The eco-friendly technique of metal leaching, called, bioleaching is gaining huge interest in bio-hydrometallurgical process. This method is a promising technology for extraction of several valuable metals. In this investigation, biological extraction of arsenic (As) from realgar was focused. The bacteria, Leptospirillum ferriphilum was used as microbial agent. The shaker flask experiments were done at 180 rpm, with 0.3% (w/v) pulp density. The temperature and initial pH were maintained at 313 K and 1.5, respectively. The concentration of Fe(II) was varied from 1−12 g/L. The influence of Fe(II) bioleaching of arsenic was studied. The experiment results explicated that arsenic bioleaching were greatly affected by Fe(II) concentration used in the media. After 30 days, it was observed that 75.42% of arsenic was extracted at maximum when using 6 g/L of Fe(II). Kinetics on the bioleaching data were observed to be the maximum rate constant value (1.4643 d 1 ) was attained while using the optimum concentration of Fe(II). Shrinking core model (SCM) was used to identify the rate-limiting step. From the study, it was observed that the bioleaching rate of arsenic was controlled by diffusion through ash layer.