Uneingeschränkter Zugang

Antifungal Activity and Physicochemical Properties of a Novel Antimicrobial Protein AMP-17 from Musca domestica

LONG-BING YANG

LONG-BING YANG

  • School of Basic Medical Sciences, Guizhou Medical UniversityGuiyang, China
  • Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
YANG, LONG-BING
, 
GUO GUO

GUO GUO

  • School of Basic Medical Sciences, Guizhou Medical UniversityGuiyang, China
  • Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
GUO, GUO
, 
XIN-YU ZHAO

XIN-YU ZHAO

  • School of Basic Medical Sciences, Guizhou Medical UniversityGuiyang, China
  • Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
ZHAO, XIN-YU
, 
PEI-PEI SU

PEI-PEI SU

  • School of Basic Medical Sciences, Guizhou Medical UniversityGuiyang, China
  • Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
SU, PEI-PEI
, 
PING FU

PING FU

School of Basic Medical Sciences, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
FU, PING
, 
JIAN PENG

JIAN PENG

Research Center of Medical Biotechnology Engineering, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
PENG, JIAN
, 
JIANG-FAN XIU

JIANG-FAN XIU

School of Basic Medical Sciences, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
XIU, JIANG-FAN
 und 
BO-YAN LI

BO-YAN LI

  • Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical UniversityGuiyang, China
  • School of Food Science, Guizhou Medical UniversityGuiyang, China
Suche nach diesem Autor auf
Sciendo|Google Scholar
LI, BO-YAN
  
03. Sept. 2019
Antifungal Activity and Physicochemical Properties of a Novel Antimicrobial Protein AMP-17 from Musca domestica's Cover Image
Über diesen Artikel
Vorheriger Artikel
Nächster Artikel
Zitieren
COVER HERUNTERLADEN
Artikel-Kategorie: original-paper
Online veröffentlicht: 03. Sept. 2019
Seitenbereich: 383 - 390
Eingereicht: 15. Juni 2019
Akzeptiert: 04. Aug. 2019
DOI: https://doi.org/10.33073/pjm-2019-041
Schlüsselwörter
© 2019 Anton Pyskun et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Time- kill curves of the AMP-17 against the fungal species tested. The growth of five fungi was monitored in the presence of 100 μg/ml AMP-17. Growth of C. albicans (A), C. tropicalis (B), C. krusei (C), C. parapsilosis (D), and C. neoformans with AMP-17 were monitored by reading the optical density (OD562) of the cultures. 100-μg/ml fluconazole was used as a positive control for fungi. Sabouraud’s dextrose broth (0 μg/ml AMP-17) was used as a negative control. The OD562 values of the remaining viable cells were monitored in 96-well plates at different times.
Time- kill curves of the AMP-17 against the fungal species tested. The growth of five fungi was monitored in the presence of 100 μg/ml AMP-17. Growth of C. albicans (A), C. tropicalis (B), C. krusei (C), C. parapsilosis (D), and C. neoformans with AMP-17 were monitored by reading the optical density (OD562) of the cultures. 100-μg/ml fluconazole was used as a positive control for fungi. Sabouraud’s dextrose broth (0 μg/ml AMP-17) was used as a negative control. The OD562 values of the remaining viable cells were monitored in 96-well plates at different times.

Fig. 2.

Influences of various NaCl or MgCl2 concentrations on the antifungal activity of AMP-17. The MIC value of AMP-17 against C. albicans was determined under the influence of different concentrations of NaCl or MgCl2 according to the conditions recommended by CLSI (2008).
Influences of various NaCl or MgCl2 concentrations on the antifungal activity of AMP-17. The MIC value of AMP-17 against C. albicans was determined under the influence of different concentrations of NaCl or MgCl2 according to the conditions recommended by CLSI (2008).

Fig. 3.

Influences of trypsin/pepsin/proteinase K on AMP-17 when used against C. albicans. Fifty μl of the proteases (at a concentration of 1 mg/ml) and 50 μl AMP-17 (at a concentration of 37.5 μg/ml) were mixed in a 96-well plate and placed in a 37°C water bath for 2, 20, 40, 60, 80, 100, and 120 min. Then, it was subjected to CFU enumeration after being treated with 100 μl of the fungal suspension for 18 hours. PBS was used as a negative control. Compared with PBS control, *P < 0.05; **P < 0.01.
Influences of trypsin/pepsin/proteinase K on AMP-17 when used against C. albicans. Fifty μl of the proteases (at a concentration of 1 mg/ml) and 50 μl AMP-17 (at a concentration of 37.5 μg/ml) were mixed in a 96-well plate and placed in a 37°C water bath for 2, 20, 40, 60, 80, 100, and 120 min. Then, it was subjected to CFU enumeration after being treated with 100 μl of the fungal suspension for 18 hours. PBS was used as a negative control. Compared with PBS control, *P < 0.05; **P < 0.01.

Fig. 4.

Hemolytic activities of AMP-17 in vitro. The diluted human red blood cells (100 μl) were incubated with AMP-17 at 37°C for 1 h and spun at 225×g for 10 minutes. The supernatants (100 μl) were transferred to a 96-well plate for measurement of the absorbance of the supernatants at 450 nm by the ultra-microplate spectrophotometer. 0.1% Triton-X 100 and 0.01M PBS were used as a positive control and a negative control, respectively. Fig. 4A is based on Fig. 4B calculated by the hemolysis rate formula. Fig. 4B was the original experimental diagram.
Hemolytic activities of AMP-17 in vitro. The diluted human red blood cells (100 μl) were incubated with AMP-17 at 37°C for 1 h and spun at 225×g for 10 minutes. The supernatants (100 μl) were transferred to a 96-well plate for measurement of the absorbance of the supernatants at 450 nm by the ultra-microplate spectrophotometer. 0.1% Triton-X 100 and 0.01M PBS were used as a positive control and a negative control, respectively. Fig. 4A is based on Fig. 4B calculated by the hemolysis rate formula. Fig. 4B was the original experimental diagram.

MIC values of AMP-17 against the fungi species tested_

PeptideMIC (μg/ml)
C. albicansATCC10231C. kruseiIFM56881C. tropicalisIFM57016C. parapsilosisATCC22019C. neoformansIFM51426
AMP-1718.7518.7518.7518.759.375

MFC values of AMP-17 against the fungi species tested_

PeptideMIC (μg/ml)
C. albicansATCC10231C. kruseiIFM56881C. tropicalisIFM57016C. parapsilosisATCC22019C. neoformansIFM51426
AMP-1737.537.537.537.518.75

Influences of temperature and the number of freeze-thaw times on the antifungal activity of AMP-17_

Temperature (98°C)(min)Colony count(x¯ ± SD)Temperature (−80°C)(Times)Colony count(x¯ ± SD)
569.50 ± 4.95**1(240.50 ± 3.54**) × 102
206.00 ± 2.83**2(273.50±3.54**) × 102
304.50 ± 2.12**4(342.00 ± 8.49**) × 102
606.50 ± 0.71**6(357.00 ± 1.41**) × 102
909.50 ± 3.54**8(351.50 ± 6.36**) × 102
1203.50 ± 0.71**10(370.50 ± 3.54**) × 102
Negative control (ddH2O)565900.00 ± 17112.00Negative control (ddH2O)(5507.00 ± 147.08) × 102
Sprache:
Englisch
Zeitrahmen der Veröffentlichung:
4 Hefte pro Jahr
Fachgebiete der Zeitschrift:
Biologie, Mikrobiologie und Virologie
Zeitschrift RSS Feed