**Abstract**

Double refractory gold ore (DRGO) not only include ppt levels of gold grains locked in sulfide minerals but also a problematic amount of carbonaceous matter. This causes a significant recovery loss of gold during cyanidation because of the strong affinity of the Au(CN)2 − with the carbonaceous matter. Combustion decreases the carbonaceous matter content, but also emits pollutant gases like CO2, SO2 and As2O3. Therefore, environmentally-friendly solutions have been explored by using biotechnology. Due to the very small amount of the above targets in the ore, it is challenging to show evidential changes in solid-phase before and after the biomineral processing of DRGO. This chapter introduces the mineralogical and chemical changes in the various solid residues produced during a sequential biotreatment, consisting of the liberation of gold from sulfides by an iron-oxidizer and decomposition of carbonaceous matter by lignin-degrading enzymes (lignin peroxidase, manganese peroxidase, laccase) secreted from a white rot-fungus, which successfully improved of gold recovery to over 90%. In addition, further development of biotechnology in the recovery of gold from DRGO is addressed.

**Keywords:** double refractory gold ore, carbonaceous matter, enzyme reaction, QEMSCAN map, biooxidation of sulfides
