Development and application of a novel signal peptide probe vector with PGA as reporter in Bacillus subtilis WB700: twenty-four tat pathway signal peptides from Bacillus subtilis were monitored.
In this study, we have developed a novel, versatile signal peptide probe vector driven by promoter P43 in Bacillus subtilis WB700, using Penicillin G Acylase (PGA) as reporter. Twenty-four signal peptides considered belonging to twin-arginine translocation (Tat) pathway were cloned into the probe vector to direct the secretion expression of PGA, respectively. Through 6-nitro-3-phenylacetamidobenzoic acid (NIPAB) filter paper assay, four signal peptides (AmyX, AlbB, LipA, and YmzC) were chosen for further investigation. The extracellular production of PGA demonstrated that these recombinants mediated efficient secretion expression in B. subtilis WB700, in which the maximum activity reached 0.11, 0.21, 0.08, and 0.26 U/mL, respectively. Thus, we provided an efficient tool for easy detection of the signal peptides in B. subtilis, and demonstrated the efficiency of Tat pathway signal peptides via PGA secretion in B. subtilis WB700.
Zhu
Fa-Ming
FM
College of Animal Sciences, Northwest A & F University, YangLing 712100, PR China.
Ji
Sheng-Yue
SY
Zhang
Wei-Wei
WW
Li
Wang
W
Cao
Bin-Yun
BY
Yang
Ming-Ming
MM
D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D017930:Genes, Reporter; D005822:Genetic Vectors; D015335:Molecular Probes; D010399:Penicillin Amidase / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics*
D017931:DNA Primers; D015335:Molecular Probes; D021382:Protein Sorting Signals; D010399:Penicillin Amidase