Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

Products

Bacillus subtilis Food Grade Expression System

Bacillus subtilis Food Grade Expression System

Vector map of pTTB1

Vector map of pTTB1

Vector map of pTTB2

Vector map of pTTB2

Features

  • Stable high- or low-level expression without addition of any antibiotics
  • All DNA contained in the final expression system is derived from B. subtilis
  • No endotoxins are produced
  • No inclusion bodies are formed
  • Protease-deficient strain for producing secretory enzymes is available
  • Shuttle vectors: Cloning can be done with E. coli

Description

  • The Bacillus Food Grade Selection System is based on the interplay of an endogenous Bacillus toxin EndoA (expressed from vector) and its antitoxin EndoB (expressed from genome). EndoA is an endoribonuclease that specifically cleaves mRNA at a five Base U↓ACAU sequence (Pellegrini et al., 2005, Park et al., 2011). During normal growth conditions EndoA is inactivated by forming a heterohexameric complex with its cognate antitoxin EndoB (Simanshu et al., 2013). Since the antitoxin is relatively unstable, it is essential for the cell to continuously produce sufficient amounts of EndoB to inactivate the more stable toxin. These characteristics are utilized for retaining the vector within the cell.

  • The Bacillus Food Grade Expression System was created to make the advantages of a Bacillus expression system also accessible to areas where antibiotic resistance gene markers are prohibited (e.g., food and feed industry). It enables stable vector-based large scale heterologous protein production by an alternative selection, without antibiotics (Yang S., et al., 2016).

B. subtilis Host Strains

  • TEA: trpC2 ydcDE::Pxyl-ycdE
    The strain B. subtilis TEA is based on B. subtilis 168 Marburg and is recommended for intracellular protein expression and pathway engineering.

  • WEA: nprE aprE epr bpr mpr::ble nprB::bsrΔvpr wprA::hyg cm::neo ydcDE::Pxyl-ycdE
    B. subtilis WEA originated from the eightfold extracellular protease-deficient strain B. subtilis WB800N and is particularly constructed for secretory protein production.

Vectors

  • pTTB1: low copy vector, constitutive promoter for expression
  • pTTB2: high copy vector, constitutive promoter for expression

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
PBS041 pTTB1 vector, lyophilized plasmid DNA 10 µg 527,00 PDF
PBS042 pTTB2 vector, lyophilized plasmid DNA 10 µg 527,00 PDF
PBS043 Bacillus subtilis strain TEA 1 ml 569,00 PDF
PBS044 Bacillus subtilis strain WEA 1 ml 569,00 PDF
  • All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.

Downloads

  • Bacillus subtilis Food Grade Expression System Handbook (PDF)

Vector Maps and Sequences

Vector map of pTTB1 (pic)
Vector map of pTTB2 (pic)

Sequence of pTTB1 (txt)
Sequence of pTTB2 (txt)

Literature

  • Park J.H. et al. (2001). Bacillus subtilis MazF-bs (EndoA) is a UACAU-specific mRNA interferase; FEBS Lett. 585, 2526-2532
  • Pellegrini O. et al. (2005). The Bacillus subtilis ydcDE operon encodes an endoribonuclease of the MazF/Penk family and its inhibitor; Mol Microbiol. 56, 1139-1148
  • Simanshu D.K. et al. (2013). Structural basis of mRNA recognition and cleavage by toxin MazF and its regulation by antitoxin MazE in Bacillus subtilis; Mol.Cell 52, 447-458
  • Yang S. et al. (2016). Construction of a novel, stable, food-grade expression system by engineering the endogenous toxin-antitoxin system in Bacillus subtilis; J. Biotechnol. 219, 40-47