Turbidimetric method was used to determine the antifungal activity of antimicrobial peptide in order to replace the time-consuming agar diffusion method. The evaluation method of antimicrobial peptide against fungi was determined by establishing a linear equation between the antimicrobial peptide concentration and absorbance value (OD600). Polymyxin sulfate was selected as the antifungal peptide, Pencillinm chrysogenum was chosen as the indicating fungus. And the concentration of indicator, the concentration of polymyxin sulfate, the ratio of indicator to polymyxin sulfate, and the reaction time were optimized. Results showed that the optimal quantitative determination conditions of polymyxin sulfate against fungi by turbidimetric method were as followed: the concentration of indicating fungi was (1-5)×106 CFU/mL, the concentration of polymyxin sulfate was 0.01-2.5 g/L (final concentration 0.01-0.5 g/L), the ratio of indicating fungi and polymyxin sulfate was 4:1, and the reaction time was 9 h. Under these conditions, the linear relationship between logarithmic value of polymyxin sulfate concentration and OD600 was fine with R2 above 0.99. Furthermore, Aspergillus niger, Aspergillus flavus and Mucor racemosus were selected to verify the method. The results showed that the turbidimetric method was feasible with high reproducibility, good accuracy and precision. The determination time was shortened to 9 h with turbidimetric method. This study provided a foundation for the quantitative determination of antimicrobial peptide.
WANG Zhixin
,
HONG Dan
,
LU Leizhen
,
ZHOU Jingbo
,
LIU Yang
,
NING Yawei
,
MA Aijin
,
JIA Yingmin
. Determination of the antifungal activity of polymyxin sulfate by turbidimetric method[J]. Food and Fermentation Industries, 2020
, 46(7)
: 21
-27
.
DOI: 10.13995/j.cnki.11-1802/ts.022897
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