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Identification of substrate-binding and selectivity-related residues of maltooligosyltrehalose synthase from the thermophilic archaeonSulfolobus solfataricusATCC 35092

Authors
Journal
Enzyme and Microbial Technology
0141-0229
Publisher
Elsevier
Publication Date
Volume
56
Identifiers
DOI: 10.1016/j.enzmictec.2014.01.003
Keywords
  • Maltooligosyltrehalose Synthase
  • Hydrogen Bond
  • Site-Directed Mutagenesis
  • Selectivity
  • Computer Simulation
  • Sulfolobus
Disciplines
  • Biology

Abstract

Abstract Maltooligosyltrehalose synthase (MTSase) is a key enzyme in the synthesis of trehalose. Computer simulations using AutoDock and NAMD were employed to assess the substrate-binding and selectivity-related residues of MTSase. We introduced mutations at residues D411, D610, and R614 to determine the substrate-binding residues of Sulfolobus solfataricus ATCC 35092 MTSase, and introduced mutations at residues P402, A406, and V426 to investigate the enzyme's selectivity-related residues. Kinetic studies of D411A, D610A, and R614A MTSases reveal significant reductions in catalytic efficiency and cause increase in the transition-state energy of mutant MTSases, indicating that residues D411, D610, and R614 form hydrogen bonds to the substrate. Compared with wild-type MTSase, the hydrolysis: transglycosylation selectivity ratio was significantly decreased for P402Q and significantly increased for A406S MTSases, while the ratio for V426T MTSase showed little change. The results suggest that P402 and A406 residues are selectivity-related.

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