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Fig. 2 | AMB Express

Fig. 2

From: Conformational determinants necessary for secretion of Paecilomyces thermophila β-1,4-xylosidase that lacks a signal peptide

Fig. 2

Construction of X12345 mutants and their secretion in E. coli BL21(DE3). a Predicted structure of X12345. The 3D structure of X12345 was predicted by Swiss-Model using the template arabinan-specific α-1, 2-arabinofuranosidase from Cellvibrio japonicas (PDB code, 3QED; 2.99-Å resolution), and colored from the N-terminus (blue) to the C-terminus (red). b Blade organization of the full-length X12345 and its mutants and their secretion in BL21(DE3). The regions of the 1 (blue), 2 (aqua), 3 (green), 4 (yellow) and 5 (red) blades were determined according to the predicted X12345 structure. Each gene was cloned into pET-28 (a) and expressed in BL21(DE3). +, +++ and −, secretion, high-level secretion and no secretion of target protein by BL21(DE3) after 24-h induction, respectively. The relative activity of purified mutants were also shown. ND, not detected. c Subcellular location of mutants of X12345 in BL21(DE3) after lactose induction for different time periods. The lane loadings are directly comparable. SF soluble fraction, IF insoluble fraction, C cells, M medium, P1 periplasm fraction (hypertonic solution fraction), P2 periplasm fraction (hypotonic solution fraction), S spheroplasts

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