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subtilis are secreted into the medium, there are also successful examples of cytoplasmic protein production in B. Although most of the proteins that are commercially produced by B. Therefore, the secreted proteins can be purified easily from the medium in their active form, which simplifies the downstream processing considerably. subtilis lacks an outer membrane and is able to secrete proteins directly into the medium. subtilis is also capable of producing large quantities of proteins. subtilis has excellent fermentation capacities that are equal to, if not better, than those of E. coli, and of Gram-negative bacteria in general, contains lipopolysaccharide (LPS or endotoxin), which is highly pyrogenic and needs to be totally removed before the produced proteins can be used for clinical purposes.ī. To acquire the protein, the inclusion bodies need to be separated from the cell and the proteins subsequently need to be recovered from the inclusion bodies. coli the produced proteins usually accumulate within the cells where they have a high potential to aggregate, resulting in the formation of inclusion bodies. Advantages of its use as a production host for proteins are that it can be grown easily in large fermentations, and that it is genetically amenable and able to produce large quantities of proteins. 1.2 Bacillus subtilis as a host for protein productionĬurrently the most commonly used bacterial host for industrial production of heterologous proteins is Escherichia coli. subtilis and clinically relevant Gram-positive pathogens also make this organism highly relevant for research on potential targets for novel antimicrobials and anti-infectives. Importantly, the relatively close relationships between B. subtilis are available, representing a rich source of information for research on Bacillus species. Detailed data sets on the transcriptome, proteome, secretome and metabolome of B.
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subtilis is one of the best understood of all living organisms, and it has become the paradigm for research on Gram-positive bacteria. A subsequent international project has led to the identification of all essential genes in B. This has led to the publication of the entire annotated genome sequence in 1997. In 1990 a European-Japanese research collaboration was started, with the aim to sequence the entire genome of B. This is one of the prime reasons why bacilli have been extensively used in both applied and fundamental scientific research for more than 50 years. subtilis is genetically highly amenable as it develops genetic competence for DNA binding and uptake. Therefore, this organism is widely regarded as a prolific "cell factory" for industrial enzymes and biopharmaceuticals. subtilis is able to produce and secrete large quantities of proteins into the culture medium.
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subtilis and related Bacillus species are being used in industry for a wide range of different applications. Many different enzymes, like proteases and amylases, originating from B. subtilis is best known as a source of useful enzymes and fine biochemicals, and as an attractive host for the production of heterologous proteins. subtilis dates back more than a thousand years, when it was already used to produce natto, a Japanese food product consisting of fermented soybeans. subtilis has even been awarded GRAS (Generally Recognized As Safe) status by the US Food and Drug Administration. subtilis that limit the production of high quality protein complexes and membrane proteins, and to enhance those systems that facilitate assembly of these proteins.īacillus subtilis is a sporulating rod-shaped Gram-positive bacterium (Fig. It will therefore be a major challenge for future research to identify and modulate quality control systems of B. Mechanisms underlying degradation of secretory proteins are nowadays well understood and often controllable. While inactivation of quality control systems has the potential to improve protein production yields, this could be achieved at the expense of product quality. Paradoxically, cellular quality control systems also represent bottlenecks for the production of various heterologous proteins at significant concentrations. subtilis results from the action of cellular quality control systems that efficiently remove misfolded or incompletely synthesized proteins. The high quality of proteins produced with B. subtilis can be exploited for protein complexes and membrane proteins to facilitate their functional and structural analysis. Therefore, one may anticipate that the high protein production potential of B. Most of these proteins are secreted into the growth medium, but successful examples of cytoplasmic protein production are also known. The Gram-positive bacterium Bacillus subtilis is an important producer of high quality industrial enzymes and a few eukaryotic proteins.