Chitin is a natural biopolymer that is found in shrimp shells and can be processed into Nacetylglucosamine which is extensively used as a dietary supplement to treat osteoarthritis, back pain and knee pain. This research was conducted to determine the optimum pH, temperature, substrate concentration and incubation period for the production of Nacetylglucosamine using crude and semi-pure intracellular chitinase enzyme extracted from Mucor circinelloides. Chitin was isolated from Penaeus monodon shell through demineralization and deproteinization process. Optimum pH was determined by measuring chitinase activity at various pH (3, 4, 5, 6, 7, 8 and 9). Optimum temperature was determined by measuring chitinase activity at various temperature (30 2)oC, 40oC, 50oC, 60oC, 70C and 80oC). Different substrate concentration (0.5%, 1.0%, 1.5% and 2.0%) and incubation period(2, 4, 6 and 24 hours) were used to determine the optimum condition to produce Nacetylglucosamine. The results showed that the crude intracellular chitinase enzyme had an optimum pH of 5 with activity of 4.16 ±0.07 U/mL and optimum temperature of 60oC with activity of 4.22 ± 0.07 U/mL. The optimum substrate concentration obtained was 0.5% and the optimum incubation period obtained was 6 hours with GlcNAc concentration of 961.6667 ±9.1287 ppm. The semi-pure intracellular chitinase enzyme had an optimum pH of 4 with activity of 4.75 ± 0.09 U/mL and optimum temperature of 50oC with activity of 5.03 ± 0.08 U/mL. The optimum substrate concentration obtained was 1.5% and the optimum incubation period obtained was 4 hours with GlcNAc concentration of 1150.5556 ± 12.5462 ppm.