Guo, J;
Coker, AR;
Wood, SP;
Cooper, JB;
Keegan, RM;
Ahmad, N;
Muhammad, MA;
... Akhtar, M; + view all
(2018)
Structure and function of the type III pullulan hydrolase from Thermococcus kodakarensis.
Acta Crystallographica: Section D - Structural Biology
, 74
pp. 305-314.
10.1107/S2059798318001754.
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Abstract
Pullulan-hydrolysing enzymes, more commonly known as debranching enzymes for starch and other polysaccharides, are of great interest and have been widely used in the starch-saccharification industry. Type III pullulan hydrolase from Thermococcus kodakarensis (TK-PUL) possesses both pullulanase and α-amylase activities. Until now, only two enzymes in this class, which are capable of hydrolysing both α-1,4- and α-1,6-glycosidic bonds in pullulan to produce a mixture of maltose, panose and maltotriose, have been described. TK-PUL shows highest activity in the temperature range 95–100°C and has a pH optimum in the range 3.5–4.2. Its unique ability to hydrolyse maltotriose into maltose and glucose has not been reported for other homologous enzymes. The crystal structure of TK-PUL has been determined at a resolution of 2.8 Å and represents the first analysis of a type III pullulan hydrolyse. The structure reveals that the last part of the N-terminal domain and the C-terminal domain are significantly different from homologous structures. In addition, the loop regions at the active-site end of the central catalytic domain are quite different. The enzyme has a well defined calcium-binding site and possesses a rare vicinal disulfide bridge. The thermostability of TK-PUL and its homologues may be attributable to several factors, including the increased content of salt bridges, helical segments, Pro, Arg and Tyr residues and the decreased content of serine.
| Type: | Article |
|---|---|
| Title: | Structure and function of the type III pullulan hydrolase from Thermococcus kodakarensis |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1107/S2059798318001754 |
| Publisher version: | https://doi.org/10.1107/S2059798318001754 |
| Language: | English |
| Additional information: | This version is the version of record. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | Science & Technology, Life Sciences & Biomedicine, Physical Sciences, Biochemical Research Methods, Biochemistry & Molecular Biology, Biophysics, Crystallography, pullulan hydrolase, Thermococcus kodakarensis, protein crystallography, structural biology, ALPHA-AMYLASE FAMILY, PROTEIN THERMAL-STABILITY, ION-PAIR NETWORKS, CRYSTAL-STRUCTURE, MALTOGENIC AMYLASE, ACTIVE-SITE, BACILLUS-STEAROTHERMOPHILUS, MOLECULAR REPLACEMENT, PYROCOCCUS-FURIOSUS, ANGSTROM RESOLUTION |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Wolfson Inst for Biomedical Research |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10049183 |
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