Takara Recombinant protein expression: Brevibacillus expression systems

Overview

• 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

More Information

Components

Components for the expression systems are provided in their respective user manuals.

Note: 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 in the table below. High expression levels have been achieved for a variety of proteins (enzymes, antigens, and cytokines) regardless of their genetic origin (bacterial, archaeal, or 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.

Proteins	Origins	Production (g/l)	Product citations
Enzymes
α-amylase	B. licheniformis	3.7
Sphingomyelinase	B. cereus	3.0
Xylanase	B. halodurans	0.2
CGTase	B. macerans	1.5	Yamamoto et al. 2011
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	Sugimoto et al. 2011
Antigens
Surface antigen	E. rhusiopathiae	0.9
Surface antigen	T. pallidum	0.8
Cytokines
EGF	Human	1.5	Mizukami et al. 2010
NGF	Mouse	0.2
IFN-γ	Chicken	0.5	Yamamoto et al. 2009
TNF-α	Bovine	0.4
GM-CSF	Bovine	0.2
GH	Flounder	0.2

References

Takagi, H., Kadowaki, K. & Udaka, S. Screening and characterization of protein-hyperproducing bacteria without detectable exoprotease activity. Agric. Biol. Chem. 53, 691–699 (1989).

Product citations

Mizukami, M., Hanagata, H. & Miyauchi, A. Brevibacillus Expression System: Host-Vector System for Efficient Production of Secretory Proteins. Curr. Pharm. Biotechnol. 11, 251–258 (2010).

Sugimoto, S. et al. Cloning, expression and purification of extracellular serine protease Esp, a biofilm-degrading enzyme, from Staphylococcus epidermidis. J. Appl. Microbiol. 111, 1,406–1,415 (2011).

Takano, T. et al. Expression of the Cycledextrin Glucanotransferase Gene of Bacillus macerans in Bacillus brevis. Biosci. Biotech. Biochem. 56, 808–809 (1992).

Teramura, N. et al. 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, 3,049–3,056 (2011).

Tojo, H. et al. Production of human protein disulfide isomerase by Bacillus brevis. J. Biotechnol. 33, 55-62 (1994).

Yamagata, H., Nakahama, N., Suzuki, Y., Tsukagoshi, N. & Udaka, S. Use of Bacillus brevis for efficient synthesis and secretion of human epidermal growth factor. Proc. Natl. Acad. Sci. USA. 86, 3,589–3,593 (1989).

Yamamoto, A., Fujino, M., Tsuchiya, T. & Iwata, A. Recombinant canine granulocyte colony-stimulating factor accelerates recovery from cyclophosphamide-induced neutropenia in dogs. Vet. Immunol. Immunopathol. 142, (2011).

Yamamoto, A., Iwata, A., Saito, T., Watanabe, F. & Ueda, S. Expression and purification of canine granulocyte colony-stimulating factor (cG-CSF). Vet. Immunol. Immunopathol. 130, 221–225 (2009).

Yashiro, K. et al. High-Level Production of Recombinant Chicken Interferon-γ by Brevibacillus choshinensis. Protein Expr. Purif. 23, 113–120 (2001).

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.