Molecular Biotechnology

, Volume 39, Issue 3, pp 225–230 | Cite as

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

  • Fa-Ming Zhu
  • Sheng-Yue Ji
  • Wei-Wei Zhang
  • Wang Li
  • Bin-Yun Cao
  • Ming-Ming Yang
Research

Abstract

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.

Keywords

PGA B. subtilis WB700 Tat signal peptide Secretion expression 

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Copyright information

© Humana Press Inc. 2008

Authors and Affiliations

  • Fa-Ming Zhu
    • 1
    • 2
  • Sheng-Yue Ji
    • 1
    • 2
  • Wei-Wei Zhang
    • 2
  • Wang Li
    • 1
    • 2
  • Bin-Yun Cao
    • 1
  • Ming-Ming Yang
    • 1
    • 2
  1. 1.College of Animal Sciences Northwest A & F UniversityYangLingP.R. China
  2. 2.Yangguang-Guangji Medical R&D Co. LtWuhanP.R. China

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