Ayder Hot Spring located in North-East of Turkey was investigated with respect to presence of thermophilic microorganisms. Based on phenotypic characteristics, all of the five aerobic isolates belonged to the genus Bacillus and grew optimally at 50-60 degrees C on nutrient agar. According to nutritional characteristics, total protein profiles, % G+C ratio and sequence of 16S rDNA gene, these isolates were identified as strains of Bacillus flavothermus with some differences from the reference strain. Their a-amylase and peroxidase activities were also investigated. These results showed that alpha-amylase activities of new Bacillus flavothermus strains differ from Bacillus flavothermus DSM 2641 and we found that Bacillus flavothermus DSM 2641 has low while 3 of our new isolates (AB03, ABD04 and AB05) have high thermostable peroxidase activity.