Karakterisasi gen nifD bakteri metanotrof asal sawah

Abstract

Methanotrophic bacteria are methane oxidizing bacteria. Some methanotrophs able of fix nitrogen using nitrogenase enzyme. Nitrogenase is composed of two multisubunit metallo protein. Component I contains the active site to nitrogen reduction, encoded by the nifD and nifK genes. Methanotrophic isolates of BGM 3 and BGM 9 had high ammonium accumulation cultured in Nitrate Mineral Salts (NMS) free nitrogen medium. nifD gene fragmens from BGM 3 and BGM 9 were amplified and sequenced. Sequences analyzed using bioinformatics tools such as Blast-N, Blast-X, Expasy Translate Tools, Conserved Domain (CDD) and Scan PROSITE. Phylogenetic construction made using MEGA 4 software. Sequence analysis showed that the nifD sequences of BGM 3 and BGM 9 most closely related to nifD fragmen of Xanthobacter autotropicus PY2. The Identity values of nifD sequences was 92% and 91% respectively. nifD genes from these isolates encode FeMo protein, which was 4Fe4S and 2Fe2S clusters. The nifD gene of BGM 3 and BGM 9 suspected to have same ancestor with the nifD gene of Xanthobacter autotropicus PY2 and Bradyhizobium japonicum USDA 110.