Actinomycetes that are antagonistic to pathogenic fungi(especially candida albicans) were screened from the soil in land that had been planted cotton perennially and had become hardened in shijiazhuang suburb, and from soil in heavy metal-contaminated areas, laboratory chemicals-contaminated areas, and industrial sewage districts. High-throughput screening was performed through still suspension micro- culture technology. The impact of time, temperature, and rotational speed of fermentation in incubator shaker, and the pH, carbon source, nitrogen source, and other physical and chemical factors of fermentation media on the inhibitory effect of antagonistic actinomycetes were investigated. The optimum condition of antagonistic actinomycetes fermentation in incubator was also explored. The results indicated that the fermentation products of antagonistic actinomycetes had inhibitory effect on c. albicans, staphylococcus aureus and other microbes；the optimum condition of antagonistic actinomycetes fermentation was determined to be fermentated at 29 ℃ and 220 r／min for 96 h, with the pH of the fermentation medium adjusted to 7.1, and with starch and corn syrup as the best carbon source and nitrogen source, respectively.