Paenibacillus alvei MP1 as a Producer of the Proteinaceous Compound with Activity against Important Human Pathogens, Including Staphylococcus aureus and Listeria monocytogenes †
Abstract
:1. Introduction
2. Results
2.1. Strain Growth Condition Associated with an Active Compound Production
2.2. Inhibitory Spectrum of P. alvei MP1 Cell-Free Precipitates against Indicator Strains
2.3. Effects of Enzymes and Temperature on Antimicrobial Activity of P.alvei MP1 Cell-Free Precipitates
2.4. Solid Phase Extraction (SPE)
2.5. Direct Detection of Antimicrobial Activity on Tricine SDS-PAGE Gels
2.6. High-Performance Liquid Chromatography (HPLC)
3. Discussion
4. Materials and Methods
4.1. Strain Growth Conditions Associated with an Active Compound Production
4.2. Partial Purification of the Bioactive Compound with Ammonium Sulfate
4.3. Effects of Proteolytic enzymes and Temperature on Antimicrobial Activity of the Compound
4.4. Solid-Phase Extraction with tC18 Column
4.5. Agar-Overlay Inhibition Assay on SDS-PAGE Gels
4.6. High-Performance Liquid Chromatography (HPLC)
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Hours of Experiment [h] | Two-fold Serial Dilutions of Cell-Free Supernatant Spotted onto TSA Soft Agar, with S. aureus L1- 0030 as Indicator Strain | Antibacterial Activity [AU/mL] | Inhibition Zones Diameter Ø [cm] |
---|---|---|---|
9 | | 0 | - |
12 | | 20 | 1.9 |
15 | | 20 | 1.7 |
18 | | 20 | 1.9 |
21 | | 40 | |
24 | | 20 | 2.0 |
27 | | - | - |
30 | | - | - |
Two-fold Serial Dilutions – 50 µL Spots | Inhibition Zone Diameter Ø [cm] |
---|---|
20% (NH4)2SO4 | |
| |
Precipitate Activity—160 AU/mL | |
40% (NH4)2SO4 | |
| |
Precipitate Activity—1280 AU/mL | |
50% (NH4)2SO4 | |
| |
Precipitate Activity—640 AU/mL | |
60% (NH4)2SO4 | |
| |
Precipitate Activity—640 AU/mL | |
70% (NH4)2SO4 | |
| |
Precipitate Activity—20 AU/mL |
Indicator Strain | |
---|---|
Two-fold serial dilutions – 50 µL spots | |
L. monocytogenes FSL – X1-0001 | E. coli O157: H7 |
| |
(160 AU/mL) | (160 AU/mL) |
Inhibition zone diameter Ø [cm] | |
L. monocytogenes FSL – X1-0001 | E. coli O157: H7 |
| |
Ammonium Sulfate Saturation [%] | Indicator Strain - S. aureus L1- 0030 |
---|---|
Two-fold Serial Dilutions – 50 µL Spots | |
40 | |
50 | |
60 | |
Proteinase K Test |
---|
|
60 °C for 30 min. | 80 °C for 30 min | 100 °C for 30 min |
---|---|---|
| | |
Incubation Time [h] | Indicator Strain | ||
---|---|---|---|
L. monocytogenes FSL – X1 - 0001 | S. aureus L1 - 0030 | E. coli O157: H7 | |
7 | | | |
24 | |
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Pajor, M.; Xiong, Z.R.; Worobo, R.W.; Szweda, P. Paenibacillus alvei MP1 as a Producer of the Proteinaceous Compound with Activity against Important Human Pathogens, Including Staphylococcus aureus and Listeria monocytogenes. Pathogens 2020, 9, 319. https://doi.org/10.3390/pathogens9050319
Pajor M, Xiong ZR, Worobo RW, Szweda P. Paenibacillus alvei MP1 as a Producer of the Proteinaceous Compound with Activity against Important Human Pathogens, Including Staphylococcus aureus and Listeria monocytogenes. Pathogens. 2020; 9(5):319. https://doi.org/10.3390/pathogens9050319
Chicago/Turabian StylePajor, Magdalena, Zirui Ray Xiong, Randy W. Worobo, and Piotr Szweda. 2020. "Paenibacillus alvei MP1 as a Producer of the Proteinaceous Compound with Activity against Important Human Pathogens, Including Staphylococcus aureus and Listeria monocytogenes" Pathogens 9, no. 5: 319. https://doi.org/10.3390/pathogens9050319