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

2020 ◽  
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 The Temperature of Growth and Sporulation Modulates the Efficiency of Spore-Display in Bacillus Subtilis

2020 ◽  
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. subtillis builds structurally and functionally different spores when grown at different temperatures, based on that 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 to control 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.

Temperature–pH effect upon germination of bacterial spores

1976 ◽  
Vol 22 (2) ◽  
pp. 322-323 ◽  
Author(s):  
Yuzaburo Ishida ◽  
Takashi Ishido ◽  
Hajime Kadota
Keyword(s):  
Bacillus Subtilis ◽  
Spore Germination ◽  
Incubation Temperature ◽  
Ph Effect ◽  
Bacterial Spores ◽  
Optimum Ph ◽  
Different Temperatures

Using several kinds of criteria for the germination of bacterial spores, germination–pH curves were drawn for Bacillus subtilis spores observed at different temperatures. The experiments revealed that optimum pH for spore germination was markedly changed by changing the incubation temperature; the optimum pH for germination was 7.4 at 37 °C and 5.4 at 10 °C. A possible mechanism involved in this phenomenon is discussed.

The Study of Bacillus Subtils Antimicrobial Activity on Some of the Pathological Isolates

2019 ◽  
Vol 9 (02) ◽  
Author(s):  
Hussein A Kadhum ◽  
Thualfakar H Hasan2
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Antimicrobial Activity ◽  
Bacillus Subtilis ◽  
Bacterial Growth ◽  
Broad Spectrum ◽  
Inhibitory Activity ◽  
Bacterial Pathogens ◽  
Inhibitory Ability ◽  
Selection Of

The study involved the selection of two isolates from Bacillus subtilis to investigate their inhibitory activity against some bacterial pathogens. B sub-bacteria were found to have a broad spectrum against test bacteria such as Staphylococcus aureus and Pseudomonas aeruginosa. They were about 23-30 mm and less against Klebsiella sp. The sensitivity of some antibodies was tested on the test samples. The results showed that the inhibitory ability of bacterial growth in the test samples using B. subtilis extract was more effective than the antibiotics used.

Nucleotide sequence of the chromosomal gene coding for the aminoglycoside 6-adenylyltransferase from Bacillus subtilis Marburg 168

Gene ◽  
1989 ◽  
Vol 78 (2) ◽  
pp. 377-378 ◽  
Author(s):  
Ohmiya Keiichi ◽  
Tanaka Takaaki ◽  
Noguchi Norihisa ◽  
O'Hara Koji ◽  
Kono Megumi
Keyword(s):  
Bacillus Subtilis ◽  
Nucleotide Sequence ◽  
Chromosomal Gene ◽  
Gene Coding

scholarly journals Meiosis-Based Laboratory Evolution of the Thermal Tolerance in Kluyveromyces marxianus

2022 ◽  
Vol 9 ◽  
Author(s):  
Li Wu ◽  
Yilin Lyu ◽  
Pingping Wu ◽  
Tongyu Luo ◽  
Junyuan Zeng ◽  
...  
Keyword(s):  
Kluyveromyces Marxianus ◽  
Thermal Tolerance ◽  
Genetic Recombination ◽  
Original Strain ◽  
Heterologous Proteins ◽  
Diploid Strain ◽  
Nonsense Mutations ◽  
Laboratory Evolution ◽  
The One ◽  
Selection Of

