Open AccessPurification, characterization and functional analysis of an endo‐arabinanase (AbnA) from Bacillus subtilis
Author(s) -
Leal Teresa Fontes,
SáNogueira Isabel
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2004.10.003
Subject(s) - bacillus subtilis , periplasmic space , escherichia coli , biochemistry , recombinant dna , bacillaceae , enzyme , bacillales , signal peptide , biology , hydrolysis , residue (chemistry) , chemistry , gene , bacteria , genetics
Bacillus subtilis synthesizes at least one arabinanase encoded by the abnA gene that is able to degrade the polysaccharide arabinan. Here, we report the expression in Escherichia coli of the full‐length abnA coding region with a His 6 ‐tag fused to the C‐terminus. The recombinant protein was secreted to the periplasmic space and correctly processed by the E. coli signal peptidase. The substrate specificity of purified AbnA, the physico‐chemical properties and kinetic parameters were determined. Functional analysis studies revealed Glu 215 as a key residue for AbnA hydrolytic activity and indicated that in addition to AbnA B. subtilis secretes other enzyme(s) able to degrade linear 1,5‐α‐ l ‐arabinan.