Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Cell membrane or cell surface is target
Patent
1996-09-17
1999-02-23
Hutzell, Paula K.
Chemistry: molecular biology and microbiology
Treatment of micro-organisms or enzymes with electrical or...
Cell membrane or cell surface is target
4351723, A61K 39395
Patent
active
058742670
DESCRIPTION:
BRIEF SUMMARY
This application is a national stage application of PCT/EP95/00147.
The present invention relates to vaccines and proteins, rDNA molecules encoding protein expression and presentation systems for the production and presentation of the said proteins, expression vectors therefor, and hosts transformed therewith, as well as methods involved therewith.
The traditional view on the native polymeric organization of the bacterial cell wall has changed dramatically over recent years with the development of new techniques for electron microscopic analysis. The classical idea that the cell membrane(s) is(are) covered by a peptide-glycan-containing matrix does not hold any longer. Besides additional surface structures such as capsules, sheaths, slimes or fimbriae, proteinaceous surface arrays or S-layers are being recognized as a main constituent of the bacterial cell wall (Sleytr and Messner, 1988).
S-layers are a common feature in archaebacterial surfaces (Konig, 1988). In some species such as Halobacterium salinarum or Thermoproteus spp. the proteinaceous S-layer even forms the sole cell wall. At present S-layers are being detected with increasing frequency in a large range of gram-positive and gram-negative eubacteria. Surface arrays are composed of protein or glycoprotein subunits that are arranged into a paracrystalline two-dimensional array, displaying hexagonal, tetragonal or oblique symmetry. Self-assembly of the S-layer is an inherent property of the subunit and is the result of non-covalent protein-protein interactions mediated through salt bridging by divalent metal cations (Mg.sup.2+ or Ca.sup.2+). Non-covalent interactions with components of the underlying cell envelope are thought to be responsible for its positioning at the outermost surface.
Despite the cloning and the characterization of several genes encoding S-layer proteins (SLP's), their function still remains speculative. A variety of functions have been attributed to surface arrays. They might serve as a protective barrier against degradative enzymes or predators, such as Bdellovibrio or help in maintaining bacterial cell shape and form. In some bacterial pathogens, S-layers have been identified as important virulence factors. Although S-layers have several physical features in common, general conclusions on their function cannot yet been drawn.
SLP's are thus present in a large number of archaebacteria, as well as gram-positive and, to a lesser extent, gram-negative bacteria. SLP's form a main constituent of the cell wall, being capable of self-assembly into arrays (crystalline arrays) at the outermost surface of the cell wall. SLP's are continuously and spontaneously produced in larger amounts than any other class of protein in the cell.
SLP's are expressed and either presented or secreted by systems therefor within cells. The genes of these SLP system(s) include: strong promoter sequence(s), a signal peptide coding sequence which is located downstream of the promoter sequence(s), a SLP coding sequence and a transcription termination sequence. The SLP coding sequence is located downstream from the signal peptide coding sequence, having its 5'-terminus operatively linked to the 3'-terminus of the signal peptide coding sequence.
As described herein, an SLP presentation system is distinguished from an SLP secretion system. In the former, the SLP's are bound-up in the cell wall of a host where they are thus presented. In the latter, the SLP's are either produced in the cytoplasm (intracellular production) or secreted into the surrounding medium (extracellular secretion).
The SLP expression and secretion systems of several bacteria have been well-characterized. Among these are those SLP expression and secretion systems of bacteria of the genus Bacillus. Bacilli are well-known as abundant producers of SLP's.
More particularly, the SLP expression and secretion system of the species Bacillus brevis has been extensively studied for its potential use in expressing and extracellularly secreting large quantities of predetermined proteins. B. brevis is ab
REFERENCES:
patent: 5043158 (1991-08-01), Sleytr et al.
Howard et al, J of Bacteriology (1982) pp. 748-757.
Andrei Lupas et al., "Domain Structure of the Acetogenium kivui Surface Layer Revealed by Electron Crystallography and Sequence Analysis," Journal of Bacteriology, Mar. 1994, pp. 1224-1233, vol. 176, No. 5.
Markus Matuschek et al., "Pullulanase of Thermoanaerobacterium thermosulfurigenes EM1 (Clostridium thermosulfurogenes): Molecular Analysis of the Gene, Composite Structure of the Enzyme, and a Common Model for its Attachment to the Cell Surface," Journal of Bacteriology, Jun. 1994, pp. 3295-3302, vol. 176, No. 11.
Norihiro Tsukagoshi et al., "Efficient Synthesis and Secretion of a Thermophilic .alpha.-Amylase by Protein-Producing Bacillus brevis 47 Carrying the Bacillus stearothermophilus Amylase Gene," Journal of Bacteriology, Dec. 1985, pp. 1182-1187.
H. Konig, "Archaeobacterial Cell Envelopes," Can. J. Microbioil., vol. 34, 1988, pp. 395-406.
Hideo Yamagata et al., "Cloning and Characterization of the 5' Region of the Cell Wall Protein Gene Operon in Bacillus brevis 47," Journal of Bacteriology, Mar. 1987, pp. 1239-1245.
N. Tsukagoshi, "Characterization and Application of S-Layer Protein Gene for Production of Foreign Proteins in a Protein-Producing Bacillus brevis 47," Department of Food Science and Technology, Nagoya University, pp. 145-148.
Takahiro Adachi et al., "Multiple and Tandemly Arranged Promoters of the Cell Wall Protein Gene Operon in Bacillus brevis 47," Journal of Bacteriology, Feb. 1989, pp. 1010-1016.
Makoto Takao et al., "Production of Swine Pepsinogen by Protein-Producing Bacillus brevis carrying Swine Pepsinogen cDNA," Appl. Microbiol Biotechnol, 1989, vol. 30, pp. 75-80.
Hideo Yamagata et al., "Use of Bacillus brevis for Efficient Synthesis and Secretion of Human Epidermal Growth Factor," Proc. Natl. Acad. Sci. USA, vol. 86, May 1989, pp. 3589-3593.
Akio Tsuboi et al., "In Vitro Reconstitution of a Hexagonal Array with a Surface Layer Protein Synthesized by Bacillus subtilis Harboring the Surface Layer protein Gene from Bacillus brevis 47," Journal of Bacteriology, Dec. 1989, vol. 171, No. 12, pp. 6747-6752.
Maohong Tang et al., "Cloning of the Crystalline Cell Wall Protein Gene of Bacillus licheniformis NM 105," Journal of Bacteriology, Dec. 1989, vol. 171, No. 12, pp. 6637-6648.
Ron D. Bowditch et al., "Cloning and Sequencing of the Gene Encoding a 125-Kilodalton Surface-Layer Protein from Bacillus sphaericus 2362 and of a Related Cryptic Gene," Journal of Bacteriology, Aug. 1989, vol. 17, No. 8, pp. 4178-4188.
Mingqiang Qiao et al., "New Plasmid Vector and Host System for Genetic Engineering in Bacillus sphaericus," Institute for Molecular Biology, Nankai University, Plasmid, May 28, 1991, pp. 237-241.
D. Speck et al., "Isolation of Bacillus sphaericus Biotin Synthesis Control Mutants; Evidence for Transcriptional Regulation of bio genes," Gene 108, 1991, pp. 39-45.
Lisa D. Taylor et al., "Transformation of an Entomopathic Strain of Bacillus Sphaericus by High Voltage Electroporation," FEMS Microbiology Letters 66 (1990), pp. 125-128.
Deblaere Rolf Y.
Desomer Jan
Dhaese Patrick
Bakalyar Heather A.
Hutzell Paula K.
Solvay ( Societe Anonyme)
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