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Products >  Protein_Research >  Protein_Folding_and_Expression >  Brevibacillus_Expression_System_II

Brevibacillus Expression System II

Brevibacillus choshinensis is a Gram positive bacterium which has exceptional capacity for heterologous protein expression. The Brevibacillus Expression System II, enables highly efficient production of target protein in secreted form1. This system allows high yield of active proteins and is well-suited for expression of eukaryotic proteins. The Brevibacillus system is nearly free of proteases, which facilitates production of intact protein products.

The Brevibacillus system facilitates disulfide bond formation (commonly required in proteins of eukaryotic orign). In addition, B. choshinensis serves as an excellent host for intracellular protein production, frequently producing intracellular proteins in soluble form in the cytoplasm without forming inclusion bodies. The Brevibacillus system often works better than E. coli for expression of particular targets.

Utilizing His-tag containing vectors (pNC-HisE, pNC-HisF, pNC-HisT, pNI-His) allows effective purification of the expressed target protein. Tags can be removed by protease treatment following purification.
At-A-Glance Documents Images & Data

Features

  • Efficient production of secreted or intracellular target proteins
  • Produces negligible amounts of extracellular protease - products remain intact in culture medium
  • Unlike E. coli, produces 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

Examples of protein production using this system are shown in in the table below. High expression levels have been achieved with a variety of proteins (enzymes, antigens and cytokines) regardless of gene origin (bacteria, archaea and eukaryotes). In particular, secretory proteins of eukaryotic origin typically have a structure that makes use of S-S bonds for obtaining activity, and it is generally difficult to produce them using other prokaryotic expression systems. The characteristic secretory production of Brevibacillus has been demonstrated to make high efficiency production possible, even of proteins with S-S bonds.

Table 1. Some examples of successful production of heterologous proteins using B. choshinensis host-vector system
Proteins Origins Production (g/L) References
Enzymes
α-amylase B. licheniformis 3.7  
Sphingomyelinase B. cereus 3.0  
Xylanase B. halodurans 0.2  
CGTase B. macerans 1.5 2
Chitosanase B. circulans 1.4  
Hyper thermo-stable protease A. pernix 0.1  
Hyper thermo-stable nuclease P. horikoshii 0.7  
PDI human 1.0 3
Antigens
Surface antigen E. rhusiopathiae 0.9  
Surface antigen T. pallidum 0.8  
Cytokines
EGF human 1.5 4
IL-2 human 0.6 5
NGF mouse 0.2  
IFN-γ chicken 0.5 6
TNF-α bovine 0.4  
GM-CSF bovine 0.2  
GH flounder 0.2

References

  1. H. Takagi, K. Kadowaki and S. Udaka., (1989) Agric. Biol. Chem.,  53 (3), 691-699.
  2. T. Takano, A. Miyauchi, H. Takagi, K. Kadowaki,  K. Yamane, S. Kobayashi., (1992) Biosci. Biotech. Biochem., 56  (5), 808-809.
  3. H. Tojo, T. Asano, K. Kato, S. Udaka, R. Horinouchi, A.  Kakinuma., (1994) J. Biotechnol., 33 (1), 55-62.
  4. H.  Yamagata, K. Nakahama, Y. Suzuki, A. Kakinuma, N. Tsukakoshi, S. Udaka., (1989)  Proc. Natl. Acad. Sci. USA., 86, 3589-3593.
  5. Y. Takimura, M.  Kato, T. Ohta, H. Yamagata, S. Udaka., (1997) Biosci. Biotechnol.  Biochem., 61 (11), 1858-1861.
  6. K. Yashiro, J. W. Lowenthal, T.  E. O' Neil, S. Ebisu, H. Takagi, (2001) Expression and Purification,  23, 113-120.
  7. N. Teramura et al. (2011) Cloning of a novel collagenase gene from the gram-negative bacterium Grimontia (Vibrio) hollisae 1706B and its efficient expression in Brevibacillus choshinensis. J. Bacteriol. 193 (12), 3049-3056.
  8. A. Yamamoto et al. (2011) Recombinant canine granulocyte colony-stimulating factor accelerates recovery from cyclophosphamide-induced neutropenia in dogs. Vet. Immunol. Immunopathol. 142 (3-4), 271-275.
  9. S. Sugimoto et al. (2011) Cloning, expression and purification of extracellular serine protease Esp, a biofilm-degrading enzyme, from Staphylococcus epidermidis. J. Appl. Microbiol. 111 (6), 1406-1415.
  10. M. Mizukami et al. (2010) Brevibacillus expression system: host-vector system for efficient production of secretory proteins. Curr. Pharm. Biotechnol. 11 (3), 251-258.
  11. A. Yamamoto et al. (2009) Expression and purification of canine granulocyte colony-stimulating factor (cG-CSF). Vet. Immunol. Immunopathol. 130 (3-4), 221-225.
 



 
 
Products
Cat. # Product Package Size Price License # of Units Select
HB116 ◊Brevibacillus Choshinensis Competent Cells 10 x 100 uL $330.00 License Statements
HB200 ◊Brevibacillus Expression System II 1 Kit $1,200.00 License Statements
HB123 ◊pNC-HisE DNA 10 ug $370.00 License Statements
HB122 ◊pNC-HisF DNA 10 ug $370.00 License Statements
HB121 ◊pNC-HisT DNA 10 ug $370.00 License Statements
HB112 ◊pNCMO2 DNA 10 ug $370.00 License Statements
HB131 ◊pNI DNA 10 ug $370.00 License Statements
HB132 ◊pNI-His DNA 10 ug $370.00 License Statements
HB111 ◊pNY 326 DNA 10 ug $370.00 License Statements
HB114 ◊pNY326-BLA DNA 1 ug $210.00 License Statements
 

 

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