Aims: To isolate new micro-organisms with R-stereospecific amidase activity and to examine their potential as biocatalysts in enantioselective hydrolysis of 2,2-dimethylcyclopropanecarboxamide (1). Methods and Results: A novel R-stereospecific amidase-producing strain ZJB-07021 was isolated through a sophisticated colorimetric screening method. Based on morphology, physiological tests, Biolog system (GP2) and 16S rRNA sequence, the new isolate was identified as Brevibacterium epidermidis. After 70 min of bioconversion at 35 degrees C, kinetic resolution of (R,S)-1 by the amidase afforded (S)-1 in 41.1% yield (> 99% ee) and (R)-2 in 49.9% yield (69.7% ee) with an average E-value of 23. The enantioselectivity was found to be temperature dependent and enhanced from 12.6 at 45 degrees C to 65.9 at 14 degrees C. Conclusions: A novel bacterial strain of B. epidermidis ZJB-07021 producing R-stereospecific amidase was isolated and characterized. The isolate exhibited high E values for kinetic resolution of racemic-1 to (S)-1. Significance and Impact of the Study: To our knowledge, this was the first report on the species B. epidermidis that harboured R-stereospecific amidase. Strain ZJB-07021 could be further improved as a suitable biocatalyst for the stereoselective bioconversion of racemic-1 after optimization of culture and biotransformation process.