scholarly journals Secretion of heterologous proteins in Bacillus subtilis can be improved by engineering cell components affecting post-translocational protein folding and degradation

2005 ◽  
Vol 99 (2) ◽  
pp. 363-375 ◽  
Author(s):  
M. Vitikainen ◽  
H.-L. Hyyrylainen ◽  
A. Kivimaki ◽  
V.P. Kontinen ◽  
M. Sarvas
Keyword(s):  
Protein Folding ◽  
Bacillus Subtilis ◽  
Heterologous Proteins ◽  
Cell Components

Chaperone-mediated protein folding enhanced D-psicose 3-epimerase expression in engineered Bacillus subtilis

2021 ◽  
Vol 103 ◽  
pp. 65-70
Author(s):  
Jing Chen ◽  
Hongbei Wei ◽  
Yan Guo ◽  
Qiufeng Li ◽  
Huiyi Wang ◽  
...  
Keyword(s):  
Protein Folding ◽  
Bacillus Subtilis

scholarly journals The temperature of growth and sporulation modulates the efficiency of spore-display in Bacillus subtilis

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Claudia Petrillo ◽  
Stefany Castaldi ◽  
Mariamichela Lanzilli ◽  
Anella Saggese ◽  
Giuliana Donadio ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Spore Production ◽  
Bacterial Spores ◽  
Heterologous Proteins ◽  
Dna Coding ◽  
Gene Coding ◽  
Different Temperatures ◽  
Mucosal Vaccines ◽  
Heterologous Antigens ◽  
Selection Of

Abstract Background Bacterial spores displaying heterologous antigens or enzymes have long been proposed as mucosal vaccines, functionalized probiotics or biocatalysts. Two main strategies have been developed to display heterologous molecules on the surface of Bacillus subtilis spores: (i) a recombinant approach, based on the construction of a gene fusion between a gene coding for a coat protein (carrier) and DNA coding for the protein to be displayed, and (ii) a non-recombinant approach, based on the spontaneous and stable adsorption of heterologous molecules on the spore surface. Both systems have advantages and drawbacks and the selection of one or the other depends on the protein to be displayed and on the final use of the activated spore. It has been recently shown that B. subtilis builds structurally and functionally different spores when grown at different temperatures; based on this finding B. subtilis spores prepared at 25, 37 or 42 °C were compared for their efficiency in displaying various model proteins by either the recombinant or the non-recombinant approach. Results Immune- and fluorescence-based assays were used to analyze the display of several model proteins on spores prepared at 25, 37 or 42 °C. Recombinant spores displayed different amounts of the same fusion protein in response to the temperature of spore production. In spores simultaneously displaying two fusion proteins, each of them was differentially displayed at the various temperatures. The display by the non-recombinant approach was only modestly affected by the temperature of spore production, with spores prepared at 37 or 42 °C slightly more efficient than 25 °C spores in adsorbing at least some of the model proteins tested. Conclusion Our results indicate that the temperature of spore production allows control of the display of heterologous proteins on spores and, therefore, that the spore-display strategy can be optimized for the specific final use of the activated spores by selecting the display approach, the carrier protein and the temperature of spore production.

scholarly journals Extracellular Expression of a Functional Recombinant Ganoderma lucidium Immunomodulatory Protein by Bacillus subtilis and Lactococcus lactis

2007 ◽  
Vol 74 (4) ◽  
pp. 1039-1049 ◽  
Author(s):  
Chuan M. Yeh ◽  
Chun K. Yeh ◽  
Xun Y. Hsu ◽  
Qiu M. Luo ◽  
Ming Y. Lin
Keyword(s):  
Bacillus Subtilis ◽  
Lactococcus Lactis ◽  
Mononuclear Cells ◽  
Heterologous Proteins ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Th1 And Th2 Cytokines ◽  
Factor Alpha ◽  
Immunomodulatory Protein ◽  
Overlapping Extension Pcr

ABSTRACT Bacillus subtilis and Lactococcus lactis are ideal hosts for the production of extracellular heterologous proteins of major commercial importance. A recombinant gene for the novel Ganoderma lucidium immunomodulatory protein LZ-8, recombinant LZ-8, was designed encoding the same amino acid sequence but using the preferred codons for both strains and was synthesized by overlapping extension PCR. Using the signal peptide (SP) from subtilisin YaB (SPYaB), recombinant LZ-8 was expressed extracellularly in Bacillus subtilis and Lactococcus lactis. In the absence of SPYaB, recombinant LZ-8 was expressed extracellularly in B. subtilis, but not in L. lactis. The three expressed recombinant LZ-8s showed different capacities for modulating the production of Th1 and Th2 cytokines by peripheral blood mononuclear cells and of tumor necrosis factor alpha by a macrophage cell line.

