Biological activity of components, protein concentration of althernaria alternate lysates by different methods / 基础医学与临床

ABSTRACT

Objective To find the best methods to make the extraction of alternaria alternate. Methods Alternaria alternate was harvested from air and identified, cultured at 26 ℃ for four weeks. Protein was extracted by different methods and its concentration was determined using the Bradford assay, its bioactivity was tested by RAST inhibition, and residual proteins were analyzed by SDS-PAGE.Results The extraction methods like grinder(a), grinder plus ultrasonic(b), ground with liquid nitrogen(c) and liquid nitrogen plus ultrasonic(d) were applied in the extraction procedure. The concentrations of protein extractions were as follows (g/mL): a was 0.44?0.04;b was 0.75?0.03;c was 0.72?0.03 and d; 1.29?0.05. From SDS-PAGE, we found that the most component in the extraction was that obtained by liquid nitrogen plus ultrasonic. From RAST, 50% inhibition was as follows(g/mL): a was 8.5, b was 9.4, c was 7.0,d was 3.7. Conclusion Ground with liquid nitrogen plus disintegration by ultrasonic was the most effective method of extraction.