To investigate the mechanism of lead biosorption and biotransformation in lead-resistant bacteria in microcosmic scale, synchrotron radiation micro X-ray fluorescence (μ-SRXRF ) and X-ray absorption near edge structure ( XANES) was used to determine the element distribution characteristics and speciation of lead ( Pb) in lead-resistant bacteria strain isolated from farmland soil samples taken from a lead-zinc mine in Lanping county, Yunnan province in China. The isolated species was identified to be Arthrobacter sp. The concentration of Pb biosorpted in lead-resistant bacteria was directly determined byμ-SRXRF, and the results showed that the highest concentration was 5925μg/g and the bioaccumulate factor was 14 . 8 . Speciation of Pb in lead-resistant bacteria B2, LB ( Lysogeny Broth) medium and soil were also presented after using Pb LⅢXANES. Compared with Pb LⅢ XANES peak shape and peak position between standard samples, it was demonstrated that the Pb in lead-resistant bacteria was mainly formed by amorphous forms like PbS (58. 0%), (C17H35COO)2Pb (22. 2%), Pb5(PO4)3Cl (19. 8%) rather than organic Pb speciation which was exist in LB medium. Therefore, the isolated lead-resistant bacteria could biotransform Pb to sulfuric compounds. The biotransformation regular could be of great interest for other investigators as reference in study of bioavailability impact factor of heavy metals.