Crystal structure and substrate interactions of an unusual fungal non-CBM carrying GH26 endo-β-mannanase from Yunnania penicillata

Pernille von Freiesleben, Olga V. Moroz, Elena Blagova, Mathias Wiemann, Nikolaj Spodsberg, Jane W. Agger, Gideon J. Davies, Keith S. Wilson, Henrik Stålbrand, Anne S. Meyer*, Kristian B. R. M. Krogh

*Corresponding author for this work

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Abstract

Endo-β(1 → 4)-mannanases (endomannanases) catalyse degradation of β-mannans, an abundant class of plant polysaccharides. This study investigates structural features and substrate binding of YpenMan26A, a non-CBM carrying endomannanase from Yunnania penicillata. Structural and sequence comparisons to other fungal family GH26 endomannanases showed high sequence similarities and conserved binding residues, indicating that fungal GH26 endomannanases accommodate galactopyranosyl units in the -3 and -2 subsites. Two striking amino acid differences in the active site were found when the YpenMan26A structure was compared to a homology model of Wsp.Man26A from Westerdykella sp. and the sequences of nine other fungal GH26 endomannanases. Two YpenMan26A mutants, W110H and D37T, inspired by differences observed in Wsp.Man26A, produced a shift in how mannopentaose bound across the active site cleft and a decreased affinity for galactose in the -2 subsite, respectively, compared to YpenMan26A. YpenMan26A was moreover found to have a flexible surface loop in the position where PansMan26A from Podospora anserina has an α-helix (α9) which interacts with its family 35 CBM. Sequence alignment inferred that the core structure of fungal GH26 endomannanases differ depending on the natural presence of this type of CBM. These new findings have implications for selecting and optimising these enzymes for galactomannandegradation.
Original languageEnglish
Article number2266
JournalScientific Reports
Volume9
Number of pages14
ISSN2045-2322
DOIs
Publication statusPublished - 2019

Cite this

von Freiesleben, Pernille ; Moroz, Olga V. ; Blagova, Elena ; Wiemann, Mathias ; Spodsberg, Nikolaj ; Agger, Jane W. ; Davies, Gideon J. ; Wilson, Keith S. ; Stålbrand, Henrik ; Meyer, Anne S. ; Krogh, Kristian B. R. M. / Crystal structure and substrate interactions of an unusual fungal non-CBM carrying GH26 endo-β-mannanase from Yunnania penicillata. In: Scientific Reports. 2019 ; Vol. 9.
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title = "Crystal structure and substrate interactions of an unusual fungal non-CBM carrying GH26 endo-β-mannanase from Yunnania penicillata",
abstract = "Endo-β(1 → 4)-mannanases (endomannanases) catalyse degradation of β-mannans, an abundant class of plant polysaccharides. This study investigates structural features and substrate binding of YpenMan26A, a non-CBM carrying endomannanase from Yunnania penicillata. Structural and sequence comparisons to other fungal family GH26 endomannanases showed high sequence similarities and conserved binding residues, indicating that fungal GH26 endomannanases accommodate galactopyranosyl units in the -3 and -2 subsites. Two striking amino acid differences in the active site were found when the YpenMan26A structure was compared to a homology model of Wsp.Man26A from Westerdykella sp. and the sequences of nine other fungal GH26 endomannanases. Two YpenMan26A mutants, W110H and D37T, inspired by differences observed in Wsp.Man26A, produced a shift in how mannopentaose bound across the active site cleft and a decreased affinity for galactose in the -2 subsite, respectively, compared to YpenMan26A. YpenMan26A was moreover found to have a flexible surface loop in the position where PansMan26A from Podospora anserina has an α-helix (α9) which interacts with its family 35 CBM. Sequence alignment inferred that the core structure of fungal GH26 endomannanases differ depending on the natural presence of this type of CBM. These new findings have implications for selecting and optimising these enzymes for galactomannandegradation.",
author = "{von Freiesleben}, Pernille and Moroz, {Olga V.} and Elena Blagova and Mathias Wiemann and Nikolaj Spodsberg and Agger, {Jane W.} and Davies, {Gideon J.} and Wilson, {Keith S.} and Henrik St{\aa}lbrand and Meyer, {Anne S.} and Krogh, {Kristian B. R. M.}",
year = "2019",
doi = "10.1038/s41598-019-38602-x",
language = "English",
volume = "9",
journal = "Scientific Reports",
issn = "2045-2322",
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von Freiesleben, P, Moroz, OV, Blagova, E, Wiemann, M, Spodsberg, N, Agger, JW, Davies, GJ, Wilson, KS, Stålbrand, H, Meyer, AS & Krogh, KBRM 2019, 'Crystal structure and substrate interactions of an unusual fungal non-CBM carrying GH26 endo-β-mannanase from Yunnania penicillata', Scientific Reports, vol. 9, 2266. https://doi.org/10.1038/s41598-019-38602-x

