B.subtilis Secretory Protein Expression System

B. subtilis Secretory Protein Expression System

Bacillus subtilis has become an increasingly popular host for recombinant protein expression. With its ability to secrete protein directly into culture media, amenability to medium- and large-scale fermentation, lack of codon bias, and designation by the U.S. Food and Drug Administration as an organism that is Generally Regarded As Safe (GRAS), it's no wonder that the majority of industrially-produced enzymes are expressed in Bacillus species such as B. subtilis. Optimization of secretion, however, can be necessary to achieve highest yields. To address this, the B. subtilis Secretory Protein Expression System from Takara Bio allows rapid development of a library of B. subtilis clones, each bearing a pBE-S construct in which the ORF for your protein of interest is fused with sequences for 173 unique signal peptides. Perform a downstream assay to identify and select clones which secrete the highest amount of functional protein into the culture media, and you can quickly identify the signal peptide that results in efficient expression of your desired secreted protein.

Features

Includes pBE-S DNA, an E. coli/B. subtilis shuttle vector with B. subtilis-derived subtilisin (aprE) promoter, secretory signal peptide (aprE SP), Multiple Cloning Site, and 3' (C-terminal) His-tag sequence

Supplied with SP DNA Mixture, a library of DNA sequences encoding 173 unique secretory signal peptides that can be inserted upstream of your target gene

Fully compatible with In-Fusion cloning kits and systems to allow rapid and easy construct generation

Includes B. subtilis strain RIK1285

Applications

Expression of soluble, recombinant protein secreted directly into the culture media
Protein expression in a host amenable to medium- and large-scale fermentation in addition to small-scale culturing
Expression of proteins with complex structure, such as proteins with disulfide (S-S) bonds
Generation of target protein in a host that is considered to be Generally Regarded As Safe (GRAS) by the U.S. Food and Drug Administration
Useful for producing easily purified recombinant protein: with proper in-frame cloning, a C-terminal His tag can aid purification from culture media

Components


SP DNA Mixture (0.032 pmol/µL)	45 µL
pBE-S DNA (0.5 µg/µL)	20 µg
B. subtilis RIK1285 (Glycerol Stock)	100 µL x 2

Notes:

The System provides sufficient reagents for 10 library development reactions.
Genotype of B. subtilis RIK1285:Marburg 168 derivative: trpC2, ys1, aprEdelta3, nprR2, nprE18

Storage

SP DNA mixture and pBE-S DNA: –20°C

B. subtilis

pBE-S Vector information

For vector map, see the User Manuals under "Documents". For vector sequence files, see "Resources."

Related In-Fusion Cloning products

For rapid and easy cloning, use the In-Fusion Cloning Systems or Kits. Products in the In-Fusion portfolio vary by geographic location; see the In-Fusion product pages from Clontech for more information about products available in your region.