Abstract: Objective:Screening and isolating producing-protease thermophiles from hot springs se-enriched in Yichun and taken the species classification and identification. Methods:By Skim milk plates and 55℃,the producing-protease thermophiles were isolated and taken the morphological observation,physiological and biochemical characterististics,growth characteristics,and the sequence analysis based on 16SrRNA gene,to identify their species and phylogenetic relationships. Results:23 strains of producing-protease thermophiles were isolated from the sample. The growth characteristics experiment of 23 strains of producing-protease thermophiles show that the pH range for growth was 5.0~9.0,the temperature range for growth was 25~65℃,optimum at 45~55℃(except N27). The physiological and biochemical and 16S rRNA gene sequence analysis showed that these strains belong to Anoxybacillus,Aneurinibacillus,Bacillus and Brevibacillus 4 genera. Among them,2 strains Bacillus licheniformis,4 strains Brevibacillus,borstelensis).in the 7 strains Anoxybacillus,N9、N13、N18、N19、N28 were identified as Anoxybacillus rupiensis,and N2,N12 as Anoxybacillus sp,The remaining 10 strains(except N8)were Aneurinibacillus thermoaerophilus,accounting for 39.1% of the total number of all isolated strains. It could be regarded as an dominant group of producing-protease thermophiles in the hot spring. Conclusion:Four genera of producing-protease thermophiles were obtained,which enriched the diversity of producing-protease thermophiles.