Cloning and Characterization of Lipase Gene from A Local Isolate of Pseudoxanthomonas Sp.

Yogi Yopa Kristia¹, Syifa F Syihab¹ and Akhmaloka^1,2

¹Biochemistry Research Group, Faculty of Mathematics and Natural Sciences. Institut Teknologi Bandung, Jl. Ganesha no. 10 Bandung 40132, Indonesia.

²Departement of Chemistry, Faculty of Sciences and Computer. Universitas Pertamina, Jakarta, Indonesia.

Corresponding Author E-mail: loka@chem.itb.ac.id

DOI : http://dx.doi.org/10.13005/bbra/2471

ABSTRACT: Lipase gene from Pseudoxanthomonas sp. was cloned through in vitro amplification from total chromosomal DNA. The gene was sequenced and characterized, coding for 312 amino acid residues. Homological analysis showed that the gene has 98% similarity to lipolytic gene from Uncultured Pseudomonas sp (GenBank No. AKA58891.1). Further analysis appeared that the sequences showed similar unique motifs of lipase sub-family I.1, such as pentapeptide (GHSHG) motif, tetrapeptide (GMLG) motif, and catalytic triad. In additional, 3D structure analysis based on crystal structure of Pseudomonas aeruginose (PDB ID 1ex9) showed that both structure of lipases are similar except on the conformation of catalytic residue of His²⁷⁷ showing to shift more far away compared to that the control.

KEYWORDS: Cloning; Sequencing; Characterization P. seudoxanthomonas sp.; Thermostable lipase;

[ HTML Full Text]