The Sphingomonas paucimobilis β-glucosidase Bgl1 is encoded by the bgl1 gene, associated with an 1308bp open reading frame. The deduced protein has a potential signal peptide of 24 amino acids in the N-terminal region, and experimental evidence is consistent with the processing and export of the Bgl1 protein through the inner membrane to the periplasmic space. A His6-tagged 44.3kDa protein was over-produced in the cytosol of Escherichia coli from a recombinant plasmid, which contained the S. paucimobilis bgl1 gene lacking the region encoding the putative signal peptide. Mature β-glucosidase Bgl1 is specific for aryl-β-glucosides and has no apparent activity with oligosaccharides derived from cellulose hydrolysis and other saccharides. A structure-based alignment established structural relations between S. paucimobilis Bgl1 and other members of the glycoside hydrolase (GH) family 1 enzymes. At subsite —1, the conserved residues required for catalysis by GH1 enzymes are present in Bgl1 with only minor differences. Major differences are found at subsite +1, the aglycone binding site. This alignment seeded a sequence-based phylogenetic analysis of GH1 enzymes, revealing an absence of horizontal transfer between phyla. Bootstrap analysis supported the definition of subfamilies and revealed that Bgl1, the first characterized β-glucosidase from the genus Sphingomonas, represents a very divergent bacterial subfamily, closer to archaeal subfamilies than to others of bacterial origin.
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March 2003
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Research Article|
March 15 2003
Sphingomonas paucimobilis beta-glucosidase Bgl1: a member of a new bacterial subfamily in glycoside hydrolase family 1
Ana Rita MARQUES;
Ana Rita MARQUES
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
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Pedro M. COUTINHO;
Pedro M. COUTINHO
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
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Paula VIDEIRA;
Paula VIDEIRA
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
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Arsénio M. FIALHO;
Arsénio M. FIALHO
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
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Isabel SÁ-CORREIA
Isabel SÁ-CORREIA
1
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
1To whom correspondence should be addressed (e-mail isacorreia@ist.utl.pt).
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Publisher: Portland Press Ltd
Received:
August 08 2002
Revision Received:
November 13 2002
Accepted:
November 22 2002
Accepted Manuscript online:
March 15 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 370 (3): 793–804.
Article history
Received:
August 08 2002
Revision Received:
November 13 2002
Accepted:
November 22 2002
Accepted Manuscript online:
March 15 2003
Citation
Ana Rita MARQUES, Pedro M. COUTINHO, Paula VIDEIRA, Arsénio M. FIALHO, Isabel SÁ-CORREIA; Sphingomonas paucimobilis beta-glucosidase Bgl1: a member of a new bacterial subfamily in glycoside hydrolase family 1. Biochem J 15 March 2003; 370 (3): 793–804. doi: https://doi.org/10.1042/bj20021249
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