Oligomeric and functional properties of a debranching enzyme (TreX) from the archaeon Sulfolobus solfataricus P2 = Sulfolobus solfataricus에서 유래한 탈가지 효소 TreX의 올리고머 및 기능적 특성

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Title
Oligomeric and functional properties of a debranching enzyme (TreX) from the archaeon Sulfolobus solfataricus P2 = Sulfolobus solfataricus에서 유래한 탈가지 효소 TreX의 올리고머 및 기능적 특성
Author(s)
J T Park; H S Park; H K Kang; J S Hong; H Cha; Eui-Jeon Woo; J W Kim; M J Kim; W Boos; S Lee; K H Park
Bibliographic Citation
Biocatalysis and Biotransformation, vol. 26, no. 1-2, pp. 76-85
Publication Year
2008
Abstract
A gene, treX, encoding a debranching enzyme previously cloned from the trehalose biosynthesis gene cluster of Sulfolobus solfataricus P2 was expressed in Escherichia coli as a His-tagged protein and the biochemical properties were studied. The specific activity of the S. solfataricus debranching enzyme (TreX) was highest at 75°C and pH 5.5. The enzyme exhibited hydrolysing activity toward α-1,6-glycosidic linkages of amylopectin, glycogen, pullulan, and other branched substrates, and glycogen was the preferred substrate. TreX has a high specificity for hydrolysis of maltohexaosyl α-1,6-β-cyclodextrin, indicating the high preference for side chains consisting of 6 glucose residues or more. The enzyme also exhibited 4-α-sulfoxide-glucan transferase activity, catalysing transfer of α-1,4-glucan oligosaccharides from one chain to another. Dimethyl sulfoxide (10%, v/v) increased the hydrolytic activity of TreX. Gel permeation chromatography and sedimentation equilibrium analytical ultracentrifugation revealed that the enzyme exists mostly as a dimer at pH 7.0, and as a mixture of dimers and tetramers at pH 5.5. Interestingly, TreX existed as a tetramer in the presence of DMSO at pH 5.5-6.5. The tetramer showed a 4-fold higher catalytic efficiency than the dimer. The enzyme catalysed not only intermolecular trans-glycosylation of malto-oligosaccharides (disproportionation) to produce linear α-1,4-glucans, but also intramolecular trans-glycosylation of glycogen. The results presented in this study indicated that TreX may be associated with glycogen metabolism by selective cleavage of the outer side chain.
Keyword
associationdebranching enzymedimethyl sulfoxidedisproportionationglycogen breakdownsulfolobus solfataricus P2
ISSN
1024-2422
Publisher
T&F (Taylor & Francis)
DOI
http://dx.doi.org/10.1080/10242420701806652
Type
Article
Appears in Collections:
Division of Biomedical Research > Disease Target Structure Research Center > 1. Journal Articles
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