Optimum Condition of Crude and Semi Pure Extracellular Chitinase Enzyme from Providencia Stuartii for the Production of N-Acetylglucosamine = Kondisi Optimum Enzim Kitinase Ekstraseluler Kasar dan Semi Murni dari Providencia Stuartii untuk Produksi Nasetilglukosamin

Cynthia, Cynthia (2019) Optimum Condition of Crude and Semi Pure Extracellular Chitinase Enzyme from Providencia Stuartii for the Production of N-Acetylglucosamine = Kondisi Optimum Enzim Kitinase Ekstraseluler Kasar dan Semi Murni dari Providencia Stuartii untuk Produksi Nasetilglukosamin. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Shrimp shells are composed of chitin. In this research, shrimp shells of (Penaeus monodon) were used to produce chitin. One of chitin derivatives, glucosamine, is commonly available in the form of N-acetylglucosamine (NAG) and can be produced by enzymatic fermentation of chitin using chitinolytic microorganism. This research was conducted to determine the optimum fermentation condition, specifically pH, temperature, substrate concentration and incubation time, to produce NAG using crude and semi pure extracellular chitinase enzyme from Providencia stuartii. The pH used in the fermentation were 3, 4, 5, 6, 7, 8 and 9 while the temperature used were 30, 40, 50, 60, 70 and 80°C. The result of crude chitinase enzyme showed that the optimum pH was 5 with 40°C temperature, with enzyme activity of 3.23 ± 0.06 U/ml and 3.42 ± 0.06 U/ml, respectively. While the semi pure chitinase enzyme has the optimum pH of 7 and 40°C temperature, with enzyme activity of 4.74 ± 0.06 U/ml and 4.44 ± 0.06 U/ml, respectively. The selected optimum pH and temperature obtained was then used for the substrate concentration and incubation time optimization. The substrate concentration of chitin were 0.5, 1, 1.5 and 2% with incubation time of 2, 4, 6 and 24 hours. Best fermentation was obtained at 1% substrate concentration and 6 hours of incubation for both crude and semi pure extracellular chitinase enzyme, with the amount of NAG concentration produced were 933.89 ± 12.55 ppm and 1050.56 ± 12.54 ppm, respectively.

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