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 efficiently (Takagi et al. 1989) 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.
Brevibacillus choshinensis is a Gram-positive bacterium well-suited for heterologous protein expression. The Brevibacillus Expression System II generates secreted target proteins efficiently (Takagi et al. 1989) 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.
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
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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.
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