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Characterization and regulation of a bacterial sugar phosphatase of the haloalk...
Godinho, Lia M.; Sá-Nogueira, Isabel deFEBS journal, Volume 278, Issue 14, pages 2511-2524, July 2011 ; AraL from Bacillus subtilis is a member of the ubiquitous haloalkanoate dehalogenase superfamily. The araL gene has been cloned, over-expressed in Escherichia coli and its product purified to homogeneity. The enzyme displays phosphatase activity, which is optimal at neutral pH (7.0) and 65 C. Substrate screening and kinetic analysis showed AraL ...
A multitask ATPase serving different ABC-type sugar importers in Bacillus subtilis
Sá-Nogueira, Isabel de; Ferreira, Mário JoséJournal of Bacteriology (Out 2010) 5312-5318 ; Bacillus subtilis is able to utilize arabinopolysaccharides derived from plant biomass. Here, by combining genetic and physiological analyses we characterize the AraNPQ importer and identify primary and secondary transporters of B. subtilis involved in the uptake of arabinosaccharides. We show that the ABC-type importer AraNPQ is involved in the uptake of α-1,5-ar...
Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degrada...
Sá-Nogueira, Isabel de; Inácio, José Manuel; Correia, Isabel LopesMicrobiology 154 (2008) 2719-2729 ; Bacillus subtilis produces alpha-l-arabinofuranosidases (EC 3.2.1.55; AFs) capable of releasing arabinosyl oligomers and l-arabinose from plant cell walls. Here, we show by insertion-deletion mutational analysis that genes abfA and xsa(asd), herein renamed abf2, encode AFs responsible for the majority of the intracellular AF activity in B. subtilis. Both enzyme activities we...
Characterization of the abn2(yxiA) encoding a Bacillus subtilis GH43 arabinanas...
Sá-Nogueira, Isabel de; Inácio, José ManuelJournal of Bacteriology (Junho 2008) 4272-4280 ; The extracellular depolymerization of arabinopolysaccharides by microorganisms is accomplished by arabinanases, xylanases, and galactanases. Here, we characterize a novel endo-alpha-1,5-l-arabinanase (EC 3.2.1.99) from Bacillus subtilis, encoded by the yxiA gene (herein renamed abn2) that contributes to arabinan degradation. Functional studies by mutational anal...
Overproduction, crystallization and preliminary X-ray characterization of Abn2,...
Sá-Nogueira, Isabel de; Sanctis, Daniele de; Bento, Isabel; Inácio, José Manuel; Custódio, Sónia; Carrondo, Maria ArméniaActa Crystallographica F64 (2008) 636-638 ; Two Bacillus subtilis extracellular endo-1,5-alpha-L-arabinanases, AbnA and Abn2, belonging to glycoside hydrolase family 43 have been identified. The recently characterized Abn2 protein hydrolyzes arabinan and has low identity to other reported 1,5-alpha-L-arabinanases. Abn2 and its selenomethionine (SeMet) derivative have been purified and crystallized. Crystals ap...
trans-Acting factors and cis elements involved in glucose repression of arabina...
Sá-Nogueira, Isabel de; Inácio, José ManuelJournal of Bacteriology (Nov 2007) 8371-8376 ; In Bacillus subtilis, the synthesis of enzymes involved in the degradation of arabinose-containing polysaccharides is subject to carbon catabolite repression (CCR). Here we show that CcpA is the major regulator of repression of the arabinases genes in the presence of glucose. CcpA acts via binding to one cre each in the promoter regions of the abnA and xsa genes a...
Probing key DNA contacts in AraR-mediated transcriptional repression of the Bac...
Sá-Nogueira, Isabel de; Franco, Irina Saraiva; Mota, Luís Jaime; Soares, Cláudio ManuelNucleic Acid Research (2007) Vol.37 N. 14 4755-4766 ; In the absence of arabinose, the AraR transcription factor represses the expression of genes involved in the utilization of arabinose, xylose and galactose in Bacillus subtilis. AraR exhibits a chimeric organization: the N-terminal DNA-binding region belongs to the GntR family and the C-terminal effector-binding domain is homologous to the GalR/LacI family....
Functional domains of Bacillus subtilis transcription factor AraR and identific...
Sá-Nogueira, Isabel de; Franco, Irina Saraiva; Mota, Luís Jaime; Soares, Cláudio ManuelJournal of Bacteriology (Apr 2006) 3024-3036 ; The Bacillus subtilis AraR transcription factor represses at least 13 genes required for the extracellular degradation of arabinose-containing polysaccharides, transport of arabinose, arabinose oligomers, xylose, and galactose, intracellular degradation of arabinose oligomers, and further catabolism of this sugar. AraR exhibits a chimeric organization comprising a...
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