Abstract
A hyperthermostable glycoside hydrolase family 51 (GH51) α-l-arabinofuranosidase from Thermotoga petrophila RKU-1 (TpAraF) was cloned, overexpressed, purified and characterized. The recombinant enzyme had optimum activity at pH 6.0 and 70°C with linear α-1,5-linked arabinoheptaose as substrate. The substrate cleavage pattern monitored by capillary zone electrophoresis showed that TpAraF is a classical exo-acting enzyme producing arabinose as its end-product. Far-UV circular dichroism analysis displayed a typical spectrum of α/β barrel proteins analogously observed for other GH51 α-l-arabinofuranosidases. Moreover, TpAraF was crystallized in two crystalline forms, which can be used to determine its crystallographic structure.
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Acknowledgments
This research was supported by grants from Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP, 08/58037-9) to FMS and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, 478059/2009-4) to MTM. RAP has funding from the Department of Energy, awards 06103-OKL and ZDJ-7-77608-01.
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The first two authors have contributed equally to this work.
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dos Santos, C.R., Squina, F.M., Navarro, A.M. et al. Functional and biophysical characterization of a hyperthermostable GH51 α-l-arabinofuranosidase from Thermotoga petrophila . Biotechnol Lett 33, 131–137 (2011). https://doi.org/10.1007/s10529-010-0409-3
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DOI: https://doi.org/10.1007/s10529-010-0409-3