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Structure and function of the type III pullulan hydrolase from Thermococcus kodakarensis

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. Green open access

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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
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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