Cephalosporin C (CPC) is a major precursor of cephalosporin antibiotics which is produced by a group of deuteromycets. After biosynthesis in optimized fermentation media, CPC is recovered and converted to a variety of potent cephalosporin antibiotic drugs. This study was performed to determine the mycoflora producing CPC in various parts of Iran soil. Soil samples were collected and cultured on selective media and Acremonium sp. were isolated. The presence of CPC antibiotic in the fermentation broths of these species was determined by a combination of biological, HPLC and mass spectrometry methods. From 350 fungi isolated, 21 (6%) were Acremonium spp, which one strain was able to produce CPC. This species was recognized as Acremonium persicinum according to its macroscopic and microscopic criteria. It is possible to apply different characteristics of the isolated species in strain optimization processes such as protoplast fusion.