Isolation and Identification of Manolytic Bacteria from Pari Island

Abstract

Indonesia's tropical marine biodiversity of microorganisms have high potential and commercial value. This study focused on the isolation and identification of potential microbes which produce mannanase enzyme to hydrolyze mannan and eventually produce manno-oligosaccharides. Sampling was conducted in Pari island at Jakarta Bay area, the screening and observation of mannanase enzyme activity from those marine biodiversity have been conducted. The research succeed to collect 20 bacteria that could produce mannanase enzyme, three of them have the most high activity from qualitative analysis based on mannolytic index using congo red method for Pari 3, 4 dan 5 such as 4,33, 2,40, dan 2,60. Further quantitative analysis for these bacteria were conducted such as activity of mannanase using dinitrosalicylic acid method and resulted the highest activity is Pari 3 with 2,474 U/mL and Pari 4 with 1,193 U/mL both in the second day of fermentation, whereas Pari 5 with 1,087 U/mL in the third day of fermentation. Analysis 16S rRNA gene were showed that Pari 3, 4, and 5 are domain Bacteria, filum Firmicutes, class Bacilli, ordo Bacillaceae and genus Bacillus. For the spesific identification are using phylogenetic tree and concluded that Pari 3 has similar sequence 91.1% with Bacillus safensis and 92% Bacillus pumilus, from the both result showed the genetic relationship which is less than 97% in similarity therefore it was suggested that the bacteria Pari 3 is a new strain and can be further investigated. Whereas The results of Pari 4 identification has similar sequence 94.1% with Bacillus anthacis and 92.2% with the Bacillus cereus in the genetic relationship. The results of Pari 5 identification has similar sequence 97.1% with Bacillus anthracis and 95.2% with the Bacillus cereus in the genetic relationship.