62 BIOTECHNOLOGY
Fig. 2. High-density expression of GFP in B. megaterium.
Photo courtesy of Prof. Dr. M. Rohde, TU Braunschweig, Germany..
transconjugation. Using His- and Strep-affinity tags, the intra- or extracellularly produced proteins can easily be purified in one-step procedures.
Different gene expression systems based on the xylose controlled promoter P(xylA) and various phage RNA polymerase driven systems (for instance, T7) enable B. megaterium to produce up to 1.25 g of recombinant protein per litre(4)
.
Lactococcus lactis, a food-grade bacterial host that has also been validated as suitable for the production of GRAS substances, is used in the production of foreign proteins that are applied to the food and pharmaceutical industry.
Lactococcus lactis is a further Gram-positive lactic acid bacterium that, in addition to its traditional use in food fermentations, is increasingly used in modern biotechnological applications.
optimisation enhanced the expression yields significantly (Dr. Nguyen, Duc Hoang, pers. comm.). A recent
L Westers, H Westers, and WJ Quax Bacillus subtilis as cell factory for pharmaceutical proteins: a biotechnological approach to optimize the host organism. Biochim Biophys Acta, Nov 2004; 1694(1-3): 299-
REFERENCES: 1
310. 2
Phan, T.T.P., Nguyen, H.D. and Schumann, W. (2006). Novel plasmid- based expression vectorsfor intra- and extracellularproduction of recombinantproteins in Bacillus subtilis. Protein Expr. Purif.; 2006
Apr; 46(2): 189-95 3
30. 4
Iwanicki A, Piątek I, Stasiłojć M, Grela A, Lęga T, Obuchowski M, Hinc K. A system of vectors for Bacillus subtilis spore surface display.
Microb Cell Fact. 2014 Feb 24;13(1):30. doi: 10.1186/1475-2859-13-
Simon Stammen, Britta Katrin Müller, Claudia Korneli, Rebekka Biedendieck, Martin Gamer, Ezequiel Franco-Lara, and Dieter Jahn High-Yield Intra- and Extracellular Protein Production Using Bacillus
megaterium Appl. Envir. Microbiol., Jun 2010; 76: 4037 - 4046. 5
Mierau, I. and M. Kleerebezem (2005). “10 years of the nisin-
controlled gene expression system (NICE) in Lactococcus lactis.” Applied Microbiology and Biotechnology 9: 1-13
www.scientistlive.com Arne Schulz is with MoBiTec in Germany.
www.mobitec.com
publication has shown that the surface of spores of Bacillus subtilis can be used as, for example, biocatalysts
or antigen-carriers in vaccine formulations(3)
.
A widely used alternative Te Gram-positive bacterium Bacillus megaterium (see Fig. 1) represents an increasingly used alternative for high yield intra- and extracellular protein production. During the past two decades, multiple tools, including gene expression plasmids and production strains, have been developed. Introduction of free replicating and integrative plasmids into B. megaterium is possible via protoplasts transformation or
In the past 25 years great progress has been made in the development of genetic engineering tools and the molecular characterisation of this species. Most notable is the versatile and tightly controlled gene expression system, based on the auto-regulation mechanism of the bacteriocin nisin – the NIsin Controlled gene Expression system, called NICE.
Tis system has become one of the most successful and widely used food-grade tools for regulated gene expression in Gram-positive bacteria(5)
. For more information ✔ at
www.scientistlive.com/eurolab
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