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Products >  Protein_Research >  Protein_Folding_and_Expression >  Brevibacillus_Expression_System_II

Recombinant Protein Expression: Brevibacillus Expression System II

Brevibacillus choshinensis is a Gram-positive bacterium well-suited for heterologous protein expression. The Brevibacillus Expression System II generates secreted target proteins efficiently1 and is ideal for eukaryotic recombinant protein expression, resulting in a high yield of active protein. The Brevibacillus system is also almost completely free of proteases, allowing for the production of intact protein products.

The Brevibacillus system facilitates disulfide bond formation, which is often required for activity in proteins of eukaryotic origin. In addition, B. choshinensis serves as an excellent host for intracellular protein production, producing soluble intracellular proteins in the cytoplasm without the formation of inclusion bodies. In fact, the Brevibacillus Expression System II often works better than comparable E. coli-based systems for the expression of certain recombinant protein targets.

Using His-tag vectors (pNC-HisE, pNC-HisF, pNC-HisT, pNI-His) allows for quick and easy purification of the expressed target proteins. These tags can be removed by protease treatment following purification.

  At-A-Glance   Documents   Images & Data

Features

  • Efficient production of secreted or intracellular recombinant target proteins
  • Negligible amounts of extracellular protease - products remain intact in culture medium
  • Unlike E. coli, no endotoxins
  • Proteins are produced in active form
  • Easy to culture, handle and sterilize

Components

HB200

Brevibacillus Expression System1 kit
Expression vector
» pNY326 DNA10 µg
» pNCMO2 DNA10 µg
Control vector
» pNY326-BLA DNA1 µg
Competent Cells
» Brevibacillus choschinesis competent cells100 µL x 10 tubes

Notes

This product requires completion of a License Agreement before purchase.

Examples of Proteins Expressed using the Brevibacillus Expression System II

Examples of expressed proteins using the Brevibacillus Expression System II are shown below (see Table 1). High expression levels have been achieved for a variety of proteins (enzymes, antigens and cytokines) regardless of their genetic origin (bacterial, archaeal and eukaryotic). Because secreted eukaryotic proteins often depend on intact disulfide bonds for activity, it is generally difficult to produce active proteins using typical prokaryote-based expression systems. However, due to the secretory advantages of the Brevibacillus Expression System II, high expression levels of active recombinant protein are possible even for proteins with extensive disulfide bonds.

Table 1. Examples of successful heterologous protein expression using the B. choshinensis host expression system
ProteinsOriginsProduction (g/L)References
Enzymes
α-amylaseB. licheniformis3.7 
SphingomyelinaseB. cereus3.0 
XylanaseB. halodurans0.2 
CGTaseB. macerans1.52
ChitosanaseB. circulans1.4 
Hyper thermo-stable proteaseA. pernix0.1 
Hyper thermo-stable nucleaseP. horikoshii0.7 
PDIhuman1.03
Antigens
Surface antigenE. rhusiopathiae0.9 
Surface antigenT. pallidum0.8 
Cytokines
EGFhuman1.54
NGFmouse0.2 
IFN-γchicken0.55
TNF-αbovine0.4 
GM-CSFbovine0.2 
GHflounder0.2

References

  1. Takagi, H. et al. (1989) Agric. Biol. Chem. 53(3):691-699.
  2. Takano, T. et al. (1992) Biosci. Biotech. Biochem. 56(5):808-809.
  3. Tojo, H. et al. (1994) J. Biotechnol. 33(1):55-62.
  4. Yamagata, H. et al. (1989) Proc. Natl. Acad. Sci. USA. 86:3589-3593.
  5. Yashiro, K. et al. (2001) ProtExpression and Purification  23:113-120.

Examples of Use

  1. Teramura, N. et al. (2011) J. Bacteriol. 193(12):3049-3056.
  2. Yamamoto, A. et al. (2011) Vet. Immunol. Immunopathol. 142(3-4):271-275.
  3. Sugimoto, S. et al. (2011) J. Appl. Microbiol. 111(6):1406-1415.
  4. Mizukami, M. et al. (2010) Curr. Pharm. Biotechnol. 11(3):251-258.
  5. Yamamoto, A. et al. (2009) Vet. Immunol. Immunopathol. 130(3-4):221-225.
 



 
 
Products
Cat. # Product Contents Size Price License Units Select
HB116 ◊Brevibacillus Choshinensis Competent Cells 10 x 100 uL $337.00 License Statements
HB200 ◊Brevibacillus Expression System II 1 Kit $1,224.00 License Statements
HB123 ◊pNC-HisE DNA 10 ug $377.00 License Statements
HB122 ◊pNC-HisF DNA 10 ug $377.00 License Statements
HB121 ◊pNC-HisT DNA 10 ug $377.00 License Statements
HB112 ◊pNCMO2 DNA 10 ug $377.00 License Statements
HB131 ◊pNI DNA 10 ug $377.00 License Statements
HB132 ◊pNI-His DNA 10 ug $377.00 License Statements
HB111 ◊pNY 326 DNA 10 ug $377.00 License Statements
HB114 ◊pNY326-BLA DNA 1 ug $214.00 License Statements
 

 

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