scholarly journals Bacillus subtilis as a tool for vaccine development: from antigen factories to delivery vectors

2005 ◽  
Vol 77 (1) ◽  
pp. 113-124 ◽  
Author(s):  
Luís C.S. Ferreira ◽  
Rita C.C. Ferreira ◽  
Wolfgang Schumann
Keyword(s):  
Bacillus Subtilis ◽  
Vaccine Development ◽  
Genetically Modified ◽  
Antigen Expression ◽  
Heterologous Proteins ◽  
Unstable Gene ◽  
The Status ◽  
History Of ◽  
Delivery Vehicles ◽  
Close Relatives

Bacillus subtilis and some of its close relatives have a long history of industrial and biotechnological applications. Search for antigen expression systems based on recombinant B. subtilis strains sounds attractive both by the extensive genetic knowledge and the lack of an outer membrane, which simplify the secretion and purification of heterologous proteins. More recently, genetically modified B. subtilis spores have been described as indestructible delivery vehicles for vaccine antigens. Nonetheless both production and delivery of antigens by B. subtilis strains face some inherent obstacles, as unstable gene expression and reduced immunogenicity that, otherwise, can be overcome by already available gene technology approaches. In the present review we present the status of B. subtilis-based vaccine research, either as protein factories or delivery vectors, and discuss some alternatives for a better use of genetically modified strains.

scholarly journals Optimization of the Cell Wall Microenvironment Allows Increased Production of Recombinant Bacillus anthracis Protective Antigen from B. subtilis

2002 ◽  
Vol 68 (1) ◽  
pp. 227-234 ◽  
Author(s):  
Joanne E. Thwaite ◽  
Les W. J. Baillie ◽  
Noel M. Carter ◽  
Keith Stephenson ◽  
Mark Rees ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Cytoplasmic Membrane ◽  
Protective Antigen ◽  
Teichoic Acid ◽  
Heterologous Proteins ◽  
Native Conformation ◽  
Anionic Polymer ◽  
Before And After ◽  
The Stability

ABSTRACT The stability of heterologous proteins secreted by gram-positive bacteria is greatly influenced by the microenvironment on the trans side of the cytoplasmic membrane, and secreted heterologous proteins are susceptible to rapid degradation by host cell proteases. In Bacillus subtilis, degradation occurs either as the proteins emerge from the presecretory translocase and prior to folding into their native conformation or after the native conformation has been reached. The former process generally involves membrane- and/or cell wall-bound proteases, while the latter involves proteases that are released into the culture medium. The identification and manipulation of factors that influence the folding of heterologous proteins has the potential to improve the yield of secreted heterologous proteins. Recombinant anthrax protective antigen (rPA) has been used as a model secreted heterologous protein because it is sensitive to proteolytic degradation both before and after folding into its native conformation. This paper describes the influence of the microenvironment on the trans side of the cytoplasmic membrane on the stability of rPA. Specifically, we have determined the influence of net cell wall charge and its modulation by the extent to which the anionic polymer teichoic acid is d-alanylated on the secretion and stability of rPA. The potential role of the dlt operon, responsible for d-alanylation, was investigated using a Bacillus subtilis strain encoding an inducible dlt operon. We show that, in the absence of d-alanylation, the yield of secreted rPA is increased 2.5-fold. The function of d-alanylation and the use of rPA as a model protein are evaluated with respect to the optimization of B. subtilis for the secretion of heterologous proteins.

The attachment of IgG to cell components: a reconsideration of Brambell’s receptor hypothesis of protein transmission

1974 ◽  
Vol 187 (1087) ◽  
pp. 209-219 ◽  
Keyword(s):  
Yolk Sac ◽  
Heterologous Proteins ◽  
Neonatal Rats ◽  
Rabbit Igg ◽  
Optimum Ph ◽  
Cell Components ◽  
Small Intestinal

Nuclear, mitochondrial and microsomal fractions were prepared from rabbit yolk-sac splanchnopleur and rabbit chorio-allantoic placenta, and tested in vivo and in vitro for attachment of labelled rabbit and bovine IgG. Similar fractions were prepared from small intestinal scrapings of neonatal rats, and tested in vivo and in vitro for attachment of labelled rat and sheep IgG. In all cases the attachment of the homologous and heterologous proteins was substantially equal. Digestion experiments on the labelled organelle pellets of rabbit yolk-sac were carried out with the proteases of the macerated tissue (the cell sap or final supernatant) at optimum pH. The nuclear pellet gave rise to a higher proportion of breakdown products derived from bovine than from rabbit IgG, showing the existence of a mechanism whereby selection between the two proteins might be brought about.

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