To study the biochemical and microbiological properties during co-composting of chicken feather and spent mushroom substrates, chicken feather by incubation of multiple microorganisms were co-composted for 28 days with Pleurotus geesteranus-degraded Eucalyptus sawdust (PE), P. geesteranus-degraded weed tree sawdust (PW) and Ganoderma lucidum-degraded weed tree sawdust (GW), respectively. Characteristics of physico-chemical, enzymatic and microbiological parameters during composting were investigated in terms of temperature, pH, moisture, activities of CMCase, and succession of bacteria and fungus. Results showed that the peak temperature in PEC was 59 °C on 5 days. No ascaris egg was detected in the final composts of PEC, PWC and GWC. The pH of compost in PEC and GWC were about 8.5, which was higher than pH of compost in PWC 7.4. The activities of CMCase in PEC were significantly higher than PWC and GWC during composting. The OM and C/N on 28 days decreased compared to the raw materials. The PCR-DGGE profiles showed significant differences in structure of bacteria and fungus after composting. Four prominent species of bacteria were replaced by uncultured bacterium and the prominent species of fungus were thermophiles and cellulolytic strains on 28 days. Correlation properties were similarly shared by the physico-chemical parameter and bacteria as well as fungus.