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A Sequential Isolation and Isolate Identification of Thermophilic Oil Degrading Bacteria from Bangko Reservoir

Megga Ratnasari Pikoli, Pingkan Aditiawati & Dea Indriani Astuti

Abstract. A study on the isolation of thermophilic oil-degrading bacteria from the crude oil (petroleum) of Banko reservoir has been conducted on a sequential way. The conventional isolation procedure did not isolate the bacteria completely. The first isolation stage was performed directly on the crude oil sample by using Stone Mineral Salt Solution plus yeast extract (SMSSe), as basal medium, at 50C. The second and third isolation stages used the same source, basal medium, and condition as the first stage, but were enriched with a depleted-oil obtained from the crude oil degraded from the previous stage's mixed culture bacteria. Four bacterial isolates were recovered from the first isolation stage, identified as Bacillus polymyxa, B. Licheniformis, Bacillus sp.1, and Pseudomonas aeruginosa. The second stage gave another three different bacterial isolates, identified as Bacillus sp.2, B. stearothermophillus, and B. brevis; whereas the third stage gave only one isolate namely Bacillus coagulans. These isolates could be applied for the degradation of crude oil.

Keywords: bacteria; crude oil; degradation; isolation; sequential isolation

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