societal applications. Here, we sought to probe the branch-point accommodation of the Alicyclobacillus sp. CAZy family GH13 α−amylase, prompted by our observation of a molecule of glucose in the position that may represent a branch point in an acarbose complex solved at 2.1 {\AA} resolution. Limit digest analysis, by 2D NMR, using both pullulan (a regular linear polysaccharide of α−1,4, α−1,4, α−1,6 repeating trisaccharides) and amylopectin starch showed how the Alicyclobacillus sp enzyme could accept α−1,6 branches in, at least, -2, +1 and +2 subsites consistent with 3-D structures with glucosyl moieties in +1 and +2 subsites and the solvent exposure of the -2 6-hydroxyl group. Together the work provides a rare insight into branch point acceptance in these industrial catalysts.",
keywords = "AliC GH13 α-analyse, Starch brancing points, Glycoside hydrolases, Pullulan, Carbohydrate-active enzymes, Alicyclobacillus",
author = "Jon Agirre and Moroz, {Olga V.} and Sebastian Meier and Jesper Braak and Astrid Munch and Tine Hoff and Carsten Anderson and Wilson, {Keith S.} and Davies, {Gideon J.}",
year = "2019",
doi = "10.1107/S2059798318014900",
language = "English",
volume = "75",
pages = "1--7",
journal = "Acta crystallographica Section D: Structural biology",
issn = "2059-7983",
publisher = "International Union of Crystallography",
number = "1",

}

Structure of the AliC GH13 α−amylase from Alicyclobacillus sp, reveals accommodation of starch branching points in the α−amylase family. / Agirre, Jon; Moroz, Olga V.; Meier, Sebastian; Braak, Jesper; Munch, Astrid; Hoff, Tine ; Anderson, Carsten; Wilson, Keith S.; Davies, Gideon J.

In: Acta crystallographica Section D: Structural biology , Vol. 75, No. 1, 2019, p. 1-7.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Structure of the AliC GH13 α−amylase from Alicyclobacillus sp, reveals accommodation of starch branching points in the α−amylase family

AU - Agirre, Jon

AU - Moroz, Olga V.

AU - Meier, Sebastian

AU - Braak, Jesper

AU - Munch, Astrid

AU - Hoff, Tine

AU - Anderson, Carsten

AU - Wilson, Keith S.

AU - Davies, Gideon J.

PY - 2019

Y1 - 2019

N2 - α−amylases are glycoside hydrolases that break the α−1,4 bonds in starch and related glycans. The degradation of starch is rendered difficult by varying degrees of α−1,6 branch points and their possible accommodation within the active centre of α−amylase enzymes. Given the myriad industrial uses for starch and thus also for α−amylase-catalysed starch degradation and modification, there is considerable interest in how different α−amylases might accommodate these branches thus impacting on the potential limit dextrins and societal applications. Here, we sought to probe the branch-point accommodation of the Alicyclobacillus sp. CAZy family GH13 α−amylase, prompted by our observation of a molecule of glucose in the position that may represent a branch point in an acarbose complex solved at 2.1 Å resolution. Limit digest analysis, by 2D NMR, using both pullulan (a regular linear polysaccharide of α−1,4, α−1,4, α−1,6 repeating trisaccharides) and amylopectin starch showed how the Alicyclobacillus sp enzyme could accept α−1,6 branches in, at least, -2, +1 and +2 subsites consistent with 3-D structures with glucosyl moieties in +1 and +2 subsites and the solvent exposure of the -2 6-hydroxyl group. Together the work provides a rare insight into branch point acceptance in these industrial catalysts.

AB - α−amylases are glycoside hydrolases that break the α−1,4 bonds in starch and related glycans. The degradation of starch is rendered difficult by varying degrees of α−1,6 branch points and their possible accommodation within the active centre of α−amylase enzymes. Given the myriad industrial uses for starch and thus also for α−amylase-catalysed starch degradation and modification, there is considerable interest in how different α−amylases might accommodate these branches thus impacting on the potential limit dextrins and societal applications. Here, we sought to probe the branch-point accommodation of the Alicyclobacillus sp. CAZy family GH13 α−amylase, prompted by our observation of a molecule of glucose in the position that may represent a branch point in an acarbose complex solved at 2.1 Å resolution. Limit digest analysis, by 2D NMR, using both pullulan (a regular linear polysaccharide of α−1,4, α−1,4, α−1,6 repeating trisaccharides) and amylopectin starch showed how the Alicyclobacillus sp enzyme could accept α−1,6 branches in, at least, -2, +1 and +2 subsites consistent with 3-D structures with glucosyl moieties in +1 and +2 subsites and the solvent exposure of the -2 6-hydroxyl group. Together the work provides a rare insight into branch point acceptance in these industrial catalysts.

KW - AliC GH13 α-analyse

KW - Starch brancing points

KW - Glycoside hydrolases

KW - Pullulan

KW - Carbohydrate-active enzymes

KW - Alicyclobacillus

U2 - 10.1107/S2059798318014900

DO - 10.1107/S2059798318014900

M3 - Journal article

C2 - 30644839

VL - 75

SP - 1

EP - 7

JO - Acta crystallographica Section D: Structural biology

JF - Acta crystallographica Section D: Structural biology

SN - 2059-7983

IS - 1

ER -