Fig. 1
The construction of the constitutive expression vectors for N-terminal lipoprotein anchoring of mannanase (ManB) in L. plantarum. a Schematic overview of the pSIP409 expression vector, the starting point for construction. The sppK and sppR genes encode the proteins in the two-component regulatory system. The gene of interest is placed under control of a strong inducible bacteriocin promoter PsppQ-sakacin Q. The multiple cloning site (MCS), the replicon region consists of two determinants pUC(pGEM)-ori for E. coli and 256rep for L. sakei and L. plantarum. The selection marker is the erythromycin resistance gene (ery), the lollipops indicate transcription terminators, L indicates the SalI-linker sequence. b Schematic overview of the constitutive expression cassette for N-terminal lipoprotein anchoring of mannanase (ManB) with BglII and EcoRI cloning sites. The inserts containing the manB sequence were fused with a myc tag for protein detection. Ppgm and PslpA: the promoters of a phosphoglycerate mutase (pgm) from L. acidophilus NCFM and a S-layer protein SlpA of L. acidophilus ATCC 4356, respectively; SP1261 and Lp_1261: a signal peptide and a lipoprotein anchor; SPase: lipobox withSignal Peptidase II cleavage site (SPase); L: linker (SalI site)