Is Bacillus an Alternative Expression System?
In this review we consider the possible use of Bacillus subtilis as a host for the production of heterologous proteins. There are several potential advantages to be gained from the use of this organism, particularly its efficient secretion of proteins into the growth medium. Although it is unlikely to become the first choice host for the production of certain types of protein, for example potentially therapeutic mammalian proteins, which often undergo specific and necessary post-translation modifications, there are certainly some important groups of proteins, for example industrial enzymes, for which the Bacilli are already heavily used. The market for such enzymes is likely to grow very rapidly as recombinant DNA methods allow for the production of proteins from diverse and often little characterised microorganisms. Exploitation of these natural products will depend upon the development of versatile and robust expression systems, and there is growing evidence that Bacillus will be a useful system. On the other hand, Bacillus has certain undesirable properties as a host, such as the elaboration of proteases, which can cause loss of product by degradation. If such problems can be overcome, and in this review we assess the technical difficulties involved, then B. subtilis may turn out to be the host of choice for the production of many of the new industrial enzymes.
KeywordsSucrose Fermentation Albumin Carbohydrate Recombination
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- 2.Errington, J., Generalized cloning vectors for Bacillus subtilis. In Vectors: a Survey of Molecular Cloning Vectors and their Uses, ed. R. L. Rodriguez & D. T. Denhardt. Butterworths, Boston, 1988, pp. 345–62.Google Scholar
- 13.Diderichsen, B., A genetic system for stabilization of cloned genes in Bacillus subtilis. Bacillus Molecular Genetics and Biotechnology applications, ed. A. T. Ganesan & J. A. Hoch. Academic Press, Orlando, 1986, pp. 35–46.Google Scholar
- 16.Errington, J., A general method for fusion of the Escherichia coli lacZ gene to chromosomal genes in Bacillus subtilis. J. Gen. Microbiol., 132 (1988) 2953–66.Google Scholar
- 22.Young, M. & Hranueli, D., Chromosomal gene amplification in Gram-positive bacteria. In Recombinant DNA and Bacterial Fermentation, ed. J. A. Thompson. CRC, Boca Raton, Florida 1988, pp. 157–200.Google Scholar
- 23.Joyet, P., Levin, D., de Louvencourt, L., Le Reverent, B., Heslot, H. & Aymerich, S., Expression of thermostable alpha-amylase gene under the control of levansucrase inducible promoter from Bacillus subtilis. In Bacillus Molecular Genetics and Biotechnology Applications, ed. A. T. Ganesan & J. A. Hoch. Academic Press, Orlando, 1986, pp. 479–91.Google Scholar
- 25.Ruppen, M., Band, L. & Henner, D. J., Efficient expression of human growth hormone in Bacillus subtilis. In Bacillus Molecular Genetics and Biotechnology Applications, ed. A. T. Ganesan & J. A. Hoch. Academic Press, Orlando, 1986, pp. 423–32.Google Scholar
- 30.Priest, F. G., Extracellular Enzymes, Van Nostrand Reinhold (UK) Co. Ltd, Wokingham, Berkshire, UK, 1984.Google Scholar
- 42.Chang, S., Gray, O., Ho, D., Kroyer, J., Chang, S. Y, McLaughlin, J. & Mark, D., Expression of eukaryotic genes in Bacillus subtilis using signals of penP, p. 159–169. In Molecular Cloning and Gene Regulation in Bacilli, ed. A. T. Ganesan, S. Chang & J. A. Hoch. Academic Press, New York, 1982.Google Scholar
- 45.Honjo, M., Akaoka, A., Nakayama, A., Shimada, H. & Furutani, Y., Construction of the secretion vector containing the prepro structure coding region of the Bacillus amyloliquefaciens neutral protease gene and secretion of Bacillus subtilis α-amylase and human interferon ß in Bacillus subtilis. J. Biotechnol., 3 (1985) 73–84.CrossRefGoogle Scholar
- 46.Stephens, M., Rudolph, C., Hannett, N., Stassi, D. & Pero, J., Secretion vector for Bacillus subtilis. International Patent Application PCT/US8600636, 1986.Google Scholar
- 50.Piggot, P. J. & Coote, J. G., Genetic aspects of bacterial endospore formation. Bact. Revs., 40 (1976) 908–62.Google Scholar
- 51.Bruckner, R. & Doi, R. H., Meeting abstract at 4th International Conference on the Genetics and Biotechnology of the Bacilli, San Diego, CA, June 1987.Google Scholar