Functional and biophysical characterization of a hyperthermostable GH51 α-L-arabinofuranosidase from Thermotoga petrophila
Overview of dos Santos CR et al.
| Authors | dos Santos CR Squina FM Navarro AM Oldiges DP Paes Leme AF Ruller R Mort AJ Prade R Murakami MT |
|---|---|
| Affiliation | Laboratório Nacional de Biociências (LNBio) Centro Nacional de Pesquisa em Energia e Materiais Rua giuseppe maximo scolfaro 10000 Campinas SP 13083-970 Brazil. |
| Journal | Biotechnol Lett |
| Year | 2010 |
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.