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KMID : 0545120130230050661
Journal of Microbiology and Biotechnology
2013 Volume.23 No. 5 p.661 ~ p.667
Isolation and Biochemical Characterization of Bacillus pumilus Lipases from the Antarctic
Arild Ranlym Arifin

Kim Soon-Ja
Yim Joung-Han
Antonius Suwanto
Kim Hyung-Kwoun
Abstract
Lipase-producing bacterial strains were isolated from Antarctic soil samples using the tricaprylin agar plate method. Seven strains with relatively strong lipase activities were selected. All of them turned out to be Bacillus pumilus strains by the 16S rRNA gene sequence analysis. Their corresponding lipase genes were cloned, sequenced, and compared. Finally, three different Bacillus pumilus lipases (BPL1, BPL2, and BPL3) were chosen. Their amino acid sequence identities were in the range of 92-98% with the previous Bacillus pumilus lipases. Their optimum temperatures and pHs were measured to be 40oC and pH 9. Lipase BPL1 and lipase BPL2 were stable up to 30oC, whereas lipase BPL3 was stable up to 20oC. Lipase BPL2 was stable within a pH range of 6-10, whereas lipase BPL1 and lipase BPL3 were stable within a pH range of 5-11, showing strong alkaline tolerance. All these lipases exhibited high hydrolytic activity toward pnitrophenyl caprylate (C8). In addition, lipase BPL1 showed high hydrolytic activity toward tributyrin, whereas lipase BPL2 and lipase BPL3 hydrolyzed tricaprylin and castor oil preferentially. These results demonstrated that the three Antarctic Bacillus lipases were alkaliphilic and had a substrate preference toward short- and mediumchain triglycerides. These Antarctic Bacillus lipases might be used in detergent and food industries.
KEYWORD
Bacillus pumilus, lipase, Antarctic
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