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"N-asetilglukosamin merupakan monosakarida derivat glukosa yangmemiliki banyak fungsi dan terdapat secara luas dalam sistem tubuh manusia. Nasetilglukosamin dimanfaatkan secara luas baik dibidang kesehatan maupunkosmetik. Produksi N-asetilglukosamin secara enzimatis menggunakan kitinaseyang salah satunya dapat diisolasi dari bakteri relatif lebih ramah lingkungan danmenghasilkan rendemen yang tinggi. Penelitian ini bertujuan untuk mendapatkankondisi optimal produksi N-asetilglukosamin secara hidrolisis enzimatikmenggunakan kitinase yang diisolasi dari bakteri. Seleksi dilakukan terhadapsembilan kultur koleksi BPPT untuk mendapatkan isolat potensial yang dapatmenghasilkan kitinase dengan aktivitas terbaik dan dapat menghidrolisis kitinmenjadi N-asetilglukosamin dengan rendemen tertinggi. Diantara isolat tersebut,BPPT CC 2 menunjukkan aktivitas kitinase terbaik serta dapat menghidrolisis substrat koloidal kitin dan menghasilkan N-asetilglukosamin dengan rendemen tertinggi. Produksi N-asetilglukosamin menggunakan kitinase BPPT CC 2dioptimasi pH, suhu, konsentrasi substrat dan enzim serta lamanya inkubasi.Rendemen N-setilglukosamin tertinggi sebanyak 99,41% didapatkan darihidrolisis 3% substrat koloidal kitin dengan 0,2 U enzim pada kondisi pH 6,0 dansuhu 500C selama 5 hari. Hasil ini mengindikasikan bahwa kitinase dari BPPT CC2 dapat digunakan untuk biokonversi kitin menjadi N-asetilglukosamin denganrendemen yang tinggi untuk kepentingan industri.

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AbstractN-acetylglucosamine is a monosaccharide derivative of glucose thatserve a number of functions and are widely distributed throughout the humanbody system. N-acetylglucosamine posses benefit as a nutritional supplement fortherapeutic usage and also in cosmetics. Enzymatic hydrolysis using chitinaseisolated from bacterial, as one of the enzyme source, produce high yield N- acetylglucosamine and environmental friendly. This research is aimed to achieveoptimum condition for N-acetylglucosamine production by enzymatic hydrolysisusing bacterial isolated chitinase. Nine isolates from BPPT culture collection wasselected to get the most potential isolate, which produced chitinase with bestactivity and able to hydrolyze colloidal chitin resulting high yield of N-acetylglucosamine. Among those isolates, isolate BPPT CC 2 showed the bestchitinase activity and able to hydrolyze colloidal chitin substrate resulting highyield of N-acetylglucosamine. Production of N-acetylglucosamine with BPPT CC2 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 N-acetylglucosamine was obtained by hydrolysis of 3% substrate colloidal chitin with 0,2 U chitinase after 5 days of incubation on pH 6,0 and temperature of 500C. This result suggest that crude chitinase produced by BPPTCC 2 could be useful for bioconversion of chitin into high yield N-acetylglucosamine for industrial application."

"ABSTRAKKitinase merupakan enzim yang berguna dalam degradasi kitin dan dapat dimanfaatkan dalam pengolahan limbah perikanan, biokontrol terhadap jamur fitopatogen, biopestisida, dan produksi protein sel tunggal. Penelitian mengenai karakterisasi kitinase kasar dari Trichoderma amazonicum LP3 dilakukan menggunakan substrat koloidal kitin. Penelitian bertujuan untuk mengetahui produksi dan karakterisasi kitinase kasar yang dihasilkan oleh T. amazonicum LP3. Penentuan aktivitas kitinase dilakukan dengan metode kolorimetri dan produk akhir hasil hidrolisis dianalisis berdasarkan metode Reissig.Hasil penelitian menunjukkan aktivitas kitinase dari T. amazonicum LP3 tertinggi pada inkubasi hari kedua sebesar 8,22 U/mL. Karakterisasi kitinase kasar menunjukkan aktivitas optimum pada pH 5, suhu 50°C, dan stabil selama 3 jam kondisi pH 5 dan selama 1 jam kondisi suhu 50°C. Ion logam Ca2+, Co2+, Fe2+, Mg2+, Zn2+, dan EDTA 1 mM bersifat sebagai aktivator, sedangkan ion logam Hg2+ dan Cu2+ bersifat sebagai inhibitor.

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ABSTRACT

Chitinase enzyme is an effective tool for chitin degradation and can be used in fishery waste management, biocontrol of phytopathogenic fungi, biopesticide, and single cell protein production. A research on the characterization of crude chitinase from Trichoderma amazonicum LP3 has been investigated using colloidal chitin as substrate. This research aims to investigate the production and characterization of crude chitinase from T. amazonicum LP3. Chitinase activity was assayed using the colorimetric method and the end products of the reaction were analyzed by the Reissig method.

The highest activity of chitinase (8,22 U/mL) was observed after 2 days incubation. Characterization of crude chitinase showed optimum activity at pH 5, temperature 50°C, stable for 3 hours at pH 5 and 1 hour at 50°C. The activities were increased when treated by Ca2+, Co2+, Fe2+, Mg2+, Zn2+, and EDTA 1 mM, whereas activities were inhibited when treated by Hg2+ dan Cu2+."