In this study, lignocellulose-degrading microbes were added to cattle manure and bagasse co-compost through initial- and two-stage inoculations. A comparison was made between the effects of the two inoculations on compost humification parameters, and an investigation was conducted into the dynamic succession of the microbial community, microbial interactions, and amino acid metabolism to uncover the underlying mechanisms. The results showed that two-stage inoculation increased the humus (HS) and humic acid (HA) contents to 86.59 mg/g and 25.80 mg/g, respectively, and achieved a germination index (GI) of 128.77%. At the genus level, it stimulated the growth of Corynebacterium, Thermobifida, and Aspergillus during the high-temperature period, and Luteimonas, Pseudomonas, Actinomadura, and Rhizopus during the maturity period. Two-stage inoculation increased the stability of the bacterial network and microbial cooperation within the fungal network. Additionally, from the cooling to the maturity period, it boosted ten amino acid synthesis pathways. In conclusion, two-stage inoculation is an effective method to promote the maturation and stabilization of co-compost.