Cloning and Sequence Analysis of Lipase Gene from DMS3 Isolate

M. P. Widhiastuty¹, Setyanto Tri Wahyudi^1,2, Maelita Ramdani Moeis³, Fida Madayanti1 and Akhmaloka¹*

¹Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jln Ganesha 10, Bandung, Indonesia. ²Biophysics Division, Department of Physics, Bogor Agricultural University, Bogor, Indonesia. ³Microbiology, Genetics and Molecular Biology Research Group, School of Life Sciences and Technology, Institut Teknologi Bandung, Jln Ganesha 10, Bandung, Indonesia.

ABSTRACT: Genes encoding a lipase from DMS3, isolated from Kawah Domas hotspring, Indonesia was cloned and characterised. The lipase gene was cloned based on PCR amplification from genomic DNA. Two set of primers was designed to amplify the lipase gene. An open reading frame of 1248 bps encoding polypeptide of 416 amino acid residues, has been amplified and sequenced. The sequence of amplicon showed high homology (99%) with lipase from Geobacillus thermoleovorans. Detail comparison among three highest homology of lipase showed that there are some variation of amino acid residues. However the subtitution of amino acid residue in lipase ITB3.1, especially at amino acid residue 194 from Asp to Asn is predicted to showh no significant different for the characteristic of lipase ITB3.1 compared to that the other lipase from family 1.5.

KEYWORDS: Lipase; Kawah Domas Hot Spring; PCR Cloning; Geobacillus thermoleovoran

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