Judul: Activity of chitooligomers produced by enzymatic hydrolisis upon proliferation of lymphocyte and cancer cells
Abstrak: Local chitin waste from crab industries can be used as a source for production of chitooligomers which has important biological activity. The aims of this research were to evaluate activities chitooligomers produced by enzymatic hydrolysis upon proliferation of lymphocyte and cancer cells. The chitosanase enzyme was obtained from thermophilic bacterium Bacillus licheniformis MB2 isolated from Tompaso Manado. The medium for producing the enzyme contained 1% colloidal chitosan and the enzyme was harvested after seven days of incubation at 55oC, The free cell supernatant were heated at 60 oC for 20 minutes. The heat stable protein enzyme was coagulated with 80% saturated ammonium sulphate and purified using hydrophobic interaction chromatography with butyl sepharose gel. The enzyme of 0.005, 0.0085, 0.10 dan 0,17 IU/mg chitosan were use on 1% soluble chitosan substrate with 85 and 90 % degree deacetylation to produce chitooligomers through incubation at one and three hours. The reaction products were analyzed and fractionated using HPLC. The effect of chitooligomers hidrolysate on normal cells and cancer cells was evaluated using lymphocyte cells previously isolated from human blood and cancer cells line : KR4 (lymphablastoid B), K-562 (chronic myelogenous leukimia), HeLa (epythel carcinoma cervix), and A549 (lung carcinoma ). The inhibition of cancer cells proliferation was visualized by spectrophotometer after addition of MTT reagent. At concentration of 17 ìg/ml the chitooligomers inhibited the cancer cells KR4 by 12.27%, K562 by 20.58%, HeLa by 31.72%, and A549 by 22.70%. In general, hidrolysate and fractionated chitooligomers (trimer to hexamer) showed better anticancer activity than 2-Bromo deoxy uridine used as positive control at similiar concentration. The inhibition 60% of lymphocyte cells was found when treated with chitooligomers hidrolysate from chitosan with 70% degree deacetylation. Fractionated chitooligomers (trimer to hexamer) 20% inhibited the lymphocytes. However, chitooligomers hidrolysate from chitosan with 85 and 90% degree deacetylation enhanced lymphocyte proliferation about 184.54%. K562, HeLa and A549 cancer cells treated with chitooligomers appeared to undergo apoptosis by 35%, 27%, and 8% respectively as shown by fluorescent microscopy. Disrupted membrane was found on cells treated with chitooligomers as observed by scanning electron mycroscope.
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Judul: Aktivitas Biologi Oligomer Hidrolisat Kitin yang Diproduksi Menggunakan Enzim Kitinase dari Isolat SSA2B4.1 (Bacillus cereus SW41) pada Sel Limfosit dan Sel Lestari Kanker.
Abstrak: Local chitin waste from crab industries can be used as a source for production of oligomer which has important biological activity. The aims of this research were to evaluate activities of oligomer produced by enzymatic hydrolysis upon proliferation of lymphocytes and cancer cells. the chitinase enzyme was obtained from thermophilic bacterium Bacillus cereus SW41 isolated from South Sulawesi. The medium for producing the enzyme contained 1% colloidal chitin and the enzyme was harvested after four days of incubation, the free cell supernatant were heated at 60oC for 20 minutes. The heat stable protein enzyme was coagulated with 30% saturated ammonium sulphate and purified using hydrophobic interaction chromatography with butyl sepharose gel. The enzyme of 0,005, and 0,0085 IU/mg chitinase were use on 1% chitin colloidal substrate. the reaction products were analyzed and fractionated using HPLC. Cytotoxic assay to determine the lethal concentration 50 (LC50) was done by BSLT method. The effect of oligomers hidrolyzate on lymphocyte proliferative activity and inhibition of cancer cells were conducted by MTT method. The oligomer hidrolyzate proceed by with or without lyophilization at concentration of 62,5 and 125 μg/ml were able to increase lymphocytes proliferation (3-22%). Antiproliferation activity of oligomer chitin hydrolyzate was detected in all tested cancer cell lines with the highest activity occurred in MT2 cell ranging from 17-48% followed by Raji Cell (17-43%), and HeLa cell (5-33%). Based on cell mentioned above, we concluded that oligomer chitin hydrolyzate could be use as an anti cancer cell proliferation and farther study is needed to develop before commercially use.
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Judul: Kebutuhan pemandu wisata di Taman Nasional Gunung Gede Pangrango
Abstrak: Pramuwisata atau lebih dikenal sebagai pemandu wisata berasal dari bahasa Sansekerta yaitu pramu, wis dan ata. Pramu berarti pelayanan atau orang yang me- layani, wis artinya tempat, dan ata berarti banyak, sehingga batasan pramuwisata adalah orang yang bertugas memberikan bimbingan, penerangan dan petunjuk ten- tang obyek wisata (Direktorat Jenderal Pariwisata, 1991), yang berarti orang yang bertugas memberikan pemanduan wisata. Pemanduan wisata alam di taman nasional merupakan kegiatan yang bertu- juan memberikan pelayanan kepada wisatawan dalam menikmati obyek-obyek wisata alam yang terdapat di kawasan taman nasional, sehingga wisatawan dapat menya- dari, menghargai dan mengerti tentang kawasan yang dikunjunginya serta membantu wisatawan untuk memperoleh tambahan pengalaman, kenyamanan dan kepuasan. Selain itu pemanduan wisata alam merupakan sarana untuk mencapai salah satu tujuan taman nasional yaitu perlindungan dan pelestarian obyek-obyek yang ada, karena pemandu wisata dapat menjelaskan dan menginformasikan secara tepat, be- nar, dan menarik tentang kawasan dan obyek-obyek di taman nasional, yang mem- buat pengunjung dapat mengerti dan memahami bahwa kawasan taman nasional yang dikunjunginya itu merupakan tempat istimewa yang memerlukan perlakuan khusus. Salah satu taman nasional yang telah melaksanakan kegiatan pemanduan wisa- ta alam adalah Taman Nasional Gunung Gede Pangrango. Namun pengelolaan kegi- atan pemanduan tersebut dirasakan masih sangat kurang serta belum terarah dan tera- tur, sehingga diperlukan usaha-usaha untuk mengatasi hal tersebut yang berupa penyediaan tenaga-tenaga pemandu yang profesional dan pengelolaan kegiatan pemanduan secara profesional. ...
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