Structure of a GH51 α- L -arabinofuranosidase from Meripilus giganteus: Conserved substrate recognition from bacteria to fungi

TY - JOUR

T1 - Structure of a GH51 α- L -arabinofuranosidase from Meripilus giganteus

T2 - Conserved substrate recognition from bacteria to fungi

AU - McGregor, Nicholas G.S.

AU - Turkenburg, Johan P.

AU - Mørkeberg Krogh, Kristian B.R.

AU - Nielsen, Jens Erik

AU - Artola, Marta

AU - Stubbs, Keith A.

AU - Overkleeft, Herman S.

AU - Davies, Gideon J.

PY - 2020/11/1

Y1 - 2020/11/1

N2 - α-l-Arabinofuranosidases from glycoside hydrolase family 51 use a stereochemically retaining hydrolytic mechanism to liberate nonreducing terminal α-l-arabinofuranose residues from plant polysaccharides such as arabinoxylan and arabinan. To date, more than ten fungal GH51 α-l-arabinofuranosidases have been functionally characterized, yet no structure of a fungal GH51 enzyme has been solved. In contrast, seven bacterial GH51 enzyme structures, with low sequence similarity to the fungal GH51 enzymes, have been determined. Here, the crystallization and structural characterization of MgGH51, an industrially relevant GH51 α-l-arabinofuranosidase cloned from Meripilus giganteus, are reported. Three crystal forms were grown in different crystallization conditions. The unliganded structure was solved using sulfur SAD data collected from a single crystal using the I23 in vacuo diffraction beamline at Diamond Light Source. Crystal soaks with arabinose, 1,4-dideoxy-1,4-imino-l-arabinitol and two cyclophellitol-derived arabinose mimics reveal a conserved catalytic site and conformational itinerary between fungal and bacterial GH51 α-l-arabino­furanosidases.

AB - α-l-Arabinofuranosidases from glycoside hydrolase family 51 use a stereochemically retaining hydrolytic mechanism to liberate nonreducing terminal α-l-arabinofuranose residues from plant polysaccharides such as arabinoxylan and arabinan. To date, more than ten fungal GH51 α-l-arabinofuranosidases have been functionally characterized, yet no structure of a fungal GH51 enzyme has been solved. In contrast, seven bacterial GH51 enzyme structures, with low sequence similarity to the fungal GH51 enzymes, have been determined. Here, the crystallization and structural characterization of MgGH51, an industrially relevant GH51 α-l-arabinofuranosidase cloned from Meripilus giganteus, are reported. Three crystal forms were grown in different crystallization conditions. The unliganded structure was solved using sulfur SAD data collected from a single crystal using the I23 in vacuo diffraction beamline at Diamond Light Source. Crystal soaks with arabinose, 1,4-dideoxy-1,4-imino-l-arabinitol and two cyclophellitol-derived arabinose mimics reveal a conserved catalytic site and conformational itinerary between fungal and bacterial GH51 α-l-arabino­furanosidases.

KW - arabinofuranosidases

KW - cyclophellitol

KW - glycoside hydrolases

KW - iminosugar

KW - sulfur SAD

UR - http://www.scopus.com/inward/record.url?scp=85095402335&partnerID=8YFLogxK

U2 - 10.1107/S205979832001253X

DO - 10.1107/S205979832001253X

M3 - Article

C2 - 33135683

AN - SCOPUS:85095402335

SN - 2059-7983

VL - 76

SP - 1124

EP - 1133

JO - Acta Crystallographica Section D: Structural Biology

JF - Acta Crystallographica Section D: Structural Biology

IS - 11

ER -