Crystal structure and substrate interactions of an unusual fungal non-CBM carrying GH26 endo-β-mannanase from Yunnania penicillata. / von Freiesleben, Pernille; Moroz, Olga V.; Blagova, Elena; Wiemann, Mathias; Spodsberg, Nikolaj; Agger, Jane W.; Davies, Gideon J.; Wilson, Keith S.; Stålbrand, Henrik; Meyer, Anne S.; Krogh, Kristian B. R. M.

In: Scientific Reports, Vol. 9, 2266, 2019.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Crystal structure and substrate interactions of an unusual fungal non-CBM carrying GH26 endo-β-mannanase from Yunnania penicillata

AU - von Freiesleben, Pernille

AU - Moroz, Olga V.

AU - Blagova, Elena

AU - Wiemann, Mathias

AU - Spodsberg, Nikolaj

AU - Agger, Jane W.

AU - Davies, Gideon J.

AU - Wilson, Keith S.

AU - Stålbrand, Henrik

AU - Meyer, Anne S.

AU - Krogh, Kristian B. R. M.

PY - 2019

Y1 - 2019

N2 - Endo-β(1 → 4)-mannanases (endomannanases) catalyse degradation of β-mannans, an abundant class of plant polysaccharides. This study investigates structural features and substrate binding of YpenMan26A, a non-CBM carrying endomannanase from Yunnania penicillata. Structural and sequence comparisons to other fungal family GH26 endomannanases showed high sequence similarities and conserved binding residues, indicating that fungal GH26 endomannanases accommodate galactopyranosyl units in the -3 and -2 subsites. Two striking amino acid differences in the active site were found when the YpenMan26A structure was compared to a homology model of Wsp.Man26A from Westerdykella sp. and the sequences of nine other fungal GH26 endomannanases. Two YpenMan26A mutants, W110H and D37T, inspired by differences observed in Wsp.Man26A, produced a shift in how mannopentaose bound across the active site cleft and a decreased affinity for galactose in the -2 subsite, respectively, compared to YpenMan26A. YpenMan26A was moreover found to have a flexible surface loop in the position where PansMan26A from Podospora anserina has an α-helix (α9) which interacts with its family 35 CBM. Sequence alignment inferred that the core structure of fungal GH26 endomannanases differ depending on the natural presence of this type of CBM. These new findings have implications for selecting and optimising these enzymes for galactomannandegradation.

AB - Endo-β(1 → 4)-mannanases (endomannanases) catalyse degradation of β-mannans, an abundant class of plant polysaccharides. This study investigates structural features and substrate binding of YpenMan26A, a non-CBM carrying endomannanase from Yunnania penicillata. Structural and sequence comparisons to other fungal family GH26 endomannanases showed high sequence similarities and conserved binding residues, indicating that fungal GH26 endomannanases accommodate galactopyranosyl units in the -3 and -2 subsites. Two striking amino acid differences in the active site were found when the YpenMan26A structure was compared to a homology model of Wsp.Man26A from Westerdykella sp. and the sequences of nine other fungal GH26 endomannanases. Two YpenMan26A mutants, W110H and D37T, inspired by differences observed in Wsp.Man26A, produced a shift in how mannopentaose bound across the active site cleft and a decreased affinity for galactose in the -2 subsite, respectively, compared to YpenMan26A. YpenMan26A was moreover found to have a flexible surface loop in the position where PansMan26A from Podospora anserina has an α-helix (α9) which interacts with its family 35 CBM. Sequence alignment inferred that the core structure of fungal GH26 endomannanases differ depending on the natural presence of this type of CBM. These new findings have implications for selecting and optimising these enzymes for galactomannandegradation.

U2 - 10.1038/s41598-019-38602-x

DO - 10.1038/s41598-019-38602-x

M3 - Journal article

VL - 9

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 2266

ER -