This research is aimed to achieve optimum condition for N-acetylglucosamine (NAG) production  by enzymatic hydrolysis using chitinase preparation of Bacillus sp. BPPTCC-2 indigenous strain. Production of NAG with BPPTCC-2 chitinase was optimized by adjusting its pH, temperature, substrate and enzyme concentration, and also time of hydrolysis.  The yield of 99,41% as maximum production of NAG was obtained by hydrolysis of 3% substrate colloidal chitin with 0,2 U chitinase after 5 d of incubation on pH 6,0 and temperature of 50⁰C. This result suggest that crude chitinase produced by Bacillus sp. BPPTCC-2 could be useful for bioconversion of chitin into high yield NAG for industrial application