Kluyveromyces marxianus is the fastest-growing eukaryote and a promising host for producing bioethanol and heterologous proteins. To perform a laboratory evolution of thermal tolerance in K. marxianus, diploid, triploid and tetraploid strains were constructed, respectively. Considering the genetic diversity caused by genetic recombination in meiosis, we established an iterative cycle of “diploid/polyploid - meiosis - selection of spores at high temperature” to screen thermotolerant strains. Results showed that the evolution of thermal tolerance in diploid strain was more efficient than that in triploid and tetraploid strains. The thermal tolerance of the progenies of diploid and triploid strains after a two-round screen was significantly improved than that after a one-round screen, while the thermal tolerance of the progenies after the one-round screen was better than that of the initial strain. After a two-round screen, the maximum tolerable temperature of Dip2-8, a progeny of diploid strain, was 3°C higher than that of the original strain. Whole-genome sequencing revealed nonsense mutations of PSR1 and PDE2 in the thermotolerant progenies. Deletion of either PSR1 or PDE2 in the original strain improved thermotolerance and two deletions displayed additive effects, suggesting PSR1 and PDE2 negatively regulated the thermotolerance of K. marxianus in parallel pathways. Therefore, the iterative cycle of “meiosis - spore screening” developed in this study provides an efficient way to perform the laboratory evolution of heat resistance in yeast.

scholarly journals Enhanced Spore Production of <i>Bacillus subtilis</i> Grown in a Chemically Defined Medium

2014 ◽  
Vol 04 (08) ◽  
pp. 444-454 ◽  
Author(s):  
S. M. S. Monteiro ◽  
J. J. Clemente ◽  
M. J. T. Carrondo ◽  
A. E. Cunha
Keyword(s):  
Bacillus Subtilis ◽  
Defined Medium ◽  
Spore Production ◽  
Chemically Defined Medium

scholarly journals Interactions among CotB, CotG, and CotH during Assembly of the Bacillus subtilis Spore Coat

2004 ◽  
Vol 186 (4) ◽  
pp. 1110-1119 ◽  
Author(s):  
Rita Zilhão ◽  
Mónica Serrano ◽  
Rachele Isticato ◽  
Ezio Ricca ◽  
Charles P. Moran ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Reverse Genetics ◽  
Spore Coat ◽  
Yeast Two Hybrid ◽  
Bacillus Subtilis Spore ◽  
Yeast Two Hybrid System ◽  
Ordered Assembly ◽  
Heterologous Antigens ◽  
Two Hybrid ◽  
Good Agreement

ABSTRACT Spores formed by wild-type Bacillus subtilis are encased in a multilayered protein structure (called the coat) formed by the ordered assembly of over 30 polypeptides. One polypeptide (CotB) is a surface-exposed coat component that has been used as a vehicle for the display of heterologous antigens at the spore surface. The cotB gene was initially identified by reverse genetics as encoding an abundant coat component. cotB is predicted to code for a 43-kDa polypeptide, but the form that prevails in the spore coat has a molecular mass of about 66 kDa (herein designated CotB-66). Here we show that in good agreement with its predicted size, expression of cotB in Escherichia coli results in the accumulation of a 46-kDa protein (CotB-46). Expression of cotB in sporulating cells of B. subtilis also results in a 46-kDa polypeptide which appears to be rapidly converted into CotB-66. These results suggest that soon after synthesis, CotB undergoes a posttranslational modification. Assembly of CotB-66 has been shown to depend on expression of both the cotH and cotG loci. We found that CotB-46 is the predominant form found in extracts prepared from sporulating cells or in spore coat preparations of cotH or cotG mutants. Therefore, both cotH and cotG are required for the efficient conversion of CotB-46 into CotB-66 but are dispensable for the association of CotB-46 with the spore coat. We also show that CotG does not accumulate in sporulating cells of a cotH mutant, suggesting that CotH (or a CotH-controlled factor) stabilizes the otherwise unstable CotG. Thus, the need for CotH for formation of CotB-66 results in part from its role in the stabilization of CotG. We also found that CotB-46 is present in complexes with CotG at the time when formation of CotB-66 is detected. Moreover, using a yeast two-hybrid system, we found evidence that CotB directly interacts with CotG and that both CotB and CotG self-interact. We suggest that an interaction between CotG and CotB is required for the formation of CotB-66, which may represent a multimeric form of CotB.

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