title: Functional and biophysical characterization of a hyperthermostable GH51 α-L-arabinofuranosidase from Thermotoga petrophila.
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.
authors.affiliation: Laboratório Nacional de Biociências (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Rua giuseppe maximo scolfaro, 10000, Campinas, SP, 13083-970, Brazil.
authors.forename: Camila Ramos
authors.initials: CR
authors.lastname: dos Santos
authors.forename: Fábio Márcio
authors.initials: FM
authors.lastname: Squina
authors.forename: Andréia Meza
authors.initials: AM
authors.lastname: Navarro
authors.forename: Daiane Patrícia
authors.initials: DP
authors.lastname: Oldiges
authors.forename: Adriana Franco
authors.initials: AF
authors.lastname: Paes Leme
authors.forename: Roberto
authors.initials: R
authors.lastname: Ruller
authors.forename: Andrew John
authors.initials: AJ
authors.lastname: Mort
authors.forename: Rolf
authors.initials: R
authors.lastname: Prade
authors.forename: Mário Tyago
authors.initials: MT
authors.lastname: Murakami
mesh_terms: D001089:Arabinose / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology*; D002942:Circular Dichroism; D003001:Cloning, Molecular; D003460:Crystallization; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity
chemical_list: D004269:DNA, Bacterial; D011994:Recombinant Proteins; D001089:Arabinose; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase