Phylogenetic identification of cellulolitic and butanol production bacteria based on 16S rDNA from Ranu Pani lake,east java, Indonesia

Global trend today is how to develop alternative fuel to transportation; one of them is butanol production from cellulose. The objective of this study was to determine of bacterial density and diversity, cellulolytic activity, butanol production and genetic relation from Ranu Pani Lake sediment based on 16S rDNA sequences. The method in this study included sample collection, isolation, enumeration and characterization, cellulolytic test (semi quantitative and quantitative), butanol production, DNA isolation, 16S amplification, sequencing and data analysis. Highest cell density is RP 2.1 (3.6 x 10³ CFU.g^-1) and bacterial diversity is 0.854. RP 3.2 and RP 7.1 isolate has highest semi-quantitative cellulolytic test (1005.84 and 992.42 mm²). Quantitative cellulolytic test was showed RP 2.2 isolate have highest total cellulolytic activity (0.224 ± 0.01 FPU.ml^-1) and resulted 1.21 mg.ml^-1 reducing sugar concentrations. High butanol concentration was resulted by RP 2.2 (10.34 ± 0.002 g.l^-1 after 3 days fermentation), RP 3.2 and RP 7.2 (10.113 ± 0.012 g.l^-1 after 5 days and 9.627 ± 0.006 g.l^-1 after 3 days fermentation). RP 2.2 was identical with Paenibacillus polymyxa DYJL38 (99.6 %), whereas RP 3.2 was identical with Bacillus sp. X15, Bacillus sp. NG06 and Bacillus methylotrophicus LZ043 (99.8 %). RP 7.2 was identical with Bacillus methylotrophicus JF38, Bacillus methylotrophicus -Y48, Bacillus methylotrophicus R13, Bacillus methylotrophicus SDS11 and Bacillus amyloliquefaciens subsp. plantarum L12 (99.8 %).