Three different media (chitin, TSB, and LB) were used to incubate Paenibacillus lautus JS-1. P. lautus JS-1 exhibited a larger clear zone in the colloidal chitin agar medium. At day 7, following incubation in the TSB medium, JS-1 strain cell growth was maximum. The P. lautus JS-1 culture contained the highest amount of protein at seven days after incubation in the TSB medium. At day 5, a crude enzyme extracted from chitin culture broth demonstrated the highest chitinase activity of 2.4 U/mL. Chitinase isozymes (CTI) from three distinct culture media were manifested on SDS-PAGE gels as six bands (76, 64, 52, 40, 30, and 28 kDa). Chitosanase isozymes (CTSI) from colloidal chitin and TSB medium were expressed as two bands (41 and 27 kDa), whereas CTSI from LB medium was expressed as a single band (27 kDa). Chitin-oligosaccharides [(GlcNAc)4, (GlcNAc)5, and (GlcNAc)6] were degraded by crude chitinase extracted from three culture broths into smaller chitin-oligosaccharides or N-acetyl-glucosamine. Chitinase cleavage was performed in an endo-manner. As a result of investigating the decomposition of colloidal chitin by TLC medthod, (GlcNAc)2 and (GlcNAc)3 were produced as major products.