Karakterisasi dan identifikasi bakteri pendegradasi 2-(thiocyanomethylthio) benzothiazole (TCMTB)

SUSILAWATI, Lela, Drs. Langkah Sembiring, M.Sc.,Ph.D

2007 | Tesis | S2 Biologi

Penelitian ini bertujuan untuk mengetahui kemampuan isolat yang diisolasi dari limbah industri penyamakan kulit dalam mendegradasi TCMTB. Isolat bakteri diseleksi berdasarkan tingkat toleransinya terhadap senyawa TCMTB dengan berbagai konsentrasi (20, 30, 40, 50 dan 60 ppm) menggunakan metoda paper disk. Isolat terpilih diseleksi lebih lanjut kemampuan degradasinya berdasarkan kadar ammonia dan sulfat yang dilepaskan pada medium uji. Aktivitas biodegradasi dianalisis berdasarkan residu TCMTB dalam medium menggunakan HPLC. Isolat unggul yang memiliki kemampuan mendegradasi TCMTB tinggi, dikarakterisasi dan diidentifikasi secara fenotipik dan molekular dengan analisis sekuen 16S rDNA sehingga hubungan kekerabatan antara isolat unggul dengan strain acuan dapat diketahui. Berdasarkan hasil penelitian diperoleh empat isolat yang menunjukkan kemampuan toleransi tinggi terhadap konsentrasi TCMTB yang berkisar antara 30-40 ppm yaitu PK1, PK2, PK4 dan PK6. Berdasarkan hasil analisis penghasilan ammonia dan sulfat, terpilih tiga isolat yakni PK2, PK4 dan PK6 yang menunjukkan penghasilan kadar ammonia tinggi berkisar 0,51-0,55 ppm dan kadar sulfat yang berkisar 0,90-1,14 ppm. Aktivitas degradasi isolat PK2 lebih tinggi dibandingkan isolat PK4 dan PK6 dengan persentase penurunan konsentrasi TCMTB sebesar 40,8% sedangkan isolat PK4 dan PK6 masing-masing sebesar 27,3% dan 27,4%. Berdasarkan karakteristik fenotipik dan molekular (sekuen 16S rDNA), isolat PK2 diidentifikasi sebagai anggota genus Pseudomonas.

The objective of this research was to investigate the capabilities of bacteria isolated from industrial tanning waste to degrade TCMTB. The bacteria was firstly screened, based on their tolerance to various concentration of TCMTB using paper disk method. Then, those strains were further analyzed in terms of their ability to produce ammonia (NH4 +) and sulphate (SO4 2-). Degradation activity was measured based on remaining residue of TCMTB analyzed using HPLC. The best strain that showed the highest activity in degradation of TCMTB then were characterized and identified based on phenotypic and 16S rDNA sequence analysis. The result of the experiments showed that four selected strains among seven were choosen based on their high tolerance to various concentration of TCMTB, namely PK1, PK2, PK4 and PK6. All four strains showed the ability to produce ammonia and sulphate but three of which, namely PK2, PK4 and PK6 showed the higher capability to degrade TCMTB. One particular strain (PK2) was observed to degrade TCMTB 40.8% within 7 days, but the others were less than 30%. Based on the phenotypic characteristics and 16S rDNA sequence analysis, the best strain (PK2) was identified to be member of genus Pseudomonas.

Kata Kunci : Biodegradasi,TCMTB,Limbah Industri Kulit, Biodegradation, TCMTB, Industrial tanning waste, PCR, Sequencing.