The humic acid (HA) obtained with the activity of three bacterial strains that transformed a lignite-type low rank coal (LRC) and released humified organic matter (HOM) was characterized. The HA obtained with the activity of Bacillus mycoides, Microbacterium sp. and Acinetobacter baumannii were compared with HA obtained with the traditional alkaline extraction method using NaOH 0.5 M (HA-NaOH). These characterizations included the elemental composition (C, H, N, O), Fourier transform infrared (FTIR) analysis, cross polarization-magic angle spinning-nuclear magnetic resonance (13C-CPMAS-NMR), tetramethylammonium hydroxide (TMAH)-thermochemolysis followed by gas chromatography-mass spectrometry (GC-MS) and size exclusion chromatography. The two evaluated HA types showed differences in the elemental composition, aromaticity degree, content of aliphatic groups, molecular weight, polydispersity, profile of the spectra 13C-CPMAS-NMR and products of TMAH-thermochemolysis. The results indicate that the process of biotransformation of LRC with bacterial activity results in the structural transformations of HA, which leads to the formation of HA with a lower degree of aromaticity, more of a hydrophilic tendency, lower oxygen content, enriched with nitrogenated functional groups and polar aliphatic chains, as compared with HA-NaOH. The HA generated with the three bacterial strains exhibited a high structural similarity to each other; however, some differences were evident in the type of metabolites generated by the TMAH-thermochemolysis of the HA obtained with B. mycoides, as well as a higher polydispersity for the HA generated with A. baumannii.