| Abstrak/Abstract |
Abstract
N-acetylglucosamine (NAG) is one of the chitin's monomer products, which has been widely used as bioactive compounds such as anti-tumor, anti-microbial, and antioxidant. Micromonospora sp. AR17 isolated from the liquid waste of the shrimp freezing industry at PT Toxindo Prima has the potential to produce chitinase. This study aims to determine the activity, purity level, and molecular weight of the purified chitinase from Micromonospora sp. AR17 determines the concentration of NAG produced by purified chitinase that has been characterized. Chitinase was produced by fermentation in colloidal chitin broth at a temperature of 40 oC, pH 7, for 7 days. Chitinase activity was checked every 24 hours to obtain optimum fermentation time. The supernatant with optimal fermentation time continued to the purification stage, including ultrafiltration, ammonium sulfate precipitation, dialysis, ion-exchange chromatography, and gel filtration chromatography. Chitinase activity, purity level, and protein fraction by SDS-PAGE were checked in every stage. The purified enzyme was further characterized for optimum time, pH and temperature to produce NAG. The results showed that the fourth day was the optimal time for chitinase production, with chitinase activity of 0.0040 U/mL and the NAG concentration of 7.62 μg/mL. The purification step successfully increased the purity by 6.82 times with chitinase specific activity value of 1.4648 U/mg. SDS-PAGE of the enzyme showed a molecular weight of 12.60 kDa and 17.32 kDa. Production of NAG with purified chitinase from Micromonospora sp. AR17 produced a NAG concentration of 32.472 μg/mL with an incubation time of 30 minutes, pH 7, and temperature of 40 oC.
Keywords: chitinase; Micromonospora sp, AR17; N-acetylglucosamine; purification. |
